DISSERTATIONES BIOLOGICAE UNIVERSITATIS TARTUENSIS 34 EVOLUTIONARY RELATIONSHIPS IN SOME CETRARIOID GENERA (LICHENIZED ASCOMYCOTA) ANDRES SAAG TARTU 1998 DISSERTATIONES BIOLOGICAE UNIVERSITATIS TARTUENSIS 34 DISSERTATIONES BIOLOGICAE UNTVERSITATIS TARTUENSIS 34 EVOLUTIONARY RELATIONSHIPS IN SOME CETRARIOID GENERA (LICHENIZED ASCOMYCOTA) ANDRES SAAG TARTU UNIVERSITY PRESS Chair o f Mycology, Institute of Botany and Ecology, University o f Tartu, Tartu, Estonia The dissertation is accepted for the commencement o f the degree o f Doctor philosophiae in botany and mycology at the University o f Tartu on December 3, 1997 by the Doctoral Committee o f the Faculty o f Biology and Geography o f the University o f Tartu. Opponent: Ph.D. Urmas Kdljalg (Institute o f Zoology and Botany) Commencement: room 207, Lai 40, Tartu; on M arch 26, 1998. Publication of this dissertation is financed by the University of Tartu. © Andres Saag, 1998 Tartu Ulikooli Kirjastuse trukikoda Tiigi 78, EE 2400, Tartu Tellimus nr. 59 CONTENTS LIST OF ORIGINAL PUBLICATIONS............... ........................................... 6 List of other relevant publications................................................................. 7 ABSTRACT............................................................................................................ 8 INTRODUCTION.................................................................................................. 9 Historical background...................................................................................... 10 MATERIAL AND M ETH O D S........................................................................... 12 RESULTS AND DISCUSSION Phylogenetic affinities in the whole group o f cetrarioid lichens ( I) ......... 13 Evolutionary relationships in the genera Asahinea and Cetrelia (II, HI).. 19 Evolutionary relationships in the genera Cetreliopsis and Nephromopsis (IV, V )........................................................................................................... 25 Evolutionary relationships in the group o f cetrarioid lichens with globose ascospores (VI, VII, VIII, IX )................................................................... 29 CONCLUSIONS.................................................................................................... 39 REFERENCES........................................................................................................ 40 EVOLUTSIOONILISED SEOSED MONEDES TSETRARIOIDSETES PEREKONDADES (LIHHENISEERUNUD KOTTSEENED). Kokkuvote............................................................................................................... 44 ACKNOW LEDGEMENTS................................................................................. 45 PUBLICATIONS 47 LIST OF ORIGINAL PUBLICATIONS This thesis is based on the following publications, which are referred to in the text by their Roman numerals. I Saag, A. & Randlane, T. 1995. Phylogenetic affinities o f cetrarioid lichens. — Cryptogamic Botany 5(2): 128-136. II Randlane, T. & Saag, A. 1989. Chemical variation and geographical distribution o f Asahinea chrysantha (Tuck.) Cuib. & C. Culb. — Lichenologist 21: 303-311. III Randlane, T. & Saag, A. 1991. Chemical and morphological variation in the genus Cetrelia in the Soviet Union. — Lichenologist 23: 113-126. IV Randlane, T., Thell, A., & Saag, A. 1995. New data about the genera Cetrariopsis, Cetreliopsis and Nephromopsis (Fam. Parmeliaceae, lichenized Ascom ycotina). — Cryptogam ie, Bryologie-Lichenologie 16: 35-60. V Randlane, T. & Saag, A. Synopsis of the genus Nephromopsis (Fam. Parmeliaceae, lichenized Ascomycota). — Cryptogamie, BryologieLichenologie. (Accepted for publication.) VI Randlane, T., Saag, A., Thell, A. & Kamefelt, I. 1994. The lichen genus Tuckneraria Randlane & Thell — a new segregate in the Parmeliaceae. — Acta Botanica Fennica 150: 143-151. VII Thell, A., Randlane, T., Kamefelt, T., Gao, X.-Q. & Saag, A. 1995. The lichen genus Allocetraria (Ascomycotina, Parmeliaceae). — In: Daniels, F. J., Schulz, M. & Peine, J. (eds.). Flechten Follmann. Contributions to lichenology in honour o f Gerhard Follmann. University o f Cologne, Germany, 3 5 3 -3 7 0 . VIII Thell, A., Goward, T., Randlane, T., Kamefelt, E. I. & Saag, A. 1995. A revision o f the North American Lichen genus Ahtiana (Parmeliaceae). — Bryologist 98(4): 596-605. IX Saag, A., Randlane, T. & Thell, A. Phylogenetic analysis o f cetrarioid lichens with globose ascospores. (Submitted.) 6 List of other relevant publications Randlane, T. & Saag, A. 1991a. Chemical variation and geographical distribution o f Asahinea chrysantha. — In: N. S. Golubkova (ed.), The problems o f experimental lichenology in the USSR. Leningrad, 58-65 (in Russian). Randlane, T. & Saag, A. 1991b. Some chemosystematical data about the lichen genus N ephrom opsis in the USSR. — F olia C ryptogam ica E stonica 28: 26-30. Randlane, T. & Saag, A. 1992a. Additional data about the genus Nephromopsis (Lichens, Parmeliaceae). — M ycotaxon 44(2): 485-489. Randlane, T. & Saag, A. 1992b. New combinations of some cetrarioid lichens (Parmeliaceae). — Mycotaxon 44(2): 491-493. Randlane, T. & Saag, A. 1992c. Genus Cetrelia Culb. et Culb. in URSS. — Novitates Systematicae Plantarum non vascularium 28: 118-133 (in Russian). Randlane, T. & Saag, A. 1992d. Tuckermannopsis americana contra Cetraria ciliaris in Russia. — Folia Cryptog. Estonica 29: 33-36. Randlane, T., Saag, A. & Kondratyuk, S. 1992. Genus Cetrelia Culb. et Culb. in the Ukraine. — Ukrainian Botanical Journal 48( 1): 4-44 (in Ukrainian). Randlane, T. & Saag, A. 1993. World list o f cetrarioid lichens. — Mycotaxon 47: 395-403. Kondratyuk, S., Randlane, T., Saag, A. & Oxner, A. 1993. Genus Cetrelia W. Culb. et C. Culb. — In: A. Oxner, Flora o f the Lichens o f Ukraine 2. Kiev, Naukova Dumka, 214-221 (in Ukrainian). Kamefelt, I., Thell, A., Randlane, T. & Saag, A. 1994. The genus Flavocetraria Kamefelt & Thell (Parmeliaceae, Ascomycotina) and its affinities. — Acta Botanica Fennica 150: 79-86. Randlane, T., Saag, A. & Thell, A. 1997. A second updated world list of cetrarioid lichens. — Bryologist 100(1): 109-122. Randlane, T. & Saag, A. Changes in systematics o f cetrarioid lichens. — Sauteria. (Submitted.) 7 ABSTRACT The group of cetrarioid lichens (fam. Parmeliaceae, lichenized Ascomycota) comprises 131 species in 22 genera. Phylogenetic relationships within some of the cetrarioid genera — Ahtiana, Allocetraria, Asahinea, Cetrelia, Cetreliopsis, Dactylina, Ess linger iana, Nephromopsis, Tuckneraria, and Tuckermannopsis — are treated in more detail in this thesis. Three chemotypes have been recognized in Asahinea chrysantha, one of them is supposed to represent the primitive chemistry o f the genus. Chemosystematical and geographical studies reveal that a possible centre o f speciation o f Asahinea is in the Russian Far East or Japan. Treatment o f chemo- and morphotypes in genus Cetrelia is presented; a table is composed where vacant squares mark the species that are theoretically possible in the genus. The role of “primary” and “secondary” species in Cetrelia according to the “species-pairs” theory is discussed. Species from the genera Cetreliopsis, Nephromopsis, and Cetrariopsis are thoroughly characterized and compared. Both species o f Cetrariopsis are transferred to the genus Nephromopsis, as no difference in the morphology, anatomy, and chemistry o f these two genera (except for their dissimilar positions o f apothecia) could be detected. The group o f cetrarioid lichens, characterized by subglobose to globose ascospores in narrowly clavate asci, is cladistically analyzed. The present state of taxonomy in the whole heterogenous group o f cetrarioid lichens is discussed. One new genus — Tuckneraria Randlane & Thell — is presented; four new species are described: Cetrelia orientalis Randlane & Saag, C. pseudocollata Randlane & Saag, Tuckneraria ahtii Randlane & Saag, Cetreliopsis papuae Randlane & Saag. A number of new combinations have also been proposed. INTRODUCTION The family Parmeliaceae, which consists o f c. 60 genera and about 1000 species, has been colloquially divided into three simple but rather ill-defined morphological categories: alectorioid, parmelioid and cetrarioid. Alectorioid lichens are beard-like or truly fruticose, pendant or caespitose. Parmelioid lichens are clearly foliose, possessing laminal apothecia and pycnidia, and are usually more or less closely adnate to the substrate. Cetrarioid lichens, on the other hand, form quite a vague and undelimited group between these two, having a strap-shaped, subfruticose or ascending foliose thallus, possessing mainly marginal apothecia and pycnidia. However, this arrangement is based mainly on morphology and, as most morphological characters, often displays a large degree o f variability. Evidently, these familiar categories (alectorioid, cetrarioid and parmelioid) have little to do with phylogeny. It was convincingly shown already at the beginning of the nineties (Kamefelt et al. 1992) that there is no doubt about the cetrarioid group o f lichens being polyphyletic. Therefore, all these species cannot be placed in one or two genera as it was done, for instance, by the Russian lichenologist Ksenya Rassadina (1950) or by the Finnish scientist Veli Rasanen (1952). Today we can count more than 130 cetrarioid lichen species which are divided between 22 genera. The aim o f the present thesis is to investigate evolutionary relationships of species in some cetrarioid genera (Ahtiana, Allocetraria, Asahinea, Cetrelia, Cetreliopsis, Nephromopsis, Tuckneraria) and to contribute to the arranging of their taxonomy according to the phylogeny. The critical revision o f all the species in the above-listed genera was carried out to this purpose. The thesis is based on nine original papers prepared by a team o f authors. Twelve more papers, written mainly by the author and his supervisor, are also closely related to the subject under investigation. Studies on the systematics of cetrarioid lichens in the University of Tartu were initiated in the late eighties by the author and his supervisor Tiina Randlane. At first our work focussed on some genera only {Asahinea, Cetrelia), with a special emphasis on chemical and geographical data. In 1992 we met with Ame Thell and Dr. Ingvar Kamefelt, lichenologists from the Department o f Systematic Botany, University o f Lund (Sweden), who were carrying out similar studies on the same group o f species paying extra attention to the anatomical characters. By that time it was evident that the so-called cetrarioid lichens form a heterogenous and informal assemblage which was in urgent need o f more thorough investigation. During the following years several papers were published by the working team consisting o f Am e Thell, Tiina Randlane, and the author. Working tasks were divided among the members of the team as 3 9 follows: identification and investigation of the morphological characters in all studied taxa — T. Randlane; the anatomical characters — A. Thell; the chemical characters — the author. The author of the thesis is also responsible for the ideas concerning the evolution and phylogenetic affinities o f the species. The present dissertation is based on the papers that introduce numerous original data on morphology, anatomy, and chemistry o f cetrarioid lichens; still, the main purpose o f all these studies has been the revision o f systematic arrangement based on evolutionary relationships. Historical background Already the early taxonomical history o f cetrarioid lichen species reveals numerous confusions connected with these taxa. Carl Linne, who recognized only one genus among lichens (Lichen) in his classical “Species Plantarum” (1 753), p rese n ted five species {L. fa h lu e n s is , L. islandicus, L. n ivalis, L. juniperinus, and L. glaucus) that were later included in the group called cetrarioid lichens. The genus Cetraria was originally described by Eric Acharius in his “Methodus Lichenum” (1803), where he comprised eight taxa: C. islandica, C. cucullata, C. nivalis, C. lacunosa, C. fa lla x , C. glauca, C. sepincola, and C. juniperina, four o f them already mentioned by Linne. The fifth species described by Linne {L. fahluensis) was incorporated in Cetraria only by Tuckennan almost eighty years later (1882). Two names mentioned by Acharius — C. fa lla x and C. glauca — later turned out to be synonyms. One more Cetraria species — C. ciliaris — was additionally introduced by Acharius some years after describing the genus (1810). Today only one of these early taxa — the type species C. islandica — has remained in the genus Cetraria according to the modem taxonomy. William Nylander was the first lichenologist who started to split the genus Cetraria. He recognized five species in Cetraria, 25 species in the newly described Platisma (“Platysma”), and one species in Dactylina (both in 1860). Some other close genera were segregated during the following years {Nephromopsis in 1891 by Muller Argoviensis; Tuckermannopsis in 1933 by Gyelnik), but most o f the specialists still recognized one big Cetraria. The genus was considered the largest by Ksenya Rassadina (1950), who incorporated 76 species in it. The modem process of dissecting the polymorphic genus Cetraria was initiated by Dr-s William and Chicita Culbersons, who presented three new genera — Asahinea, Cetrelia, and Platismatia — in the 1960-ies (Culberson & Culberson 1965, 1968). These genera, as well as Masonhalea (Kamefelt 1977), Parmelaria (Awasthi 1987), Ahtiana (Goward 1985) etc., are now accepted by everybody 10 without a doubt. We are quite sure that the same will happen to most o f the new cetrarioid genera, which are predominantly described on the complex of different characters with special emphasis on the anatomical characters. Four years ago 14 genera o f cetrarioid lichens could be listed (Randlane & Saag 1993); at present there are 131 species divided between 22 genera (Randlane et al. 1997); in addition, four former Cetraria species have been transferred to M elanelia (Thell 1995). The list o f separating all these cetrarioid genera in chronological order is presented in Table 1. Table 1. Describing of cetrarioid genera in chronological order. Year of description Name of the genus and the author(s) Number of species today 1794 1803 Cornicularia Hoffin. Cetraria Ach. 1860 1891 1933 1965 1968 1968 1977 1981 1981 1985 1987 1991 1991 1993 1993 1993 1993 1994 1994 1996 Dactylina Nyl. Nephromopsis Mull. Arg. Tuckermannopsis Gyeln. Asahinea W. L. Culb. & C. F. Culb. Cetrelia W. L. Culb. & C. F. Culb. Platismatia W. L. Culb. & C. F. Culb. Masonhalea Kamefelt Cetreliopsis M. J. Lai Esslingeriana Hale & M. J. Lai Ahtiana Goward Parmelaria D. D. Awasthi Allocetraria Kurok. & M. J. Lai Coelopogon Brusse & Kamefelt Arctocetraria Kamefelt & Thell Cetrariella Kamefelt & Thell Nimisia Kamefelt & Thell Vulpicida J.-E. Mattsson & M. J. Lai Flavocetraria Kamefelt & Thell Tuckneraria Randlane & Thell Kaernefeltia Thell & Goward 1 33 (16 “true” Cetrarias + 17 uncertain species) 2 11 11 11 2 17 10 5 1 3 2 10 2 2 2 6 2 5 2 MATERIAL AND METHODS Herbarium material from B, BM, CANB, DUKE, COLO, E, FH, G, GZU, H, KW, LD, LE, M, MB, PC, S, TAIM, TBA, TNM, TNS, TU, UPS, US, WU, and private herbaria of A. Aptroot, D. D. Awasthi, J. A. Elix, and M. J. Lai has been used for this study. Anatomical studies of cortical and reproductive structures were carried out by Dr. Ame Thell in the University of Lund (Sweden) using methods decribed in publication VII. The morphology was studied by T. Randlane and partly by the author using a stereomicroscope Technival 2. Chemical data are based on both “spot tests” and thin layer chromatographic analyses (TLC and HPTLC). The “spot tests” were made with 10% KOH, p-phenylendiamine in ethanol and sodium hypochlorite. The TLC and HPTLC analyses were carried out by the author, mainly according to the standardized TLC methods described by Culberson & Kristinsson (1970), Culberson (1972, 1974), and White & James (1985). The acetone extracts were run in solvent systems A, B, C and G (Culberson et al. 1981). The plates were developed with 10% H2S 0 4 either in H20 or in C2H 5OH. The major secondary compounds were identified in most specimens, the minor constituents were identified only when the spots were sufficiently well developed (mainly in the species of Cetrelia). The cladistical analyses presented in publication I were carried out using the program PAUP version 3.0s (Swofford 1991) on a Macintosh Dei personal computer. The following option settings were used: character-state optimization — DELTRAN; MULPARS; MAXTREE = 1000, 3900; heuristic search; trees rooted using outgroup method; for functional outgroup rooting a monophyletic sister group used; addition sequence — simple; 3 trees held at each step during stepwise addition; tree-bisection-reconnection (TBR) branch-swapping performed; characters reweighted by maximum value of rescaled consistency indices. Strict consensus trees of both original data based cladograms and after characters reweighted were retained. Computer programs for phylogenetic analysis PAUP 3.1.1 (Swofford 1993) and MacClade 3.04 (Maddison & Maddison 1992) were used in the cladistical analyses presented in publication IX. Both programs were run on a Macintosh Color Classic. Using the program PAUP 3.1.1, the follwing heuristic search settings were applied: character-state optimization - ACCTRAN; MULPARS; MAXTREE = 1000, 3000, 6000; addition sequence — simple; 1 tree held at each step during stepwise addition; tree-bisection-recollection (TBR) branchswapping performed; multi-state taxa interpreted as uncertainty; characters weighted equally. The same settings were used in the successive approximations character weighting method. Tree support was investigated using Bremer support and bootstrap in PAUP 3.1.1 and by using the program Parsimony Jackknifer 4.22 (Farris 1995). 12 RESULTS AND DISCUSSION Phylogenetic affinities in the whole group of cetrarioid lichens (I) The group of cetrarioid lichens has always been an artificial conglomerate of species arranged in some heterogenous genera (e.g. Cetraria, Nephromopsis, Parmelia, Tuckermannopsis). To delimit the objects o f the present study, a world list o f cetrarioid lichens was compiled at first (Randlane & Saag 1993). This was the first attempt after 40 years (Rassadina 1950; Rasanen 1952) to sum up all the species closely related to the genus Cetraria Ach. The generic location and valid epithet o f that time was indicated for each of the 120 species. It became evident that the generic location of 27 species was not acceptable and information available about nine species was unsufficient. The list served as a starting point for the following studies. It was evident that the group under discussion was not monophyletic. The first review about the phylogeny o f the group o f cetrarioid lichens was presented by K am efelt et al. (1992), where cladistic analyses were carried out on 50 species as terminal taxa. This analysis was based mainly on the anatomical characters that had been originally revised. M orphological characters, as well as chemical characters, were represented comparatively poorly. In addition, the list of chemical characters was compiled in such a way, that the weight o f some of them was clearly overrated. As a result, the analysed taxa were grouped in three separate aggregates described by important anatomical characters: taxa with an “apical ring structure”; taxa with “uniseriate asci”; taxa with “broadly clavate asci” (Fig. 1). At about the same time (in 1991-92) the cladistic analyses on the same group of lichens were independently carried out also by us. The results were presented in 1AL 2 symposium in 1992 but, unfortunately, were published only three years later (I). O f about 120 known cetrarioid lichen species, 83 were chosen for the analysis to evaluate the systematic arrangement of the taxa. 42 morphological, anatomical, and chemical characters, traditionally in wide use, were used altogether. Still, some of the common characters applied usually in handbooks o f macrolichens (e.g. colour o f the thallus, shape and arrangement o f lobes etc.) were not used here because o f difficulties in defining them properly. Morphological and chemical characters (character states) were identified or verified originally, while descriptions of anatomical characters were based mainly on literature data. The characters were grouped as follows: I thallus structures (9 characters), II apothecia and ascospores (8 characters), III conidiomata and pycnospores (6 characters), IV secondary chemical compounds (19 characters). In the selection of characters we kept to the idea that the taxa should not be described by the data o f reproductive structures only but using also the thalline and especially the chemical characters. The possibly special role of secondary metabolites in the evolution o f lichens has been referred to by J. Poelt 4 13 in his species-pairs theory (Poelt 1970, 1972). It seems plausible that the formation o f biochemical pathways producing lichen substances has taken place earlier, during sexual stages, rather than through mutations in asexual “secondary taxa” (Bowler & Rundel 1975). The genus Cetrelia was selected as the outgroup for the present cladistic analysis; more detailed survey about Cetrelia species is presented in the next chapter. --------------------------------------_________________________ ------- ------------------------------,----------------------------I___________________ __ _______________________ 1______________________ c E Nephromopsis stracheyi Nephromopsis pallescens M asonhalea richardsonii Cetrariella delisei Cetrariella fastigiata Arctocetraria andrejevii Arctocetraria nigricascens Flavocetraria cucullata Flavocetraria nivalis Cetraria kamczatica Cetraria nigricans Cetraria aculeata Cetraria ericetorum Cetraria islandica Cetraria laevigata Ahtiana aurescens Ahtiana pallidula Allocetraria oakesiana Tuckermannopsis chlorophylla Tuckneraria taureri Tuckermannopsis inermis Allocetraria everniella Tuckermannopsis orbata Tuckermannopsis platyphylla Esslingeriana idanoensis Tuckermannopsis americana Tuckermannopsis ciliaris Cetraria fenaleri Cetraria sepincola M elanelia commixta I Melanelia commixta n Melanelia commixta HI M elanelia hepatizon Cornicularia normoerica Bryoria abbreviata Kaernefeltia californica Kaernefeltia merrilli Vulpicida < vuipiciaa canadensis Vutpicida viridis ■ Vulpicida itubulosus Vutpicida Vutpicida tilesii Vutpicida juniperinus Vutpicida pinastri Fig. 1. One of 16 equally parsimonous cladograms. Length 192, consistency index 0.32, retention index 0.68 (Kamefelt et al. 1992, names of terminal taxa according to the modem taxonomy). Three separate analyses were carried out using the same data basis, which was arranged differently for each analysis. In the first analysis, major secondary metabolites were treated as separate characters, totally independent from each other. Evolutionary affinities shown in this analysis should be especially critically evaluated. One of the most important characters of higher taxonomical units — biochemical connections between different lichen compounds — is completely ignored here. That problem is avoided in the second and third 14 analyses, where the lichen substances were not represented as independent characters but grouped into biochemically related sets according to their chemical structure and possible biochemical pathways. The disadvantage of this method is the inability to determine affinities within subclades (several polytomies in cladograms) due to lack o f detailed chemical data. In addition, in the third analysis, some characters treated as binary at first were coded as multistate. Methodically, the third analysis should be evaluated as the most correct. The strict consensus tree o f the third analysis (Fig. 2) was constructed of 3900 equally parsimonous cladograms (after reweighting the characters). Evolution for considerable number o f species (from genera Nephromopsis, Cetreliopsis, Cetrelia) is unresolved (showing several polytomies in the respective part o f the consensus tree). These are just the taxa that were not included in the cladistic analyses by Kamefelt et al. (1992). The rest o f the species are grouped in such a way, which shows considerable similarity with the main three assemblages pointed out in the analysis by our Swedish colleagues. The most clearly separated clade in our analysis (Fig. 2) includes 14 species from the genera Ahtiana, Allocetraria, Esslingeriana, and Tuckermannopsis. This clade completely corresponds to the group described by Kam efelt et al. (1992) as “taxa with uniseriate asci including mainly Tuckermannopsis and related entities”. Three further neighbouring clades in our analysis consist o f species of Kaernefeltia; Platismatia and Vulpicida; M elanelia and Cetraria fendleri group. Two genera, Platismatia and Vulpicida, form separate subclades while species of Melanelia and Cetraria fendleri group are united in one subclade. The whole assemblage is referred to in the Swedish analysis as “taxa with broadly clavate asci”. Membership o f Vulpicida, Melanelia and Kaernefeltia species in this group was pointed out by Kamefelt et al. (1992), while Platismatia was added according to our results. One more clade shows good separation of several species. The clade comprises the Cetraria islandica group, species of Flavocetraria, Arctocetraria, Masonhalea and some other small genera. This grouping is called “taxa with an apical ring structure including mainly the Cetraria islandica group” in the Swedish survey and consists in general o f the same species as in our consensus tree. Resuming the results o f two cladistic analyses carried out independantly in Lund and in Tartu, three separate groupings could be defined in both of them. Similar results are particularly remarkable because o f the fact that the two analyses were based on different data matrices, with emphasis on anatomical characters in Swedish study, and morphological and chemical characters in our analysis. Still, during the preparation o f data matrix it became evident that the descriptions of several cetrarioid species were unsufficient or even eironous, especially with regard to anatomy. This is probably the reason why several rare and poorly investigated species from genera Nephromopsis, 15 Ahtiana attrescens Allocetraria oakesiana Ahtiana pallidula Ahtiana sphaerosporella Tuckermannopsis americana Tuckermannopsis ciliaris Tuckermannopsis microphyllica Tuckermannopsis platyphylla Esslingeriana idahoensis Tuckermannopsis chlorophylla Tuckermannopsis inermis Allocetraria stracheyi Tuckermannopsis subalpina Tuckermannopsis orbata Kaernefeltia merrillii Platismatia erosa Platismatia formosana Platismatia glauca Platismatia Tacunosa Platismatia norvegica Platismatia herrei Platismatia stenophylla Platismatia tuckermanni Vulpicida tubulosus Vulpicida tilesii Vulpicida canadensis Vutpn Vuh r^E € Melanelia agnata Melanelia commixta Melanelia hepatizon Cetraria fenaleri Cetraria weberi Tuckermannopsis platyphylloides Cetraria sepincola Arctocetraria andrejevii Masonhalea richarasonii Cetrariella delisei Cetrariella fastigiata Flavocetraria nivalis Cetraria arenaria Cetraria ericetorum Cetraria islandica Cetraria laevigata Cetraria rassadinae Cetraria kamzatica Kaernefeltia californica Cetraria nigricans Arctocetraria nigricascens Cetraria odonteua Flavocetraria cucullata Allocetraria ambigua Nephromopsis ectocarpisma Nephromopsis endocrocea Nephromopsis komarovii Tuckneraria laureri Nephromopsis pallescens Tuckneraria pseadocomplicata Nephromopsis rugosa Nephromopsis stracheyi Nephromopsis yunnanensis Cetreliopsis endoxanthoides Allocetraria globulans Nephromopsis omata Cetreliopsis asahinae Cetreliopsis rhytidocarpa Nephromopsis wallichiana Cetrelia nuda Cetrelia cetrarioides Cetrelia monachonim Cetrelia chicitae Cetrelia olivetorum Cetrelia collata Cetrelia davidiana Cetrelia delavciyana Cetrelia japoni'ca Cetrelia pseudolivetorum Cetrelia sanguinea Asahinea chrysantha Asahinea scholanderi Parmelaria thomsonii Cetrelia braunsiana Fig. 2. Strict consensus tree constructed from 3900 equally parsimonous cladograms. Secondary compounds as representatives of biochemical groupings. Length of shortest trees 39307, consistency index 0.576, retention index = 0.863. (I, names of terminal taxa according to the modem taxonomy). 16 Cetreliopsis etc. remained unresolved in our cladistic analysis. Both analyses carried out six years ago on a great number o f cetrarioid lichens demonstrated clearly the necessity of additional original studies on this group. The authors o f both papers agreed that the results presented were introductory only and the taxonomical changes, although inevitable in many cases, could be done after further studies. Today the polyphyletic origin o f cetrarioid lichens is generally acknowledged. Three evolutionary lines referred to above, based mainly on reproductive structures and structural characters, have been recognized for the group. Two of the three lines are presumed to be monophyletic (Thell 1996). The first line includes species from seven genera: Arctocetraria, Cett'aria s. str., Cetreliopsis, Coelopogon, Flavocetraria, Masonhalea, and Nephromopsis. They are all characterized by the ellipsoid ascospores in narrowly clavate asci with a small axial body. An amyloid ring structure in tholus is a significant character that is present in Cetraria, Flavocetraria and occasionally in Nephromopsis. The second evolutionary line comprises six genera: Ahtiana, Allocetraria, Dactylina, Esslingeriana, Tuckneraria, and Tuckermannopsis. These species are anatomically characterized by subglobose to globose ascospores in narrowly clavate asci (earlier called “uniseriate asci”). This evolutionary line is also presumably monophyletic (Thell 1996). More detailed survey about the phylogenetic affinities inside the group is presented in the last part o f this chapter (p., 29). The third line is represented by several genera and informal groups: Asahinea, Cetraria fendleri group, Cetrariella, Cetrelia, Cornicularia, Kaernefeltia, M elanelia commixta group, Nimisia, Parmelaria, Platismatia,and Vulpicida. Ellipsoid ascospores in broadly clavate asci with a broad axial body are characteristic to these species. This heterogenous grouping is evidently paraphyletic in the previous treatments of cetrarioid taxa. A number of parmelioid lichens probably also belong here. The present systematic arrangement o f cetrarioid lichens is introduced in the second updated world list (Randlane et al. 1997). The list includes 131 cetrarioid lichen species placed into 22 genera. Six new genera {Arctocetraria Kam efelt & Thell, Cetrariella Kamefelt & Thell, Flavocetraria Kam efelt & Thell, Kaernefeltia Thell & Goward, Nimisia Kam efelt & Thell, and Tuckneraria Randlane & Thell) have been separated since 1993, when the first world list of cetrarioid lichens was published (Randlane & Saag 1993). Splitting o f big genera and describing o f several new small genera has been the obvious tendency in systematics o f lichens during the last decade. In cetrarioid group of lichens this tendency is caused mainly by recent detailed studies of thallus structures and, especially, o f ascomatal anatomy, accompanied by the revisionary work o f morphological and chemical characters. Separation o f most 5 17 o f the new cetrarioid genera has been based on the complex o f various characters with the certain emphasis on the anatomical characters (Randlane & Saag, in press). Nowadays the anatomical features especially o f the inner structures o f reproductive organs are considered more conservative and, therefore, suitable for delimitation at higher taxonomic levels (Hafellner 1984, Kamefelt & Thell 1992). Still, the process o f presenting several new genera has caused countless misunderstandings when using the nomenclature o f cetrarioid lichens, and also objections in principle. We support the opinion that the m ain disagreem ent in this debate is not the grouping o f taxa but their ranking (Elix 1993), although in some cases the separation o f new genera has not been quite sufficiently founded (e.g. Cetrariopsis). In some cases lower taxonomic rank might be more suitable (e.g. subgenus for Arctocetraria and Flavocetraria inside the genus Cetraria). The description of many small genera is rather the first stage in the contemporary process of arrangement of systematics. In the group o f cetrarioid lichens, this stage is mainly finished by now. The following stage will probably include proposals to join some closely related genera. The incorporating Coelocaulon species into Cetraria (Kamefelt et al. 1993) and Cetrariopsis species into Nephromopsis (Randlane et al. 1997) are examples of this tendency already. This is accompanied by proposing numerous new combinations to establish a more convenient position for the species earlier included in big and ill-defined genera such as Cetraria, Nephromopsis, Tuckermannopsis. As a result, a number of cetrarioid genera that had originally been described deficiently, as monotypic or including a few species only, are now thoroughly characterized and comprise several taxa clearly related to each other. This is the case with Ahtiana (described as monotypic in 1985, now including 3 species), Allocetraria (described in 1990 with 3 species, now 10 species), Cetreliopsis (described as monotypic in 1981, now 5 species). During the preparation of this thesis, the following new taxa have been described: Cetrelia orientalis Randlane & Saag (III), Cetrelia pseudocollata Randlane & Saag (III), Tuckneraria ahtii Randlane & Saag (VI), Cetreliopsis papuae Randlane & Saag (IV). A number o f new combinations have also been proposed (only valid combinations are listed here): Nephromopsis isidioidea (Rasanen) Randlane & Saag (Randlane & Saag 1992a), Nephromopsis yunnanensis (Nyl.) Randlane & Saag (Randlane & Saag 1992a), Tuckneraria pseudocomplicata (Asahina) Randlane & Saag (VI), Cetreliopsis endoxanthoides (D. D. Awasthi) Randlane & Saag (IV), Cetreliopsis laeteflava (Zahlbr.) Randlane & Saag (IV), Nephromopsis laii (Thell & Randlane) Saag & Thell (Randlane et al. 1997). The present state o f taxonomy in the heterogenous group o f cetrarioid lichens is far from being final. There are ten species described in Cetraria that we have not seen or for which the available m aterial is sterile and too poor to decide on generic location. These species are: C. albopunctata, C. annae, 18 C. antarctica, C. dermatoidea, C. hypotrachyna, C. kurokawae, C. microphylla, C. nova-zelandiae, C. subscutata, and C. zisangensis. Furthermore, the present generic location o f seven species from the genus Cetraria (C. coralligera, C. fendleri, C. leucostigma, C. melaloma, C. sepincola, C. subfendleri, C. weberi) is not acceptable in our opinion, but it is not clear where they belong (Randlane et al. 1997). Although the systematics in this group has changed considerably during the last decades, it is not truly phylogenetic yet. The contribution of cladistic analyses o f the whole cetrarioid conglomerate to the taxonomy o f this group has been limited primarily because of the difficulties in defining the monophyletic entities. Evolutionary relationships in the genera A sahinea and Cetrelia (II, III) Both genera — Asahinea W. L. Culb. & C. F. Culb. and Cetrelia W. L. Culb. & C. F. Culb. — represent the third, heterogenous evolutionary line of cetrarioid lichens characterized by ellipsoid ascospores in broadly clavate asci. The description o f these genera (in addition to Platismatia W. L. Culb. & C. F. Culb.) in the 60-ies (Culberson & Culberson 1965, 1968) acted as a starting point to the modern splitting of the big genus Cetraria. Segregation of Asahinea from the so-called “parmelioid Cetrariae” (Cetrelia and Platismatia according to the present taxonomy), as well as from Cetraria s. str., was based on the complex o f different characters: prosoplectenchymatous upper cortex, absence of rhizines, laminal up to marginal position of apothecia and pycnidia, imperforation o f the disc, contents of four aromatic compounds in the medulla and arctic-montane pattern o f distribution (Culberson & Culberson 1965). Today the interpretation o f the structure o f the cortex has changed considerably. In earlier papers the terms “prosoplectenchyma” and “paraplectenchyma” referred to the form of cell lumina but since studies of thallus structures were carried out by Hale (1976), these terms are usually applied to indicate the hyphal orientation in the cortex. Thus, according to Culberson & Culberson (1965, 1968) the upper cortex o f Asahinea, Cetrelia, and Platismatia was classified as prosoplectenchymatous, although not characterized by parallel periclinal orientation o f the hyphae. According to the present terminology, all these genera have a pachydermatous paraplectenchymatic cortex with randomly oriented cells (IV). Originally three species were included in Asahinea — A. chrysantha (Tuck.) W. L. Culb. & C. F. Culb., A. scholanderi (Llano) W. L. Culb. & C. F. Culb., and A. kurodakensis (Asahina) W. L. Culb. & C. F. Culb. A. chrysantha is easily distinguished from other members of the genus by its yellow upper cortex and absence of isidia. The range o f it is more or less circumpolar. A. scholanderi was desribed as a Cetraria by Llano (1951) from Alaska. Later, Oxner & Rassadina (1960) described a new species from Asia, Cetraria saviczii. On the 19 basis of the diagnosis Krog (1962) supposed that C. saviczii might be identical with C. scholanderi, a view now widely accepted. A. kurodakensis was the least widely distributed o f the three, supposedly endemic to Japan (Asahina 1953). The morphological and chemical similarity o f A. kurodakensis and A. scho­ landeri was pointed out for the first time in our paper (II) on the basis of original descriptions; unfortunately we had not seen the type materials at that time. The careful comparison of the isotype o f A. scholanderi from Alaska with the lectotype of A. kurodakensis from Japan some years later drew to the conclusion that these taxa are conspecific (Gao 1991). Comparative studies on chemistry, morphology, and geographical distribution of Asahinea chrysantha and A. scholanderi were carried out by us to discuss the phylogenetic affinities o f these taxa (II, Randlane & Saag 1991a). Composition of secondary metabolites in this genus is of special importance. It was generally believed that atranorin and usnic acid occur in the upper cortex and alectoronic acid in the medulla of A. chrysantha as constant metabolites, although usnic acid is sometimes lacking from Japanese m aterial (Yoshim ura 1979). A fourth compound, a-ccllatolic acid, was reported as constant for the Japanese specimens. Some other substances (e.g. P-alectoronic and |3-collatolic acids) have also been announced in the material from the Russian Far East (Krivoschchekova et al. 1983). These results indicate that chemically interesting populations o f A. chrysantha occur in Japan and in the Far East of Russia. Sixty-two samples o f A. chrysantha from the territory o f the former Soviet Union were studied chemically by the author. Atranorin (in the cortex) and alectoronic acid (in the medulla) are the only compunds identified by us as occuring in all samples. Usnic and a-collatolic acids occur in the species in three combinations: I. a-collatolic acid alone; n. a-collatolic acid with usnic acid; El. usnic acid alone. Thus there are three distinct chemotypes of A. chrysantha in the former Soviet Union, each with its own particular geographical distribution. The presence or absence of a-collatolic acid does not seem to correlate with any morphological characters (colour o f upper and lower cortices, width o f brown edges on the lower surface, size of pseudocyphellae, degree o f surface reticulation). The yellow colour of the upper surface depends on the quantity of usnic acid deposited in the cortex. Samples without usnic acid (chemotype I, segregated also as Asahinea culbersoniorum Trass) are uniformly grey. Such specimens, and samples containing usnic acid in small amounts (chemotypes II and III partly) are distinguished only with difficulty. Chemotypes II and III are very variable in the colour of the upper cortex. Treatment o f a single chemotype on the level of independent species (Trass 1992) has not been generally accepted. The geographical distribution o f chemotypes o f A. chrysantha is o f special interest. The least num erous o f the three is chemotype I which is known from Sikhote-Alin (Prim orje region, Russian Far East), Badzhal (Khabarovsk region, Russian Far East) and Japan. Its chem istry (atranorin, 20 alectoronic and a-collatolic acids) is the same as that of A. scholanderi. Chem otype I o f A. chrysantha is supposed — upon biogenetic relationships of lichen products — to represent the basic, primitive chemistry of Asahinea (II). In chemotype II, usnic acid is added to the original complex of substances. The distributional area of chemotype II shows a disjunction being found in Taimyr, Siberia, the Far East, Japan and North America, not however, in Tschukotka or Kamschatka. The lack of a-collatolic acid in chemotype HI represents a second qualitative change in the chemical composition of A. chrysantha. Chemotype HI is the most widely distributed ranging from Scandinavia and Kola over Tschukotka to North America. The different chemotypes of A. chrysantha and their relative distributions are shown schematically in Fig. 3. Combining the distributional patterns of the three chemotypes and assumptions about an original or primitive suite of secondary metabolites in Asahinea, we suggest that a possible centre of speciation of the genus was in Russian Primorye or Japan. This is the only region where chemotypes I and II of A. chrysantha coexist and where A. scholanderi also occurs, while chemotype DI — the most derived chemotype in the genus — is absent. 1 2 3 4 5 Fig. 3. Relative range of Asahinea scholanderi and chemotypes of Axhrysantha in Russia. 1 = chemotype HI; 2 = chemotype II & A. scholanderi', 3 = chemotype III & A. scholanderi; 4 = chemotypes I & II & A. scholanderi (proposed centre of speciation of the genus); 5 = chemotypes II & III & A. scholanderi. The genus Cetrelia was separated from the genera Cetraria and Parmelia on the complex of characters where the medullary compounds had a special importance — all the Cetrelia species produce aromatic substances (orcinol depsides and depsidones) only, and never contain aliphatic fatty acids which 6 21 occur in all taxa o f the close genus Platismatia (Culberson & Culberson 1968). The so-called chemosyndromic variation was demonstrated analysing the medullary chemistry o f Cetrelia species. Instead o f accumulating only one or two m edullary constituents, as previously believed, the species o f Cetrelia syntesize lichen substances in characteristic biogenetically related sets called chemosyndromes. In each chemosyndrome one or two compounds are regularly the m ajor components, and the minor constituents o f one chemosyndrome may become m ajor constituents o f other chemosyndromes (Culberson & Culberson 1976). 15 species were originally included in the genus, o f w hich the m ajority are distributed in eastern and south-eastern Asia. Several species had so far been found also on the territory o f the form er Soviet U nion but no special chemical or distributional studies o f this material had been carried out. Altogether 244 specimens all over the world (including 203 samples from the Soviet Union) were analysed chemically by the author (III, Randlane & Saag 1992c). These studies led to the specification o f the species treatment in the genus Cetrelia. A type o f chemical species consept has traditionally been accepted within this group o f lichens. Different species with similar or even totally identical morphology may differ from each other only by the medullary constituents (Culberson & Culberson 1976). We have proposed to call the morphological groups separated by the Culbersons “morphotypes”. The terms “morphotype” and “chemotype” have been given precise meanings and they are applied to populations o f undetermined taxonomic rank or o f no taxonomic value (Hawskworth 1974). Although these terms are usually used for marking infraspecific variations, they can sometimes also be applied to supraspecific groupings, as the concept of species in different lichen genera varies considerably. Generally, each species in the genus Cetrelia belongs to one morphotype and to one chemotype. Thus the combination o f morpho- and chemotypes makes it possible to characterize concisely all the species known (Table 2). This makes it possible to use the table as an identification guide to the species. Besides its practical use, this table has also some theoretical value. Vacant squares in the table mark the species that are theoretically possible in the genus. This suggestion is confirmed by the fact that there were two specimens found in our analyses that did not belong to any species hitherto described, but still fitted the vacant places in the table. Since we think that material showing new combinations o f known morpho- and chemotypes deserves the rank of species, the two new species, Cetrelia orientalis Randlane & Saag and Cetrelia pseudocollata Randlane & Saag, were described (III). In Cetrelia the highest number of combinations of morpho- and chemotypes (i.e. species) are represented in eastern and south-eastern Asia. From among five morphotypes, only one (cetrarioides) is widely distributed in the world, whereas the areas of non-sorediate morphotypes (isidiata, sinensis, collata, and davidiana) 22 Table 2. Morpho- and chemotypes o f Cetrelia (new species are in bold italics). Morphotypes and their diagnostic characters Chemotypes and their major components I II m IV V Alectoronic and Microphyllinic a. Olivetoric a. Anziaic a. Perlatolic a. a-collatolic a-s VI Imbricaric a. C. chicitae C. monachorum Cetrarioides Thallus sorediose C. olivetorum C. cetrarioides Isidiata Thallus isidiate C. braunsiana C. isidiata Sinensis Thallus with lobules C. orientalis C. japonica C, pseudolivetorum C. sinensis Collata Thallus without vegetative propagules; large pseudocyphellae C. nuda Davidiana Thallus without vegetative propagules; small pseudocyphellae C. pseudocollata C. collata C. davidiana C. sanguinea C. delavayana C. alaskana fall only into East and Southeast Asia. Lichens that have developed soredia can expand and effectively enlarge their area of distribution contrary to species that use only ascospores. The complicated process of resynthesis of a lichen thallus from a germinated ascospore and a free-living photobiont first depends upon the distribution area of algae (Tehler 1982). The other forms of vegetative propagules should theoretically also have the ability to colonize habitats in which the photobiont cannot thrive in its free-living form. In the genus Cetrelia, isidia and lobules still appear to be almost as ineffective in colonizing new territories as the spores. Thus only four sorediate species are found also in Europe and North America, in addition to Asia. The centre of speciation in the genus is presumably located in eastern and southern Asia as can be seen from the vast majority of the combinations of chemo- and morphotypes. The first scheme dealing with the connections between species of Cetrelia was proposed by Poelt (1970, 1972) and related to his discussion on “species-pairs” . Our scheme includes all the combinations of chemo- and morphotypes known in the genus up to now (Fig.4). The horizontal axis on the scheme shows the direction of chemical evolution and the vertical axis represents the variety of reproductive propagules. We presume that the formation and evolution of biochemical pathways producing the complex of lichen substances has taken place during sexual stages in the evolution of lichens, rather than through mutations in asexual stages (Bowler & Rundel 1975). The genus Cetrelia fits into the theory of development of “species-pairs” rather well, since every chemosyndrome is represented not only by the “secondary” but also by the “primary” species. This avoids the necessity of referring to some hypothetical ancestral taxa. All “primary” species with sexual reproduction appear to be quite rare with a very restricted distribution. However, not only pairs of species but also triplets or even tetrades of species can be observed in the genus Cetrelia (HI). A similar pattern has been reported only in a few other cases, for example in Parmelia and Physcia (Hawksworth & Hill 1984). The taxonomic treatment proposed by Tehler (1982), to recognize fertile and sterile counterparts as infraspecific taxa with the rank of forma, could be acceptable theoretically but not in practice. “Primary” species differ from the “secondary” species essentially in their morphology, and the “secondary” species of one column differ from each other as well. The morphological similarity between the taxa of the same morphotype is certainly great, but they all have presumably quite a different origin and cannot belong to one species. Otherwise we would be “back to the traditional typological species concept adamantly blind to everything but morphology” (Culberson 1986). 24 I II III IV V VI Direction of chemical evolution (chemotypes) Fig. 4. Evolutionary relationships between Cetrelia species. Restricted to eastern and southeastern Asia (£f), limited distribution outside eastern and southeastern Asia (HI), wide distribution both in- and outside eastern and southeastern Asia d ) . 1 = C- nuda\ 2 = C. orientalise 3 = C. braunsiana; 4 = C. chicitae; 5 = C. pseudocollata; 6 = C. japonica; 1 - C. davidiana', 8 = C. pseudolivetorunr, 9 = C. olivetorum; 10 = C. sanguinea, 11 = C. isidiata, 12 = C. delavayana; 13 = C. cetrarioides', 14 = C. collata', 15 = C. alaskana; 16 = C. sinensis; 17 = C. monachorum The frequency and distribution of the examined species in the genus Cetrelia, serve, in our opinion, as good examples of some of the theoretical assumptions discussed above. All the sorediate species, although common and widely distributed, may be considered “evolutionary blind alleys”. The organism loses its genetic flexibility by having an asexual propagation system superior to the sexual process (Tehler 1982). On the other hand, the sexual species, although occasionally less successful in dispersing, have retained their ability for further speciation by the process of gene recombination. Evolutionary relationships in the genera Cetreliopsis and Nephromopsis (IV, V) These genera — Cetreliopsis M. J. Lai and Nephromopsis Miill. Arg. — represent the first evolutionary line of cetrariod lichens characterized by the ellipsoid ascospores in narrowly clavate asci with a small axial body. The g;enus Nephromopsis was described already in 1891 by M uller Argoviensis to accomodate N. stracheyi which was said to have a thallus similar 7 25 to Cetraria but the position o f apothecia like in Nephroma. In contemporary lichenology the genus was not generally recognized until 1981, when Lai resurrected the treatment o f the species with cetrarioid thallus, nephromoid apothecia, and pseudocyphellae on the lower surface in a separate genus under the name Nephromopsis. Still, the evaluation o f important characters on the genus level has changed considerably. Today, as was remarked above, the anatomical features o f the thallus and especially o f the inner structures of ascomata are considered more conservative and, consequently, suitable for delimitation at generic level. Mainly on these grounds, the genus Tuckneraria) was separated from Nephromopsis (VI), and some taxa were transferred from Nephromopsis to Allocetraria (VII) or Cetreliopsis (IV). The m onotypic genus Cetrariopsis was separated from Cetraria s. lat. by Kurokawa in Decem ber 1980 (Kurokaw a 1980). Less than a m onth later Lai (1981) proposed the genus Ahtia to accomodate the same species — Cetraria wallichiana (Taylor) Mull. Arg. Two essential characters were pointed out by both authors to describe the new genus -— the numerous, small, laminal apothecia and the prosoplectenchymatic upper cortex. The structure o f the cortex, however, was a m isinterpretation by Kurokawa and Lai (see the discussion on p. 19). According to the contemporary knowledge and terminology, the species of Cetrariopsis have a pachydermatous paraplectenchymatic cortex with randomly oriented cells like Asahinea, Cetrelia, and Platismatia. Still, Cetrariopsis is probably not closely related to those entities as it has been supposed earlier (Kurokawa 1980, Kamefelt et al. 1992, Elix 1993). The position of apothecia on the lower side of the thallus has been one of the most attractive and significant characters in defining the genus Nephromopsis since Muller Argoviensis, while the laminal position o f apothecia over the upper surface remains the only true feature for the genus Cetrariopsis (Kurokawa 1980). The position o f the apothecia is indeed a striking character, but a more thorough examination shows that specimens with clearly laminal apothecia usually have fruiting bodies along the margins as well. Furthermore, on many specimens with mostly marginal apothecia both submarginal or even truly laminal ascomata also occur. Our studies have shown that there is not much difference in the morphology, anatomy, and chemistry of these two genera except for their dissimilar positions of apothecia (IV). Therefore we have proposed transferring the two species of Cetrariopsis (C. pallescens and C. laii) to the genus Nephromopsis (Randlane et al. 1997). The genus Nephromopsis is defined today by the following characters: a large foliose thallus; apothecia marginal on the lower surface o f the thallus, or submarginal and laminal on the upper surface; presence of pseudocyphellae only on the lower surface; exciple usually three-layered but sometimes the 26 middle layer is not distinctly developed and is seen as two-layered in a few species; narrowly clavate asci and oblong ascospores; bifusiform pycnoconidia; usnic acid in the cortex; various fatty acids and/or some orcinol depsidesdepsidones in the medulla. According to our present knowledge (V) the genus includes eleven species: N. endocrocea Asahina N. isidioidea (Rasanen) Randlane & Saag N komarovii (Elenkin) J. C. Wei N. laii (Thell & Randlane) Saag & Thell N. morrisonicola M. J. Lai N. nephromoides (Nyl.) Ahti & Randlane, N. ornata (Mull. Arg.) Hue N. pallescens (Schaer.) Park N. rugosa Asahina N. stracheyi (Bab.) Mull. Arg. N. yunnanensis (Nyl.) Randlane & Saag. The genus Cetreliopsis was described by Lai (1981) in his paper on cetrarioid lichens in East Asia to settle the Cetraria rhytidocarpa-comiplQx as he called it. The group included Cetraria rhytidocarpa Mont. & Bosch from Java, Cetraria straminea Vain, from the Philippines, and Cetraria laeteflava Zahlbr. from Taiwan. Lai synonymized all the three species on morphological and chemical grounds. He also pointed out the affinities of Cetrelia with Nephromopsis combining the names o f these genera for the new taxon. Our studies on Cetreliopsis have shown that this genus is well limited and clearly separated from other cetrarioid lichens. The identifying characters o f the genus Cetreliopsis are: large foliose or subfruticose thallus; marginal or submarginal apothecia; presence of pseudocyphellae on both surfaces of the thallus; large, ellipsoid ascospores in rather broadly clavate asci and a two-layered exciple; content of fumarprotocetraric and protocetraric acids as major compounds in the medulla (PD+ red) and usnic acid in the cortex. We cannot agree with Lumbsch (Eriksson & Hawksworth 1988) who proposed to include C. rhytido­ carpa in Cetrelia as a separate subgenus. He recognized mainly chemical differences between Cetrelia and Cetreliopsis and considered their morphology very similar. We find not only essential chemical differences but also morpho­ logical and anatomical differences between these two entities (IV: 47). The genus Cetreliopsis is probably more closely connected to Nephromopsis than to Cetrelia. According to our present knowledge the genus cannot be treated as monotypic. Three new combinations have been proposed; furthermore, one new species and one new subspecies have been described (IV). We do not support in all parts the wide species treatment of Cetreliopsis rhytidocarpa proposed by 27 Lai (1981) and prefer to keep the sorediate Taiwan material as a separate species (C. laeteflava). The following five species are included in Cetreliopsis: C. asahinae (Sato) Randlane & Thell C. endoxanthoides (D. D. Awasthi) Randlane & Saag C. laeteflava (Zahlbr.) Randlane & Saag C. papuae Randlane & Saag C. rhytidocarpa (Mont. & Bosch) M. J. Lai. In our opinion these two genera — Nephromopsis (incl. Cetrariopsis) and Cetreliopsis — form a group of closely related taxa inside the first evolutionary line of cetrarioid lichens. Comparison of characters (Table 3) shows considerable similarity in general habit and anatomy of the thallus, inner structures of the ascocarps, conidial characters and cortical substances. All the 16 species of this generic complex are distributed in the mountainous forests of eastern and southeastern Asia only. Still, Cetreliopsis differs from Nephromopsis in some special morphological (pseudocyphellae on both surfaces), anatomical (asci rather broadly clavate, ascospores ellipsoid), and medullary chemical characters (absence of orcinol depsides and depsidones, and presence of B-orcinol depsidones — fumarprotocetraric acid and related substances in all species), and is therefore maintained as a separate genus (V). Table 3. Comparison of characters in Cetreliopsis and Nephromopsis. Character Cetreliopsis Nephromopsis Thallus foliose foliose Upper and lower cortex 1-layered, paraplectenchymatous 1-layered, paraplectenchymatous Pseudocyphellae on both surfaces on lower surface Soredia may be present absent Marginal cilia may be present absent Apothecia marginal and submarginal on upper surface marginal on lower surface, submarginal and laminal on upper surface Exciple 2-layered usually 3-layered, sometimes 2-layered 28 Ascus shape rather broadly clavate narrowly clavate Ascospores ellipsoid, 6-12 x 4-7 |im oblong, 5-10 x 2,5-5 jim Axial body 2,5-4 |im 0,5-4 jim Ring structure absent present in two species Pycnidia laminal or marginal, immersed or on projections laminal or marginal, immersed or on projections Pycnoconidia bifusiform, 5 x 1-2 jim bifusiform, 4-5 x 1-1,5 fim Cortical substances usnic acid usnic acid Medullary substances: a) fatty acids present present alectoronic a., physodic a., olivetoric a. b) orcinol depsides & depsidones c) (3-orcinol depsidones fumarprotocetraric a., protocetraric a., physodalic salazinic a., Cph-1 endocrocin, secalonic acids A and C d) secalonic acids Evolutionary relationships in the group of cetrarioid lichens with globose ascospores (VI, VII, VIII and IX) The species belonging to the second evolutionary line o f cetrarioid lichens are treated in more detail here. This evolutionary line, anatomically characterized by subglobose to globose ascospores in narrowly clavate asci, is considered monophyletic (Thell 1996). This assumption is supported by two earlier cladistic analyses on several various species o f cetrarioid lichens, carried out independently in Lund (Kamefelt et al. 1992) and in Tartu (I). In both analyses the clade including species with globose ascospores was clearly separated. The group comprised at first (Thell 1996) five genera — Ahtiana, Allocetraria, Esslingeriana, Tuckneraria, and Tuckermanniopsis; we also add the genus Dactylina here because o f its obvious affinities to Allocetraria. Four genera o f 8 29 the six, Ahtiana, Allocetraria, Dactylina, and Tuckneraria, have been thoroughly revised lately (VI, VH, VIH, Kamefelt & Thell 1996). The sole species o f the monotypic genus Esslingeriana was described completely by Esslinger (1971). Tuckermannopsis is the only taxon still in need o f revision. Nevertheless, the data characterizing these taxa are scattered in many papers and evolutionary affinities between all the species included have not been evaluated yet. Phylogenetic analysis o f the group was carried out using cladistic parsimony methods. Taxa analysed. 30 species from the six above-listed genera were applied as terminal taxa (DC). For choosing the outgroup we had, in principle, two alternatives: a taxon either from the group close to Cetraria (the first evolutionary line of cetrarioid lichens) or from the group related to the so-called “parmelioid Cetrariae” (the third evolutionary line). The form o f asci — narrowly clavate — is the same in the ingroup and in the group close to Cetraria, while the third group has typically broadly clavate asci. Therefore, the parmelioid and allied genera were discarded at first. Two genera o f those seven included in the first evolutionary line — Cetraria s. str. and Flavocetraria — were seriously considered as possible outgroups. Ascocarps o f Flavocetraria are characterized by ellipsoid ascospores, narrowly clavate asci with a small axial body, and presence o f an amyloid ring structure. These characters are similar to Cetraria s. str. and differ from those o f the ingroup. Still, the stucture o f upper and lower cortices and secondary chemistry o f Flavocetraria is more similar to the studied taxa. Finally, Flavocetraria cucullata was selected as outgroup for one series of analyses. Later the species from the third evolutionary line were also evaluated as possible samples for outgroup. In earlier analyses (Kamefelt et al. 1992, I) C etraria fe n d le ri appeared as one representative o f the sister group to the assam blege w hich is treated as ingroup here. A natom ically, ascocarps o f C. fendleri are characterized by ellipsoid ascospores, broadly clavate asci, a large axial body, and absence o f an amyloid ring structure. The ingroup differs from the species under discussion in two former and is similar in two latter characters. Therefore, C. fendleri was additionally chosen as outgroup. Both selected species — Flavocetraria cucullata and Cetraria fendleri — were used to root the trees in separate series o f analyses. Characters. All in all, 36 morphological, anatomical, and chemical characters were used for the analysis. Lichen substances were not treated independently from each other but grouped into biochemically related sets, as suggested in our previous study (I). 30 Character states were coded as 0 ,1 , 2, 3 and 4; all multistate characters were treated as unordered. The characters and character states were the following. 1. Form o f thallus: adnate (0), ascending (1) 2. Symmetry o f thallus: dorsiventral (0), radial-symmetrical (1) 3. Interior o f thallus: of densely arranged hyphae (1), o f loosely arranged hyphae (1), becoming hollow (3) 4. Form o f lobes: length o f lobes ~ width o f lobes (0), lobes longer than wide (1) 5. Width o f lobes: up to 3 mm (0), up to 6 mm (1), up to 12 mm (2) 6. Colour o f upper surface: yellow (0), brown (1), grey (2) 7. Colour o f lower surface: whitish (0), yellow (1), brown (2), black (3) 8. Pseudocyphellae on upper side: absent (0), present (1) 9. Pseudocyphellae on lower side: absent (0), present (1) 10. Form o f pseudocyphellae on lower side: spots (0), lines (1) 11. Cilia: absent (0), present (1) 12. Rhizines: absent (0), present (1) 13. Soredia: absent (0), present (1) 14. Isidia: absent (0), present (1) 15. Structure o f the cortex (orientation of hyphae): both corteces paraplectenchymatous, i.e. hyphae randomly oriented (0), upper cortex palisade plectenchym atous, i.e. hyphae anticlinally oriented (1), both corteces palisade (2), both corteces prosoplectenchym atous, i.e. hyphae parallel to cortex (3) 16. Cell wall thickness (compared to cell lumina): leptodermatous (0), pachydermatous (1) 17. Position o f apothecia: marginal only (0), marginal to submarginal (1), laminal (2), terminal (3) 18. Ascus shape: broadly clavate (0), narrowly clavate (1) 19. Ascus form: melanelia type (0), tuckermannopsis type (1), cetraria type (2) 20. Tholus: small (0), large (1) 21. Axial body: medium [3-5 fxm] (0), broad [>5 jam] (1), narrow [<3 jam] (2), very narrow [<1 jam] (3) 22. Shape o f ascospores: globose to subglobose (0), broadly ellipsoid (1), ellipsoid (2) 23. Length or diameter of ascospores: short [6 fim] (0), long [>6 jim] (1) 24. Position o f pycnidia: marginal only (0), marginal and laminal (1), laminal only (2) 25. Emergency o f pycnidia: emergent (0), immersed (1) 26. Shape of pycnoconidia: bacillariform (0), oblong citriform (1), dumb­ bell shaped incl. disc-bar shaped (2), filiform (3), sublageniform (4) 27. Length o f pycnoconidia: short [<7 jum] (0), medium [7-10 jam] (1), long [>10 jam] (2) 28. Usnic acid: absent (0), present (1) 29. Atranorin: absent (0), present (1) 30. Fatty acids: absent (0), present (1) 31 31. Substance o f fatty acids: lichesterinic-protolichesterinic type acids (0), caperatic acid (1), rangiformic acid (2), unidentified (3) 32. Secalonic acids: absent (0), present (1) 33. Orcinol depsides and depsidones: absent (0), present (1) 34. Substance o f orcinol depsides and depsidones: alectoronic and collatolic acids (0), physodic acid (1), olivetoric acid (2), microphyllinic acid (3), gyrophoric acid (4) 35. (3-orcinol depsidones: absent (0), present (1) 36. substance of P-orcinol depsidones: fumarprotocetraric acid (0), physodalic acid (1) The data matrix and terminal taxa are presented in publication IX. Results. Two series o f separate analyses were carried out using different outgroups (Cetraria fendleri and Flavocetraria cucullata) (IX). W ith C etraria fe n d le ri as outgroup, 36 equally parsim onous trees w ere obtained (heuristic search, all characters w ith equal w eights, length o f trees = 1 2 1 steps, Fig. 5). The strict consensus tree and the 50% majority-rule consensus of 36 trees were also retained. The successive approximations character weighting method produced 108 equally parsimonous trees and, after the second reweightening o f characters, 81 equally parsimonous trees were --------------------------------------------------- ------------------------------------------------------------------ Cetraria fendleri I------------- Ahtiana pallidula I I------- Ahtiana aurescens ____ '------- Ahtiana sphaerosporella --------------------------------------------------------------------------------------------------------- Tuckermannopsis inermis ___ ____________________ ___________________________________________ Tuckermannopsis platyphylla ____ ______________________________________ ____________________ Esslingeriana idahoensis Tuckermannopsis microphyllica Tuckermannopsis ciliaris Tuckermannopsis americana Tuckermannopsis gilva Tuckermannopsis orbata Tuckermannopsis platyphylloides j ------ Tuckermannopsis chlorophylla *------ Tuckermannopsis ulophylloides Tuckermannopsis subalpina Tuckneraria laureri Tuckneraria togashii Tuckneraria pseudocomplicata Tuckneraria ahtii Tuckneraria laxa j ------- Allocetraria globulans '------- Allocetraria oakesiana Allocetraria ambigua Allocetraria stracheyi A llocetrariaflavonigrescens Allocetraria sinensis Allocetraria endochrysea Allocetraria madreporiformis Dactylina arctica Dactylina ramulosa Fig. 5. One of 36 equally parsimonous cladograms (based on characters with equal weights; Cetraria fendleri used to root trees). Length 121, consistency index 0.405, retention index 0.654. 32 obtained (strict consensus tree on Fig. 6). Successive weighting is particularly useful when applied to data sets with much homoplasy (Tehler & Egea 1997). Character reweighting by maximum value o f rescaled consistency indices generated remarkably low weights for some conspicuous morphological characters often used in key-books, such as presence o f isidia and soredia. This is quite acceptable, in our opinion, as these asexual structures are certainly derived but they cannot be treated as shared characters. In other words, we suppose that sorediate and isidiate species did not share sorediate or isidiate ancestors but descended from sexual species instead. The most highly weighted characters are either connected w ith the anatomy o f the thallus (characters 2, 3, 15), anatomy o f the ascocarps (18, 19, 20) or medullary secondary compounds (31, 34, 36). Cetraria fendleri Tuckermannopsis inermis Ahtiana aurescens Ahtiana pallidula Ahtiana sphaerosporella Esslingeriana idahoensis Tuckermannopsis platyphylla Tuckermannopsis microphyllica Tuckermannopsis ciliaris Tuckermannopsis americana Tuckermannopsis gilva Tuckermannopsis chlorophylla Tuckermannopsis orbata Tuckermannopsis platyphylloides Tuckermannopsis ulophylloides Tuckermannopsis subalpina Tuckneraria laureri Tuckneraria togashii Tuckneraria pseudocomplicata Tuckneraria ahtii Tuckneraria laxa Allocetraria globulans Allocetraria oakesiana Allocetraria ambigua Allocetraria stracheyi Allocetraria jlavonigrescens Allocetraria sinensis Allocetraria endochrysea Allocetraria madreporiformis Dactylina arctica Dactylina ramulosa Fig. 6. Strict consensus tree constructed from 108 equally parsimonous cladograms (based on characters reweighted; Cetraria fendleri used to root trees). Length of shortest trees 35 723, consistency index 0.644, retention index 0.800. Some clades, which are seen on one o f the equally parsimonous cladograms of the initial analysis (Fig. 5), are distinct also in the following analyses (Fig. 6). The biggest clade consists o f eight Allocetraria and two Dactylina species. Next clade includes five Tuckneraria species. Four species o f Tuckermannopsis (the so-called T. ciliaris group) also form a separate clade, while the other members o f that genus are solved differently in different analyses. The fourth clade, including three Ahtiana species, is supported by 92% o f trees (characters with equal weights); trees produced by the successive approximations character weighting method always show the same Ahtiana clade. 9 33 The Bremer support test was carried out next. In this test all the trees are kept successively one step longer than the shortest tree, until all the groups are lost in consensus. Bremer’s length difference has also been referred to as the decay test or decay index (Parmasto 1996, Tehler & Egea 1997). In our analysis, inclusion o f trees one step longer than the shortest tree, causes the majority of the tree to collapse into a polytomy, and only the Allocetraria-Dactylina clade remains separately. The latter begins to decay by the Bremer support value of three. With Flavocetraria cucullata as outgroup, 1557 equally parsimonous trees (heuristic search, all characters with equal weights, length of trees = 1 2 3 steps) were obtained. Both strict consensus tree and 50% m ajority-rule consensus (Fig. 7) were saved. The clade of Ahtiana, consisting o f three species, is supported by 78% o f trees. The clade of Allocetraria (including Dactylina), which was strongly supported in the first analysis, collapses into a polytomy. Still, two species of Dactylina form a separate clade with two Allocetraria species (A. endochrysea and A. madreporiformis) in the strict consensus tree. The clade including Tuckneraria species is not supported by this analysis either. One new clade, which consists o f 11 species o f Tuckermannopsis and one species o f Esslingeriana, is composed in all cladograms. When characters were reweighted by maximum value o f rescaled consistency indices, five equally parsimonous trees were obtained. Strict consensus tree (Fig. 8) o f them is essentially different from that achieved by analysis, where characters were with equal weights (Fig. 7); besides, some groups similar to the clades mentioned in the first series of analysis (with Cetraria fendleri as an outgroup), are formed. These are: the clade of three species o f Ahtiana; the clade o f eight species of Allocetraria and two species of Dactylina; the clade o f four species of Tuckermannopsis (T. ciliaris group). Tuckneraria species do not form a clade in this analysis. In addition, analyses were carried out using the parsimony jackknifer pro­ gramme Jac, to identify well-supported monophyletic groups. In Jac, the data are resampled with a jackknifing technique, i. e. in every replicate c. 66% o f the characters are chosen at random, without replacement for parsimony analysis. The resampling procedure can be repeated up to 10 000 times, as it was also done in the present study. The objective of the jackknife method in Jac is the same as that o f bootstrapping in PAUP (Tehler & Egea 1997), but considered much faster than other available techniques. The resulting tree (not presented) shows that all the clades described above are not supported by this very efficient procedure, except for a group consisting of two Dactylina species, and another group comprising Allocetraria globulans and A. oakesiana. 34 Flavocetraria cucullata Ahtiana aurescens Ahtiana pallidula Ahtiana sphaerosporella AUocetraria ambigua Allocetraria flavonigrescens AUocetraria globulans Allocetraria oakesiana Allocetraria sinensis AUocetraria stracheyi Tuckneraria ahtii Tuckneraria laureri Tuckneraria laxa Tuckneraria pseudocomplicata Tuckneraria togashii Allocetraria endochrysea Allocetraria madreporiformis Dactylina arctica Dactylina ramulosa Tuckermannopsis orbata Tuckermannopsis platyphylloides Tuckermannopsis chlorophylla Tuckermannopsis ulophylloides Tuckermannopsis inermis Tuckermannopsis subalpina Esslingeriana idahoensis Tuckermannopsis platyphylla Tuckermannopsis microphyllica Tuckermannopsis ciliaris Tuckermannopsis americana Tuckermannopsis gilva 98 68 100 90 100 , Flavocetraria cucullata Allocetraria ambigua Allocetraria stracheyi Allocetraria globulans Allocetraria oakesiana AUocetraria flavonigrescens Allocetraria sinensis AUocetraria endochrysea Allocetraria madreporiformis Dactylina arctica Dactylina ramulosa Tuckneraria laxa Tuckneraria ahtii Tuckneraria laureri Tuckneraria pseudocomplicata Tuckneraria togashii Ahtiana aurescens Ahtiana pallidula Ahtiana sphaerosporella Tuckermannopsis orbata Tuckermannopsis platyphylloides Tuckermannopsis chlorophylla Tuckermannopsis ulophylloides Tuckermannopsis inermis Tuckermannopsis subalpina Esslingeriana idahoensis Tuckermannopsis platyphylla Tuckermannopsis microphyllica Tuckermannopsis ciliaris Tuckermannopsis americana Tuckermannopsis gilva Fig. 7. Strict consensus tree and 50% majority-rule consensus tree constructed from 1557 equally parsimonous cladograms (based on characters with equal weights; Flavocetraria cucullata used to root trees). Length of shortest trees 123, consistency index 0.415, retention index 0.650. 35 Bootstrapping (500 replicates) was used as well — on the same purpose. Three small groups were supported by this method: Allocetraria globulans A. oakesiana\ Tuckermannopsis americana - T. ciliaris - T. gilva; Allocetraria endochrysea - A. madreporiformis together with the two Dactylinas as a subclade. Flavocetraria cucullata Allocetraria endochrysea Allocetraria madreporiformis Dactylina arctica Dactylina ramulosa Allocetraria sinensis Allocetraria flavonigrescens Allocetraria ambigua Allocetraria stracheyi Allocetraria globulans Allocetraria oakesiana Tuckneraria laxa Tuckneraria ahtii Tuckneraria pseudocomplicata Tuckneraria laureri Tuckneraria togashii Ahtiana aurescens Ahtiana pallidula Ahtiana sphaerosporella Tuckermannopsis orbata Tuckermannopsis platyphylloides Tuckermannopsis ulophylloides Tuckermannopsis inermis Tuckermannopsis subalpina Tuckermannopsis chlorophylla Esslingeriana idahoensis Tuckermannopsis platyphylla Tuckermannopsis microphyllica Tuckermannopsis ciliaris Tuckermannopsis americana Tuckermannopsis gilva Fig. 8. Strict consensus tree constructed from 5 equally parsimonous cladograms (based on characters reweighted; Flavocetraria cucullata used to root trees). Length of shortest trees 43 164, consistency index 0.658, retention index 0.808. Discussion. The aim of the present phylogenetic analysis was to search for the monophyletic groups inside the ingroup and check the correspondence of the present taxonomy to the probable evolution of taxa involved. Originally monotypic genus Ahtiana was segregated from Parmelia s. lat. on the basis o f emergent pycnidia, globose ascospores, leptodermatous cortex, and presence of medullary caperatic acid (Goward 1985). Despite its parmelioid habit with laminal apothecia and pycnidia, Ahtiana sphaerosporella was shown to be closely allied to Cetraria pallidula. Today the genus includes three species. In our present analysis, it appears as a separate clade in most of the cladograms of characters with equal weights and also in the successive weighting strict consensus trees o f both series o f analyses (with different outgroups). This fact supports the latest changes in the system atics o f the genus — transferring C. pallidula and C. aurescens to the originally monotypic Ahtiana (VIII). 36 The genus Allocetraria, at first including only three species from high altitudes in south-east Asia, was introduced by Kurokawa & Lai (1991). It was separated from Cetraria because of the dichotomously branched lobes, the special appearance o f pseudocyphellae, the palisade plectenchymatous cortex, and on the unique chemistry. The authors did not pay attention to ascomatal and pycnidial characters. Later studies o f these structures confirmed the necessity of a separate genus (VII). Today, ten species are combined in the genus; eight of them were included in our analyses. We have not seen the herbarium material of two taxa (A. denticulata and A. isidiigera), and their descriptions (Hue 1899; Kurokawa & Lai 1991) are too poor to present them properly in the data matrix. The value o f these taxa is rather uncertain — both species are known from the type localities only; in addition, they are sterile according to literature. The paraphyletic nature o f the genus with respect to two species o f Dactylina is obvious. The closeness of these two genera was noticed only recently when two former Dactylinas — A. madreporiformis and A. endochrysea — were transferred to Allocetraria (Kamefelt & Thell 1996). Presence of filiform pycnoconidia and asci with extremely broad axial body are the essential characters for separating Allocetraria. Dactylina is distinguished by the radialsymmetrical thallus becoming hollow, the terminal position o f apothecia, and also the secondary chemistry. The palisade plectenchymatous arrangement o f cortical hyphae, which is rather unusual in the group of cetrarioid lichens, is characteristic of both Allocetraria and Dactylina. The splitting o f the species involved into two separate genera is not supported by our present study. At the same time, the analyses based on morphological, anatomical, and chemical characters only do not show enough confidence when using more severe methods, such as jackknifing or bootstrapping. We share the opinion recently approved by the symposium on taxonomy, evolution and classification o f lichens and related fungi (January 9-11, 1998, London), that on such occasions quick changes in nomenclature are not advisable. At the present stage o f lichenological studies, additional phylogenetic analyses using the modem molecular data should be carried out, before proposing extensive nomenclatural changes. Tuckneraria includes five species. M ost o f them were transferred from Nephromopsis to the newly described genus because o f im portant ana­ tom ical characters (globose ascospores, Tuckerm annopsis-type asci, small axial body etc.) (VI). Today the genera Nephromopsis and Tuckneraria are even considered to represent different evolutionary lines (Thell 1996) of cetrarioid lichens. The idea of close affinities o f Tuckneraria and Ahtiana has also been proposed (VIII). The monophyletic origin o f the genus Tuckneraria is supported in one series of our analyses (Cetraria fendleri used to root trees). In the other series {Flavocetraria cucullata used to root trees) the species of Tuckneraria do not form a separate clade but are branched out successively. Genus Tuckermannopsis was described by Gyelnik (1933) in a very short manner: “Affinis generi Nephromopsi sed thallus subtus pseudocyphellis 10 37 deficientibus”. Today much more is known about the genus but the correct description is still not presented. According to different authors (Lai 1981, Hale in Egan 1987, Kurokawa 1991, Weber in Egan 1991) various species have been transferred to Tuckermannopsis', many of them have later been combined again into other genera {Ahtiana, Allocetraria, Kaernefeltia, Melanelia, Vulpicida). At present it is generally accepted that globose ascospores in narrowly clavate asci, large axial body, dumb-bell shaped pycnoconidia, and moderately small foliose brown to greenish thallus (absence of usnic acid in the cortex) are the important characters in delimiting the genus. Eleven species are now recognized in Tuckermannopsis. Our phylogenetic analyses reveal further problems within this taxon. Monophyletic origin can be declared only for the so-called Tuckermannopsis ciliaris group (T. ciliaris is also the type species o f the genus). The clade consisting of four close species is strongly supported in all parsimonous trees. On the whole, the genus Tuckermannopsis, in its generally accepted treatment, should be considered paraphyletic. For instance, E. idahoensis, the sole species of the genus Esslingeriana, is predominately connected with Tuckermannopsis platyphylla and this pair o f species always branches out next to Tuckermannopsis ciliaris group. Other members o f the genus do not form a distinguished group. In our opinion, any further taxonomical rearrangements in this genus are not justified before additional — preferably molecular — research has been carried out. 38 CONCLUSIONS 1. The group o f cetrarioid lichens (fam. Parmeliaceae, lichenized Ascomycota), comprising 131 species in 22 genera, is polyphyletic. Three evolutionary lines, based mainly on reproductive structures and anatomical characters, have been recognized for the group. Phylogenetic relationships within some cetrarioid genera representing different evolutionary lines have been treated in more detail. 2. Chemosystematical and geographical studies in genus Asahinea reveal that a possible centre o f speciation o f the genus is in the Russian Far East or Japan. Three chemotypes have been recognized in Asahinea chrysantha: I — atranorin, alectoronic acid, and a-collatolic acid; II — atranorin, alectoronic acid, a-collatolic acid, and usnic acid; III — atranorin, alectoronic acid, and usnic acid. Chemotype I of A. chrysantha is the same as that o f A. scholanderi’, this is supposed to represent the basic, primitive chemistry o f the genus. 3. A type o f chemical species concept has traditionally been accepted in genus Cetrelia. We propose to treat every species in this genus as a combination o f morpho- and chemotypes, and present an appropriate table where vacant squares mark the species that are theoretically possible in the genus. As a result, two new species — Cetrelia orientalis Randlane & Saag and C. pseudocollata Randlane & Saag — are described. The role of “primary” and “secondary” species in the systematics o f the genus according to the “species-pairs” theory is discussed. 4. Altogether 16 species from the genera Cetreliopsis, Nephromopsis, and Cetrariopsis are thoroughly characterized and compared. In our opinion two of these taxa — Nephromopsis and Cetreliopsis — are separate, although closely related units, while the species o f Cetrariopsis are transferred to the genus Nephromopsis. One new species (Cetreliopsis papuae Randlane & Saag) is described. 5. The group o f cetrarioid lichens comprising six genera (Ahtiana, Allocetraria, Dactylina, Esslingeriana, Tuckneraria, and Tuckermannopsis), characterized by subglobose to globose ascospores in narrowly clavate asci, is phylogenetically analysed. Genus Tuckneraria is originally described and two other genera — Ahtiana and Allocetraria — are thoroughly revised. Cladistic analyses refer to further necessary changes in the systematics o f the group. 6. The present state o f taxonomy in the heterogenous group o f cetrarioid lichens is far from being final. Splitting o f big genera and describing o f several new small genera has been the obvious tendency in systematics o f lichens during the last decade. This process has caused difficulties when using the nomenclature and also objections in principle. We support the opinion that the main disagreement in this debate is not the grouping o f taxa but their ranking. At the present stage o f lichenological studies, additional phylogenetic analyses using molecular data should be carried out, before proposing extensive nomenclatural changes. 39 REFERENCES Acharius, E. 1803. Methodus lichenum. Stockholm. Acharius, E. 1810. Lichenographia universalis. Gottingen. Asahina, Y. 1953. Lichenes Japoniae novae vel minus cogitae 11. — Journal of Japanese Botany 28: 6-12. Awasthi, D. D. 1987. a new position for Platysma thomsonii Stirton. — Journal of the Hattori Botanical Laboratory 63: 367-372. Bowler, P. A. & Rundel, P. W. 1975. Reproductive strategies in lichens. — Botanical Journal of Linnean Society 70: 325-340. Culberson, C. F. 1972. Improved conditions and new data for the identification of lichen products by a standardized thin-layer chromatographic method. — Journal of Chromatography 72: 123-125. Culberson, C. F. 1974. Conditions for the use of Merck silica gel 60 F 254 plates in the standardized thin-layer chromatography technique for lichen products. — Journal of Chromatography 97: 107-108. Culberson, C. F. & Culberson, W. L. 1976. Chemosyndromic variation in lichens. — Systematic Botany 1: 325-329. Culberson, C. F., Culberson, W. L. and Johnson, A. 1981. A standardized TLC analysis of fl-orcinol depsidones. — Bryologist 84: 16-29. Culberson, C. F. & Kristinsson, H. 1970. A standardized method for the identification of lichen products. — Journal of Chromatography 46: 85-93. Culberson, W. L. & Culberson, C. F. 1965. Asahinea, a new genus in the Parmeliaceae. — Brittonia 17: 182-190. Culberson, W. L. & Culberson, C. F. 1968. The lichen genera Cetrelia and Platismatia (Parmeliaceae). — Contributions from the United States National Herbarium 34: 449-558. Culberson, W. L. 1986. Chemistry and sibling speciation in the lichen forming fungi: ecological and biological considerations. — Bryologist 89: 123-131. Egan, R. S. 1987. A fifth checklist of the lichen-forming, lichenicolous and allied fungi of the continental United States and Canada. — Bryologist 90: 77-173. Egan, R. S. 1991. Changes to the “Fifth checklist of the lichen-forming, lichenicolous and allied fungi of the continental United States and Canada”. — Bryologist 94: 396-400. Elix, J. A. 1993. Progress in the generic delimitation of Parmelia sensu lato Lichens (Ascomycotina: Parmeliaceae) and a synoptic key to the Parmeliaceae. — Bryologist 3: 359-383. Eriksson, O. E. & Hawksworth, D. L. 1988. Notes on ascomycete systematics. Nos. 733-803. — Systema Ascomycetum 7: 103-117. Esslinger, T. 1971. Cetraria idahoensis, a new species of lichen endemic to western Norh America. — Bryologist 74: 364-369. Farris, J. S. 1995. Guide to the parsimony Jackknifer, version 4.22. Computer program distributed by the Natural History Museum, Stockholm. 40 Gao, X. 1991. Studies in species of the lichen genus Asahinea. — Nordic Journal of Botany 11: 483-485. Goward, T. 1985. Ahtiana, a new genus in the Parmeliaceae. — Biyologist 88: 367-371. Gyelnik, V. 1933. Lichenes varii novi critique. — Acta pro Fauna et Flora Universalis, Ser. 2,1: 3-10. Hafellner, J. 1984. Studien in Richtung einer natiirlichen Gliederung der Sammelfamilien Lecanoraceae und Lecideaceae. — Beiheft 79 zur Nova Hedwigia. Festschrift J. Poelt, 241-371. Hale, M. E. 1976. Lichen structure viewed with scanning electron microscope. — In: D. H. Brown et al (eds.) Lichenology: progress and problems. London, Academic Press, 1-15. Hawksworth, D. L. 1974. Mycologist’s handbook. Kew, Commonwealth Mycological Institute. Hawksworth, D. L. & Hill, D. J. 1984. The lichen forming fungi. Glasgow, Blackie. Hue, A.-M. 1899. Lichenes extra-europaei. — Nouvelles Archives du Museum (Paris) I, 4: 27-220. Kamefelt, I. 1977. Masonhalea, a new genus in the Parmeliaceae. — Botaniska Notiser 130: 101-107. Kamefelt, I., Mattsson, J.-E. & Thell, A. 1992. Evolution and phylogeny of cetrarioid lichens. — Plant Systematics and Evolution 183: 113-160. Karnefelt, I., Mattsson, J.-E. & Thell, A. 1993. The lichen genera Arctocetraria, Cetraria and Cetrariella (Parmeliaceae) and their presumed evolutionary affinities. — Bryologist 96: 394-404. Kamefelt, I. & Thell, A. 1992. The evaluation of characters in lichenized families, exemplified with the alectorioid and some parmelioid genera. — Plant Systematics and Evolution 180: 181-204. Kamefelt, I. & Thell, A. 1996. A new classification for the Dactylina/Dufourea complex. — Nova Hedwigia 91: 595-605. Kamefelt, I., Thell, A., Randlane, T. & Saag, A. 1994. The genus Flavocetraria Karnefelt & Thell (Parmeliaceae, Ascomycotina) and its affinities. — Acta Botanica Fennica 150: 79-86. Kondratyuk, S., Randlane, T., Saag, A. & Oxner, A. 1993. Genus Cetrelia W. Culb. et C. Culb. — In: A. Oxner, Flora of the Lichens of Ukraine 2. Kiev, Naukova Dumka, 214-221 (in Ukrainian). Krivoschchekova, O. E., Stepanenko, L. S., Mischenko, N, P.& Maksimov, O. B. 1983. Aromatic metabolic substances of lichen fam. Parmeliaceae I. Depsidones. — Khimija prirodnyh sojedinenii 1: 13-19 (in Russian). Krog, H. 1962. A contribution to the lichen flora of Alaska. — Arkiv fur Botanik 4: 489-513. Kurokawa, S. 1980. Cetrariopsis, a new genus in Parmeliaceae, and its distribution. — Memoirs of the National Science Museum (Tokyo) 13: 139-142. Kurokawa, S. 1991. Japanese species and genera of the Parmeliaceae. — Journal of Japanese Botany 66:152-159. 41 Kurokawa, S. & Lai, M.-J. 1991. Allocetraria, a new genus in the Parmeliaceae. — Bulletin of the National Science Museum (Tokyo), Ser. B, 17: 59-65. Lai, M-J. 1981 [1980]. Studies on the cetrarioid lichens in Parmeliaceae of east Asia. I. — Quarterly Journal of the Taivan Museum 33: 215-229. Linnaeus, C. 1753. Species Plantarum 2. Stockholm. Llano, G. A. 1951. A contribution to the lichen flora of Alaska. — Journal of the Washington Academy of Sciences 41: 196-200. Maddison, W. P. & Maddison, D. R. 1992. MacClade: Analysis of Phylogeny and Character Evolution, version 3. Sinauer Associates, Sunderland. Muller Argoviensis, J. 1891. Lichenologische Beitrage 35. — Flora 74: 371-382. Nylander, W. 1860. Synopsis methodica lichenum. I. 2. Paris. Oxner, A. N. & Rassadina, K. A. 1960. Ad genus Cetraria ex URSS novitates. — Notulae Systematicae et Sectione Cryptogamica Instituti Botanici nomine V. L. Komarovii AcademiaeScientiarum URSS 13: 5-14 (in Russian). Parmasto, E. 1996. Biostistemaatika teooria ja meetodid. Tartu. Poelt, J. 1970. Das Konzept der Artenpaare bei der Flechten. — Deutsche Botanische Gesellschaft. Neue Folge 4: 187-198. Poelt, J. 1972. Die taxonomische Behandlung von Artenpaaren den Flechten. — Botaniska Notiser 125: 77-81. Randlane, T. & Saag, A. 1991a. Chemical variation and geographical distribution of Asahinea chrysantha. — In: N. S. Golubkova (ed.), The problems of experimental lichenology in the USSR. Leningrad, 58-65 (in Russian). Randlane, T. & Saag, A. 1991b. Some chemosystematical data about the lichen genus Nephromopsis in the USSR. — Folia Cryptogamica Estonica 28: 26-30 Randlane, T. & Saag, A. 1992a. Additional data about genus Nephromopsis (Lichenes, Parmeliaceae). — Mycotaxon 44(2): 485-489. Randlane, T. & Saag, A. 1992b. New combinations of some cetrarioid lichens (Parmeliaceae). — Mycotaxon 44(2): 491-493. Randlane, T., Saag, A. 1992c. Genus Cetrelia Culb. et Culb. in URSS. — Novitates Systematicae Plantarum non vascularium 28: 118-133 (in Russian). Randlane, T. & Saag, A. 1992d. Tuckermannopsis americana contra Cetraria ciliaris in Russia. — Folia Cryptogamica Estonica 29: 33-36. Randlane, T. & Saag, A. 1993. World list of cetrarioid lichens. — Mycotaxon 47:395 - 403. Randlane, T. & Saag, A. Changes in systematics of cetrarioid lichens. Sauteria. (Submitted.) Randlane, T., Saag, A. & Kondratyuk, S. 1992. Genus Cetrelia Culb. et Culb. in the Ukraine. — Ukrainian Botanical Journal 48(1): 41-44 (in Ukrainian). Randlane, T., Saag, A. & Thell, A. 1997. A second updated world list of cetrarioid lichens. — Bryologist 100(1): 109-122. Rasanen, V. 1952. Studies on the species of the lichen genera Cornicularia, Cetraria and Nephromopsis. — Kuopion Luonnon Ystavain Yhdistyksen Julkaisuja B 2: 1-53. 42 Rassadina, K. A. 1950. Cetraria in the U.S.S.R. — Plantae Cryptogamae 5: 171-304 (in Russian). Swofford, D. L. 1991. PAUP: Phylogenetic analysis using parsimony, version 3.0s. Computer program distributed by the Illinois History Survey. Swofford, D. L. 1993. PAUP: Phylogenetic analysis using parsimony, version 3.1.1. Computer program distributed by the Illinois Natural History Survey. Tehler, A. 1982. The species pair concept in lichenology. — Taxon 31:708-714. Tehler, A. & Egea, J. M. 1997. The phylogeny of Lecanactis (Opegraphaceae). — Lichenologist 29: 397-414. Thell, A. 1995. A new position of the Cetraria commixta group in Melanelia (Ascomycotina, Parmeliaceae). Nova Hedwigia 60: 407-422. Thell, A. 1996. Anatomy and taxonomy of cetrarioid lichens. Summary of doctoral dissertation. Department of Systematic Botany, Lund University. Trass, H. 1992. A new species of Asahinea ( Ascomycotina, Parmeliaceae) — Folia Cryptogamica Estonica 29: 31-32. Tuckerman, E. 1882. A synopsis of North American lichens. I. Boston. Yoshimura, I. 1979. Lichen flora of Japan in colour. Osaka, Hoikusha Publishing Co. White, F. I. & James, P. W. 1985. A new guide to microchemical techniques for the identification of lichen substances. — British Lichen Society Bulletin 57 (suppl.): 1-41. 43 EVOLUTSIOONILISED SEOSED MONEDES TSETRARIOIDSETES PEREKONDADES (LIHHENISEERUNUD KOTTSEENED) Kokkuvote Tsetrarioidsete samblike ruhm (suguk. Parmeliaceae, lihheniseerunud kottseened), mis sisaldab praegu teadaolevatei andmetel 131 liiki 22 pere-konnast, on uldtunnustatult polufuleetiline. Vaadeldavas riihmas on taheldatud kolme evolutsioonilist suunda, mille piiritlemine pohineb eelkoige samblike anatoomilistel tunnustel ja viljakehade siseehitusel. Kaesolevasse doktoritoosse on koondatud uurimused, mis kasitlevad uksikasjalikumalt fulogeneetilisi seoseid jargmistes tsetrarioidsetes samblikuperekondades: Ahtiana, Allocet­ raria, Asahinea, Cetrelia, Cetreliopsis, Dactylina, Esslingeriana, Nephro­ mopsis, Tuckneraria ja Tuckermannopsis. Kemosiistemaatiline ja geograafiline analiiiis voimaldab teha oletusi perekonna Asahinea liikide tdenaose tekkekeskme kohta. Liigis A. chrysantha on maaratud kolm kemotuupi: I kemotliup sisaldab atranoriini, alektoroonhapet ja a-kollatoolhapet; II — atranoriini, alektoroonhapet, a-kollatoolhapet ja usniinhapet; III — atranoriini, alektoroonhapet ja usniinhapet. A. chrysantha I kemotiiup esindab sama samblikuainete komplekti, mis esineb ka liigis A. scho­ landeri. Lahtudes samblikuainete keemilisest koosseisust ja nende tekke biosiinteesi radadest, voib vaita, et see kemotuiibi keemiline koostis on vaadeldavas perekonnas iirgseim. Vottes arvesse eri kemotuupide levikut, tuleb perekonna Asahinea liigitekke tsentriks pidada Venemaa Kaug-Ida voi Jaapanit. Perekonna Cetrelia liikide puhul on samuti oluliseks tunnuseks sambliku­ ainete koosseis. Kaesolevas toos vaadeldakse Cetrelia liike kui kemo- ja morfotiiupide kombinatsioone ning esitatakse vastav tabel, mille tiihjad ruudud viitavad teoreetiliselt voimalikele liikidele selles perekonnas. Tabelist lahtudes kirjeldataksegi kaks teadusele uut liiki. Kasitletakse ka liikide paaride teooriat ning selle rakendamise tulemusi perekonnas Cetrelia. Esitatakse iiksikasjalik tilevaade perekondade Cetreliopsis, Nephromopsis ja Cetrariopsis liikidest. Tunnuste vSrdlemisel selgub, et pole olulisi erinevusi perekondade Nephromopsis ja Cetrariopsis vahel (v.a. apoteetsiumite erinev asend tallusel). Seetottu ei peeta vajalikuks perekonna Cetrariopsis tunnustamist iseseisva taksonina. Perekondi Cetreliopsis ja Nephromopsis kasitletakse erinevate, kuid fulogeneetiliselt lahedaste riihmadena. Morfoloogiliste, anatoomiliste ja keemiliste tunnuste alusel analiiiisitakse kladistiliselt ruhma, kuhu kuulub kuus perekonda {Ahtiana, Allocetraria, Dactylina, Esslingeriana, Tuckneraria ja Tuckermannopsis), et anda hinnang selle ruhma praegusele taksonoomiale. Selgub, et vajalikud on susteemi ja nomenklatuuri taiendavad muudatused, kuid neid ei peeta voimalikuks enne molekulaarseid uuringuid. Seniste uurimuste kaigus on kirjeldatud iiks uus perekond: Tuckneraria Randlane & Thell, ning neli uut liiki: Cetrelia orientalis Randlane & Saag, C. pseudocollata Randlane & Saag, Tuckneraria ahtii Randlane & Saag, Cetreliopsis papuae Randlane & Saag. Samuti on esitatud mitu uut kombinatsiooni. 44 AKNOWLEDGEMENTS I wish to express my gratitude to my colleague and supervisor Tiina Randlane, who has initiated this study and shared all the difficulties in earring out the research. I appreciate highly the contribution of my second co-author and good friend Arne Thell from the University o f Lund whose original studies in anatomy of cetrarioid lichens has led to essential changes in the systematics of this group. Special thanks are due to Erast Parmasto for the help with cladistic analyses. I would like to acknowledge the group o f lichenologists in Tartu as well as the whole staff o f the Institute o f Botany and Ecology, University of Tartu, for the working conditions and general support. Many lichenologists are recognized for their consultations, discussions, and improvements to manuscripts: Teuvo A M (Helsinki), William Culberson (Durham, USA), Ingvar Kamefelt (Lund), Ming Jou Lai (Taiwan), Jan-Eric Mattsson (Uppsala), Anders Tehler (Stockholm), Josef Poelt (Graz). Yuri Kotlov and Mikhail Zhurbenko (St. Petersburg), Roland Moberg (Uppsala), Harrie Sipman (Berlin), Roland Skyten (Helsinki) and Goran Thor (Stockholm) are thanked for kindly arranging study visits to their herbaria. I am also deeply grateful to Gertrud Dahlgren, Cecilia Lonnqvist and Asta and Sture Thell for their permanent hospitality during our visits to Lund and Eslov. Triin Randlane is thanked for revising the English text. This study has been supported financially by the Estonian Science Foundation (grant 401), International Science Foundation (grants LCZ 000 and LLO 100) and the Swedish Institute. 12 45 PUBLICATIONS I 13 Saag, A. & Randlane, T. 1995. Phylogenetic affinities o f cetrarioid lichens. — Cryptogamic Botany 5(2): 128-136. C rypt. Bor. 5, 1 2 8 - 1 3 6 (1995) CryptogamicBotany Phylogenetic affinities of cetrarioid Lichens Andres Saag and Tiina Randlane Institute of Botany and Ecology, Tartu University, Lai Street 38, Tartu, Estonia EE-2400 SUMMARY O f about 120 known cetrarioid lichen species, 83 were chosen for cladistic analysis to evaluate the present systematic arrangement of these taxa. 42 classically accepted morphological, anatomical and chemical characters were examined using original and literature data. According to the analysis, several recently described cetrarioid genera (e.g. Cetrelia, Platismatia, Masotthalea, Cetrariopsis, Vulpicida) appear to be monophyletic and thus acceptable. Species of Nephromopsis are very closely related to the monospecific genus Cetreliopsis. Neither the brown fruticose Cetraria species group nor the genus Tuckermannopsis could be treated as monophyletic. Introduction The genus Cetraria has been since its description by E. Acharius in 1803 a heterogeneous assemblage of poorly related taxa. From eight species originally included in Cet­ raria, now only the type species - C. islandica —has been left in the genus. C. cucullata and C. nivalis are proposed to be included in Allocetraria [17]; C. lacunosa, C. glauca and C, fallax represent species of Platismatia [6]; C. sepincola belongs to the poorly defined genus Tuckermannopsis [7]; and C. juniperina is a member of the newly described Vulpicida [12]. In the contemporary taxonomy of iichenized fungi, about 120 species are known as cet­ rarioid [16], i.e. Cetraria-like. While the original delimita­ tion of the genus Cetraria is insufficient, it is still more complicated to define the colloquial term “cetrarioid”. The total list of cetrarioid lichens [18] presents all species that ever have been included in the genus Cetraria, and the allied taxa. The list serves as a starting point for the pre­ sent paper. It is evident that the group of species under discussion is not: monophyletic. 14 different genera have been newly described or resurrected to accommodate the species [18]. Still, the present systematic arrangement of these taxa is not convincing in all aspects. The objective of the present paper was to evaluate the traditional classifica­ tion of this group on the basis of cladistic analysis and get new ideas for further studies. © 1995 by G ustav Fischer V erlag, S tu ttg art Material and Methods The data matrix for the cladistic analysis was compiled on the basis of both literature (mainly on anatomical and partly chemi­ cal characters) and original studies (morphological and chemical data). Although the reliability of the information in the literature may be doubtful in some cases, it was not practical to investigate all the characters of all species newly. Herbarium material has been examined from H , KW , LD, LE, TB, TU and UPS using microscope MBS-9. Chemical analyses were performed according to the standardized TLC methods [3, 4]. Cladistic analyses were carried out using the program PAUP version 3.0s [19] on a Macintosh llci personal computer. The following option settings were used: character-state optimization - DELTRAN; MULPARS; M A X T R E E = 1000, 3900; heuristic search; trees rooted using outgroup method; for functional outgroup rooting a mono­ phyletic sister group used; addition sequence -simple; 3 trees held at each step during stepwise addition; tree-bisection-reconnection (TBR) branch-swapping performed; characters reweighted by maximum value of rescaled consistency indices. Strict consensus trees of both original data based cladograms and after characters reweighted were retained. Three analyses were carried out independently on the same data basis which was arranged differently for each analysis. In the first analysis the major secondary compounds of lichens were treated as separate characters (characters 26, 32 and 39 exclud­ ed). In the second analysis the lichen substances were not rep­ resented as independent characters (characters 27-29, 33-38, 40—42 exluded) but grouped into bigger units (fatty acids, orcinol series products, (3-orcinol series products) according to their 0 9 3 5 -2 1 4 7 /9 5 /0 2 1 6 -0 2 2 4 $ 3,50/0 Phylogeny o f c etrario id lichens • 129 Table 1. 1.) Data matrix and terminal taxa. 000000000111111111122222222223333333333444 123456789012345678901234567890123456789012 Ahtiana sphaerosporella Allocetraria ambigua A. cucullata A. nivalis A. stracheyi Asahinea chrysantha A. scholanderi Cetraria agnata C. andrejevii C. arenaria C. caiifornica C. commixta C. delisei C. ericetorum C. fastigiata C. inermis C. islandica C. kamczarica C. laevigata C. nigricans C. nigricascens C. odontella C. rassadinae C. subalpina C. weberi Cetrariopsis wallichiana Cetrelia braunsiana C. cetrarioides C. chicitae C. collata C. davidiana C delavayana C. japonica C. monachorum C. nuda C. olivetorum C. pseudolivetorum C. sanguinea Cetreliopsis rhytidocarpa Esslingeriana idahoensis Masonhalea richardsonii Nephromopsis asahinae N. ectocarpisma N. endocrocea N. endoxanthoides N. globulans N. komarovii N . laureri N. ornata N . pallescens N. pseudocomplicata N . rugosa N . stracheyi N. yunnanensis Parmelaria thomsonii Platismatia erosa P. formosana P. glauca P. herrei P. lacunosa P. norvegica P. stenophylla 000000100100101011010001011000000000000000 2010001000101100001? ? ? ?1010010000000000000 201100101010110000110001010011000000000000 200100100010110000110001000000000000000000 201000101010101010100011000011000000000000 110000000110210110100101100000010010000000 looooioooucuoiiiiooiooiooooooiooiooooooo 010000100110110111010000000000000000000000 200100001010210010110100010100000000000000 201100001010110000100100010010000000000000 310000010010110000110000000000000000000000 001000101010110110101000000000010010000000 200100000010110000110000000000010000100000 201000001010110000100100010010000000000000 200100000010110000110000000000010000100000 101000100010101010100100010010000000000000 201100001010210000100100010010000000001100 2000000000100100001?0?00010010000000000000 201000001010110000100100010010000000001100 200000011010110000100100010010000000000000 200000011010110000110000010100000000000000 201000011010010010110000010010000000000000 201100001010110000100100010010000000000000 100000001010101010100100010010000000000000 000001100110110011110000000000010001000000 10010010010001010? ? ? ? ? ?1010010010010000000 110001100100010120110000100000010010000000 110110100100010120110000100000011000000000 110110100100110120110000100000010010000000 110100100100210120110000100000011000000000 110100100100210121110000100000010001000000 110100100100110120110000100000011000000000 110101100100110120110000100000010000010000 110110100100010120110000100000011000000000 110100100100210i 11 110000100000010010000000 110110100100110120110000100000010001000000 110101100100210120110000100000010001000000 110100100100210120100100100000010100000000 1101101010011101101? ?? P1010010000000001100 100000100110101011110000110001000000000000 200100000010110010100100000000010010000000 1101001010011101111??? ?1000000000000001100 1001001000011100001? ? ? ?1010000000000000000 1001001010011100001100000100U1000000000000 1101001010011100101????0010011000000001100 1001001010011010101????1010011000000000000 10010010100111000? ? ? ? ? ?1010010000000001000 100110100001110000110001010010000000000000 1001001010012010101? ? ? ?1000001000000001100 1001001000010100001? ? ? ?1010010000000000000 1001001010012100001? ? ? ?1000000010010000000 1001001010012100011? ? ? ?1000000010001000000 100100100001210000110001000U00010101000000 n01001010011100011????1010010000000000000 100000110100210220100100100000010010000000 110101100010110000101000111000000000000000 100100100010110000101000111000000000000000 100011100010111000101000111000000000000000 100001100010111010101000111000000000000000 100000100010210001101000111000000000001100 110001100010210000101000111000000000000000 100000100010111010101000111000000000000000 130 • A ndreas Saag and T im a R an d lan e OOOOOOOOO1 1 1 1 1 1 1 1 1 1 2 2 2 2 2 2 2 2 2 2 3 3 3 3 .3 3 3 3 3 3 4 4 4 123456789012345678901234567890123456789012 P. tucketm anni Tuckerm annopsis americana T. aurescens T. chlorophylla T. ciliaris T. fcndleri T. hep3tizon T. merrillii T. microphyllica T. oakesiana T. orbata T. pallidula T. platyphylla T. platyphylloides T. sepincola Vulpicida canadensis V. juniperinus V. pinastri V. tilesii V. tubulosus V. viridis 1 0 0 0 0 0 1 0 0 1 1 0 1 1 0 0 1 1 1 0 1 0 0 0 1 1 lOOOOOOOOO'OOOOOO 100000111010101011110000100000010010000000 1000001000101010111? ? ? ?1011000000000000000 100011100010101010110000010010000000000000 100000111010101011110000100000010001000000 000000100010110010110000010010000000000000 001000101110110111110000000000000000001011 100000100110110011110000000000000000000000 100000111010101010100100100000010000010000 1000101000101010101?? ??1011001000000000000 100000111010101011110000010010000000000000 1000001000101010101????1011000000000000000 10000110001010101 1 110000110010000000000000 100000101010110001110000010010000000000000 100000100010110000110000010010000000000000 1 0 0 0 0 0 1 0 0 1 1 0 1 10 0 1 1 0 0 1 0 0 1 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 10 0 0 0 0 10 0 0 10 1 10 0 1 110 10 0 1 0 0 0 0 0 0 10 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 1 0 1 1 0 0 10 10 1 0 0 10 0 0 0 0 0 10 0 0 0 0 0 0 0 0 0 0 200000101010110010101001000000100000000000 2 ,3 0 0 0 0 0 1 0 1 0 1 0 1 1 0 0 1 1 1 0 1 0 0 1 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 100000100110110011101001000000100000000000 chemical structure and possible biochemical pathways. Data mat­ rix for these too analysis is presented in table 1. In the third analysis som e characters treated as binary at first {2 -3 , 3—4, 10—12, 1 4 -1 5 , 1 8 -1 9 , 2 0 -2 3 ) were coded now as multistate while the chemical characters were arranged as in the second analysis. Taxa analysed All the cetrarioid lichens, about 120 species, were taken into account at first. Then 10 species were excepted because o f insuffi­ cient inform ation. A lso, 28 species which are sterile or for which reproductive structures have not been sufficiently studied were also excluded from the further work. Finally, the follow ing 83 cetrarioid lichen species were included in the cladistic analysis as terminal taxa o f the ingroup: A htia n a sphaerosporella, A llo cet­ raria am bigua, A . cucullata, A . nivalis, A . stracheyi, Asahinea chrysantha, A . scholanderi, Cetraria agnata, C. andrejevii, C. arenaria, C. californica, C. c o m m ix ta , C. delisei, C. ericetorum , C. fastigiata, C. inerm is, C. islandica, C. kam czatica, C. laeviga­ ta, C. nigricans, C. nigricascens, C. odontella, C. rassadiiuie, C. subalpina, C. weberi, C etrariopsis wallichiana, Cetrelia braunsiana, C. cetrarioides, C. chicitae, C. collata, C. davidiana, C. delavayana, C. japonica, C. m on a ch o ru m , C. nuda, C. o livetorum , C. pse udolivetorum , C. sanguinea, Cetreliopsis rhytidocarpa, E s­ slingeriana idahoensis, M asonhalea richardsonii, N ep h ro m o p sis asahinae, N . ectocarpism a, N . endocrocea, N . e n d o x a n th o id es, N . globulans, N . ko m a ro v ii, N . laureri, N . ornata, N . pallescens, N . pseudocom plicata, N . rugosa, N . stracheyi, N . yunnanensis, Parmelaria th o m so n ii, P latism atia erosa, P. fo rm o sa n a , P. glauca, P. herrei, P. lacunosa, P. norvegica, P. stenophylla, P. tuckerm anni, T u ckerm annopsis am ericana, T . aurescens, T. chlorophylla, T. ciliaris, T. fendleri, T, h e p atizo n , T . m errillii, T . m icro p h yl­ lica, T. oakesiana, T. orbata, T. pallidula, T. pla typ h ylla , T. p la ­ typhylloides, T. sepincola, Vulpicida canadensis, V. juniperinus, V. pinastri, V. tilesii, V. tubulosus, V. viridis. 14 Table 1. (2.) Characters In all, 42 morphological, anatomical and chemical characters, traditionally in wide use, were chosen for the analysis of cet­ rarioid lichens. Still, some of the common characters applied usu­ ally in handbooks of macrolichens (e.g. colour of the thallus, shape and arrangement of lobes etc.) were not used here because of difficulties of defining them properly. Multistate characters were treated as unordered except the characters of numeral measurements (nos. 13, 16, 17) which were ordered. I THALLUS STRUCTURES 1. Thallus habit a) foliose thallus closely adnate to the substrate (0) b) foliose thallus loosely adnate to the substrate (1) c) foliose erect thallus (2) d) fruticose thallus (3) 2. Pseudocyphellae on upper cortex (absent-0, present-1) 3. Pseudocyphellae marginal (absent-0, present-1) 4. Pseudocyphellae on lower cortex (absent-0, present-1) In the third analysis the characters 2-4 are coded as multis­ tate characters pseudocyphellae on upper cortex (absent-0, laminal-1, marginal-2) and pseudocyphellae on lower cortex (absent-0, laminal-1, marginal-2). 5. Soredia (absent-0, present-1) 6. Isidia (absent-0, present-1) 7. Rhizines (absent-0, present-1) 8. Cilia (absent-C, present-1) 9. Marginal projections (absent-0, present-1) II APOTHECIA A N D ASCOSPORES 10. Apohtecia lamina! on the upper surface (absent-0, present-1) 11. Apothecia marginal on the upper surface (absent-0, present-1) 12. Apothecia on the lower surface (absent-0, present-1) In the third analysis the characters 10-12 are connected to Phylogeny o f c etrario id lichens • 131 13. 14. 15. 16. 17. one multistate character (apothecia laminal on the upper surtace-0, apothecia marginal on the upper surface-1, apo­ thecia on the lower surface-2). Diameter of the apothecia a) d < 2 mm (0) b) 2 mm < d < 10 mm (1) c) d > 10 mm (2) Spores ellipsoid (absent-0, present-1) Spores globose (absent-0, present-1) In the third analysis the characters 14,15 are connected to one multistate character (spores ellipsoid-0, globose-1). Medium length of the spores a) 1 < 8 um (0) b) 8 um < 1 < 20 urn (1) c) 1 > 20 um (2) Medium width of the spores a) w < 4,5 um (0) b) 4,5 um < w < 8 um (1) c) w > 8 um (2) III C O N ID IO M A T A AND PYCNOSPORES (absent-0, present1) 18. Pycnidia laminal 19. Pycnidia marginal In the third analysis characters 18, 19 are connected to one muitisrace character (pycnidia laminal-0, marginal-1). 20. Conidia bifusiform 21. Conidia sublageniform 22. Conidia bacillariform 23. Conidia filiform In the third analyis characters 20-23 are connected to one multistate character (conidia bifusiform-0, sublageniform-1, bacillariform-2, filiform-3). IV SECONDARY CHEM ICAL C OM POU NDS (absent-0, pre­ sent-1) 24. Usnic acid 25. Atranorin 26. Harry acids 27. C.'aperatic acid 28. Rangiformic and/or norrangitormic acids 29. Lichesterinic and/or protolichesterinic acids 30. Anthraquinones 31. Pulvinic acid derivates 32. Orcinol series products 33. Imbriearic and/or perlatolic acids 34. Anziaic acid 35. Alectoronic and/or collatolic acids 36. Olivetoric and/or physodic acids 37. Hiascic and/or gyrophoric acids 38. Microphyllinic acid 39. p-orcinol series products 40. Protocetraric and/or fumarprotocetraric acids 41. Notstictic and/or salazinic acids 42. Stictic acid Outgroup In the selection of characters we kept to the idea that the taxa should be described not by the data of reproductive structures only but equally also by the thalline and especially by chemical characters. The possible special role of secondary metabolites in the evolution of lichens has been referred by J. Poelt in his species-pairs theory. It seems plausible that the formation of bio­ chemical pathways producing lichen substances has taken place earlier, during sexual stages rather than through mutations in asexual “secondary taxa” [2]. This concept has been applied in the dissection of evolution of the genus Cetrelia [14]. Structure of medullary secondary metabolites as well as morphological and anatomical characters of all Cetrelia species refer to the mono­ phyletic origin of that group. The genus is probably related to other cetrarioid lichens and may represent the more primitive trend. This is why the genus Cetrelia (C. braunsiana, C. cetra­ rioides, C. chicitae, C. collata, C. davidiana, C. delavayana, C. ja- potiica, C. monacborum, C. nuda, C. olivetorum, C. pseudolivetorum and C. sanguinea ) was selected as the sister group (outgroup) for the present cladistic analysis. Results The strict consensus trees were constructed of 1000 (fig. 1 and 2) and 3900 (fig. 3) equally parsimonous cladog­ rams after reweighting of characters. In the first analysis (secondary metabolites treated as separate characters) the unrooted trees could not be rooted such that specified ingroup was monophyletic. Parmelaria tbomsonii together with Asahinea chrysantha and A. scholanderi shows close relations to outgroup (12 species of Cetrelia). Genus N e­ phromopsis together with Cetreliopsis rhytidocarpa and Cetrariopsis wallichiana form a clade supported by the presence of usnic acid. Another branch including all the rest of the species is supported by the thallus habit, lack of the pseudocyphellae on the lower cortex and the marginal position of apothecia on the upper surface. Two genera, Platismatia and newly described Vulpicida (the group of species with yellow medulla, containing pulvinic acid de­ rivates) can be created homogeneous. The subclade of A l­ locetraria ambigua, A. cucullata and A. nivalis is related to the group of brown “fruticose” Cetraria species. Genus Tuckermannopsis is clearly heterogeneous. In the second analysis (fig. 2) lichen substances were grouped according lo their chemical .structure. Parmelaria thomsonii, Cetrariopsis wallichiana, Asahinea chrysantha together with A. scholanderi and Masonhalea richardsonii are the closest to the outgroup. The complex o f brown “fruticose” Cetraria species is evidently not homogeneous. The affinities between C. rassadinae, C. nigricans, C. is­ landica - C. laevigata, C. ericetorum, C. arenaria, C. andrejevii and the rest of the group remain dissolved. Taking secondary metabolites of these species into account as ad­ ditional information might be of use here. Three Allocet­ raria species (A. ambigua, A. cucullata and A. nivalis) form a separate subclade as well as genera Platismatia and Vulpicida. In the last polytomy three main entities can be noticed: genus Nephrom opsis (13 species) together with Cetreliopsis rhytidocarpa, Tuckermannopsis ciliaris group together with Esslingeriana idahoensis, and a subclade consisting of species from quite different genera. In the third analysis lichen substances were grouped according to their chemical structure and homologous character states as multistate characters. The unrooted trees could not be rooted such that specified ingroup was monophyletic. The affinities of the proposed ingroup and Parmelaria thomsonii together with Asuhinea chrysantha and A. scholanderi remain dissolved. The species of genus Platismatia and genus Vulpicida form one clade consisting 132 ■ A ndreas Saag and T iina R andlane T u c k e ra a n n o p s is T . o a k e s ia n a • p a lli T u c k e ra a n n o p s jis a s e r ic a n a T. c ilia r is y iS S J & d a te T u c k e r a a n r io p s i? c h l o r o p n y l la C e t r a r ia i n e r i i s T u c k e n ia n n o p s is . lic r o p h y l1 ic a C e t r a r ia s u b a lp in a A llo c e tr a r ia s tr a c fi T_____k u c k ee rrms aa nn nn op pp ss i s o r b a t a £ t: saffiltaito J e C. C. C. L. C. I. K a C. I. s e p in c o U t r a r la 'a n flr e je v ii a r g n a ria e r ic e t o r u * la e v ig a ta n ig r ic a n s i s la n d ic a r a s s a d in a e s a n h a le a r i c h a r d s o n ii o d o n te lla c a littr m c a C . ( le lis e i L \ r a s tig ia ta L , k a a c z a t jc a A l lo c e t r a r i a a ib ig u a A . c g c u lja ta A . n iv a lis P la tis a a tia e ro s a F . n o r v e g ic a P . fo rro s a n a P . g la u c a P . f ie r r e i, P . s t e n o p h y lla P . la c u n o s a P . tu c k e ra a n n i V u lp jc id ja tu b u ltH u s 'v , t i l e s u V, e n t a s is V . v ir id is V . ju n ip ^ r in a s V . p in a s tn C e t r a r ia c o f fla jx t j T u c k e ra a n n o p s is r e n d le r i C e t r a r ia w e b e ri iu c k e r n a n n c p s is h e p a t iz o n C e t r a r ia a g n a ta . N e p n r o fflo p s I? a s a h in a e L_i— - 1—■ J— _|— CH i —r - c z ------ B N . e c t o c a r p is a a N . e n d o c ro c e a N . p se u d o c o m p 1 ic a ta N . ru g o s a N . s tra c h e y i N . k o s a r o v ii N . la y r e r i N . p a lle s c e n s N . y u n n a n e n s is f t. e n d c x a n th a id e s N . g lo b u ia n s N . o rn a ta C e t r e ijo p s js r h y h d p c a r p a C e t r a n o p s is t ia llic h ia n a Q e t r e lia n u d a L . d a v id ia n a C . c e t r a r io id e s U . a o n a c h o ru m C . c o jla ta C , d e ia v a y a n a C, c M a ta e C . o h v e to r u # C , s a n g u in e a A s a h in e a c h r y s a n th a A . s c h o la n d e r i P a r n e la r ia t h m s o n n Q e t r e l ia b ra u n s ia n a L . ja p o m c a u p a e u d o h v e t o r u iB Fig. 1. Strict consensus tree constru cted from 1000 equally p a rsim o n o u s cladogram s. Secondary c o m p o u n d s treated as sep arate characters. Length o f sh o rte st tree = 2S544. C onsistency index = 0 .4S0. R etention index = 0 .878. Phylogeny o f c e trario id lichens • 133 V k e r # a n n p p s i5 a u re s c e n s ; ia n a s p h a e r o s p o r e lla . i h r w o p s i s a s a h in a e t e t r e h o p s is r n y tid o c a r p a N e p h r p o o p s i? e m k w a n t h o id e s N . e c t o c a r p is a a w re n ille s c e n s , . , ieuQocooplicata o rn a ta llo b u la n s . , .: e r » a n n o p s i5 o a k e s ia n a m io c e c r a r ia s t r a c h e y i T u c k e r m a n n o p s is p a l l i a u a C e t r a r ia i n e r i i s u c ^ e r n a n n o p s is c h l o r o p h y l l a T . a m e ric a n a T . c ilia r is E i: . > s l i n q e r i a n a . i d a h o e n s is u c k e r la n n o p s is o r b a ta : r a r ia c o a c ix t a u c t ir s a n n o p s is h e p a t iio n f e n ijle r i . u d tra n a x e b e ri . T u c k e r m a n n o p s is p l a t y p h y l l o i d e s T . s e p in c o la A l lo c e t r a r i a a a b ig u a A . c u c u lla t a r a s s a d in a ? . . . . is p n h a le a , r i c h a r d s o n ii . I s a n in e a c h r y s a n t h a A . s c h o la n d e r i .e tr a r io p s is n a lh c b ia n a sa r # e U n a t h o n s o n n J e t r e lla . o r a u n s ia n a !! ^seu3o^iyetoru« Fig. 2. Strict consensus tree co n stru c te d from 1000 equally p arsim o n o u s d a d o g ra m s. S econdary co m p o u n d s as representatives o f biochem ical groupin g s. L ength o f sh o rtest tre e fo u n d = 2 4 6 6 0 . C onsistency index - 0 .3 8 1 . R etention index O .b /l. 134 • A ndreas Saag a n d T iin a R and lan e T u c k e r i& a n n o p s is a u r e s c e n s T . o a k e s ia n a lana^sp^lerDspore} u c fc e r® .. c ilia r is ;, . la aaencana E s s lin g e r ia n a .id a h o e n s is T u c k e r a a n n o p s is c h lo r o p h y lls ' - ‘ ■ . r a r ia i n e r a i s . ....o c e tr a n a s tra c h e y i C e t r a r ia s u b a jp in a , T u c k e r ffla n n o p s is o r b a ta T w r r illii .a tis a a t ia e ro s a fp re o s a n a g la u c a Ia c u n o s a . j n o r v e g ic a P. h e rre i ,, P . s t e n o p h y lla fe ia ra u » . V. V. V. V. [. le t ile s ii . c a n a d e n s is jy n ip e r jn u s p in a s tr i v ir id is tr a r ia a g n a ta c o a s ix ta . .. lu d e r a a n n o p s is h e p a t n o n f. fe n d le r i, . C e t r a r ia w e b e rx , , T u c k e r * a n n o p s is p l a t y p h y U o i d e s T . s e p in c o la C e t r a r ia a n d r e ie v n .. n a s o n h a le a r ic n a r d s o n n C e t r a r ia d e lis e i C . f a s t ig i|t a A llo c e tr a r ia m y a lls C e t r a r ia a r e n a n a e r ic e t p r u a u . is la n d ic a C , la e v ig a ta ra s s a a in a e l . k a a q a tic a C. c a h to rm c a C . n ig r ic a n s C . m g r ic a s c e n s A llD c e iir a r ia c u c u lla t a A . a jib ig u a . N e p n r o a o p s is e c t o c a r p is a a N . e n d o c ro c e a N. k o a a ro v u It N . p s e u d o c o s p h c a ta N . ru g o s a N . s tr a c h e y i. N . y u n n a r e r is is N . e n d p x a n t h o id e s N . g lo b u la n s N . o rn a ta N . a s a h in a ? C e t r e lio p s is r h v h d p c a r p a L e t r a r io p s is H a lh c n ia n a C e t r e lia n u d a C . c e t r a r io id e s C i ip n a c h o r u f i C . c h ic ita e E: s u a r C C C C . d a v id ia n a . d e ia v a y a n a . ja p o n ic ? . p s e u d o h v e to ru s i f t. s c h o la n d e r i .. P a r a e la r ia th o s s o n n C e t r e l ia b ra u n s ia n a Fig. 3. Strict consensus tree co n stru c te d fro m 3 9 0 0 equally p a rsim o n o u s cladogram s. Secondary c o m p o u n d s as representatives o f biochem ical groupings. H o m ologou s c h a ra c te r states a re c o ded as m u ltistate ch aracters. Length o f s h o rtest tree = 3 9 3 0 7 . C onsistency index = 0.5 7 6 . R etention index = 0 .8 6 3 . 15 Phylogeny o f c etrario id lichens • 135 of two subclades. AUocetraria ambigua, A. cucullata and A. nivalis together with Masonhalea richardsonii are re­ lated to the group of brown “ fruticose” Cetraria species. Genus Tuckermannopsis is clearly heterogeneous, the af­ finities between the species of genus Nephrom opsis remain dissolved in this analysis. Discussion The question of monophyly or polyphyly seems to be essential in the interpretation of the results obtained. The monophyletic evolution of all cetrarioid taxa analysed here is not probable. Therefore, it would not be justified to make extensive rearrangements in the system on the basis of either of the present analyses only. Evolutionary af­ finities of separate entities shown in the first analysis (fig. 1), where the major secondary metabolites are treated tot­ ally independently from each other, should be evaluated especially critically. One of the most important characters of higher taxonomical units - biochemical connections be­ tween different lichen compounds — is completely ignored here. That problem is avoided in the second and third analysis (fig. 2 and 3) where the lichen subsrances are grouped into biochemically related sets. The inability to determine affinities within subclades (several polytomies on cladograms of the second analysis) due to the lack of detailed chemical data is the disadvantage of that method. O n all cladograms of the first and second analysis, all the 13 species of Nephromopsis form a monophyletic enti­ ty, closely related to Cetreliopsis rhytidocarpa. The latter is the species of a monotypic genus separated from other cetrarioid lichens by its complicated chemistry (usnic acid, fatty acids as well as derivatives of p-orcinol series) and some morphological characters (pseudocyphellae on both thallus surfaces). M . J. Lai, the author of the genus, says its relationships to Cetrelia and N ephrom opsis are obvious [11]. We are inclined to consider the affinities with Cet­ relia not to be tight and include the species into the genus Nephromopsis as done by Zahlbruckner in 1928. The species seems to be quite close to N. asahinae (both con­ tain fumarprotocetraric acid, have pseudocyphellae also on the upper cortex, etc.). The problem of synonymity of C. rhytidocarpa with Cetraria straminea Vainio and Cetrai ia laeteflava Zahlbr. established by Lai, needs addition­ al study. Our analyses surely support the recent inclusion of several species (N. endoxanthoides [15], N . komarovii [20] N. laureri [9], N. pallescens [13], N. yunnanensis [15]) into this genus. Thus the separation of Cetreliopsis rhytidocarpa as an independent genus is not supported by the present cladistic srudy, but some other taxa such as Pannclaria tbonisonii , C'.etrariupsis wallichiana and Masonbalea richardsonii form clearly separate clades (fig. 2). This does not con­ tradict the concept of treating them as monotypic genera as has been proposed for some time ago [1, 8, 10]. The genus Platismatia and the recently defined genus Vulpicida are both monophyletic entities according to the present analyses. This similarity is achieved due to the fact that all the members of these genera contain the same cortical and medullary substances (atranorin and caperatic acid in Platismatia species and usnic acid together with pinastric and vulpinic acids in the species of Vulpicida). Consequently there are no differences between our two analyses for these genera, and the consensus trees are prac­ tically identical for that part. Relationships inside the heterogeneous group of brown “ fruticose” Cetraria species are better resolved in the first analysis where the detailed data of secondary metabolites enable the finding of some more mutual affinities between the species, and in the third analysis. C. inermis and C. subalpina cannot be included in this complex. The pair of C. delisei - C. fastigiata and C. kamczatica appear to be the most separated taxa from the group. The latter has also affinities to the genus of Allocetraria (A. ambigua, A. cucullata and A. nivalis). The position of Allocetraria stracheyi far from the other close species on all cladograms is the most incom­ prehensible result of the present study. The heterogeneous nature of the genus Tuckermannop­ sis is certain even if the species with pulvinic acid derivates are excluded. Two main entities can be noticed. The group of T. ciliaris, the so-called “true” Tuckermannopsis con­ sists of T. americana, T. ciliaris, T. microphyllica, T. platy­ phylla and somewhat surprisingly also of Esslingeriana idahoensis. Another entity in the genus includes the yel­ lowish species with usnic acid in the cortex — T. pallidula, T. oakesiana, T. aurescens and perhaps also Ahtiana sphaerosporella. This group is well delimited on the cladograms of the first and third analysis and quite scat­ tered on the second one shewing affinities partly with such taxa as Allocetraria stracheyi, Cetraria inermis - C. subal­ pina and partly with the genus Nephromopsis. Several Tuckermannopsis species (T. sepincola, T. platyphyl­ loides, T. merrillii, T. fendleri and T. weberi) are not close­ ly related to any of the examined taxa, and consequently this genus is in urgent need of revision. Acknowledgements We are most grateful to Prof. Erast Parmasto (Institute of Zoology and Botany, Tartu) for the possibility to use his computer and programs and for his numerous advices. We are also indebted in many ways to Prof. Teuvo Ahti (University of Helsinki), Dr. Ming-Jou Lai (Fu-jen Univer­ sity, Taipei), Dr. Gertrud Dahlgren, Mrs. Cecilia Lonnqvist (University of Lund) and the group of lichenologists from the Departement of Systematic Botany, University of Lund headed by Dr. Ingvar Karnefelt. We thank the curators of the herbaria H , KW, LD, LE, TB and UPS for their help. References 1 Avvasthi, D. D. (1987): A new position for Plalysma thornsonii Stirton. Journ. Hattori Bot. Lab. 63, 367-3/2. 2 Bowler, P. A. and Rundel, P. W . (1975): Reproductive strategies in lichens. Bot. Journ. Lin. Soc. 70, 325—340. 136 • A ndreas Saag and T iina R andlane 3 Culberson, C. F. (1972): Improved conditions and new data for the identification of lichen products by a standardized thin-layer chromatographic method. Journ. Chromatogr. 72, 123-125. •4 Culberson, C. I\and Ammaim, K. (1979): Standardmelhodc zur Oilnnsol)ichIth rom;i 1<>^iapliic von i'lechtonsubstaii/.on. Herzogia S, 1-24. 5 Culberson, C. F. and Culberson, W. L. (1976): Chemosyndromic variation in lichens. Syst. Bot. 1, 325-339. 6 Culberson, W . L. and Culberson, C. F. (1968): The lichen genera Cetrelia and Platismatia (Parmeliaceae). Contr. U.S. Nat. Herb. 34, 449-558. . 7 Egan, R. S. (1987): A fifth checklist of the lichen-forming, lichenocolous and allied fungi of the continental United States and Canada. Bryologist 90, 77-173. 8 Kurokawa, S. (1980): Cetrariopsis , a new genus in the Par­ meliaceae, and its distribution. Mem. Nat. Sc. Mus. (Tokyo) 13, 139-142. 9 Kurokawa, S. (1991): Japanese species and genera of the Par­ meliaceae. Journ. Jap. Bot. 66, 152-159. 10 Karnefelt, I. (1977): Masonhalea, a new lichen genus in the Parmeliaceae. Bot. Not. 130, 101-107. 11 Lai, M.-J. (1980): Studies on the cetrarioid lichens in Par­ meliaceae of East Asia (I). Quart. Journ. Taiwan Mus. 33, 215-229. 12 Mattsson, J.-E. and Lai, M.-J. (1993): Vulpicida, a new genus in Parmeliaceae (Lichenized Ascomycetes). Mycotaxon 49, 425-428. 13 Park, Y. S. (1990): The macrolichen flora of South Korea. Bryologist 9.1, 105—160. 14 Randlane, T. and Saag, A. (1991): Chemical and morpho­ logical variation in the genus Cetrelia in the Soviet Union. Lichenologist 23, 113-126. 15 Randlane, T. and Saag, A. (1992a): Additional data about the genus Nephromopsis (Lichenes, Parmeliaceae). Mycot­ axon 44, 485—489. 16 Randlane, T. and Saag, A. (1992b): Comparison of cetrarioid lichen genera. In: Karnefelt, I. (ed.): Second International Lichenological Symposium IAL 2 Abstracts, p. 17. Dept, of Syst. Bot. Univ. Lund, Lund. 17 Randlane, T. and Saag, A. (1992c): New combinations of some cetrarioid lichens (Parmeliaceae). Mycotaxon 44, 491493. 18 Randlane, T. and Saag, A. (1993): List of cetrarioid lichens. Mycotaxon 47, 395-403. 19 Swofford, D. L. (1991): PAUP: Phylogenetic analysis using parsimony, version 3.0s. Computer program distributed by the Illinois History Survey. 20 Wei, J.-C. (1991): A enumeration of lichens in China. Intern. Acad. Publ., Beijing. Key words: Ascomycotina, Lecanorales, Parmeliaceae, cetrarioid lichen genera, phylogeny, cladistics. Andres Saag and Tiina Randlane, Institute of Botany and Ecology, Tartu University, Lai Street 38, Tartu, Estonia EE-2400 II 16 Randlane, T. & Saag, A. 1989. Chemical variation and geographical distribution of Asahinea chrysantha (Tuck.) Culb. & C. Culb. — Lichenologist 21: 303-311. Lichenologist 21(4): 303-311 (1989) CHEMICAL VARIATION AND GEOGRAPHICAL DISTRIBUTION OF ASAHINEA CHRYSANTHA (TUCK.) CULB. & C.CULB. T. RANDLANE* and A. SAAG* Abstract: Sixty-two samples of Asahinea chrysantha from different localities in the Soviet Union were analysed by TLC. Three different chemotypes are recognised, each with a different geographical distribution. The most restricted of the three is an usnic acid deficient chemotype (I), with atranorin, alectoronic and a-collatolic acids, which is chemically the same as A. scholanderi and A. kurodakensis. Chemotype II contains usnic acid in addition to the basic chemistry, and chemotype III is acollatolic acid deficient and has the widest geographical range. A centre of speciation of Asahinea in Soviet Primorje or Japan is proposed. Introduction Cetraria Ach. s.lat., is now divided into several more restricted genera, one of which is Asahinea Culb. & C.Culb. (Culberson & Culberson 1965) which com­ prises three species: A. chrysantha (Tuck.) Culb. & C.Culb., A. scholanderi (Llano) Culb. & C.Culb and A. kurodakensis (Asahina) Culb. & C.Culb. The principal morphological characteristics of the genus are: foliose, loosely appressed thallus; grey or yellow upper cortex and black lower cortex; and absence o f rhizines. The genus shows an arctic-alpine distribution (Krog 1968). Characteristic secondary metabolites o f Asahinea are atranorin and usnic acid as the main cortical substances, and the orcinol depsides alectoronic and also usually a-collatolic acids, as diagnostic medullary substances. Some purple, lavender or pink pigments have also been reported for all three species (Culberson 1969, 1970; Culberson et al. 1977). More precise characteristics of Asahinea and related genera in the Parmeliaceae are given in Goward (1985). The Species Morphology and distribution Asahinea kurodakensis is the least widely distributed o f the three and is supposedly endemic to Japan. It has been collected only three times in the high mountains on the islands of Hokkaido and Honshu (Asahina 1953, Culberson & Culberson 1965, Yoshimura 1979). According to both morphological and chemical characteristics, A. kurodakensis is closely allied to A. scholanderi. Both are tan to blackish (lacking usnic acid in the upper cortex), produce isidia, and their upper surface lacks pseudocyphellae. Asahinea kurodakensis and A. scholanderi contain atranorin, alectoronic and a-collatolic acids. Unfortunately laboratory of Bioindication, Tartu University, 202400 Lai Street 38, Tartu, Estonian SSR, USSR. 0024-2829/89/040303 + 09 $03.00/0 © 1989 The British Lichen Society 304 THE LICHENOLOGIST Vol. 21 we have not seen a sample of A . kurodakensis, and from the literature the difference between A . kurodakensis and A . scholanderi is unclear. It is signifi­ cant that A . scholanderi has not been reported from Japan. Asahinea scholanderi was described as a Cetraria by Llano (1951). Later, Oxner & Rassadina (1960) described a new species from Asia, C. saviczii. K rog (1962) supposed, on the basis of the diagnosis, that C. saviczii m ight be identical w ith C. scholanderi, a view now widely accepted. Asahinea scholanderi is an arctic-alpine am phi-Beringian species (Krog 1968), its recorded range extending from eastern Siberia to the Behring Straits and to northern and central Alaska (Culberson & C ulberson 1965; H andbook of the lichens of the U S S R 1971). However, its distribution is considerably wider and it occurs in the Soviet U nion in the U rals (Rjabkova 1982), in T aim yr (Piin & T rass 1971), in Kamchatka (M ikulin 1987), in the Khabarovsk region, Badzhal (Randlane 1984), and in the Prim orje region, Sikhote-Alin (herbarium specimens in T U ). T hom son (1984) remarks that in N orth America its range extends eastward to Baker Lake and the m outh of the Black River. Asahinea chrysantha is readily distinguished from other m em bers of the genus by its yellow upper cortex, the presence of pseudocyphellae, and by the absence of isidia. It has a wide m orphological variation. T h e size of the white pseudocyphellae on the upper surface varies as does the degree of reticulation of the thallus. T h e lower surface is black with brown m argins, while the w idth and darkness of the margins is also variable. T h e upper surface of the thallus is usually bright to pale yellow but specimens with an almost grey or greenish grey upper cortex are also known; taxa such as C. chrysantha f. cinerascens Asahina, and f. glaucescens Oxner & Rassad. (Asahina 1934; O xner & Rassadina 1960) represent these latter extremes. T h e range of A . chrysantha is considerably broader than that o f A . scholan­ deri which occurs within the arctic-alpine distribution area of A . chrysantha. Both species may grow together. In Eurasia, A . chrysantha is known from Norway, Sweden, and Finland, in the northern parts of the U S S R (from Kola to Tschukotka and in the m ountainous regions from the U rals to Sikhote-Alin), in Mongolia, Korea, and Japan. In N orth America it is recorded from the Aleutian Islands, Alaska, the N orthw est T erritory and the southern part of Baffin Island. T h e distribution of A . chrysantha is thus ± circum polar, b u t it is m ost abundant in Siberia, the F ar East and Alaska. D istribution maps are given by Oxner & Rassadina (1960), Culberson & Culberson (1965), H akulinen & U lvinen (1966) and Thom son (1984). Chemistry It is generally believed that atranorin and usnic acid occur in the upper cortex and alectoronic acid in the m edulla of A . chrysantha as constant metabolites although usnic acid is sometimes lacking from Japanese material (Yoshim ura 1979). A fourth com pound, a-collatolic acid, is reported as constant for the Japanese specimens. Culberson & C ulberson (1965) found this substance in only one non-Japanese sample tested (Siberia, the T schita region). Subse­ quently, a-collatolic acid was m entioned as an accessory in material from Alaska (K rog 1968) and other N orth American localities (Thom son 1984). As 1989 305 Asahinea chrysantha— Randlane & Saag the Culbersons m ention, the colour of the upper cortex does not depend on the presence or absence of a-collatolic acid for it is a colourless m edullary substance, rather the intensity of the yellow colour of the upper cortex is determ ined by the am ount of usnic acid deposited there. In addition, several unidentified purple or lavender pigm ents are also known in A . chrysantha and in other species of the genus. T hese occasionally occur in the m edulla, particularly near the lower cortex in older parts of the thallus (Culberson & C ulberson 1965). Collaborators of the Pacific Institute of Bio-organic C hem istry in Vladivostok working on specimens collected from the Soviet F ar East have shown the following. Six purple and lavender coloured anthraquinone pigm ents were identified from A . chrysantha and their structures determ ined (Mischenko et al. 1980, Krivoshchekova et al. 1983b). Krivoshchekova et al. 1983a reported the presence of two new substances in A . chrysantha and A . scholanderi. J3Alectoronic acid was isolated and identified for the first time from plant m aterial, although it was earlier detected by Asahina from alkaline hydrolysis of a-collatolic acid. A specimen of A . chrysantha from M agadan was reported to contain a- and [3-alectoronic acids, usnic acid, atranorin and m ethyl-P-orcinolcarboxyiate. T h e latter substance was previously known only from Parmelia tinctorum (Culberson 1969). A specimen of Asahinea scholanderi (also from M agadan) contained a- and f3-alectoronic, a- and (3-collatolic acids, usnic acid and atranorin. T his finding of usnic acid in A . scholanderi is somewhat surprising. Stepanenko et al. (1985) examined two morphologically different specimens of A . chrysantha (Table 1). Com pounds 9-11 are anthraquinonoid pigm ents. Haem atom m ic acid and methyl-f3-orcinol-carboxylate are recorded for the first time for this species, in rather low concentrations. However, Sample I (with a yellow upper surface, from the M agadan region) did not contain a- or (3collatolic acids while Sample II (with a grey upper surface, from the Prim orje T a b le 1. Chemical compounds isolatedfrom Asahinea chrysantha* Amount (% dry wt) Number 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. Compound Usnic acid Atranorin a-Alectoronic acid P-Alectoronic acid a-Collatolic acid P-Collatolic acid Methyl-P-orcinol-carboxylate Haematommic acid Islandicin Cynodontin Asahinin *Data from Stepanenko et al. (1985). 17 Sample I Sample II 1-39 009 0-25 0-31 __ — 0-01 __ 0-001 0-001 0-001 0-22 0-25 __ __ 0-31 0-23 0-021 0003 0001 0001 0-001 306 THE LICHENOLOGIST Vol. 21 region) lacked either a- or P-alectoronic acids; the first reported absence of alectoronic acid in the genus Asahinea. These results indicate that chemically interesting populations of A . chrysantha occur in Japan and in the F ar East. Fig. 1. Distribution of Asahinea chrysantha in the USSR (from Oxner & Rassadina 1960, Culberson & Culberson 1965). Dots (62) show localities of samples analysed by TLC. The line marks the limits of its distribution. Materials and Methods In the lichen herbaria of T U and TBA material of A . chrysantha is well represented from the whole of the Soviet Union. Sixty-two samples of A . chrysantha (localities shown in Fig. 1) were studied chemically between 1981 and 1987. For comparison, a specimen from North America (Yukon) was also analysed, as were specimens of A. scholanderi. Standard methods were used (Culberson & Kristinsson 1970, C. Culberson 1972, 1974, Culberson & Amman 1979, White & James 1985). Although SILUFOL UV 254 plates were used, controls were also run on MERCK Silica gel 60 F 254 TLC and HPTLC plates. Secondary metabolites were extracted with acetone rather than chloroform/hexane as used by Stepanenko et al. (1985). Atranorin, usnic, alectoronic and collatolic acids were positively identified. Anthraquinonoid pigments and some accessory compounds were noted but not determined further. Results N ine specimens of our m aterial from the Prim orje region, Sikhote-Alin, were analysed by T L C by D r C. F. Culberson (Duke C FC no. 5703-07; 5717-20). A tranorin, alectoronic and a-collatolic acids, traces o f physodic acid, and an unidentified substance were detected in all samples. U snic acid was found in 8 specimens b u t no traces of it in one of the samples. T his prom pted us to investigate the main secondary m etabolites of A . chrysantha in specimens from as many different localities in the Soviet U nion as possible w ith special emphasis on F ar Eastern material. 1989 Asahinea chrysantha— Randlane & Saag 307 a - Alectoronic acid and atranorin. T hese substances occurred in all samples and thus are of no im portance in separating different chemotypes of A . chrysantha. Usnic acid was determ ined in the m ajority of our samples, but was not found in two specimens from the Prim orje region, Sikhote-Alin and in one from the K habarovsk region, Badzhal. Yoshimura (1979) also m entioned the occasional absence of usnic acid in Japanese material. a -Collatolic acid was detected only in specimens from the eastern part o f the Soviet U nion (excluding the Kamchatka and Tschukotka Peninsulas) up to the Baikal region, and the T aim yr Peninsula. In material from the regions of Baikal and T aimyr there were specimens of both chemotypes, w ith and w ithout a-collatolic acid. T h e three specimens w ithout usnic acid m entioned above undoubtedly contain a-collatolic acid. T h e only N orth American sample (Yukon) did not contain this substance. P-Alectoronic and P-collatolic acids were detected by chemists in Vladivostok. W orking w ith pure samples of a- and P-alectoronic, and a- and p-collatolic acids (sent by colleagues at the Pacific Institute of Bio-organic Chem istry) we observed that both P-forms ran on chromatogram s lower than the spots of the respective a-form s, while differences in R f values of a- and P-collatolic are considerably greater than those of a- and P-alectoronic acids. T h u s P-collatolic is situated only a little above a-alectoronic acid. All four spots have a similar colour. In our herbarium collections we found P-collatolic acid in only a few cases, but never P-alectoronic acid. P-Collatolic acid was detected in a sample of A . chrysantha from the Baikal region and in a sample of A . scholanderi from the M agadan region. T h e latter specimen contained both forms of collatolic acid. In vitro, P-alectoronic acid was obtained by treating pure a-alectoronic acid with alkaline solution. T he same treatm ent of pure a-collatolic acid gave us P-collatolic acid and, somewhat suprisingly, both forms of alectoronic acid in trace am ounts. All four forms of the acids were identified by T L C comparison with pure samples. T h e problem is whether the P-forms of alectoronic and collatolic acids occur in fresh material, and under w hat conditions the change from one into another m ight take place. In their treatm ent of Parmelia subgen. Amphigymnia in East Africa, Krog & Swinscow (1981: 150) record ‘ . . . in old specimens w ith alectoronic and a-collatolic acids we obtained additional spots below those of the main substances on the T L C p la te s.. . We found no corre­ lation between morphological characters and the presence or absence of such substances, and were inclined to regard them as artefacts ’. W e consider it possible that these unidentified substances are simply P-forms of alectoronic and collatolic acids. Discussion Atranorin and alectoronic acid are the only com pounds identified by us as occurring in all samples of Asahinea chrysantha. U snic and a-collatolic acids occur in the species in three combinations: (1) a-collatolic acid alone; (2) a-collatolic acid w ith usnic acid; (3) usnic acid alone. T h u s there are three distinct chemotypes of A . chrysantha in the Soviet U nion, each with its own particular geographical distribution (see T able 2 and Fig. 2). 308 THE LICHENOLOGIST Vol. 21 T a b le 2. Occurrence of chemotypes of Asahinea chrysantha Chemotype Composition of major aromatic compounds Occurrence in the Soviet Union I a-Collatolic acid (+ atranorin and a-alectoronic acid) Primorje region, Sikhote-Alin (2)*; Khabarovsk region, Badzhal (1) II Usnic acid, a-collatolic acid ( t atranorin and a-alectoronic acid) Primorje region, Sikhote-Alin (6); Khabarovsk region, Badzhal (12); Amur region (1); Tscita region (1); Baikal region (2); Taimyr (8) III Usnic acid (+ atranorin and a-alectoronic acid) Tschukotka (2); Kamchatka (3); Jakutia (1); Baikal region (5); Taimyr (12); Jamal (2); the Urals (2); the Khibins (2) *Number of analysed specimens. F ig . 2. Occurrence of chemotypes of Asahinea chrysantha in the USSR A = chemotype I, O = chemotype II, • = chemotype III. T h e systematic rank of these three chemotypes is not treated in this account. T h e presence or absence of a-collatolic acid does not seem to correlate w ith any morphological characters (colour of upper and lower cortices, w idth of brow n edges on the lower surface, size of pseudocyphellae, degree of surface reticu­ lation). T h e yellow colour of the upper surface depends on the quantity o f usnic acid deposited in the cortex. Samples w ithout usnic acid are uniform ly grey. Such specimens, and samples containing usnic acid in small am ounts, are dis­ tinguished only with difficulty from chemotypes II and III. Chem otypes II and 1989 Asahinea chrysantha— Randlane & Saag 309 II I are very variable in the colour o f their upper cortex, and there are specimens in our herbarium that are yellow in one part and almost grey in another part of the same thallus. T h e geographical distributions of chemotypes of A . chrysantha is of special interest. T h e least num erous of the three is chemotype I (w ithout usnic acid), which is known from Sikhote-Alin, Badzhal and Japan. Its chemistry (atranorin, alectoronic and a-collatolic acids) is the same as that of A . scholanderi and A . kurodakensis. In a paper concerning biogenetic relationships of lichen products in the genus Cetrelia, Culberson & C ulberson (1976) state that m icrophyllinic acid, an orcinol depside which combines two phenolic acid units with seven-carbon side chains, is the m ost prim itive product of this genus. Chem osyndrom ic variation in Cetrelia may reflect a trend towards reduced side-chain length. Alectoronic and a-collatolic acids are orcinol depsidones, both having the greatest num ber (7 + 7) of carbons in the side-chains. It is known that depsi­ dones are derived directly from depsides and hence the somewhat prim itive character of alectoronic and a-collatolic acids in Cetrelia is also evident. Such considerations support the hypothesis of a possible origin of the genus Cetrelia from an Am phigym nia-like ancestor producing alectoronic and a-collatolic acids (Culberson & Culberson 1968). As in C etrelia , all species of Parmelia subgen. Amphigymnia also produce atranorin in the upper cortex. It is signifi­ cant that the other common cortical substance, usnic acid, also occurs in some species of subgen. Amphigymnia but never in those which produce alectoronic and a-collatolic acids. T his indicates that Asahinea may also share a similar ancient chemical composition which includes atranorin in the cortex and alectoronic and a-collatolic acid in the medulla. T h u s chemotype I of A . chrysantha is thought to represent the basic, prim itive chem istry of Asahinea. Chem otype II contains, in addition to atranorin, alectoronic and a-collatolic acids, usnic acid in the cortex. T h e distributional area of this chemotype shows a disjunction, being found in Taim yr, Siberia, the F ar East, Japan and N orth America but not in Tschukotka or Kam tschatka. T h e lack of a-collatolic acid in chemotype II I represents a second qualitative change in the chemical com­ position of A . chrysantha. Chem otype I I I is the m ost widely distributed, ranging from Scandinavia and Kola over Tschukotka to N orth America. T h e different chemotypes of A . chrysantha and their relative distributions are shown schematically in Fig. 3. T h e distribution area of chemotype II is sympatric over m ost of its range with that o f chemotype III. T hus two chemo­ types (II and III) occur simultaneously in this territory and a-collatolic acid may be present or absent in specimens collected in this zone. K rog (1968: 114) considers a-collatolic acid as an accessory in Alaskan populations of A . chrysantha and T hom son (1984) in N orth Am erican populations. W e record the same for populations in the T aim yr Peninsula and the Baikal region. T h a t p art of the distribution area of chemotype II , where specimens of chemotype II I do not occur, includes the known distribution of chemotype I. Com bining the distributional patterns of the three chemotypes of A . chrysantha (Fig. 3) and assum ptions about an original or prim itive suite of secondary metabolites in Asahinea , we suggest that a possible centre of specia­ tion was in Soviet Prim orje or Japan. T his is the only region where chemotypes 18 310 THE LICHENOLOGIST Vol. 21 m F ig . 3. Relative ranges of chemotypes of Asahinea chrysantha (1-4) and A . scholanderi (5) in the USSR 1 = Chemotypes I & II, 2 = chemotype 1I, 3 = chemotypes II & III, 4 = chemotype III. I and II of A . chrysantha coexist, and where A . scholanderi and A . kurodakensis also occur. Chemotype III of ^4. chrysantha (the most derived chemotype in the genus) is absent from there. The present distribution of A . chrysantha has the limits of the ranges of chemotypes II and III, both containing usnic acid. The wider distribution of A . chrysantha compared with the known ranges of the other two species of the genus may be in part a reflection of antimicrobial properties of usnic acid, a cortical substance which is additional to the primitive assemblage of secondary metabolites characteristic for the genus Thanks are due to Prof. Hans Trass for his encouragement and supervision of this study. We are very grateful to Taimi Piin from Tallinn Botanical Garden for performing some chemical analyses and for many critical and constructive remarks. We are also indebted to collaborators from the Pacific Institute of Bio-organic Chemistry in Vladivostok for pure samples of some lichen substances. R efer en ces Asahina, Y. (1934) Aufzahlung von Cetraria Arten aus Japan I. Journal of Japanese Botany 1 0 : 481. Asahina, Y. (1953) Lichenes Japoniae novae vel minus cogitae 11. Journal of Japanese Botany 28: 6- 12 . Culberson, C. F. (1969) Chemical and Botanical Guide to Lichen Products. Chapel Hill: University of North Carolina Press. Culberson, C. F. (1970) Supplement to ‘ Chemical and Botanical Guide to Lichen Products’. Bryologist 73: 177-377. Culberson, C. F. (1972) Improved conditions and new data for the identification of lichen products by a standardized thin-layer chromatographic method. Journal of Chromatography 72:113-115. Culberson, C. F. (1974) Conditions for the use of Merck silica gel 60 F254plates in the standardized thin-layer chromatographic technique for lichen products. Journal of Chromatography 97 : 107-108. Culberson, C. F. & Ammann, K. (1979) Standardmethode zur Diinnschichtchromatographie von Flechtensubstanzen. Herzogia 5:1-24. Culberson, C. F. & Culberson, W. L. (1976) Chemosyndromic Variation in Lichens. Systematic Botany 1:325-339. Culberson, C. F. & Kristinsson, H. (1970) A standardized method for the identificnrion of lichen products. Journal of Chromatography 46: 85-93. 1989 Asahinea chrysantha— Randlane & Saag 311 Culberson, C. F., Culberson, W. L. & Johnson, A. (1977) Thermally induced chemical artifacts in lichens. Phytochemistry 16:127-130. Culberson, W. L. & Culberson, C. F. (1965) Asahinea, a new genus in the Parmeliaceae. Brittonia 17:182-190. Culberson, W. L. & Culberson, C. F. (1968) The lichen genera Cetrelia and Platismatia (Parmeliaceae). Contributions from the United States National Herbarium 34 (7): 449-558. Goward, T. (1985) Ahtiana, a new lichen genus in the Parmeliaceae. Bryologist 88:367-371. Handbook of the lichens of the USSR I (1971). Leningrad: Nauka (in Russian). Hakulinen, R. & Ulvinen, T. (1966) Asahinea chrysantha (Tuck.) Culb. et Culb. in Fennaskandien. Annales Universitatis Turkuensis (A II) 36:101-105. Krivoshchekova, O. E., Stepanenko, L. S., Mishchenko, N. P. & Maksimov, O. B. (1983a) Aromatic metabolic substances of lichen fam. Parmeliaceae, I Depsidones. Khimija prirodnyh sojedinenii 1: 13-19 (in Russian). Krivoshchekova, O. E., Stepanenko, L. S., Mishchenko, N. P., Denisenko, V. A. & Maksimov, O. B. (19836) Studies on aromatic metabolic substances of lichens fam. Parmeliaceae, II Pigments. Khimija prirodnyh sojedinenii 3:283-289 (in Russian). Krog, H. (1962) A contribution to the lichen flora of Alaska. A rkiv fu r Botanik 4:489-513. Krog, H. (1968) The macrolichens of Alaska. Norsk Polar-institutt Shrifter 144: 1-180. Krug, H. & Swinscow, T. D. V. (1981) Pannclia subgcnus Amphigynmia (lichcns) in East Africa. Bulletin of the British Museum (Natural History), Botany 9: 143—231. Llano, G. A. (1951) A contribution to the lichenflora of Alaska. Journal of the Washington Academy of Sciences 41:196-200, Mikulin, A. G. (1987) Lichenss pro paeninsula Kamczatka novi. Novitates systematicae plantarum non vascularium 24: 163-165 (in Russian). Mischenko, N. P., Stepanenko, L. S., Krivoshchekova, O. E. & Maksimov, O. B. (1980) Anthraquinones of the lichen Asahinea chrysantha. Khimija prirodnyh sojedinenii 2: 160-165 (in Russian). Oxner, A. N. & Rassadina, K. A. (1960) Ad genus Cetraria ex URSS novitates. Notulae System­ aticae e Sectione Cryptogamica Instituti Botanici nomine. V. L. Komarovii Academiae Scientiarum U R S S 13: 5-14 (in Russian). Piin, T. H. & Trass, H. H. (1971) The lichens growing on the soil surface in the vicinity of Tareya (Western Taimyr). In Biogeocenses of Taimyr tundra and their productivity (K. A. Tihomirov, ed.): 151-160. Leningrad: Nauka (in Russian, English summary). Randlane, T. V. (1984) Lichens of the goltsy belt of the Badzhal mountains (Khabarovsk territory). In Flora and groupings of lower plants in natural and anthropogenous extreme environment conditions (J. L. Martin, ed.): 120-133. Tallinn: Academy of Sciences of the Estonian SSR (in Russian, English summary). Rjabkova, K. A. (1982) Species Parmeliacearum in montibus Uralensibus inventae. Novitates systematicae plantarum non vascularium 19:149-154 (in Russian). Stepanenko, L. S., Krivoshchekova, O. E. & Mishchenko, N. P. (1985) Chemical variations of Asahinea chrysantha. Phytochemistry 24:354-355. Thomson, J. W. (1984) American arctic lichens 1. The Macrolichens. New York: Columbia University Press. Yoshimura, J. (1979) Lichen flora of Japan in colour. Osaka: Hoikusha Publishing Co. White, F. I. & James, P. W. (1985) A new guide to microchemical techniques for the identification of lichen substances. British Lichen Society Bulletin SI (suppl.): 1-41. Accepted for publication 25 June 1989 19 Randlane, T. & Saag, A. 1991. Chemical and morphological variation in the genus Cetrelia in the Soviet Union. — Lichenologist 23: 113-126. Lichenologist 23(2): 113-126 (1991) CHEMICAL AND MORPHOLOGICAL VARIATION IN THE GENUS CETRELIA IN THE SOVIET UNION T. R A N D LA N E* a n d A. SA A G * Abstract: Two hundred and three specimens belonging to the genus Cetrelia from the Soviet Union and also 41 samples from other countries have been analysed by TLC. Eight species were determined in the Soviet Union. In addition two new species, C. orietitalis and C. pseudocollata sp. nov. are described. Five morphotypes and six chemotypes have been recognized in the genus: their main characteristics are tabulated in a form that can be used for identification and a key is also included. Evolutionary relationships between the ‘primary’ and ‘secondary’ species related to ‘species-pairs’ are discussed. In tr o d u c tio n T h e genus Cetrelia was separated from the genera Cetraria and Parm elia by C ulberson & Culberson (1968). It is mainly characterized by the foliose, loosely attached thallus; ashy white or tan upper cortex and black lower cortex; sparse rhizines; laminal pseudocyphellae; marginal pycnidia; ellipsoid ascospores; atranorin as the main cortical substance, and various orcinol depsides and depsidones as diagnostic medullary substances (see G ow ard 1985). Fifteen species were previously included in the genus, of which the m ajority are d istri­ buted in eastern and south-eastern Asia. In the Soviet U nion eight species have so far been found, i.e. C. braunsiana, C. cetrarioides, C. collata , C. chicitae and C. pseudolivetorum (Kopaczevskaja et al. 1971), C. alaskana (M akarova 1980), C. olivetorum (Malysheva & Smirnov 1982) and C. japonica (Skirina 1987). H ow ever, no special chemical or distributional studies of the Soviet m aterial had been carried out. M a te ria ls a n d M e th o d s Two hundred and three specimens belonging to the genus, collected in the Soviet Union and kept in T U , LE and KW, were studied chemically in 1988-1989. For comparison, 41 specimens from Austria, Germany, Great Britain, Hungary, Italy, Norway, Poland, Portugal, Romania, Spain, Sweden and Switzerland; Canada and the USA; Japan, Mongolia, China and Tibet; the islands of Java, Sumatra and the Philippines and kept in TU, LE, LD and UPS were also analysed. The major medullary compounds were identified according to standardized methods (Culberson 1972, 1974). The minor constituents were identified only when the spots were sufficiently wrell developed. The acetone extracts were run in solvent system C only and the plates were later developed with 10°o H,SO., in ethanol. The plates were air dried after spraying with reagent and then heated at about 100-120°C for up to 15 min. ♦Laboratory of Bioindication, Tartu University, 202 400 Lai Street 38, Tartu, Estonian SSR, USSR. 0024-2829/91/020113 + 14 $03.00/0 ©1991 The British Lichen Society 114 THE LICHENOLOGIST Vol. 23 Fig. 1. Distribution of Cetrelia alaskana ( • ) , C. japonica (★), C. orientalis (■ ) and C. pseudolivetorum ( A ) in the Soviet Union. R esu lts S p ecies r e co rd ed in th e S o v ie t U n ion T h e following eight species w ith their respective major m edullary com ­ pounds were determ ined in the U S S R according to the previously accepted systematics (C ulberson & C ulberson 1976). (1) C. alaskana (Culb. & C. C ulb.) Culb. & C. Culb. Very rare in the Soviet U nion, found only in Tschukotka (Fig. 1); three specimens analysed. T h e major m edullary substance is im bricaric acid. (2) C. braunsiana (M ull. Arg.) Culb. & C. Culb. D istributed in K habarovsk and Prim orje regions in the F ar East (Fig. 2); 44 specimens examined. T h e m ajor m edullary substances are alectoronic and a-collatolic acids. C ontrary to C ulberson & C ulberson (1976) we consider physodic and 4-O -m ethylphysodic acids to be m inor com pounds. (3) C. cetrarioides (Del. ex D uby) Culb. & C. Culb. Found in the zone of mixed and deciduous forests throughout the Soviet Union (Fig. 3); 51 speci­ mens examined. T h e m ajor substance is perlatolic acid. (4) C. chicitae (C ulb.) Culb. & C. Culb. Quite common in the F ar East but very rare in the w estern districts (Fig. 2). O f the 29 specimens examined 27 are from the K habarovsk and Prim orje regions, one from Siberia (Krasnoyarski region) and one from the European part (the Crimea Peninsula). T h e m ajor substances are alectoronic and a-collatolic acids. (5) C. japonica (Zahlbr.) Culb. & C. Culb. Found only in the F ar E astPrim orje region (Fig. 1); 14 specimens examined. T he m ajor substance is m icrophyllinic acid. 1991 Cetrelia—Randlane & Saag 115 F ig . 3. D istrib u tion o f Cetrelia cetrarioides ( • ) , C. monachorum (A ) and C. olivetorum (★) in the Soviet Union. (6) C„ monachorum (Zahlbr.) Culb & C. Culb. Distributed in the zone of mixed and deciduous forests on the whole territory of the Soviet Union (Fig. 3); 29 specimens examined. The major medullary substance is imbricaric acid. 20 116 THE LIGHENOLOGIST Vol. 23 (7) C. olivetorum (Nyl.) C ulb & C. Culb. D istributed throughout the U S S R (Fig. 3); 31 samples examined. T h e major substance is olivetoric acid. (8) C. pseudolivetorum (Asah.) Culb & C. Culb. According to the literature it has been found only in the F ar East (Kopaczevskaja et al. 1971, Knjazheva 1973) (Fig. 1). T h e only specimen tested was collected in the Prim orje region. T h e major m edullary com pound is olivetoric acid. New species I n addition to the species m entioned above there is one specimen from the Khabarovsk region in the Soviet F ar East, with alectoronic and a-collatolic acids as m ajor m edullary substances, that does not seem to belong to any described species (Culberson & Culberson 1968). It is described here as C. orientalis. A nother specimen, collected by D r J. C. Wei from C hina, distributed in Lichenes Sinenses Exsiccati no. 20 as C. collata, contains m icrophyllinic acid as the m ajor m edullary com pound. I t is described here as C. pseudocollata. Furtherm ore, the species C. collata (Nyl.) Culb. & C. C ulb., which had been reported from the Prim orje region in the eastern part of the U S S R (Kopaczevskaja et al. 1971, Skirina & Knjazheva 1985, Skirina 1987), m ust be excluded from the list of the lichens of the Soviet U nion. C. collata is in fact a rare species occurring in China w ith im bricaric acid as m ajor m edullary sub­ stance b u t lacking soredia, isidia or lobules (Culberson 1965, C ulberson & Culberson 1968). N ot a single specimen examined from the U S S R corresponds to these characteristics. T his m isunderstanding is probably due to a somewhat different circum scription of Cetraria collata in the Soviet lichenological literature. It mainly included various infraspecific taxa presently treated as synonyms of other species in Cetrelia , e.g., Cetraria collata f. isidiata Asah. = Cetrelia braunsiana Culb. & C. Culb and Cetraria collata f. microphyllina (Hue) Zahlbr. = Cetrelia japonica (Zahlbr.) Culb. & C. Culb. Cetrelia orientalis Randl. & Saag sp. nov. Thallus mediocris, 10-12 cm latus; laciniae 0-5-1 -5 cm latae, marginibus dense intructae vel planta tota saepe critata lobulis, vel expansis et foliosis. Superficies superior cana vel cinerea, levis, pseudocyphellata; pseudocyphellae punctatae vel nonnihil elongatae, parvae (tenus 0-5 mm latae). Superficies inferior nigra, marginibus castanea vel pallida, levis vel subrugosa, non punctata; rhizinae nigrae. Cortex superior 16-23 (im crassus, medulla 112—162 (im crassa, cortex inferior 20-23 (im crassus. Apothecia et pycnidia ignota. Atranorinum in cortice superiore, acidum alectoronicum et acidum a-collatolicum in magna summa et acidum physodicum et acidum 4-0methylphysodicum in minima summa in medulla. Typus: USSR, the Far East, Khabarovsk region, the Badzhal Mountains, the valley of the Urmi River, mixed forest, on a fallen trunk, 2 July 1981, Randlane 223 (TU—holotypus.) (Figs 1,4) M orphology: Thallus m edium , 10-12 cm broad; lobes 0-5-1-5 cm broad, the m argins densely fringed w ith lobulae, quite broad and lobe-like so that the specimen may appear cristate. Upper surface light grey to ashy, smooth, pseudocyphellate, the pores punctiform or somewhat elongate, small (up to 0-5 m m broad). Lower surface black, m arginal zone brow n to pale, sm ooth or 1991 Cetrelia— Randlane & Saag 117 Fig. 4. Cetrelia orientalis (magnified x 1-5). little wrinkled, not punctate; rhizines black. Upper cortex 16-23 |im thick, medulla 112-116 (im thick, lower cortex 20-23 fim thick. Apothecia and pycnidia not observed. Chem istry: Atranorin in the upper cortex, alectoronic and a-collatolic acids in m ajor am ounts and physodic and 4-0-m ethylphysodic acids in m inor am ounts in the medulla. T h is species is mainly characterized on its m edullary chem istry and presence of abundant marginal lobules. It differs from C. japonica and C. pseudolivetorum distributed in the same region in the presence o f alectoronic and a-collatolic acids in the medulla as the m ajor compounds. Cetreliia pseudocollata Randl. & Saag sp. nov. Thallus mediocris; laciniae 0-7-2 0 cm latae. Superficies superior glauca vel cinerea, levis, pseudocyphellata; pseudocyphellae magnae, crebrae, saepe confluentes et ad 1 mm. Superficies inferior nigra, marginibus castanea, levis vel subrugosa, nonnihil nitidus, fere punctata; rhizinae nigrae. Cortex superior 11-20 (im crassus, medulla 70-112 |im crassa, cortex inferior 11-16 um crassus. Apothecia ignota. Pycnidia numerosa, marginalia; conidia 5-6 x 1-1 -5 (im recta, extremis nonnihil inflatis. Atranorinum in cortice superiore, acidum microphyllinicum in medulla. Typus: China, Anhui, Mountain Huang Shan, Beihai, on the bark of Quercus sp., alt. 1720 m, 1980, J. C. Wei [Lich. Sinenses Exs. no. 20] (LD—holotypus). (Fig. 5) M orphology: Thallus m edium ; lobes 0-7-2 0 cm broad. Upper surface green­ ish grey to ashy, smooth pseudocyphellate, the pores large, num erous, often becoming confluent and exceeding 1 m m . Lower surface black, m arginal zone brow n, smooth or little wrinkled, somewhat glossy, w ith single pores; rhizines black. Upper cortex l l - 2 0 |im thick, m edulla 70-112 (im thick, lower cortex 118 THE LICHENOLOGIST Vol. 23 F ig . 5. Cetrelia pseudocollata (magnified x 1-5) 11-16 |im thick. Apothecia not observed. Pycnidia num erous, m arginal, unstalked; conidia bifusiform,, the ends slightly inflated, 5-6 x 1-1-5 |im. Chem istry : Atranorin in the upper cortex, microphyllinic acid in the medulla. Cetrelia pseudocollata is mainly characterized by the m edullary chem istry. It is structurally similar to C. collata and C. nuda Culb. & C. Culb. and differs from these entities in the presence of microphyllinic acid as m edullary com­ pound. O ther taxa w ithout vegetative propagules such as C. davidiana Culb. & C. Culb., C. sanguinea Culb. & C. Culb. and C. delavayana Culb. & C. Culb. have tiny punctiform pseudocyphellae in contrast to the large irregular pseudocyphellae o f C. pseudocollata. K ey to th e S p ecies o f C e tre lia 1 Terricolous, occurring on tundra soil. T hallus sterile, lacking propa­ gules for asexual r e p r o d u c tio n ......................................... C . alaskana Corticolous or occurring on boulders in the zone of mixed or deciduous forest. Thallus usually sorediate, isidiate, w ith lobules or w ith a p o th e c ia ................................................................................................... 2 2(1) Thallus sorediate. D istributed in Europe, eastern N orth America and in E and SE A s ia ............................................................................................ 3 Thallus not sorediate. Species restricted to E and SE A s i a ................. 6 1991 Cetrelia— Randlane & Saag 119 3(2) M edulla C + pink to reddish (olivetoric a c id ) ..............C . o liv e to ru m M edulla C — ...................................................................................................4 4(3) M edulla K C + red (alectoronic and a-collatolic acids) . . C. c h ic ita e M edulla K C - .................................................................................... . . . . 5 5(4) Perlatolic acid as the major medullary com ponent (T L C needed) . . . .............................................................................................C. c e tr a r io id e s Im bricaric acid as the major m edullary com ponent ( T L C ) ................. . ......................................................................................... C. m o n a c h o r u m 6(2) Thallus with cylindrical and coralloid isidia or dorsiventral lobules, apothecia extremely r a r e ........................................................................ 7 T hallus w ithout isidia or lobules, often with a p o th e c ia .....................12 7(6) M edulla C + pink to r e d ............................................................................... 8 M edulla C — ................................................................................................... 9 8(7) Thallus with abundant m arginal dorsiventral lobules, some small isidialike structures may also be present. Olivetoric acid as the major m edullary co m p o n en t......................................... C. p s e u d o liv e to ru m T hallus with globose or coralloid isidia (may be poorly developed), dorsiventral lobules absent. Anziaic acid as the m ajor m edullary c o m p o n e n t .............................................C. is id ia ta Culb. & C. Culb. 9(7) M edulla K C + red .................................................................................. 10 M edulla K C —. Im bricaric acid as the m ajor m edullary com ponent . . .................................................................... C. sin e n sis Culb. & C. Culb. 10(9) Thallus with laminal and marginal well developed isidia. Alectoronic and a-collatolic acids as the major medullary c o m p o n e n ts ................. ............................................................................................ C. b r a u n s ia n a Thallus with mainly marginal, dorsiventral lo b u les............................11 11(10) M icrophyllinic acid as the major m edullary com ponent (T L C ) . . . . ................................................................................................... C. ja p o n ic a Alectoronic and a-collatolic acids as the major m edullary components ( T L C ) .................................................................................. C. o rie n ta lis 12(6) M edulla C + pink to red ............................................................................13 M edulla C — ................................................................................................14 13(12) Olivetoric acid as the major medullary com ponent ( T L C ) ..................... ................................................................................................C. d a v id ia n a Anziaic acid as the major m edullary com ponent (T L C ) C. s a n g u in e a 14(12) M edulla K C + r e d ...................................................................................... 15 M edulla K C — .............................................................................................16 15(14) Alectoronic and a-collatolic acids as the major m edullary components ( T L C ) ............................................................................................ C. n u d a M icrophyllinic acid as the major m edullary component (T L C ) . . . . ......................................................................................... C. p s e u d o c o lla ta 21 120 THE LICHENOLOGIST Vol. 23 16(14) Pseudocyphellae large, irregular and becoming confluent, some of them more than 1 mm diam. Im bricaric acid as the major medullary c o m p o n e n t ............................................................................... C. collata Pseudocyphellae small, punctiform , less than 1 m m in diam. Perlatolic acid as the major m edullary c o m p o n e n t ................. C. delavayana Species Concept in the Genus Cetrelia Traditionally a type of chemical species concept has been accepted within this group of lichens. Different species with similar or even totally identical morphology may differ from each other only by the medullary constituents (Culberson & Culberson 1976). We propose here to call the morphological groups separated by the Culbersons ‘m orphotypes’. T h e term s ‘m orphotype’ and ‘chem otype’ have been given precise meanings and they are applied to populations of undeterm ined taxonomic rank or of no taxonomic value (Hawksworth 1974). I f these term s are usually used for m arking infraspecific variations, then sometimes they can also be applied to supraspecific groupings, as the concept of species in different lichen genera varies widely. Generally, recognized species in the genus Cetrelia are restricted w ithin very narrow limits and do not contain essential morphological or chemical variation per se. Generally, each species w ithin the genus Cetrelia belongs to one m orphotype and to one chemotype. T hus, the combination of the m orpho- and chemotypes makes it possible to characterize concisely all 17 known species (Table 1). T his enables the table to be used as an identification guide to the species. Besides its practical use, this table also has some theoretical value. Vacant squares in the table mark the species that are theoretically possible in the genus. T his sugges­ tion is confirmed by the fact that there were two specimens found in our ana­ lyses that did not belong to any hitherto described species but fitted the vacant places in the table. Since we consider that material showing new combinations of known m orpho- and chemotypes deserves the rank of species the two new species C. orientalis and C. pseudocollata have been described above. Some hints about one more specimen fitting another vacant space T able 1 have already been published. In the description of C. sanguinea , C ulberson & Culberson (1968) noted a single Japanese collection lacking apothecia and possessing marginal lobules, somewhat rem iniscent of C. japonica. T h eir chemical studies showed that it contained anziaic acid. For that reason we believe the sample could be assigned to the m orphotype of sinensis and chemotype IV. T his place in the table is unfilled. U nfortunately we have not seen the herbarium material and therefore cannot decide its proper taxonomic position. Morphotypes M orphological groups within Cetrelia as presented by Culberson & Culberson (1976) were designated by the L atin epithets of the m ost prom inent species in the respective group. W e have adopted the same term inology here. Five m orphotypes can basically be recognized in the genus: cetrarioides, thallus with soredia; isidiata, thallus w ith isidia; sinensis, thallus w ith m arginal dorsiventral lobules; collata , thallus without soredia, isidia or lobules, often with T a b le 1. M orpho- and chem otypes o f Cetrelia ( new species are in bold italics) Chemotypes and their major components Morphotypes and their diagnostic characters Cetrarioides Thallus sorediose Isidiata Thallus isidiate Sinensis Thallus with lobules Collata Thallus without vegetative propagules; large pseudocyphellae Davidiana Thallus without vegetative propagules; small pseudocyphellae I Alectoronic and a-collatolic II Microphyllinic C. chicitae III Olivetoric IV Anziaic C. cetrarioides C. olivetorum C. braunsiana V Perlatolic VI Imbricaric C. monachorum C. isidiata C. o rie n ta lis C. japonica C.nuda C. p seu d o co lla ta C. sinensis C. pseudo-olivetorum C. collata C. davidiana C. sanguinea C. delavayana C. alaskana THE LICHENOLOGIST 122 Vol. 23 apothecia and large pseudocyphellae; davidiana , thallus w ithout vegetative propagules and frequently w ith apothecia, but pseudocyphellae small. T he division of species into m orphotypes is, however, somewhat conventional. For example we suggest that C. alaskana should be placed in the m orphotype davidiana , although it differs externally from the other species of this m orpho­ type (C. davidiana , C. delavayana, C. sanguined) to some degree. T his is prob­ ably due to the different habitat selection in C. alaskana , since it is the only terricolous species in the genus occurring on barren tundra soil. Cetrelia sanguinea belongs in its typical form to the same m orphotype but occasionally marginal lobules can also be developed, which makes it fall into the variation type of sinensis as defined above. Retaining the division of Cetrelia species into m orphotypes makes the survey of the genus simpler. Chemotypes As with the m orphotypes, the genus Cetrelia also contains a num ber of chemotypes related to the content of major m edullary substances. Species of a certain chemotype always have the same (one or two) major constituents while the complex of m inor substances may vary somewhat (in some species all the m inor substances are possibly not yet characterized). Six chemotypes have so far been recognized in our examined material (Table 2) and also in the genus as a whole. T h e num bering of the chemotypes corresponds to the trend of chemical evolution with reduced side-chain length in depsides and depsidones (Culberson & Culberson 1976). All the Cetrelia chemotypes are certainly related to each other, but the degree of connection between different chemo­ types is unequal. We propose the following scheme of chemical relationships between the chemotypes observed in Cetrelia. / K/ 'I ''I I Chemotype I is presum ably the m ost diverging type since it is the only chemotype which contains depsidones and no depsides. In addition it is the only chemotype where the detected com pounds are not represented in the other chemotypes. Both major constituents of chemotype I have a great num ber of side-chain carbons (7-7). T his is the reason why chemotype I is somewhat related to chemotype II. T h e major constituent of the latter, i.e. microphyllinic acid which combines two units of seven-carbon side chains, was considered to be the m ost prim itive of the com pounds occurring in Cetrelia (Culberson & Culberson 1976: 336). Chem otype III is a progression in the reduction of side chains of the major compound. Chemotypes IV and V, both characterized by major substances with 5 + 5 carbons in the side chain, could be interpreted as having reduced series of side chains. Chem otype VI can be 1991 123 Cetrelia■—Randlane & Saag T a b le 2. Contents of major (M ) and minor (m ) medullary substances in the chemotypes of genus Cetrelia* Medullary compounds Depsides Depsidones K 3 O </) rj* J2« •5 < L> o u *2 O ou» '•5 o/i §<u o < >» oi U J5 < 4* 3 '3 % JS aQ l-i 0 1 *3 XI +-» u 6(U J? n Cu o O <D 9 u > © u % ii O 4 y ‘C y «y *3 6 _o o <u > •5 1> y s o *cN 4 < 2 D >■> XI V 6K Q1 o 4 x> 6 ’oj s o 0 •S <u y o _u w 6V 'u u y 3 'C Qi *C TO X > o s Jo s 4 Number of carbons in side-chain Species C. chicitae C. braunsiana C. orientalis C. nuda II C. japonica C. pseudoeoilata III C. olivetorum C. pseudolivetorum C. davidiana IV C. isidiata C. sanguinea V C. cetrarioides C. delavayana VI C. monachorum C. sinensis C. collata C. alaskana 7— 7 M M M M M M M M 7— 5 m m m m 7—7 7—5 5— 5 5— 3 3— 3 m m m m m m m m m M M m m M M M m m m m m m m m m M M m m m m m m m m M M m m M m m m m m m m m m m m m m m m m m m m M M M M m m m m ‘ Data from Culberson & Culberson (1976) with some additions. interpreted as the most advanced chemosyndrome characterized by several minor substances including divaricatic acid which represents the most advanced product in the genus with two units of three-carbon side chains. It is remarkable that the similarity of different chemotypes grows with numeration. Chemotypes IV, V and VI form a relatively uniform group regarding their chemosyndromes. Discussion In the genus Cetrelia the highest number of combinations of morpho- and chemotypes (i.e. species) are represented in eastern and south-eastern Asia 22 THE LICHENOLOGIST 124 Vol. 23 S o re d ia (? ) 4 Is id ia _ L o b u le s F ra g m e n ts of - 9 • • 3 II • • • 2 6 8 o o 13 17 a T3 c o o a) CO • 15 o 16 th a llu s A p o th e c ia •» i i l • 5 I • I i 7 HI i i l 1 in <0 o a> CL w >> >> w> o .® • 10 32 i i • • .§ 12 14 a. S’ 3z: 3zr Direction of chemical evolution (chemotypes) Fig . 6. Evolutionary relationships between the Cetrelia species. Restricted to eastern and southeast­ ern Asia ( • ) , limited distribution outside eastern and southeastern Asia (o), wide distribution outside eastern and southeastern Asia (O). 1, C. nuda\ 2, C. orientalis', 3, C. braunsiana; 4, C. chicitae; 5, C. pseudocollata', 6, C.japonicw, 7, C. davidiana; 8, C. pseudolivetorum; 9, C. olivetorum', 10, C. sanguinea\ 11, C. isidiata', 12, C. delavayanaj 13, C. cetrarioides; 14, C. collata', 15, C. alaskana] 16, C. sinensis’, 17, C. monachorum. (Culberson & Culberson 1968). From among five m orphotypes, only one, namely cetrarioides, is widely distributed in the world, whereas the areas of non-sorediate m orphotypes (isidiata, sinensis, collata and davidiana) fall only into East and Southeast Asia. Lichens that have developed soredia can expand and effectively enlarge their area of distribution in contrast to species that use only ascospores. T h e complicated process of the resynthesis of a lichen thallus from a germinated ascospore and a free-living photobiont first depends upon the distribution area of algae (T ehler 1982). T h e other forms of vegative propagules should theoretically also have the ability to colonize habitats in which the photobiont cannot thrive in its free-living form. In the genus Cetrelia , isidia and lobules still appear to be almost as ineffective in colonizing new territories as the spores. T h u s, only four sorediate species (C. cetrarioides, C. monachorum, C. olivetorum and C. chicitae ) are also found in Europe and N orth America. T h e centre of speciation in the genus is presum ably located in eastern and southern Asia, as seen from the vast m ajority of the combinations of chemo- and morphotypes. Evolutionary relationships T h e first scheme dealing with the connections between species of Cetrelia was proposed by Poelt (1970, 1972) and related to his discussion on ‘speciespairs’. O ur scheme includes all the combinations of chemo- and m orphotypes known up to now in the genus (Fig. 6). T h e horizontal axis on the scheme shows the direction of chemical evolution and the vertical axis represents the variety of reproductive propagules. We presum e that the form ation and evolution of biochemical pathways producing the complex of lichen substances has taken place during sexual stages in the evolution of lichens, rather than through m utations in asexual stages (Bowler & R undel 1975). T h e genus Cetrelia fits rather well into the theory of developm ent of species pairs since every chemosyndrome is represented not only by the ‘secondary’ but also by the ‘prim ary’ 1991 Cetrelia— Randlane & Saag 125 species. This avoids the necessity of referring to some hypothetical ancestral taxa. All ‘primary’ species with sexual reproduction appear to be quite rare with a very restricted distribution. However, we observe not only pairs of species but also triplets or even tetrads of species in the genus Cetrelia. A similar pattern has been reported only in a few other cases, for example in Parmelia and Physcia (Hawksworth & Hill 1984). The taxonomic treatment proposed by Tehler (1982) to recognize fertile and sterile counterparts as infraspecific taxa with the rank of forma could theoreti­ cally be acceptable, but in practice it is not. ‘Primary’ species differ essentially from the ‘secondary’ species by their morphology as well as the ‘secondary’ species of one column from the other. It is difficult to imagine, for example, the treatment of C. chicitae, C. braunsiana} C. orientalis and C. nuda as one species. The morphological similarity is great between taxa such as C. orientalis , C. japonica, C. pseudolivetorum and C. sinensis, or C. chicitae , C. olivetorum , C. cetrarioides and C. monachorum but presumably they all have quite a different origin and cannot belong to one species. Otherwise we would be ‘ back to the traditional typological species concept adamantly blind to everything but morphology ’ (Culberson, 1986). One species, C. alaskana , with an isolated distribution, however, does not fit too well into the normal primary/secondary species relationship described above. This species is not an epiphyte, as are essentially all the other examined species, and furthermore neither soredia, isidia, lobulae nor any sexual repro­ ductive structures have been observed. Presumably it reproduces through frag­ mentation as do many other macrolichens growing on tundra soil. Therefore, we suggest that C. alaskana can also be placed in the group of ‘secondary’ species. The frequency and distribution of the examined species in the genus Cetrelia , in our opinion, serve as good examples of some of the theoretical assumptions discussed above. All the sorediate species, although common and widely distributed, may be considered ‘evolutionary blind alleys’. The organism loses its genetic flexibility by having an asexual propagation system superior to the sexual process (Tehler 1982). On the other hand, the sexual species, although occasionally less successful in dispersing, have retained their ability for further speciation by the process of gene recombination. We wish to express our gratitude to the curators in charge of the herbaria listed in the text. Edward Sakk is thanked for taking the photographs. Special thanks are due to Sergej Kondratjuk-from Kiew, Dr Gertrud Dahlgren and Jan-Eric Mattsson from Lund for general support and practical help. We are mostly grateful to Dr I. Karnefelt for revising the manuscript and making valuable suggestions for improving the text. R eferences Bowler, P. A. & Rundel, P. W. (1975) Reproductive strategies in lichens. Botanical Journal Linnean Society 70:325-340. Culberson, C. F. (1972) Improved conditions and new data for the identification of lichen products by a standardized thin-layer chromatographic method. Journal of Chromatography 72: 123-125. Culberson, C. F . (1974) Conditions for the use of Merck silica gel 60 F 254 plates in the standar­ dized thin-layer chromatography technique for lichen products. Journal of Chromatography 97:107-108. 126 THE LICHENOLOGIST Vol. 23 Culberson, C. F. & Culberson, W. L. (1976) Chemosyndromic variation in lichens. S ystem atic B o ta n y 1:325-339. Culberson, W. L. (1965) C etraria chicitae , a new and widely distributed lichen species. B ryologist 68:95-99. Culberson, W. L. (1986) Chemistry and sibling speciation in the lichen forming fungi: ecological and biological considerations. B ryologist 89:123-131. Culberson, W. L. & Culberson, C. F. (1968) The lichen genera Cetrelia and P latism atia (Parmeliaceae). Contributions from the U n ited S tates N a tio n a l Herbarium 34:449-558. Goward, T. (1985) A h tian a , a new lichen genus in the Parmeliaceae. B ryologist 88:367-371. Hawksworth, D. L. (1974) M ycologist’s handbook. Kew: Commonwealth Mycological Institute. Hawksworth, D. L. & Hill, D. J. (1984) The Lichen-forming Fungi. Glasgow: Blackie. Knjazheva, L. A. (1973) Lichens o f the southern p a rt o f Prim orje Region. Dissertation for the cand. degree. Vladivostok (in Russian). Kopaczevskaja, E. G., Makarevicz, M. F., Oksner, A. N. & Rassadina, K. A. (1971) Handbook o f the lichens o f the U S S R I (I. I. Abramov, ed.). Leningrad: Nauka (in Russian). Makarova, I. I. (1980) Species lichenum pro USSR et peninsula Czukotka novae. N o vita tes system aticae plantarum non vascularium 17:150-152 (in Russian). Malysheva, N. V. & Smirnov, A. G. (1982) K e y f o r the lichens o f the T artar A S S R . Kasan: Kasan University Press (in Russian). Poelt, J. (1970) Das Konzept der Artenpaare bei der Flechten. Deutsche Botanische Gesellschaft. Neue Folge 4:187-198. Poelt, J. (1972) Die taxonomische Behandlung von Artenpaaren den Flechten. B otaniska N otiser 125:77-81. Skirina, I. F. (1987) Lichens o f the western slopes o f the M iddle S ikh ote-A lin . Vladivostok: Pacific Institute of Geography (in Russian). Skirina, I. F. & Knjazheva, L. A. (1985) Lichens o f the eastern slopes o f the M iddle S ikh ote-A lin . Vladivostok: Pacific Institute of Geography (in Russian). Tehler, A. (1982) The species pair concept in lichenology. Taxon 31:708-714. A ccepted f o r publication 22 October 1990 IV 23 Randlane, T., Thell, A., & Saag, A. 1995. New data about the genera Cetrari­ opsis , Cetreliopsis and Nephromopsis (Fam. Parmeliaceae, lichenized Ascomycotina). — Cryptogamie, Bryologie-Lichenologie 16: 35-60. Cryptogamie, Bryol. Lichenol. 1995, 16 (1): 35-60 35 NEW DATA ABOUT THE GENERA CETRARIOPSIS, CETRELIOPSIS AND NEPHROMOPSIS (FAM. PARMELIACEAE, LICHENIZED ASCOMYCOTINA) Tiina RANDLANE1, Ame THELL2 and Andres SAAG1 1Institute of Botany and Ecology, Tartu University Lai Street 38, Tartu, Estonia EE-2400 2 Department of Systematic Botany, University of Lund Ostra Vallgatan 18-20, S-223 61 Lund, Sweden ABSTRACT - New morphological, anatomical and chemical data about the lichen genera Cetra­ riopsis, Cetreliopsis and Nephromopsis are presented. Two new species, Cetrariopsis laii Thell & Randl., Cetreliopsis papuae Randl. & Saag, and one new subspecies Cetreliopsis rhytidocarpa subsp. ilangtangi Randl. & Tell, are described and the following new combinations proposed: Cetra­ riopsis pallescens (Schaerer) Randl. & Thell, Cetrariopsis pallescens var. citrina (Taylor) Thell & Randl., Cetreliopsis asahinae (Sato) Randl. & Thell, Cetreliopsis endoxanthoides (Awasthi) Randl. & Saag and Cetreliopsis laeteflava (Zahlbr.) Randl. & Saag. Cetrariopsis comprises now two, Cetre­ liopsis five and Nephromopsis nine species. All the three genera, distributed in East and Southeast Asia, are characterized by the dorsiventral foliose thallus, presence of pseudocyphellae on the lower surface, clavate asci usually with a rather large axial body in the tholus, oblong or ellipsoid ascospo­ res and usnic acid as the cortical substance. RESUME - Des donn£es morphologiques, anatomiques et chimiques nouvelles sont presentees, concemant les genres Cetrariopsis, Cetreliopsis et Nephromopsis (lichens). Deux nouvelles esp&ces, Cetrariopsis laii Thell & Randl., Cetreliopsis papuae Randl. & Saag. et une nouvelle sous-esp&ce Cetreliopsis rhytidocarpa subsp. langtangi Randl. & Thell sont dgcrites. Les combinaisons suivantes sont proposees: Cetrariopsis pallescens (Schaerer) Randl. & Thell, Cetrariopsis pallescens var. citrina (Taylor) Thell & Randl., Cetreliopsis asahinae (Sato) Randl. & Thell, Cetreliopsis endoxan­ thoides (Awasthi) Randl. & Saag et Cetreliopsis laeteflava (Zahlbr.) Randl. & Saag. Cetrariopsis comprend maintenant deux esp£ces, Cetreliopsis, cinq et Nephromopsis, neuf. Ces trois genres, distribuds en Asie de l’Est et du Sud-Est, sont tous caracteris^s par un thalle foliac£ h sym£trie dorsiventrale, la presence de pseudocyphelles ii la face infdrieure, des asques clavules avec, dans le bouchon apical, un corps axial relativement grand, des ascospores oblongues et ellipsoidales et de l’acide usnique dans le cortex superieur. KEY-WORDS - lichenized Ascomycotina, Parmeliaceae, Cetrariopsis, Cetreliopsis, Nephromopsis, anatomy of thallus and ascocarps. INTRODUCTION 1993) A general review of cetrarioid lichens (Kamefelt et al. 1992, Randlane & Saag has clearly shown that besides the indistinctly limited genus Tuckermannopsis, T. RANDLANE, A. THELL and A. SAAG 36 the complex Cetrariopsis - Cetreliopsis - Nephromopsis is another poorly studied group of taxa. In this paper we delimit these three genera, compare their various characters and identify the generic positions of the species involved. A new genus, Tuckneraria, was segregated from Nephromopsis in one of our previous papers (Randlane et al. 1994). As we consider Tuckneraria evolutionary allied rather to Tuckermannopsis than to Nephromopsis, the species included in that entity are not treated here. MATERIAL AND METHODS More than 300 herbarium specimens from BM, CAN, FH, G, GZU, H, KW, LD, LE, M, PC, S, TAIM, TNM, TU, UPS, US and private herbaria of A. Aptroot, D.D. Awasthi and J. A. Elix were examined. Anatomical studies of cortical and reproductive structures were carried out on most of the specimens. Sections were made with a kryomat, Leitz freezing microtome and put in lactophenol cottonblue. After pretreatment with 10% KOH-solution, asci were squashed in a 0,3% Lugol’s solution. The characters were studied with a Zeiss Axioscope light microscope, and the photos were made with a Zeiss M 35 W camera. Chemical analyses were carried out according to the standardized TLC methods (Culberson & Kristinsson 1970, Culberson 1972) on about 200 specimens. The acetone extracts were run in solvent systems B, C and G (Culberson et al. 1981). RESULTS AND DISCUSSION 1. The genus Nephromopsis Mull. Arg. This genus was described already in 1891 by Muller Argoviensis to accomodate N. stracheyi, which was said to have a thallus similar to Cetreria but the position of apothecia like in Nephroma. In contemporary lichenology the genus was not generally recognized until 1980 when Lai resurrected the treatment of the species with cetrarioid thallus, nephromoid apothecia and pseudocyphellae on the lower surface, in a separate genus under the name Nephromopsis. Still, the marginal position of apothecia on the lower side of the thallus is a character that could be easily misidentified and also the anatomical differences had not been studied sufficiently in that time. In this paper we continue the revision of Nephromopsis (Randlane & Saag 1991, 1992) by presenting detailed anatomical data characterizing the whole genus, transferring some species (N. asahinae, N. endoxanlhoides, N. pallescens) to other genera (Cetreliopsis, Cetrariopsis) and complementing the description of two rarely collected species (N. morrisonicola, N. yunnanensis). The genus Nephromopsis is defined by the following characters: a large foliose thallus; marginal apothecia on the lower side of the thallus; presence of pseudocyophellae only on the lower surface; three-layered exciple; narrowly clavate asci and oblong ascospores; bifusiform pycnoconidia; usnic acid in the cortex; various fatty CETRARIOPSIS, CETRELIOPSIS AND NEPHROMOPSIS 37 acids and/or some orcinol depsides-depsidones in the medulla. The genus includes according to our present knowledge nine species: N. ectocarpisma (Hue) Gyelnik N. endocrocea Asahina N. isidioidea (Rasanen) Randl. & Saag N. komarovii (Elenkin) Wei N. morrisonicola Lai N. ornata (Miill. Arg.) Hue N. rugosa Asahina N. stracheyi (Bab.) Miill. Arg. N. yunnanensis (Nyl.) Randl. & Saag Several other species that have been treated as belonging to Nephromopsis do not correspond to these characters and were lately transferred to the other genera. Thus N. laureri (Kremp.) Kurok., N. laxa (Zahlbr.) Sato and N. pseudocomplicata (Asah.) Lai [including N. nipponensis (Asahina) Lai] differ in having marginal cilia, globose to subglobose ascospores, a two-layered exciple and were therefore removed to Tuckneraria (Randlane et al. 1994). N. globulans (Nyl. ex Hue) Lai shows clear affini­ ties (globose to subglobose ascospores, filiform pycnoconidia, presence of secalonic acid in the medulla) with the genus Allocetraria. N. asahinae (Sato) Rasanen and N. endoxanthoides (Awasthi) Randl. & Saag are transferred here to Cetreliopsis (see be­ low) due to the presence of pseudocyphellae also on the upper surface, two-layered exciple, rather broadly clavate asci and contents of fumarprotocetraric acid in the me­ dulla. N. pallescens (Schaerer) P&rk has apothecia situated laminally on the upper surface and is included on these grounds in Cetrariopsis (see below). NEPHROMOPSIS Mull. Arg., Flora 74: 374 (1891). Type species: Nephromopsis stracheyi (Bab.) Mull. Arg. - (Fig. 15). Thallus foliose, may be very large (up to 20 cm in diameter), often remarkably rugose or reticulated, greenish yellow on the upper and light to dark brown or even black on the lower surface. Isidia, soredia and true cilia absent but numerous black lamina! and marginal emergent projections bearing pycnidia at their tops are present in many species. Pseudocyphellae occur over the lower surface either in the form of small white dots or larger regular patches, they may be plain, concave or convex, usually located on the ridges of the thallus or on the special plug-like outgrowths. Rhizines sparse or numerous, of the same colour as the lower surface. Pycnidia marginal and laminal, often on emergent black projections but may also be immersed in the thallus. Cortical layers paraplectenchymatous, either of the same type as in Cetrariopsis (see below) with larger cells near the medulla or with rather equal-sized, strongly gelatinized cells also characteristic for the genus Flavocetraria (Kamefelt et al. 1994). Apothecia marginal, sometimes very large (to 32 mm in diameter), situated on the lower surface of the thallus but the disc often turns upwards which may confuse the true position of the ascocarps. Disc brown, rounded or reniform in outline, may 24 38 T. RANDLANE, A. THELL and A. SAAG have a short stalk. Exciple usually distinctly three-layered, asci narrowly clavate, 30-70 x 8-14 |im, mainly with an axial body of medium size, 2-4 (am broad, but in some spe­ cies with an amyloid apical ring structure and a very small axial body (0.5-1.2 jam broad) like in Flavocetraria (Karnefelt et al. 1994). Ascospores 6-12 x 3-6 |im. Pycnoconidia bifusiform (dumb-bell shaped), 4-5 x 1-1.5 pm. Chemical constituents: usnic acid (+/-) in the cortex; various fatty acids (lichesterinic, protolichesterinic, caperatic, nephrosteranic acids, etc.) and some orcinol depsides and depsidones (olivetoric, anziaic, physodic, conphysodic acids) in the me­ dulla. A few species contain also substances related to the anthraquinonic pigments (endocrocin, secalonic acids A and C) colouring the medulla either orange or yellow. Distribution and habitat: Eastern and Southeastern Asia. All the species within Nephromopsis with the exception of N. komarovii are corticolous on deciduous or coniferous trees in the montaneous forests. The latter grows on rocks and boulders or on the ground. Two species of nine - N. morrisonicola and N. yunnanensis - have earlier been represented by a few specimens from very limited areas only (Taiwan, Mt Morrison and China, prov. Yunnan accordingly). The descriptions of these rare lichens are com­ plemented here, since we have identified some additional specimens from other locali­ ties. Nephromopsis morrisonicola Lai - (Fig. 10) Quart. J. Taiwan Mus. 33: 223, 1980. - Type: Taiwan, Nanton Co., Mt. Morri­ son, alt. 3500-3900 m, Lai, 1978, no. 10438 (TAIM, holotype; seen). Thallus foliose, up to 8 cm in diameter; upper surface yellow, smooth in the younger parts and somewhat rugose in the center; lower surface black with brown or even whitish margins; pseudocyphellae on the underside in the form of white conspi­ cuous rounded spots, plain or slightly elevated. Rhizines sparse, black or brown, sim­ ple. Pycnidia marginal, located on the black emergent projections, may be absent on younger specimens. Both cortical layers c. 25 |im thick, composed of about three cell layers each, with the cells near the medulla somewhat larger. Lower cortex brownish pigmented. Medullary hyphae c. 5 |im in diameter. Apothecia marginal, up to 9 mm in diameter, disc brown, rounded or irregular, faced upwards. Exciple usually three-layered. The upper layer up to 50 |im thick and the middle layer, composed of cells with larger lumina, if present, 10-15 pm thick.The thickness of the lowest layer is also up to 50 jam. Asci 35-70 x 10-15 pm, axial body 2-4 |im, tholus may have an undistinct ring structure similar to that in N. endocrocea and N. ornata. Ascospores 7-12 x 3-6 pm. Pycnoconidia not seen. Chemical constituents: usnic acid (+/-) in the cortex; lichesterinic, protoliches­ terinic acids and additionally some unidentified fatty acids (+/-) in the medulla. Distribution and habitat: originally described as a Taiwanese endemic (Lai 1980). We have identified N. morrisonicola also from the Philippines, Java, Borneo and CETRARIOPSIS, CETRELIOPSIS AND NEPHROMOPSIS 39 the mainland of China. The lichen grows in comparatively high altitudes (about 2500 m and higher) on coniferous trees or shrubs. N. morrisonicola (Fig. 10) is easily recognized due to its black lower surface. Black underside is present also in N. ornata and N. endocrocea but both of them have coloured medulla. Lai (1980) took notice of the morphological resemblance between N. morrisonicola and Cetreliopsis asahinae. Still, the latter has laminal pseudocyphellae also over the upper cortex and the fumarprotocetraric acid, characteristic to the genus Cetreliopsis, is easily identified with Pd positive reaction on the medulla. Specimens examined. Taiwan. Nanton Co., Mt. Morrison, Maiyun Hostel to the peak, alt. 3500-3900 m, Lai, 1978, no 10 438 (holotype), no 10 459 (TAIM). China. Sikang, Kangting distr., Giilingkong, Gomba La, alt. 3700 m, Smith, 1934, no 14 075 (UPS). Indonesia. Borneo, Mt. Kina­ balu, Palca Cave to Lobang, Strong Clemens, 1915, no. 10 760 (FH). Mt. Kinabalu, alt. 3600 m, Samsudin, 1984, (Herb. Elix). Java. Pangerango, Schiffner, 1894, no 3004 (FH, TNM, US). Karaah Marwak, Herman, 1913 (FH). Mt. Gedah, Seifriz, 1920, nos. 77, 1952 (US). Res. Pasoeroean, Goenoeng, Ardjoena, Lalidjiewa - Welirang track, Du Rietz, 1927, no 61b\l (UPS). Philippines. Luzon, Prov. Benquet, alt. 2400 m, Curron, Zsckokke, Merritt, 1909 (H). Nephromopsis yunnanensis (Nyl.) Randl. & Saag. Mycotaxon 44: 488, 1992. Basionym: Platysma yunnanense Nyl., Lich. Novae Zelandiae: 252, 1888 - Type: China, Yunnan, alt. 1800 m, Delavay, no. 1602 (H-NYL36 134, lectotype; seen). - Synonym: Cetraria yunnannensis (Nyl.) Zahlbr., Trudy Troitskos.-Kyakthtinsk. Old. Priamursk. Otd. Imp. Russk. Geogr. Obshch. 12: 89, 1911 (1909). Thallus foliose, up to 7 cm in diameter; upper surface light greenish or yellow­ ish, strongly rugose and reticulated in well developed specimens; lower surface white to light brown, conspicuously rugose, with numerous pseudocyphellae growing on the ridges of the underside or on the special outgrowths. Rhizines sparse, pale, simple. Pycnidia extremely numerous, situated laminally and marginally (and sometimes even laminally on the lower surface) on the emergent projections which are thallus coloured and black only at their tops. The projections may easily be broken away and leave white patches without cortical layer on the upper surface. Still, these patches can neither be considered true pseudocyphellae nor soredia as described by Hue (1899). Upper and lower cortex both c. 20 fam thick, composed of rather small, equal-sized, strongly gela­ tinized cells. Medullary hyphae 4-6 pm in diameter. Apothecia marginal, up to 13 mm in diameter, disc brown, rounded. Exciple two-layered, upper and lower layer c. 40 pm each. Occasionally, a thin, third middle-layer is present with larger lumina. Asci 40-45 x 12-14 pm, axial body 4 pm, ascospores 8-9 x 4-4.5 pm. Pycnoconidia 4 x 1 pm. Chemical constituents: usnic acid in the cortex; lichesterinic and protolichesterinic acids in the medulla. Distribution and habitat: China (prov. Yunnan); on deciduous and coniferous trees between 1800 and 2800 m altitudes. The most characteristic feature of this species is the abundance of laminal pyc­ nidia on pale emergent projections (all other species in Nephromopsis bear black pro­ 40 T. RANDLANE, A. THELL and A. SAAG jections, if they have them at all); pseudocyphellae on the lower surface are typically on the ridges and outgrowths of very rugose lowerside. The latter reminds somewhat of the lower surface of Cetrariopsis pallescens and is quite different from other species in Nephromopsis. Specimens examined. China. Yunnan, Delavay, no. 1602 (H, leccotype). Yunnan, An Ning Co., An Ning, alt. 1800 m, 24°55’ N, 102°29’ E, Koponen, 1981, no 37 925 (H). Yunnan, Kon-toni, Delavay, 18.06.1888 (PC). Yunnan, Lijiang Co., Yulongshan, alt. 2600-2800 m, 270l l ’N, 100°15’ E, A M et al., 1987, no 46 370a (H, TU). Yunnan, Lijiang Co., Yulongshan, Yufeng Temple, 26°58’ N, 100°12’ E,Ahti, 1987, no. 46 303 (H). Yunnan, Ninglang Co., alt. 2700 m, VV<wg Li-Song, 1987, no. 10 373 (H). II. The genus Cetrariopsis Kurok. The monotypic genus Cetrariopsis was separated from Cetraria s. lat. by Kuro­ kawa in December 1980. Less than a month later Lai proposed the genus Ahtia to accomodate the same species - Cetraria wallichiana (Taylor) Miill. Arg. Two essential characters were pointed out by both authors to describe the new genus - the numerous, small, laminal apothecia (Fig. 4) and the prosoplectenchymatous upper cortex. The structure of the cortex, however, was a misinterpretation by Kurokawa and Lai. In earlier papers the terms «prosoplectenchyma» and «paraplectenchyma» referred to the form of the cell lumina, but since studies of thallus structures were carried out by Hale (1976), these terms are usually applied to indicate the hyphal orientation in the cortex. Thus, according to Culberson & Culberson (1965, 1968) the upper cortex of Asahinea, Cetrelia and Platismatia was classified as prosoplectenchymatous, although not charac­ terized by a parallel periclinal orientation of the hyphae. All these genera as well as Cetrariopsis have a pachydermatous paraplectenchymatic cortex with randomly orien­ ted cells. Still, Cetrariopsis is probably not closely related to those entities as supposed earlier (Kurokawa 1980, Karnefelt et al. 1992, Elix 1993). The characteristic features of the genus Cetrariopsis are: a large foliose thallus; comparatively small, marginal as well as laminal apothecia on the upper side of the thallus; presence of pseudocyphellae only on the lower surface; two-layered exciple; narrowly clavate asci and oblong ascospores; usnic acid in the cortex; fatty acids or alectoronic acid in the medulla. This genus is considered to be a distinct taxon which is sometimes referred to as parmelioid. The laminal position of apothecia is really an easily recognized parmelioid character, while a nonpored epicortex (Elix 1993) and presence of pseudocyphellae on the lower surface remind of the cetrarioid genera. Our studies have shown that there is not much difference in anatomy of Cetrariopsis and Nephromopsis. Both genera are characterized by oblong ascospores and narrowly clavate asci with a medium sized axial body, 2-4 jim broad (Figs. 13, 15, 16). The exciple is two-layered in Cetrariopsis and three-layered in most species of Nephromopsis but sometimes the middle layer is not distinctly developed. The layer called upper excipular layer in this paper (Figs. 1, 12) is referred to by some authors as hypothecium. CETRARIOPSIS, CETRELIOPSIS AND NEPHROMOPSIS 41 1 I s i hymenium subhymenium upper excipular layer middle layer • # « lower layer algal layer Fig. I - Schematic drawing of the hymenial and excipular layers of Nephromopsis rugosa. The upper excipular layer is by some authors called hypothecium. Bar = 10 pm. The position of the apothecia is indeed a striking character but a more thorough examination shows that specimens with clearly laminal apothecia have usually fruiting bodies also along the margins. Furthermore, on many specimens with mostly marginal apothecia both submarginal or even truly laminal ones occur. Other morphological characters (reticulation of the thallus, presence of pseudocyphellae on the lower sur­ face, absence of isidia, soredia and cilia) as well as cortical and medullary chemistry do not show essentia! differences. However, we still prefer to maintain the separation of the genera Cetrariopsis and Nephromopsis upon the position of the apothecia (Cetrariopsis with laminal and marginal apothecia; Nephromopsis with marginal apo­ thecia only) until additional comparative studies have been carried out. The genus Cetrariopsis includes two species: C. pallescens (Schaerer) Randl. & Thell and the newly described C. lai Thell & Randl. CETRARIOPSIS Kurok., Mem. Natl. Sci. Mus. (Tokyo) 13:140 (1980). Type species: Cetrariopsis pallescens (Schaerer) Randl. & Thell. Thallus foliose, may be up to 16 cm in diameter but usually less than 10 cm, smooth to rugose, greenish yellow to yellow on the upper and white to yellow or brown on the lower surface. Isidia, soredia, cilia and emergent projections absent. Pseudocy­ phellae occur over the lower surface either in the form of small white dots or wider regular patches situated on the ridges of the thallus or on the special plug-like outgrowths. Rhizines sparse, white or brown. Cortical layers paraplectenchymatous, up to 30 pm thick, with larger cells near the medulla; medullary hyphae up to 6 pm thick. Apothecia 1-5 mm in diameter, laminal, submarginal or marginal but not situa­ ted on the lower surface of the thallus; disc brown, rounded or oblong and reniform, 25 42 T. RANDLANE, A. THELL and A. SAAG may be surrounded by a thin thalline margin. Exciple two-layered, upper layer c. 2030 nm and lower layer c. 30 |am thick. Asci narrowly clavate, 35-45 x 8-10 |im, axial body of medium size, 2.5-4 jam, ascospores oblong, 7-10 x 3-5 (jm. Pycnidia and pycnoconidia not seen. Chemical constituents: usnic acid (+/-) in the cortex; fatty acids (lichesterinic and protolichesterinic acids) and orcinol depsidones (alectoronic acid) in the medulla. Distribution and habitat: Eastern and Southeastern Asia. Epiphytic on deci­ duous and coniferous trees in the montaneous forests. Cetrariopsis pallescens (Schaerer) Randl. & Thell comb. nov. var. pallescens - (Fig. 4, 13) Basionym: Cetraria pallescens Schaerer, in Moritzi, Syst. Verz.: 129, 1846. Type: Java, Mt. Pangerango, Zollinger no. 449 (G, holotype; seen). - Synonyms: Platysma pallescens (Schaerer) Nyl., Mem. Soc. Sci. Nat. Cherbourg 5: 100, 1857. Nephromopsis pallescens (Schaerer) Park, Bryologist 93: 122, 1990. Sticta wallichiana Taylor in Hooker, London J. Bot. 6: 177, 1847, syn. nov. Type: Nepal, Wallich (G, no. 2003/2 - lectotype, selected here; G, no. 2003/1, PC, isolectotypes; seen). - Syn.: Parmelia wallichiana (Taylor) Nyl., Mem. Soc. Sci. Nat. Cherbourg 5: 105, 1857. - Platysma leucostigmeum Nyl. var. wallichianum (Taylor) Nyl., Syn. Meth. Lich. I: 306, 1860. - Platysma wallichianum (Taylor) Nyl., Flora 52: 443, 1869. - Cetraria wallichiana (Taylor) Miill. Arg., Flora 71: 139, 1888: Cetrariopsis wallichiana (Taylor) Lai, Quart. J. Taiwan Mus. 33: 220, 1980. Cetraria sulphurea Mont. & v.d. Bosch, Mont., Syll. Gen. Sp. Crypt.: 322, 1856, invalidly published - Type: Java, Junghuhn (not seen). Cetraria teysmanni Mont. & v.d. Bosch in Mont., Syll. Gen. Sp. Crypt.: 474, 322, 1856; in Miquel, PI. Jungh. 4: 431,1857 - Type: Java, Teismann (not seen).- Syn.: Platysma teysmanni (Mont. & v.d. Bosch) Nyl., Mem. Soc. Sci. Nat. Cherbourg 5: 100, 1857. Thallus foliose, up to 16 cm diameter but usually less than 10 cm; upper sur­ face greenish yellow, lower surface white to yellow, slightly or strongly rugose. Pseu­ docyphellae present on the lower surface in the form of white small dots or white patches located on special plug-like outgrowths, sometimes surrounded by a light brown rim. Rhizines sparse, white or light brown. Soredia and cilia absent, pycnidia not observed. Upper and lower cortex 20-30 |im thick, with larger cells near the me­ dulla, medullary hyphae up to 5 |im thick. Apothecia 1-2.5 mm in diameter, laminal, submarginal and marginal, at times extremely numerous and covering the whole thallus but sometimes located mainly in the marginal parts of the upper surface and only a few apothecia can be observed as really laminal. Ascocarps clearly appear to originate superficially since very tiny apothecia may usually be found not in the thallus margins but further to the center. Disc brown, flat or slightly convex, sometimes surrounded by a thin thalline margin. Hymenium c. 50 jim high, subhymenium c. 10 jam. Exciple twolayered, both layers c. 30 pm thick. Asci narrowly clavate, 35-40 x c. 10 jim, axial body 3-4 (am, ascospores oblong, 7-10 x 3-4 (5) pm. CETRARIOPSIS, CETRELIOPSIS AND NEPHROMOPSIS 43 Chemical constituents: usnic acid in the upper cortex; alectoronic and/or lichesterinic, protolichesterinic acids in the medulla. PD-, K-, KC+ or Distribution and habitat: Nepal, India, China, Russian Far East (Primorye region), Taiwan, Java; Japan (Kurokawa 1980), South-Korea (Park 1990). Epiphytic on deciduous and coniferous trees in the montaneous forests. Cetrariopsis pallescens was earlier known as Cetrariopsis wallichiana (Taylor) Kurokawa (type from Himalaya). Cetraria pallescens Schaerer being the oldest name, a new combination in Cetrariopsis must be presented here. This species is collected in Nepal, India, Java, South-Korea, China, Taiwan, Japan and Russian Far East (Awasthi 1982, Kurokawa 1980, Lai 1980, Park 1990, Rassadina 1971, Wei 1991). Lai (1980) drew attention to some other taxa in Southeast Asia which might belong to the same group, i.e. Cetraria citrina Taylor and Cetraria teysmanni Mont. & v.d. Bosch (both types from Java). Our studies of types and other herbarium material have shown that they represent the same species. Unfortunately, we did not succeed in finding type material of C. teysmanni (= C. sulphurea Mont. & v.d. Bosch, see below). Cetraria sulphurea Mont. & v.d. Bosch (type also from Java) was described in 1856 (Montagne 1856: 322). A new name, Cetraria teysmanni Mont & v.d. Bosch, was presented for this taxon already in the same paper (Montagne 1856: 474) by the same authors: «Pag. 322, n°1189, loco: «Cetraria sulphurea», Legendum est: «Cetraria teysmanni M. et v.d. B.»». We think this a sufficient reason to consider the epithet sulphurea as invalid. Still, these authors presented a new description of Cetraria teysmanni once more a year after (Montagne & v.d. Bosch 1857: 431, Stafleu & Cowan 1981). Cetrariopsis pallescens var. pallescens is distributed mainly in Nepal, China and Russia and the specimens are greenish yellow on the upper surface and white on the lower. They have numerous laminal apothecia and a strongly rugose lower side with large pseudocyphellae on special outgrowths on central parts of the thallus (Fig. 4). Younger pseudocyphellae in more marginal positions may still be quite small and plain. Most of the specimens from these regions contain alectoronic acid in the medulla (KC+) but also material with fatty acids (KC-) is known as well as specimens with both types of medullary substances. Also a few specimens from Java and Taiwan were iden­ tified as C. pallescens var. pallescens. Some specimens from Nepal may differ considerably in their morphological features from most specimens collected in Java, usually called Cetraria citrina. The smoothness of the thallus, the colour of both cortices, the number and position of apo­ thecia, and the form of pseudocyphellae, however show a continuous variation, and no strict limits can be drawn. As we cannot be sure whether the morphological variation described above is not due to the extreme ecological conditions only, and distribution of different variants is partly overlapping, we prefer to preserve C. citrina only as a variety. Altogether 39 specimens were examined. 44 T. RANDLANE, A. THELL and A. SAAG Selected specimens examined. China. Yunnan, Lopin-chan, Lan-Kong, alt. 3200 m, Delavay, 1888 (PC). Yunnan, Lijiang County, Yulongshan, alt. 2600-2800 m, 27°H ’ N, 100°15’ E,Ahti et al., 1978, no. 46 370 (H, TU). India. Almora distr., Dhakuri ridge, alt. 9500 ft., Awasthi, 1S50, no. 641 (UPS ex Herb. Awasthi). Indonesia. Java. Pangerango, Zollinger no. 449 (G - holotype of Cetraria pallescens Schaerer). Java. Gedeh, alt. 2400 m, Kjellberg, 1929, no. 125L (S). Java. Res. Pasoeroean, Goenoeng Ardjoens, Lalidjiwa - Welirang track, Du Rietz, 1927, no. 54-la (UPS). Nepal. Langtang Area, slopes above Syarpagaon, alt. 2900 m, Poelt, 1986, no. 86-L1010 (GZU). Langtang Area, Lambatis, alt. 2760 m, G. & S. Miehe, 1986, no. 15985a (GZU). 15 km SSE Kathmandu, 2 km E Godawari, alt. 2760 m, 27°35’ N, 85°20’ E, Thor. 1979, no. 1282 (S). Russia. Primorye region, Kedrovaya Padj, Parn, 1961 (TU). Taiwan. Mt. Arisan, Sato, 1936 (S). Cetrariopsis pallescens (Schaerer) Randl. & Thell var. citrina (Taylor) Thell & Randl., comb, et stat. nov. Basionym: Cetraria citrina Taylor in Hook., London J. Bot. 6: 176, 1847. Type: Java, Hooker (BM, lectotype, selected here). - Syn.: Platysma citrinum (Taylor) Nyl., Mem. Soc. Sci. Nat. Cherbourg 5: 100, 1857. Thallus like Cetrariopsis pallescens but both surfaces uniformly light yellow, lower side more or less smooth (not rugose) and pseudocyphellae in the form of small, plain, white dots. Apothecia not numerous, mainly marginally and submarginally loca­ ted although some laminal apothecia are also present. Ascocarp anatomy like in C. pallescens var. pallescens. Chemical constituents: usnic acid in the cortex; lichesterinic, protolichesterinic acids (usually) or alectoronic acid (occasionally) in the medulla. Distribution: Java, Taiwan. This variety, known from southern regions (Taiwan and Java), is light or bright yellow in colour on both surfaces, the thallus is more or less smooth, the apothecia are not very numerous and situated mainly on the submarginal parts of the upper cortex while some truly laminal apothecia are usually present. The pseudocyphellae on the lower cortex mainly have the form of small white and plain dots but are larger someti­ mes and may be located on ridges or on almost outgrowth-like structures. Both varieties of Cetrariopsis pallescens are present on Java. A majority of the specimens (including the type material of C. pallescens var. citrina) from there contain only fatty acids in the medulla but a few specimens represent the alectoronic acid che­ motype. Thus, both varieties within C. pallescens are chemically variable and are re­ presented by specimens with alectoronic as well as lichesterinic acid in the medulla. The distribution areas of these two varieties are also partly overlapping (in Java and Taiwan). In this case we consider variety as a suitable taxonomic level. Altogether 30 specimens examined. Selected specimens examined. Indonesia. Java. Pangerango, alt. 2820 m, Schiffner, 1894, no. 2987a (FH, M). Java. Preanger, Mt. Gedeh, alt. 2300 m, Schiffner, 1894, no. 3387 (FH, M). Java. Kandang Badals, alt. 2600 in, Yates, 1927, no. 2827 (US, LD). East Java, Andjasmora-complex, alt. 2100 m, G ro en h a rt , 1937, no. 2 5 3 8 (Herb. Aptroot). Java. Res. Pasoeraean, Goenoeng Ardjoens, Tretes - Lalidjiwa track, D u R ietz, 1927, no. 1 0 5 - lb (UPS). Taiwan. Mt. Arisan, Asahina (US). CETRARIOPSIS, CETRELIOPSIS AND NEPHROMOPSIS 45 Fig. 2-4 - Morphology in Cetrariopsis. Fig. 2. Cetrariopsis laii, richly fertile part of upper surface, Russia, Primorye 17.09.1961, S. Pam (TU). Fig. 3. Lower surface of the same specimen, showing the pseudocyphellae located on dark ridges. Fig. 4. Cetrariopsis pallescens, upper surface with numerous, small, laminal apothecia, and lower surface with characteristic pseudocyphellae on pale ridges, Nepal, Langtang, Miehe 16139 (GZU). Bars in Figs. 2-4 = 1 cm, ps = pseudocephyllae. 26 46 T. RANDLANE, A. THELL and A. SAAG Cetrariopsis laii Thell & Randl., sp. nov. - (Fig. 2, 3, 16) Thallus foliaceus, ad 8 cm diametro, superficies superior virescens vel flavens, rugosa; subtus fusca ad pallide fusca, reticulosa et pseudocyphellata, rhizinata; pseu­ docyphellae praecipue in cristis. Soredia, cilia et spinulae marginales desunt, lobi lobulis marginalibus saepe ornati. Cortex superior paraplectenchymatus, Apothecia numerosa, marginalia, ad 5 mm lata; discus brunneus, oblongus. Excipulum 2-stratum. Asci clavati, 40-45 x 8-10 pm, ascosporae oblongae, 8 x 3 pm. Pycnidia et pycnoconidia non visa. Acidum usnicum in cortice superiore; acidum lichesterinicum et protolichesterinicum in medulla. Typus: Russia, Primorye region, Kedrovaya Pedj Nature Reserve, on Be tula dahurica, 17.09.1961 S. Pam (TU - holotypus, LD - isotypus). Thallus foliose, up to 8 cm in diameter, upper surface greenish yellow, modera­ tely rugose; lower surface brown to light brown and whitish on the margins, densely reticulated and with white rounded or oval pseudocyphellae situated on the ridges, sometimes surrounded with a brown rim. Soredia, cilia and marginal projections absent. Rhizines on the lower surface brown and simple. Pycnidia not seen. Upper and lower cortex c. 20 pm thick, with larger cells near the medulla. Medullary hyphae 3-6 pm thick. Apothecia numerous, marginal, sometimes situated on the secondary marginal lobules, up to 5 mm in diameter; disc brown, usually oblong or reniform; in many specimens only juvenile apothecia in the form of marginal brown lines are present. Exciple two-layered, upper layer c. 20 pm, lower layer c. 30 pm. Asci 40-45 x 8-10 pm, axial body 2.5-3 pm, ascospores oblong, 8 x 3 pm. Chemical constituents: usnic acid in the upper cortex; lichesterinic and protolichesterinic acids in the medulla. Distribution and habitat: Russian Far East (Habarovsk and Primorye regions), Japan, Vietnam, Taiwan. Epiphytic on coniferous and deciduous trees in the zone of montaneous forests. Cetrariopsis laii is named after our collegue, the Chinese lichenologist Mingjou Lai from Taiwan whose contribution to the knowledge of Asiatic cetrarioid lichens is significant. This new Cetrariopsis species is not a rare lichen in the eastern Asia but it has been confused with other taxa, labelled usually in the herbaria as Cetraria wallichiana or Cetraria pallescens but occasionally also as Nephromopsis ectocarpisma. Two for­ mer names represent, as we know now, the same species - Cetrariopsis pallescens. In Russia where the new species is most widely distributed, it has been uncorrectly iden­ tified as Cetraria pallescens while the true Cetrariopsis pallescens with numerous laminal apothecia was named Cetraria wallichiana. Consequently these two lichens were practically separated but, because of the taxonomical disorder, the other of them has remained undescribed till now. Some specimens of C. laii may be quite similar to C. pallescens var. pallescens but the apothecia of the former are situated mainly margi­ nally and submarginally, the disc is usually not rounded but reniform and sometimes only numerous juvenile apothecia are present. Other characteristic morphological featu­ CETRARIOPSIS, CETRELIOPSIS AND NEPHROMOPSIS 47 res of C. laii that separate it from C. pallescens are as follow: strongly rugose upper and reticulated lower surface; darker (brown to light brown) colour of the lower side with pseudocyphellae located conspicuously on the ridges (Fig. 2, 3); presence of se­ condary marginal lobules as a fringe along the margins in many specimens. Altogether 21 specimens from Russia, Japan, Vietnam and Taiwan were exami­ ned. Selected specimens examined. Japan. Hokkaido, Mt. Daisetu, Sato, 1936 (FH). Gotenniwa, Mt. Huzi, Asahina, 1956 (Lich. Jap. Exs. 157, H). Russia. Primorye region, island Putyana, Mt. Startseva, Chabanenko, 1982 (LD). Primorye region, Lazo Nature Reserve, Mt. Tchomaya, alt. 1300 m, Chabanenko, 1982 (LD). Primorye region, Livadiski Range, Mt. Livadiskaya, alt. 1000 m, 43°8’ N, 132°42’ E, Skirina, 1980, no. 7563 (LD). Primorye region, Sikhote-Alin Range, Mt. Eldorado, alt. 1300 m, 44°41’ N, 135°20’ E, Skirina, 1982, no. 7564 (LD). Primorye region, Mt. Pryamaya, 43°20’ N, 133°36’ E, alt. 850 m, Karnefelt, 1991, no. 910 810 (LD). Habarovsk region, Badzhal Mt. Range, river Urmi, Randlane, 1981, no. 167 (TU). Taiwan. Chiayi Co., Mt. Alishan, alt. 2275 m, Lai, 1978, no. 10 198 (US). Vietnam. Zon Kin, Phan*si-Pau, Lao Kay, 2900 m, 1929, no. 17156 (PC). III. The Genus Cetreliopsis Lai This genus was described by Lai (1980) in his paper on cetrarioid lichens in East Asia to settle the Cetraria rhytidocarpa-complex, as Lai called it. The group in­ cluded Cetraria rhytidocarpa Mont. & v.d. Bosch from Java, Cetraria straminea Vainio from the Philippines and Cetraria laeteflava Zahlbr. from Taiwan. Lai synonymized all the three species on morphological and chemical grounds. He also pointed out the affinities to Cetrelia and Nephromopsis, combining the names of these genera for the new taxon. Our studies on Cetreliopsis have shown that this genus is well limited and clearly separated from other cetrarioid lichens. The identifying characters of the genus Cetreliopsis are: large foliose or subfruticose thallus; marginal or submarginal apothe­ cia; presence of pseudocyphellae on both surfaces of the thallus; large, ellipsoid as­ cospores in rather broadly clavate asci and a two-layered exciple; content of fumarprotocetraric and protocetraric acids as major compounds in the medulla (PD + red) and usnic acid in the cortex (Figs. 5-7, 11, 14). We cannot agree with Lumbsch (in Eriksson & Hawksworth 1988) who proposed to include C. rhytidocarpa in Cetrelia as a separate subgenus. He recognized mainly chemical differences between Cetrelia and Cetreliop­ sis and considered their morphology very similar. We find not only essential chemical differences (atranorin and orcinol depsides in Cetrelia ; usnic acid, (3-orcinol depsidones and fatty acids in Cetreliopsis) but also morphological (ashy white or tan upper surface, perforate submarginal to laminal apothecia, lower cortex punctate or not in Cetrelia-, yellow upper surface with frequent black-blotted areas, marginal and entire apothecia, lower cortex with distinct pseudocypellae in Cetreliopsis) and anatomical differences (large asci with a strongly amyloid tholus and large thick-walled ascospores - 11-24 x 6-12 |am - in Cetrelia\ asci usually less amyloid and considerably smaller in Cetre­ liopsis, ascospores always smaller) between these two entities. The genus Cetreliopsis is probably more closely connected to Nephromopsis and Cetrariopsis than to Cetrelia. 48 T. RANDLANE, A. THELL and A. SAAG The confusion within the Cetraria rhytidocarpa-complex has one more aspect. Asahina (1954) used the name C. rhytidocarpa for a Japanese lichen which is totally different from the complex treated here. Specimens of Cetraria rhytidocarpa f. nipponensis Asah. in Kurokawa’s «Lichenes Rariores et Critici Exsiccati» and in all other collections from Japan belong to Tuckneraria pseudocomplicata (Randlane et al., 1994). The latter is easily distinguished from the true Cetraria rhytidocarpa complex by the lack of pseudocyphellae on the upper side and the negative PD reaction in the medulla. Anatomy of ascocarps (globose to subglobose ascospores, asci with uniseriately arranged spores and broad axial body, two-layered exciple) refers to the affinities of this taxon rather to Tuckermannopsis than to Nephromopsis or Cetreliopsis. According to our present knowledge Cetreliopsis cannot be treated as a monotypic genus. Three new combinations are proposed here; furthermore, one new species and one new subspecies is described. We do not support in all parts the wide species treatment of Cetreliopsis rhytidocarpa proposed by Lai (1980) and prefer to keep the sorediate Taiwan material as a separate species. The following five species are included in Cetreliopsis: C. asahinae (Sato) Randl. & Thell C. endoxanthoides (Awasthi) Randl. & Saag C. laeteflava (Zahlbr.) Randl. & Saag C. papuae Randl. & Saag C. rhytidocarpa (Mont. & v.d. Bosch) Lai. CETRELIOPSIS Lai, Quart. J. Taiwan Mus. 33:218 (1980). Type species; Cetreliopsis rhytidocarpa (Mont. & v.d. Bosch) Lai Thallus dorsiventral, foliose or subfruticose, straw yellow or greenish yellow on the upper surface, often with black-blotted areas and conspicuous pseudocyphellae surrounded by a black rim or laminal emergent pycnidia. The lower surface is either totally black or black in central parts and brownish on margins, pseudocyphellae are present also on the lower side usually in the form of minute white dots. Black rhizines sparse or numerous. Cilia and soredia may be present on different species. The pycni­ dia are laminal and/or marginal, on emergent projections or immersed in the thallus. The cortical tissue is present beneath the pycnidium. The upper and lower cortex are composed of the same paraplectenchymatic type as in Cetrariopsis, with large cells near the medulla. Apothecia marginal or submarginal but not nephromoid (disc clearly faced upwards), to 14 mm in diameter; disc brown, surrounded by the thalline margin. Exciple two-layered, upper layer composed of longitudinally arranged hyphae. Asci rather broadly clavate, 35-60 x 11-20 |um; axial body 2.5-4 |im. Ascospores ellipsoid, 6-12 x 4-7 jam. Pycnoconidia bifusiform (dumb-bell shaped), 5 x 1 -2 (am. Chemical constituents: usnic acid (+/-) in the cortex; fumarprotocetraric acid and other closely related (3-orcinol depsidones together with fatty acids in the medulla. CETRARIOPSIS, CETRELIOPSIS AND NEPHROMOPSIS 49 Distribution and habitat: Russian Far East, Japan, China, Vietnam, SouthKorea, India, Nepal, Taiwan, the Philippines, Borneo, Java, New Guinea. Epiphytic on coniferous and deciduous trees. Cetreliopsis asahinae (Sato) Randl. & Thell, comb. nov. - (Fig. 5, 6, 14) Basionym: Cetraria asahinae Sato, Saito Ho-on Kai Mus. Res. Bui. 11: 12, 1936. - Type: Kuril Islands, Kunashiri, Tomarimura, Yasuda 16.08.1923 (not seen). Synonym: Nephromopsis asahinae (Sato) Rasanen, Kuopion Luonnon Ystavain Yhdistyksen Julkaisuja B 2(6): 50, 1952. Thallus foliose, with ascending margins; upper surface yellowish green, lower surface black in central parts and with brown margins. Pseudocyphellae present on both surfaces, on upper surface surrounded with dark rim or black projections, on lower surface in the form of white patches located mainly on the thallus ridges. Soredia and cilia absent. Pycnidia laminal or marginal, on emergent projections. Upper cortex c. 15 pm, composed of three to four layers of cells, cells up to 5pm, the largest concentrated near the medulla; lower cortex similar to the upper but outer cells brownish pigmented; medullary hyphae c. 3 pm. Apothecia marginal or submarginal, to 14 mm in diameter, disc brown; hymenium c. 45 pm, subhymenium c. 10 pm, exciple usually two-layered, both layers 30-40 pm, sometimes with an undistinct third layer between with larger lumina, up to 10 pm thick; asci 35-40 x 12-17 pm, axial body 2.5 pm; ascospores 8-12 x 4-7 pm. Pycnoconidia c. 4 x l pm. Chemical constituents: usnic acid in the cortex (rarely absent); protocetraric (major), fumarprotocetraric acid and physodalic acid (+/-) in the medulla. Distribution: Russian Far East (Primorye region, the Kuril Island), Japan, Vietnam; China (Wei 1991), South-Korea (Park 1990), India (Cetraria rhytidocarpa sensu Awasthi, in Awasthi 1982). C. asahinae is a distinct entity and morphologically easily recognized by the pseudocyphellae on the upper cortex surrounded with a black rim and black emergent projections (Fig. 5-6). The medullary chemistry - presence of protocetraric acid as major compound - is also typical. C. asahinae has a wider distribution than other taxa in Cetreliopsis. Altogether 20 specimens were examined. Selected specimens examined. Japan. Mt. Akagi, Gunma, Sato, 1957 (H). Mt. Akagi, Yasuda, 1911, no. 47 (LD). Hokkaido, Hidaka distr., Shizukai-cho, alt. 400 m, Koponen, 1970, no. 14525 (H). Russia. Primorye region, Peter the Great Bay, Furughelm Island, 42°31’ N, 130°55’ E, Skirina, 1987, no. 6251 (LD). Primorye region, Sikhote Alin Range, Abrek Urochische, 45°5 N, 136°40’ E, Skirina, 1977, no. 8932 (LD). Primorye region, Lazo Nature Reserve, Mt. Tumannaya, alt. 160 m, Chabanenko, 1983, no. 23 (LD). Primorye region, Island Petrova, 42°45’ N, 133°48’ E, Karnefelt, 1991, no. 910 615 (LD). Primorye region, Kedrovaya Padj, Vasineva, 1954 (LE). Kuril Islands, Shikotan, Malokurilskoye, Blum, 1965 (KW). Vietnam. Zon Kiu, Phan-si-Pan, Lao Kay, alt. 2600 m, 1929, no. 17194 (PC). 27 50 T. RANDLANE, A. THELL and A. SAAG Fig. 5-8. Morphology in Cetreliopsis. Fig. 5. Cetreliopsis asahinae, upper surface, Russia, Primorye, Guriljova 1951 (TU). Fig. 6. Part of the same specimen showing the white pseudocyphellae surroun­ ded by pycnidial projections. Fig. 7. Cetreliopsis papuae, habit, Papua-New Guinea, 05. 1975, Hope, No. 483, Univ. Col. Mus (US). Fig. 8. Cetreliopsis rhytidocarpa subsp. langtangi, upper surface with conspicuous pseudocyphellae surrounded with black rims, Nepal, Langtang, Miehe 15 738 b, holotype (GZU). Bars in Figs. 5,7-8 = 1 cm, in Fig. 6 = 1 mm, p = pycnidia, ps = pseudocyphellae. CETRARIOPSIS, CETRELIOPSIS AND NEPHROMOPSIS 51 Cetreliopsis endoxanthoid.es (Awasthi) Randl. & Saag comb. nov. Basionym: Cetraria endoxanthoides Awasthi, Bull. Bot. Surv. India 24: 9, 1982. - Type: E. Nepal, Mewakhola valley, alt. 2400-2700 m, D.D. Awasthi, 28.05.1953, no. 2477 (Herb. Awasthi, holotype; not seen). - Synonym: Nephromopsis endoxanthoides (Awasthi) Randl. & Saag, Mycotaxon 44: 486,1992. Thallus foliose, greenish yellow on upper and black to dark brown on the lower side. Pseudocyphellae present on both surfaces, sparse on the upper cortex, coloured light orange and surrounded by a dark rim or black spinules; more numerous on the lower cortex, in the form of convex conspicuous white dots. Rhizines black, sparse. Cilia and soredia absent. Medulla pale orange. Pycnidia laminal and marginal, on emergent projections. Upper and lower cortex 20-25 |im, crystallized, composed of three to four layers of small, equal-sized, thick-walled cells, medullary hyphae 3-4 (im in diameter. Apothecia marginal, up to 6 mm in diameter, with dark brown disc, hymenium c. 50 |im, subhymenium to 10 (im, exciple two-layered, upper layer c. 10 |im, lower layer c. 30 |jm; asci 50 x 15 (am, ascospores 9 x 5,5 |im. Pycnoconidia compara­ tively thick, 5-6 x 1.5-2 (am. Chemical constituents: fumarprotocetraric acid (major), salazinic-like subs­ tance Cph-1 (major), protocetraric acid, salazinic acid, an unidentified fatty acid in the medulla. Distribution and habitat: Nepal. Corticolous. Awasthi (1982) mentions in the original description of the species the type collection only - E. Nepal, Mewakhola valley, alt. c. 2400-2700 m, 28.05.1953, D.D. Awasthi no. 2477 (Herb. Awasthi). We have identified one more specimen from the same locality (no. 2227, collected 27.05.0953 and determined by Awasthi at first as Cetraria asahinae and in 1980 as Cetraria rhytidocarpa) on the grounds of orange coloured medulla, form of pseudocyphellae and complex of medullary substances. Earlier (Randlane & Saag 1992) this species was considered to have phylogenetic affi­ nities with those taxa in Nephromopsis which medulla is similarly coloured (AT. endocrocea, N. omata, N. globulans). Although we were not able to identify exactly the pigment in it. Still, later studies have shown that the anthraquinones and secalonic acids occur in different genera of Parmeliaceae much more frequently than supposed and the colour of the medulla cannot be qualified as a genus character. Other medullary consti­ tuents as well as morphological and anatomical features of C. endoxanthoides undoubt­ edly unveil its close relations to Cetreliopsis. Cetreliopsis laeteflava (Zahlbr.) Randl. & Saag comb. nov. - (Fig. 11) Basionym: Cetraria laeteflava Zahlbr., Feddes, Repert. 33: 60, 1933. - Type: Taiwan, Mt. Arisan, Nimandaira, Asahina (H, US, isotype; seen). - Synonyms: Cetraria straminea Vainio var. laeteflava (Zahlbr.) Rasanen (nom. illeg., non Cetraria straminea Krempelh. ex Schwend., 1860), Kuopion Luonnon Ystavain Yhdistyksen Julkaisuja B 2(6): 50, 1952. 52 T. RANDLANE, A. THELL and A. SAAG Cetraria straminea Vainio var. sorediata Rasanen (nom. illeg.), Suom. Elain-ja Kasvit. Seuran Van. Kasvit. Julk. 3: 78, 1949 (1948). Thallus foliose, to 12 cm in diameter, greenish yellow and occasionally blackblotted on the upper side; totally black on the lower side with brown marginal parts. Thallus margins partly sorediate; cilia absent. Pseudocyphellae present on both surfa­ ces: in the form of small white and somewhat convex patches on the upper side, usually surrounded by a black rim and almost of the same size but plain dots on the lower side. Pycnidia rare, marginal, on emergent projections. Upper and lower cortex c. 30 pm thick, with the cells near the medulla clearly larger. Apothecia marginal, absent on many specimens, with disc faced upwards; hymenium including subhymenium 70-80 Urn thick, exciple two-layered, both layers c. 30 pm. Asci 50-60 x 11-17 nm, axial body 3-4 |jm, ascospores 8.5-9 x 5 pm. Pycnoconidia not observed. Chemical constituents: usnic acid in the cortex (+/-); fumarprotocetraric acid (major), salazinic-like substance Cph-1 (major), protocetraric acid, salazinic acid, lichesterinic and protolichesterinic type fatty acids. Distribution: Taiwan and the Philippines (Rasanen 1949). Corticolous in the montane districts. C. laeteflava is morphologically recognized by the presence of marginal soredia and chemically by the occurrence of both fumarprotocetraric acid and Cph-1 as major substances in the medulla. It is the only sorediate species in the genus, evidently closely related to C. rhytidocarpa, the variety of which it has been treated by Rasanen (1952) and Lai (1980). The presence of soredia is usually considered a good character at the species level (Poelt 1973). As this distinctive quality is combined here with a small chemical differentiation (Cph-1 also as a major substance), we prefer to maintain the taxon on species level as it was originally described. A taxonomical confusion with the name Cetraria straminea was caused by the fact that the epithet «straminea» has been used by two different authors, Schwendener (1860) and Vainio (1909), for two totally different lichens. Cetraria straminea Krempelh. ex Schwend., which has priority, has generally been forgotten; lately, with help from Prof. Rolf Santesson, we could make clear that this species is synonymous with Tuckneraria laureri (Randlane et al., 1994). Cetraria straminea Vainio not valid is synonymous with Cetreliopsis rhytidocarpa (Mont. & v.d. Bosch) Lai. Specimens examined. Taiwan. Nimandairana, Mt. Arisan, Asahina, 24.12.1925 (H, US, isotypes). Chiayi prov., Mt. Arisan, Sisters’ Pond, alt. 2275 m, Lai, 1978 (H). Chiayi prov., Mt. Tsutson-san, Mt. Ali, alt. 2200-2600 m, Kurokawa, 1964, no. 478 (H). Taitung prov., Mt. Lachialachiaerh, alt. 1800 m, Kurokawa, 1965, no. 2473 (H, TAIM). Cetreliopsispapuae Randl. & Saag sp. nov. - (Fig. 7) Thallus foliaceus, lobi ad 12 mm lati et 45 mm longi; margo anthracina ciliata, cilia ad 2.5 mm longa; superficies superior sulfurea et nigra, pseudocyphellata; subtus nigra, pseudocyphellata, in margine sulfurea. Pseudocyphellae ambarum superficierum nigro cinctae. Rhizinae desunt. Cortex superior paraplectenchymatus, 20-25 pm; me­ CETRARIOPSIS, CETRELIOPSIS AND NEPHROMOPSIS 53 dulla alba; cortex inferior c. 30 jum. Apothecia et pycnidia non visa. Acidum usnicum in cortice superiore; acidi fumarprotocetraricum, protocetraricum, substantia Cph-1 in medulla. Typus: Papua New Guinea, Star Mountains, eastern side of Mt. Scorpion, elev. 3600 m, in an open herbfield of Tetramolopium and Astelia, May 1975, G.S. Hope (Lich. Exsic. distributed by the University of Colorado Museum, Boulder no. 483) (US - holotypus; isotypi in several herbaria, e.g. CAN, GZU). Thallus foliose, 100-150 pm thick, consisting of separate oblong lobes with rounded margins to 45 mm long and 12 mm wide; upper surface sulphuric yellow with black patches here and there and wide pseudocyphellae often surrounded by a black rim; marginal cilia black, to 2.5 mm long; lower surface mainly black in the basal parts and sulphuric yellow black-blotted in the marginal portions; pseudocyphellae similar to those on the upper surface; rhizines absent. Upper cortex 20-25 pm, crystallized, com­ posed of c. three layers of equal-sized cells, lower cortex c. 30 pm, less crystallized than the upper cortex, composed of c. four layers of equal-sized cells; algal cells usually in clusters, medulla white, medullary hyphae c. 4 pm in diameter. Pycnidia and apothecia not observed. Chemical constituents: usnic acid in the medulla; fumarprotocetraric acid (major), protocetraric acid, salazinic-like substance Cph-1 and an unidentified fatty acid in the medulla. Distribution and habitat: Papua New Guinea; grows in high mountains (more than 3000 m), evidently on the ground in open herbfields or shrublet communities. Although the species lacks generative organs, there is no doubt about its generic position - presence of pseudocyphellae on both surfaces as well as the complex of medullary substances is typical for Cetreliopsis. Its morphology is different from the other representatives of the genus: the thallus is foliose, clearly dorsiventral, but the lobes are separated and probably growing on the ground partly upright why it could be difficult to identify on herbarium material which side is the upper (all other species in Cetreliopsis appear to be corticolous); true marginal cilia and not pycnidial projections as in other taxa are always present; relatively wide pseudocyphellae are similar on both sides of the thallus (Fig. 7). These conspicuous characters confirm the description of C. papuae as a separate species. C. papuae is known from two localities in Papua New Guinea. Another studied specimen besides the type collection: Central distr., Tapini subdistr., Mt. Strong, summit area, elev. 3500 m, scattered component of low tussock-prostrate shrublet communities, 03.05.1971, M.J.E. Coode, no. 3805 (Herb. Aptroot). Cetreliopsis rhytidocarpa (Mont. & v.d. Bosch) Lai subsp. rhytidocarpa - (Fig. 9) Basionym: Cetraria rhytidocarpa Mont. & v.d. Bosch, Miquel, PI. Jungh. 4: 430, 1857. - Type: Java, Junghuhn, Herb. v.d. Bosch (PC, lectotype; seen). - Synonym: Platysma rhytidocarpum (Mont. & v.d. Bosch) Nyl., Mem. Soc. Sci. Nat. Cherbourg 5: 28 T. RANDLANE, A. THELL and A. SAAG 54 100, 1857. - Nephromopsis rhytidocarpa (Mont. & v.d. Bosch) Zahlbr., Ann. Cryptog. Exot. 1: 208, 1928. Cetraria straminea Vainio, Philipp. J. Sci. Bot. 4: 657, 1909 (nom. illeg., non Cetraria straminea Krempelh. ex Schwend., 1860) - Type: Philippines, Luzon, Prov. Laguna, Mt. Banajao, Curran & Merritt, no. 7988 (US, isotype; seen). - Syn.: Nephromopsis straminea (Vainio) Rasanen, Kuopion Luonnon Ystdvdin Yhdistyksen Julkaisuja B 2(6): 50, 1952. Thallus foliose, to 11 cm in diameter, upper surface greenish to sulphuric yel­ low, partly, especially in margins black-blotted, with small, regular and somewhat emergent pseudocyphellae with black rim; lower surface totally black, only occasio­ nally with light brown or even whitish margins, pseudocyphellae usually in the form of small white plain punctae which sometimes may be quite large and even elevated. Soredia and cilia absent. Pycnidia marginal and laminal, on black emergent projections (in younger parts not always very emergent). Rhizines on the lower surface black and numerous. Upper cortex 20-25 jam thick, composed of about four layers of cells, usually somewhat larger near the medulla; lower cortex thin, 10-15 (im, composed of only one-two layers of cells, the outer layer brownish pigmented; medulla white, me­ dullary hyphae c. 4 jam in diameter. Apothecia marginal or submarginal, up to 10 mm in diameter, disc brown, faced upwards; hymenium c. 55 jam, subhymenium c. 10 (im, exciple two-layered, sometimes with a thin, undistinct third layer, both upper and lower layers c. 20 (im. Asci 35-60 x 12-20 |im, axial body 3-4 |im, ascospores 6.5-11 x 4-6 (im. Pycnoconidia not seen. Chemical constituents: usnic acid in the cortex; fumarprotocetraric acid (major), protocetraric acid, salazinic-like substance Cph-1, salazinic acid (+/-), and one or two lichesterinic-protolichesterinic type fatty acids (+/-) in the medulla. Distribution and habitat: the Philippines, Java, Borneo; corticolous. The material of Cetreliopsis rhytidocarpa from southern islands is typically yellowish on the upper and totally black on the lower surface while the pseudocyphellae are small and plain on upper as well as lower surface (Fig. 9). From the other species in the same genus, C. rhytidocarpa is morphologically distinguished by the white medulla (from C. endoxanthoides), absence of soredia (from C. laeteflava) and cilia (C. papuae) and lack of emergent projections around the pseudocyphellae (C. asahinae). Specimens examined. Indonesia. Borneo. Mt. Kinabalu, Paka Cave to Lohang, Strong Clemens, 1945, no. 10 755 (FH). Java. Junghuhn, Herb. v.d. Bosch (PC, lectotype). Pangerango, alt. 2500 m, Schiffner, 1894, no. 3004 (TNM). Philippines. Luzon, Prov. Laguna, Mt. Banajao, Curran & Merritt, 1907, no. 7988 (US, isotype of Cetraria straminea Vainio). Luzon, Mi. Banajao, alt. 1900 in, Robinson, 1909, no. 6586 (H ). Cetreliopsis rhytidocarpa (Mont. & v.d. Bosch) Lai subsp. langtangi Randl. & Thell subsp. nov. - (Fig. 8) Thallus foliaceus, ad 5 cm latus; superficies superior virescens vel straminea, pseudocyphellata; superficies inferior centro nigra, margine pallide fusca vel alba, CETRARIOPSIS, CETRELIOPSIS AND NEPHROMOPSIS 55 pseudocyphellata. Pseudocyphellae ambarum superficierum convexae rotundae vel irregulares, nigro cinctae. Apothecia marginalia, ad 10 mm lata, asci clavati, 45-60 x 14-20 pm; ascosporae ellipsoideae, 7.5-11 x 4-5.5 pm. Pycnidia laminalia et margina­ lia, parum emergentia. Typus: Nepal, Central Himalaya, Langtang Area, below Dotsche, alt. 2880 m, Quercus semicarpifolia forest, on branches, 08.11.1986, G. & S. Miehe, no. 15 738 b (GZU - holotypus). Thallus foliose, to 5 cm in diameter, greenish yellow on the upper surface, and from black on the central part of the lower surface to almost white on the margins. Pseudocyphellae present on both surfaces in the form of white rounded or irregular convex patches, usually with a black rim on the upper side and of the similar form or smaller on the lower side. Upper and lower cortex both 20-25 pm, composed of two layers of cells, those close to the medulla clearly larger. Medullary hyphae c. 4 pm thick. Apothecia marginal, hymenium c. 55 pm, subhymenium c. 10 pm, exciple twolayered, sometimes with a thin undistinct third layer, both upper and lower layers c. 20 jim. Asci 45-60 x 14-20 (am, ascospores 7.5-11 x 4-5.5 jam. Pycnidia marginal and laminal, slightly or more prominently emergent; pycnoconidia not seen. Chemical constituents the same as in C. rhytidocarpa subsp. rhytidocarpa. Distribution and habitat: Nepal and India ( Cetraria laeteflava sensu Awasthi in Awasthi 1992). Epiphytic in the middle altitudes (2400-3300 m) of montane areas. This subspecies, distributed in Nepal and according to the description by Awas­ thi (1982), also in India, incline to be more greenish on the upper side and black in the center to almost whitish in the marginal parts of the lower surface with large and emer­ gent pseudocyphellae. Because of the well delimited distribution this material is descri­ bed as a separate subspecies. Specimens examined. Nepal. Central Himalaya, Langtang Area, below Dotsche, alt. 2880 m, Miehe, 1986, no. 15 738b (GZU, holotype). Langtang Area, SE of Schiabon Kedo, alt. 3300 m, Miehe, 1986, no. 1185b (GZU). Langtang Area, Upper Tadi Khola, alt. 2800 m, Miehe, 1986, no. 15 822a (GZU). CONCLUSIONS All the three genera treated here - Cetrariopsis, Cetreliopsis and Nephremopsis - form a group of related taxa. It is distinguished by large foliose thalli with rugose or reticulated central parts and ascending margins (Table 1). Apothecia develop in most species marginally. Pycnidia are immersed or situated on the marginal and/or laminal emergent projections. Presence of distinct pseudocyphellae on the lower surface is a good character for identifying this group from Tuckermannopsis in a strict sense. The yellowish colour of the thallus caused by the occurrence of usnic acid in the upper cortex makes differences from Cetrelia and Platismatia. The anatomy of the thallus of all studied species is clearly uniform showing pachydermatous paraplectenchymatic 56 T. RANDLANE, A. THELL and A. SAAG Table 1. - Comparison of characters in Cetrariopsis, Cetreliopsis and Nephromopsis. Cetrariopsis Cetreliopsis Nephromopsis Thallus Upper and lower cortex Pseudocyphellae Soredia Marginal cilia Apothecia foliose 1-layered, paraplectenchymatous on lower surface absent absent foliose 1-layered, paraplectenchymatous on both surfaces may be present may be present marginal and laminal on upper surface Exciple Ascus shape Ascospores Axial body Ring structure Pycnidia 2-layered narrowly clavate oblong, 7-10 x 3-5 urn 2,5-4 um absent not seen Pycnoconidia not seen Cortical substances Medullary substances: a) fatty acids b) orcinol depsides & depsidones c) p-orcinol depsidones usnic acid marginal and sub­ marginal on upper surface 2-layered rather broadly clavate ellipsoid, 6-12 x 4-7 um 2,5-4 |im absent laminal or marginal, immersed or on projections bifusiform, 5 x 1-2 um usnic acid foliose 1-layered, paraplectenchymatous on lower surface absent absent marginal on lower surface present present present alectoronic a. - olivetoric a., physodic <T d) secalonic acids 3-layered narrowly clavate oblong, 5-10 x 2.5-5um 0.5-4 um present in two species laminal or marginal, immersed or on projections bifusiform, 4-5 x 1-1.5 um usnic acid fumarprotocetraric a., protocetraric a., physodalic a., salazinic a., Cph-1 endocrocin, secalonic a. A, secalonic a. C cortices, usually with larger cells near the medulla. Anatomical characters of the ascocarps are also similar (clavate asci with a moderately large axial body in the tholus; oblong or ellipsoid ascospores) but not identical (two-layered exciple in Cetrariopsis and Cetreliopsis and usually three-layered in Nephromopsis; distinctly broader asci and ascospores in Cetreliopsis compared to those in Cetrariopsis and Nephromopsis). Still, the shape of ascospores and asci of investigated genera presents an essential difference from Tuckermannopsis and related groups including Tuckneraria. The latter is charac­ terized by globose-subglobose ascospores which are arranged +/- uniseriately in asci with rather small tholus, very broad ocular chamber and broad axial body. The medul CETRARIOPSIS, CETRELIOPSIS AND NEPHROMOPSIS 57 Fig. 9-10 - Morphology in Cetreliopsis and Nephromopsis. Fig. 9. Upper surface of Cetreliopsis rhytidocarpa subsp. rhytidocarpa, Philippines, Luzon, 1909, Robinson 6586 (H). Fig. 10. Nephro­ mopsis morrisonicola, Philippines, Luzon, 1909, Merrill et al. 16359 (H). Bars in Figs. 9-10 = 1 cm. ps = pseudocyphellae. 29 58 T. RANDLANE, A. THELL and A. SAAG CETRARIOPSIS, CETRELIOPSIS AND NEPHROMOPSIS 59 Fig. 11-16 - Anatomy in Cetrariopsis, Cetreliopsis and Nephromopsis. Fig. 11. Cross section of an apothecium with a two-layered exciple of Cetreliopsis laeteflava, Taiwan, Kurokawa 205 (TAIM). Fig. 12. Cross section of an apothecium showing a three-layered exciple, typical for the genus Nephromopsis, N. rugosa, Russia, Primorye, Skirina 1982 (VLA). Fig. 13. Narrowly clavate ascus of Cetrariopsis pallescens, Nepal, Langtang, Miehe 16139 (GZU). Fig. 14. Somewhat broader asci and ascospores of Cetreliopsis asahinae, Russia, Primorye, 1951, Guriljova (TU). Fig. 15. Ascus of Nephromopsis stracheyi, Himalaya, Strachey & Winterbottom, 36138, isotypus (H-NYL). Fig. 16. Oblong ellipsoid ascospores of Cetrariopsis laii, Russia, Primorye 17.9.1961, S. Pam (TU). Bars in Figs. 11-16 = 10 pm, ab = axial body, th = tholus. lary chemistry of Cetrariopsis - Cetreliopsis - Nephromopsis is determined mainly by the occurrence of fatty acids. Some orcinol depsides and depsidones may be present in Cetrariopsis and Nephromopsis while (5-orcinol depsidones always occur in Cetreliop­ sis. All the 16 species of this generic complex are distributed in the montaneous forests of Eastern and Southeastern Asia only. So, we consider actually that the three genera Cetrariopsis, Cetreliopsis, and Nephromopsis - form a group of phytogenetically rela­ ted taxa. ACKNOWLEDGEMENTS. - The authors are grateful to the keepers of herbaria mentioned in the text for sending kindly the lichen specimens. Special thanks are due to Prof. Josef Poelt, Graz, for all help during the work and for lending us his Nepal materials. Thanks to Prof. Andrd Bellem&re, Dr. Marie-Agnfes Letrouit-Galinou, Paris, Prof. Rolf Santesson, Uppsala, Dr. Ingvar Karnefelt, Lund, and our collegues at the University of Lund for valuable comments and improvements on the manuscript. We are indebted to Dr. Weber, University of Colorado Museum, who had recognized Cetreliopsis papuae as a separate cetrarioid species and distributed it in his exsiccatae although the taxon was not identified. We are grateful also to Mr. Per Lassen for the help with the Latin diagnoses. The research described in this publication was made possible in part by Grant No. LCZ 000 from the International Science Foundation and also by the financial support from the Swedish Institute. REFERENCES ASAHINA Y„ 1954 - Lichenologische Notizen (§ 105-106). J. Jap. Bot. 29: 227-229. AWASTHI D.D., 1982 - Lichen genus Cetraria in India and Nepal. Bull. Bot. Surv. India 24: 1-27. CULBERSON C.F. and KRISTINSSON H., 1970 - A standardized method for the identification of lichen products. J. Chromatogr. 46: 85-93. CULBERSON C.F., 1972 - Improved conditions and new data for the identification of lichen pro­ ducts by a standardized thin-layer chromatographic method. J. Chromatogr. 72:113-115. CULBERSON C.F., CULBERSON W.L. and JOHNSON A., 1981 - A standardized TLC analysis of P-orcinol depsidones. The Bryologist 84: 16-29. CULBERSON W.L. and CULBERSON C.F., 1965 - Asahinea, a new genus in the Parmeliaceae. Brittonia 17: 182-190. CULBERSON W.L. and CULBERSON C.F., 1968 - The lichen genera Cetrelia and Platismatia (Parmeliaceae). Contr. U.S. Natl. Herb.: 449-558. 60 T. RANDLANE, A. THELL and A. SAAG ELIX J.A., 1993 - Progress in the generic delimitation of Parmelia sensu lato, lichens (Ascomyco­ tina: Parmeliaceae) and a synoptic key to the Parmeliaceae. The Bryologist 96: 359-383. ERIKSSON O.E. and HAWKSWORTH D.L., 1988 - Notes on ascomycete systematics. Nos. 733803. Syst. Ascomycetum 7: 103-117. HALE M.E., 1976 - Lichen structure viewed with scanning electron microscope. In: D.H. BROWN et al. (eds.), Lichenology: progress and problems. London, Academic Press, pp. 1-15. HUE A.-M., 1899 - Lichenes extra-Europaei. Nouv. Arch. Mus. Hist. Nat. s£r. 4,1: 27-220. KARNEFELT I., MATTSSON J.-E., THELL A., 1992 - Evolution and phylogeny of cetrarioid lichens. PI. Syst. Evol. 183:113-160. KARNEFELT I., THELL A., RANDLANE T. and SAAG A., 1994 - The genus Flavocetraria Karnefelt & Thell (Parmeliaceae, Ascomycotina) and its affinities. Ann. Bot. Fenn. 150: 7986. KUROKAWA S., 1980 - Cetrariopsis, a new genus in Parmeliaceae, and its distribution. Mem. Natl. Sci. Mus. 13: 139-142. LAl M.-J., 1980 - Studies on the cetrarioid lichens in Parmeliaceae of East Asia. Quart. J. Taiwan Mus. 33: 215-229. MONTAGNE J.P.F.C., 1856 - Sylloge generum specierumque cryptogamarum. Paris, 498 p. MONTAGNE J.P.F.C. and VAN DEN BOSCH R.B., 1857 «1855» - Lichenes javanici. In: Miquel, Plantae junghuhnianae, 4: 395-522. PARK Y.S., 1990 - The macrolichen flora of South Korea. The Bryologist 93: 105-160. POELT J., 1973 - Systematic evaluation of morphological characters. In'. V. AHMADJIAN and M.E. HALE (eds), The Lichens. New York and London, Academic Press, pp. 91-115. RANDLANE T. and SAAG A., 1991 - Some chemosystematical data about the lichen genus Nephromopsis in the U.S.S.R. Folia Cryptog. Estonica 28: 26-30. i RANDLANE T. and SAAG A., 1992 - Additional data about the genus Nephromopsis (Lichenes, Parmeliaceae). Mycotaxon 44:485-489. RANDLANE T. and SAAG A., 1993 - World list of cetrarioid lichens. Mycotaxon 47: 395-403. RANDLANE T., SAAG A., THELL A. and KARNEFELT I., 1994 - The lichen genus Tuckneraria Randlane & Thell - a new segregate in the Parmeliaceae. Ann- Bot. Fenn. 150: 143-151. RASANEN V., 1949 - Lichenes novi IV. Suom. Elain-ja Kasvit. Seuran Van. Kasvit Julk. 1948, 3: 78-79. RASANEN V., 1.952 - Studies on the species of the lichen genera Comicularia, Cetraria and Nephromopsis. Kuopion Luonnon Ystavain Yhdistyksen Julkaisuja B2: 1-53. RASSADINA K.A., 1971 - Fam. Parmeliaceae. In: Handbook of the lichens of the U.S.S.R. Lenin­ grad, Nauka, pp. 282-386 (in Russian). SCHWENDENER S., 1860 - Untersuchungen iiber den Flechtenthallus. In: Nageli, Beitr. Wiss. bot. 2:109-181. STAFLEU F.A. and COWAN R.S., 1981 - Taxonomic literature. Utrecht and the Hague, vol. 3. 980 PVAINIO E.A., 1909 - Lichenes in viciniis hibemae expeditionis -Vegae prope pagum Pitlekai in Siberia septentrionali a Dre Almquist collecti. Ark. Bot. 8. WEI J., 1991 - An enumeration of lichens in China. Beijing, International Academic Publishers, 278 p. V 30 Randlane, T. & Saag, A. Synopsis of the genus Nephrom opsis (Fam. Parmeliaceae, lichenized Ascomycota). — Cryptogamie, BryologieLichenologie. (Accepted for publication.) SYNOPSIS OF THE GENUS NEPHROMOPSIS (FAM. PARMELIACEAE, LICHENIZED ASCOMYCOTA) Tiina RANDLANE and Andres SAAG Institute o f Botany and Ecology, University o f Tartu, Lai Street 38, EE-2400 Tartu, Estonia RESUME! - Un aper?u global du genre N ephrom opsis Miill. Arg. (fam. Parmeliaceae) est presente. La cle d’identification comprend les 11 especes (N. endocrocea, N. isidioidea, N. komarovii, N. laii, N. morrisonicola, N. nephromoides, N. ornata, N. p a lle s­ cens, N. rugosa, N. stracheyi et N. yunnanensis) reconnus dans ce genre; une description detaillee et la distribution mondiale de chacune d’entre elles sont presentees pour la premiere fois. Une nouvelle combination N. nephromoides (Nyl.) Ahti & Randlane est proposee. ABSTRACT - The paper presents a global survey o f the genus N ephrom opsis Miill. Arg. (fam. Parmeliaceae). The identification key includes the 11 species now accepted in the genus (N. endocrocea, N. isidioidea, N. komarovii, N. laii, N. m orrisonicola, N. nephromoides, N. ornata, N. pallescens, N. rugosa, N. stracheyi and N. yunnanensis ); detailed descriptions and world distribution data o f each taxa are presented for the first time. A new combination N. nephromoides (Nyl.) Ahti & Randlane is proposed. KEY WORDS - lichenized Ascomycota, Parmeliaceae, C etrariopsis, N ephrom opsis, key for species, distribution maps. INTRODUCTION The genus N ephrom opsis Miill. Arg., described in 1891, was again brought to light in 1981 when Lai resurrected it in a treatment o f cetrarioid species with nephromoid apothecia and pseudocyphellae on the lower surface (Lai, 1981). Still, the evaluation of important characters on the genus level has changed considerably. Nowadays the anatomical features o f the thallus and especially o f the inner structures o f ascomata are considered more conservative and therefore suitable for delimitation at generic level. Mainly on these grounds the genus Tuckneraria (Randlane et al., 1994) was separated from N ephrom opsis and some taxa were transferred from N ephrom opsis to A llocetraria (Thell et al. , 1995b) or Cetreliopsis (Randlane et al., 1995). The position o f apothecia on the lower side o f the thallus has been one o f the most attractive and significant characters in defining the genus N ephrom opsis since Muller Argoviensis (1891), while the laminal position o f apothecia over the upper surface remains the only true feature for the genus Cetrariopsis (Kurokawa, 1980). Our studies have shown that there is not much difference in morphology, anatomy, and chemistry of these two genera except for their dissimilar positions o f apothecia (Randlane et al., 1995), Therefore we proposed transferring the two species o f Cetrariopsis (C. pallescens and C. laii) to the genus Nephrom opsis (Randlane et al. , 1997). The genus Cetreliopsis, also related to that group, differs considerably from Nephrom opsis in morphological (pseudocyphellae on both surfaces), anatomical (asci rather broadly clavate, ascospores ellipsoid) and chemical (absence o f orcinol depsides and depsidones and presence o f fi-orcinol 1 depsidones - fumarprotocetraric acid and related substances in all species) characters, and is therefore maintained as a separate genus. As a result, Nephromopsis includes now 11 species. The concise description of the genus has been published in an earlier paper (Randlane et al, 1995); in this one, an identification key is presented as well as short descriptions and distribution data for these taxa that were not treated previously. Distribution maps are provided for all species. MATERIAL AND METHODS About 380 herbarium specimens from B, BM, CANB, C OLO, E, FH, G, GZU, H, KW, LD, LE, M, PC, S, TAIM, TNM, TU, UPS, US, WU and private herbaria of A. Aptroot, D. D. Awasthi and J. A. Elix were examined. Anatomical studies of cortical and reproductive structures were carried out by Dr. Arne Thell in the University of Lund (Sweden) using methods described in Thell et al. (1995a). Chemical analyses were carried out according to the standardized TLC methods (Culberson & Kristinsson, 1970; Culberson, 1972). The acetone extracts were run in solvent systems B, C and G (Culberson et a l ., 1981). RESULTS 1. Identification key 1. Medulla coloured............................................................................................... . 2 - Medulla w hite....................................................................................................... 4 2. Lower side strongly rugose and reticulated, pseudocyphellae laminally developedon ridges and on special plug-like outgrowths. Rhizines absent............. ....................N. isidioidea - Lower side regularly reticulated, pseudocyphellae mainly developed in the marginal zone - on ridges or on the lower surface itself. Rhizines present...................................................... 3 3. Medulla pale yellow, K+ yellow (secalonic acid).....................................N. ornata - Medulla orange, K+ lilac (endocrocin)..............................................N. endocrocea 4. Apothecia small and numerous, mainly laminal......... N. pallescen s var. pallescen s - Apothecia of various size and number, mainly marginal...................................................................................................... 5 5. Lower surface black, only margins brown to pale brown................................................................................. N. m orrisonicola - Lower surface brown to whitish..............................................................................6 6. Epilithic; thallus strongly rugose, with concentric rings........................................................................... N. komarovii - Epiphytic; thallus smooth or rugose but not in concentric rings.................................................................................................. 7 7. Thallus light or bright yellow on both surfaces, more or less smooth; pseudocyphellae in the form of small flat white dots situated on the surface................. N. pallescen s var. citrina - Upper and lower surfaces of different colour, not uniformly yellow; thallus smooth to strongly rugose; pseudocyphellae different....................................................................... . 8 8. Medulla C+red (olivetoric or anziaic acid)............................................................ 9 - Medulla C - .......................................................................................................... 10 2 9. Thallus regularly reticulated; pseudocyphellae small and flat developed mainly on ridges; pycnidia on emergent projections; olivetoric acid in medulla...................................... N. rugosa - Thallus smooth or slightly wrinkled; pseudocyphellae medium to large, flat or concave, developed on the surface; pycnidia immersed; anziaic acid in medulla.............................N. stracheyi 10. Medulla KC+ red (physodic acid)............................................................ N. rugosa - Medulla KC - (lichesterinic and protolichesterinic acids)..................................... 11 11. Lower surface remarkably rugose, pseudocyphellae on ridges and plug-like outgrowths; pycnidia numerous on emergent projections, possible on both surfaces..........................N. yunnanensis - Lower surface smooth or moderately rugose, pseudo­ cyphellae either on the surface or on ridges but not on special outgrowths; pycnidia absent or marginally immersed............................................................................................. 12 12. Thallus moderately rugose, often with secondary marginal lobules; pseudocyphellae small and flat, mainly developed on ridges............................................................................ N. laii - Thallus smooth or slightly wrinkled, without secondary marginal lobules; pseudocyphellae medium to large, flat or concave, developed on the surface...................................... N. nephromoides 2. Taxonomical part N ephrom opsis Miill. Arg., Flora 74: 374,1891. Synonym: Cetrariopsis Kurok., Mem. Natl. Sci. Mus. Tokyo 13: 140,1980. Type species: Nephromopsis stracheyi (Bab.) Miill. Arg. Decription o f the genus was presented in a previous survey (Randlane et a l, 1995: 37-38). After transferring Cetrariopsis pallescens and C. laii to Nephromopsis (Randlane et al ., 1997) the description o f that genus should be emended in the following way: apothecia marginal on the lower surface o f the thallus, or submarginal and laminal on the upper surface, from small (1-2,5 mm) to very large (to 32 mm in diameter); exiple usually three-layered but sometimes the middle layer is not distinctly developed and is seen as two-layered in a few species. N ephrom opsis endocrocea Asahina, J. Jap. Bot. 11: 24, 1935. Type: Japan, insula Nippon (Honshu), Nasuzan, Faurie, 30.07.1897, no. 339 (KY, n. v.). Synonyms: C etraria endocrocea (Asahina) M. Sato in Nakai & Honda, Nov. FI. Jap. 5: 37,1939. Thallus foliose, up to 15 cm in diameter, with prolonged ascending lobes up to 8 mm wide; upper surface greenish or yellowish grey, smooth; medulla dark yellow or orange; lower surface light brown or dark brown to almost black, regularly reticulated; pseudocyphellae in the form o f minute white dots, mainly developed marginally on the lower surface itself or on ridges. Rhizines numerous to sparse, slender. Pycnidia marginal and laminal, located on the black emergent projections, sometimes very numerous. Pycnoconidia bifusiform, 5 x 1,5 fim. Apothecia marginal on the lower side o f the thallus, rounded or reniform, up to 12 mm in diameter, disc brown, faced upwards. Exciple three-layered. Asci 45 x 12 (am, axial body extremely small (0,5 jim), tholus with an amyloid ring structure, ascospores oblong, 9-10 x 4-5 jam. 31 3 Chemical constituents: usnic acid in the cortex; endocrocin and various fatty acids (e.g. lichesterinic and protolichesterinic acids) in the medulla. Medulla K+ lilac. Distribution and habitat: Japan, Russian Far East; China (Wei, 1991) (Fig. 1); corticolous on coniferous (Abies, Larix, Tsuga) or deciduous (Betula) trees at lower and medium altitudes (up to 2500 m). Morphologically quite similar to N. ornata except for the brighter colour of medulla and somewhat more delicate general habit. Altogether 32 specimens examined. Selected specimens examined. Japan. Prov. Shimotsuke, Karikomi-ko near Nikko, C. & W. Culberson, 1961, no. 11076 , 11127 (M, US). Prov. Shinano, Yatsugatake, Kurokawa, 1951 (LD). Hokkaido, Mt. Tomurausi, alt. 1000 m, Sato, 31.07.1935 (UPS). Russia. Far East, Kuril Islands, Kunashir, Goryachi Plyazh, Parmasto, 20.09.1960 (TU). Far East, Primorye Reg., Sikhote Alin, Mt. Kitovoye Rebro, 44°33’N 136°80’E, alt. 600 m, Skirina, 1982, no. 1661 (LD). Nephromopsis isidioidea (Rasanen) Randlane & Saag, Mycotaxon 44: 487,1992. Basionym: Cetraria wallichiana var. isidioidea Rasanen, Arch. Soc. Zool. Bot. Fenn. Vanamo 5, 1: 25, 1950. Type: India, E. Himalayas (West Bengal), Darjeeling Distr., Rimbick to Sandakhpoo, alt. 9000 ft., Awasthi, June 1948, no. 179 (H, holotype!). Synonym: Cetraria isidioidea (Rasanen) D. D. Awasthi, Bull. Bot. Surv. India 24: 10, 1982. Thallus foliose, ca. 5,5 cm in diameter, with lobes 1-2 cm wide; upper surface yellowish grey, strongly rugose; medulla yellow to ochraceous; lower surface dark brown to black, strongly rugose and reticulated, with special plug-like outgrowths. Rhizines absent, possibly broken from the outgrowths. White small pseudocyphellae situated on ridges and outgrowths of the lower side. Pycnidia mainly laminal but also marginal, located on numerous black emergent projections. Pycnoconidia bifusiform, 5 x 1,5 jim. Apothecia not seen. Chemical constituents: usnic acid in the cortex; secalonic acid C and fatty acids (lichesterinic and protolichesterinic acids) in the medulla. Medulla K+ reddish. Distribution and habitat: known only from the type collection from India, East Himalaya, West Bengal (Fig. 1), at altitude of about 2700 m; on a dead tree stump. The taxonomic status of N. isidioidea is not definitely clear, it is probably closely related to the other two species with coloured medulla, especially to N. ornata from which it differs in the more rugose and reticulated thallus, lack of rhizines and apothecia. Nephromopsis komarovii (Elenkin) J. C. Wei, Enumer. Lich. China: 158, 1991. Basionym: Cetraria komarovii Elenkin, Izv. Imp. S.-Peterburgsk. Bot. Sada 3: 51, 1903. Type: Russia, Irkutsk Region, in viciniis Nilova Pustyn, ad terram montis Chongoldoi montium Sajanensium, Elenkin, 1902, no. 155 (LE, holotype; FH, isotype!). Synonym: Cetraria perstraminea Zahlbr., Trudy Troitskos.-Kyakhtinsk. Otd. Priamursk. Otd. Imp. Russk. Geogr. Obshch. 12: 88, 1911 [1909]. Type: Russia, Transbaicalia, Chilgindin, Mikhno (n. v.). Thallus foliose, up to 15 cm in diameter, with rounded lobes up to 2 cm wide; upper surface from bright yellow to yellowish green, strongly rugose in a somewhat concentric pattern; medulla white; lower surface brown, smooth or slightly rugose; pseudocyphellae in the form of regular flat white patches of various size, developed laminally on the lower surface. Rhizines numerous to sparse, short and slender, light 4 Fig. 1. World distribution of N. endocrocea (#) and N. isidioidea ( k ). Fig. 3. World distribution ofN. laii. 5 brown to whitish. Pycnidia not seen. Apothecia usually marginal, occasionally laminal, rounded or reniform, up to 15 mm in diameter, disc brown, faced upwards, often only juvenile apothecia present. Exciple three-layered. Asci 30 x 9 )tim, axial body 3 ]im, ascospores oblong, 6 x 3 |xm. Chemical constituents: usnic acid in the cortex; lichesterinic and protolichesterinic acids in the medulla; fumarprotocetraric acid (Huneck et al., 1984) and stictic and constictic acids (personal comments by T. Ahti) have also been detected in some specimens. Distribution and habitat: Russia (eastern Siberia and Far East), Mongolia, China (Fig. 2); the only epilithic representative of the genus, growing mainly over rocks and boulders in shaded localities, often together with mosses; in mountane forests at lower and medium altitudes (200-3000 m); prefers humid ecotypes, e.g. valleys of mountain rivulets. Easily recognized by its characteristically rugosed upper surface which is usually intensively green up lemon yellow. Marginally situated juvenile apothecia are also frequent. Altogether 36 specimens examined. Selected specimens examined. Russia. Buryatia, eastern Sayans, Arshan, Trass, 1979 (TU). Baical Reg., Hamar-Daban, Masing, 1963 (TU). Far East, Primorye Terr., Sikhote Alin, Temei, 44°20’N 136°35’E, Skirina, 1985, no. 1661 (LD). Mongolia. Ulan-Bator, Bogd-uul in Zaisan, alt. 1600 m, Huneck, 1988, no. 88-15 (B). AraKhangai Reg., Tevshrulek, river Khuh-Sumein-gol, Biazrov, 1970, no. 7370 (LD). China. Yunnan, Setschwan Co., Hoso, alt. 2950 m, Handel-Mazzetti, 08.08.1915, no. 1364 (WU). Chili Co., Hsiao-wu-tai-shan, Smith, 1921 (COLO). Nephromopsis laii (Thell & Randlane) Saag & Thell, Bryologist 100: 111, 1997. Basionym: Cetrariopsis laii Thell & Randlane, Crytog. Bryol. Lichenol. 16: 46, 1995. Type: Russia, Primorye region, Kedrovaya Padj Nature Reserve, Pam, 17.09.1961 (TU, holotype!; LD, isotype!). Description and discussion in Randlane et al. (1995: 46-47). Distribution and habitat: Russia, China, Taiwan, Japan, India, Vietnam (Fig. 3); corticolous on coniferous {Abies, Pinus) and deciduous {Betula, Quercus) trees in mountainous forests between 600-3000 m elevation. The species has usually been compared to N. pallescens (both taxa were earlier referred to Cetrariopsis ); we now observe some similarities between N. laii and N. nephromoides, especially in the form, size and position of apothecia. The important characters to identify N. laii are the following: moderately rugose thallus with secondary lobules as a fringe along the margins; small and flat pseudocyphellae situated mainly on brown coloured ridges of the generally lighter lower surface. Altogether 24 specimens examined. Selected specimens examined. Russia. Far East, Primorye Reg., Sikhote Alin, Lazovsky Mt., alt. 950 m, 43°21’ N, 133°50’ E, Skirina, 1982, no. 1448 (LD). Primorye Reg., Sikhote Alin, Mt. Kitovoye Rebro, alt. 600 m, 44°33’ N, 136°80’ E, Skirina, 1982, no. 1445 (LD). Far East, Chabarovsk Terr., Obluchensk, Jadrino, Pam, 08.08.1961 (TU). China. Yunnan, Lijiang Co., Mt. Yulung-schan, alt. 3500 m, HandelMazzetti, no. 660 (WU). Taiwan. Chiayi Co., Mt. Alishan, alt. 2275 m, Lai, 1978, no. 10 198 (US). Japan. Nikko, Miyoshi, 1886 (FH). Goten-niwa, Mt. Huzi, Asahina, 1956, Lich. Jap. Exs. no. 157 (H). Vietnam. Zon Kin, Phan-si-Pau, Lao Kay, alt. 2900 m, collector unknown, 1929, no. 17156 (PC). 6 Nephromopsis morrisonicola M. J. Lai, Quart. J. Taiwan Mus. 33: 223,1981. Type: Taiwan, Nanton Co., Mt. Morrison, alt. 3500-3900 m, Lai, 1978, no. 10 438 (TAIM, holotype!). Description in Randlane et al. (1995: 38-40). Distribution and habitat: originally described as a Taiwanese endemic (Lai, 1981) - named after Mt. Morrison in Taiwan - but now known from a wide region in southeastern Asia: Taiwan, China, Nepal, Philippines, Indonesia (Java, Borneo, West Irian), Papua New Guinea (Fig. 4); grows as an epiphyte on coniferous {Abies) and deciduous (.Betula) trees or shrubs ( Vaccinium) at high altitudes (2400-4000 m). The taxon is easily recognized by the black underside and white medulla (N. endocrocea, N. isidioidea and N. ornata also have a dark brown to almost black lower surface but the medulla is coloured in all these taxa). Altogether 16 specimens examined. Selected specimens examined. China. Muli Kingdom, Mts. of Kopati, Djago & Muli, alt. 3100 m, Rock (B). Nepal. Khumbu Himal, Ngotung La, alt. 3410 m, Remus & Menzel, 16.04.1981, no. 239 (B). Indonesia. West Irian, Carstensz Mts., Lower Meren valley near Blue Lake, Hope, 30.12.1971, no. CGE L43 (COLO). Papua New Guinea. Southern Highlands, Mt. Giluwe, alt. 11 000 ft., McVean, 1967, no. 67142 (COLO). Nephromopsis nephromoides (Nyl.) Ahti & Randlane comb. nov. Baisionym: Platysma nephromoides Nyl., Flora 52: 442, 443, 1869. Type: India, West Bengal, Darjeeling Distr., Tongloo, alt. 10,000 ft., Hooker fit. & Thomson, no. 2080 (as '2020' in protologue) (H-NYL 36068, lectotype!; B, PC, UPS isolectotypes!). - Synonyms: Nephromopsis stracheyi var. nephromoides (Nyl.) Rasanen, Kuopion Luonnon Ystavain Yhdistyksen Julkaisuja, ser. B 2, 6: 48, 1952. Cetraria nephromoides (Nyl.) D. D. Awasthi, Bull. Bot. Surv. India 24: 11,1982. Nephromopsis stracheyi f. ectocarpisma Hue, Nouv. Arch. Mus. Hist. Nat., Ser. 4,1: 218,1899. Type: Japan, insula Yeso (Hokkaido), in sylvis Mombetsu, Faurie, 1891, no. 3521 (PC, lectotype!, selected here). - Nephromopsis ectocarpisma (Hue) Gyeln., Ann. Cryptog. Exot. 4: 173, 1931. Thallus foliose, up to 20 cm in diameter, with rounded wide lobes which may be convoluted; upper surface greenish grey, thick, smooth or slightly wrinkled; medulla white; lower surface light or yellowish brown, smooth or somewhat reticulated at the margins; pseudocyphellae conspicuous, oval or rounded, flat to concave, situated mainly on the surface, occasionally - in the marginal zone - also on ridges. Rhizines sparse, short and simple. Pycnidia rare, marginal, immersed. Pycnoconidia not seen. Apothecia usually numerous, marginal, comparatively small, up to 8 mm in diameter, disc brown, rounded or more often irregular, faced upwards. Exciple three-layered. Asci 35-40 x 10 nm, axial body small (3 (im), ascospores ellipsoid, 7-8 x 3 |um. Chemical constituents: usnic acid in the cortex; lichesterinic and protolichesterinic acids and additionally some other fatty acids, e.g. caperatic acid (+/-) in the medulla. Distribution and habitat: Japan, China, Taiwan, Vietnam, India, Nepal (Fig. 5); corticolous on trees at the altitudes between 2400 and 3600 m. The new combination Nephromopsis nephromoides has to be proposed for N. ectocarpisma because the name Platysma nephromoides Nyl. turned out not to be a nomen nudum as it was considered to be until recently (Randlane et al., 1997). Some characters of that taxon are pointed out in the same paper where the species is mentioned for the first time (Nylander, 1869: 442), but under the description of quite another species {Platysma stracheyi) (Nylander, 1869: 443). 32 7 N. nephromoides is similar to N. stracheyi in general morphology (smoothness of the thallus, form of lobes and pseudocyphellae) but is slightly smaller and thinner, also the apothecia are considerably smaller and often more numerous; the secondary compounds (and thus the C reaction on medulla) are also quite different. Sometimes may resemble N. laii but the latter has typically marginal secondary lobes and different pseudocyphellae. Altogether 50 specimens were examined. Selected specimens examined. Japan. Prov. Aomori, Osorezan, Faurie, 1902, no. 5341 (FH, PC); Prov. Nara, Yoshino, Mt. Odaigahara, Nakanishi, I960 (US); Honshu, Prov. Tottori, Mt. Daisen, Koponen, 20.07.1971, no. 21818 (H). China. Yunnan, Lijiang Co., alt. 3000 m, L. S. Wang, 06.08.1985, no. 85-354 (H). Taiwan. Hwalien Co., Mt. Kilaishan, Nakamura, 30.12.1940, no. 385 (US). India. West Bengal, Darjeeling, from Sandakhpoo to Phalut, alt. 3600 m, Awasthi & Agarwal, 16.06.1967, no. 67442 (UPS). Nepal. Prov. Helambu-Langtang, Kutumsang, 27°57’N 85°29’E, alt. 2750 m, Rettig, 21.11.1988, no. 6471a (GZU). From Rakhshe to Ethung, alt. 9000 ft., Awasthi, 16.05.1953, no. 2128 (UPS). Nephromopsis ornata (Mull. Arg.) Hue, Nouv. Arch. Mus. Hist. Nat., Ser. 4,2: 90,1900. Basionym: Cetraria ornata MOIL Arg., Nuovo Giom. Bot. Ital. 23: 122, 1891. Type: Japan, Mt. Ontake, no. 109 (n. v.). Synonyms: Nephromopsis delavayi Hue, Nouv. Arch. Mus. Hist. Nat., Ser. 4, 1: 219, 1899. - Type: China, Yunnan, Lopinchan, Lanhong, alt. 3200 m, Delavay, 31.07.1888 (DUKE, n. v.). - Syn.: Cetraria delavayi (Hue) M. Sato in Nakai & Honda, Nov. FI. Jap. 5: 48,1939. Nephromopsis endoxantha Hue, Nouv. Arch. Mus. Hist. Nat., Ser. 4, 1: 220, 1899. - Type: Japan, Togakushi, Faurie, 17.09.1898, no. 776 (KY, lectotype; DUKE, isolectotype; n. v.). - Syn.: Tuckermannopsis endoxantha (Hue) Gyeln., Acta Fauna FI. Universali, Ser. 2, Bot. 1, 5/6: 6, 1933. - Cetraria endoxantha (Hue) D. D. Awasthi, Bull. Bot. Surv. India 24: 9, 1982. Thallus foliose, up to 15 cm in diameter, with prolonged ascending lobes up to 1.5 cm wide; upper surface greenish or yellowish grey, smooth or somewhat wrinkled; medulla pale yellow; lower surface brown or dark brown to almost black, regularly reticulated; pseudocyphellae in the form of minute white dots, mainly developed marginally on the lower surface or on ridges. Rhizines sparse, slender. Pycnidia marginal and laminal, located on the black emergent projections, sometimes very numerous. Pycnoconidia bifusiform, 5 x 1 ^im. Apothecia marginal on the lower side of the thallus, rounded or reniform, up to 20 mm in diameter, disc brown, faced upwards. Exciple three-layered. Asci 40-45 x 10 |im, axial body extremely small (0,5-1,2 |im), tholus with an amyloid ring stucture, ascospores oblong, 7-9 x 4-5 (im. Chemical constituents: usnic acid in the cortex; secalonic acids A or C, traces of endocrocin and fumarprotocetraric acid (+/-), additionally some fatty acids (+/-) in the medulla. Medulla K+ deep yellow. Distribution and habitat: Japan, Russian Far East; China (Wei, 1991), Taiwan (Lai, 1981), South Korea (Park, 1990) (Fig. 6); corticolous on coniferous (Abies, Larix, Picea, Pinus, Taxus) and deciduous trees (Alnus, Betula, Padus, Phellodendron, Populus, Salix, Tilia) or bushes (Rhododendron), occasionally also on boulders covered with mosses in various types of forests at lower and medium altitudes (up to 3200 m). The most common species among Nephromopsis. 8 0 Vi QV- • )•--— .• •--- ---- r r Z .^ i IwPr ' 'A. Lv A-„.... - y 8 0 ■ic^ ' w ^77 ■ ■/■■■• p . m.m a >v l Fig. 4. World distribution of N. morrisonicola. 9 N. ornata is a distinct taxon, easily recognized by its pale yellow medulla; systematically closely related and morphologically similar to N. endocrocea (see the differences on p. 3). Altogether 78 specimens examined. Selected specimens examined. Japan. Prov. Shinano, Mt. Kita-Yokodake, Kurokawa, 1958, no. 58348 (M, US). Honshu, Prov. Kozuke, Sannoh Pass, alt. 1600 m, Degelius, 29.04.1964 (UPS). Mt. Fuji, alt. 1000-2400 m, Hulten, 05.10.1961 (S). Russia. Far East, Primorye Terr., Mt. Snezhnaya, Randlane, 17.09.1983 (TU). Primorye Terr., Sikhote Alin, Mt. Eldorado, 44°41’N 135°20’E, Skirina , 1984, no. 8995 (LD). Khabarovsk Terr., Selihin, Kabansopka, Pam, 18.08.61 (TU). Nephromopsis pallescens (Schaer.) S. Y. Park var. pallescens, Biyologist 93:122,1990. Basionym: Cetraria pallescens Schaer., in Moritzi, Syst. Verzeichn.: 129, 1845-1846. Type: Java, Mt. Pangerango, Zollinger, no. 449 (G, holotype!). Synonyms: Platysma pallescens (Schaer.) Nyl., Mem. Soc. Sci. Nat. Cherbourg 5: 100, 1858 [1857], - Cetrariopsis pallescens (Schaer.) Randlane & Thell, Crytog. Bryol. Lichenol. 16: 42,1995. Sticta wallichiana Taylor, London J. Bot. 6: 177, 1847. - Type: Nepal, Wallich (FH, holotype, n. v.; G, no. 2003/2; G, no. 2003/1, PC, isotypes!). - Syn.: Parmelia wallichiana (Taylor) Nyl., M&n. Soc. Sci. Nat. Cherbourg 5: 105, 1858 [1857]. Platysma leucostigmeum var. wallichianum (Taylor) Nyl., Syn. Meth. Lich. I: 306, 1860. - Platysma wallichianum (Taylor) Nyl., Flora 52: 443, 1869. - Cetraria wallichiana (Taylor) Miill. Arg., Flora 71: 139, 1888. - Cetrariopsis wallichiana (Taylor) Kurok., Mem. Natl. Sci. Mus. Tokyo 13: 140, 1980. - Ahtia wallichiana (Taylor) M. J. Lai, Quart. J. Taiwan Mus. 33: 220, 1981 (nom. illeg.). Cetraria sulphurea Mont. & Bosch, in Montagne, Syll. Gen. Sp. Crypt.: 322, 1856 (not validly published). - Orig. coll.: Java, Junghuhn (n. v.). Cetraria teijsmannii (“Teysmannf”) Mont. & Bosch, in Montagne, Syll. Gen. Sp. Crypt.: 474, 1856. - Type: Java, Teijsmann (n. v.). - Synonym: Platysma teijsmannii (“Teysmannr) (Mont. & Bosch) Nyl., Mem. Soc. Sci. Nat. Cherbourg 5: 100, 1858 [1857]. Description and discussion in Randlane et al. (1995: 42-44). Distribution and habitat: Nepal, India, China, Taiwan, Russian Far East, Indonesia (Java); Japan, Thailand (Kurokawa, 1980), South Korea (Park, 1990), Papua New Guinea (Streimann, 1986) (Fig. 7); epiphytic on coniferous (Larix, Pinus) and deciduous (Carpinus, Quercus) trees or shrubs (.Rhododendron) in the mountainous forests at medium and high altitudes (1200-4000 m). In its typical form, N. pallescens is very conspicuous and has always been easily recognized by numerous small apothecia laminally situated all over the thallus. In less typical cases the characteristic features of the lower side - strongly rugose and reticulated surface with pseudocyphellae on ridges and special plug-like outgrowths are also of great help in identification. Altogether 47 specimens examined. Selected specimens examined. Nepal. Mewakhola valley, alt. 8000 ft., Awasthi, 27.05.1953, no. 2237a (UPS); Langtang Area, below Dotsche, alt. 2900 m, G. & S. Miehe, 1986, no. 15 615 (GZU). India. Bhutan, Taba, Thimpu, 27°30’N 89°39’E, alt. 2500 m, Grierson & Long, 12.05.1979, no. 972 (E). China. Yunnan, Lijiang Co., Mt. Tiejia Shan, 26°56’N 100°10’E, alt. 2750 m, Moberg & Santesson, 24.09.1987, no. 8075 (UPS). Mt. Gibboh, alt. 4000 m, Rock, no. 1630 (B). Russia. Far East, Primorye Terr., Kedrovaya Padj, Ivaninnikova, 1959 (TU). 10 Nephromopsis pallescens var. citrina (Taylor) Thell & Randlane, Bryologist 100: 110, 1997. Basionym: Cetraria citrina Taylor, London J. Bot. 6: 176, 1847. Type: Java, Hooker (BM, lectotype!). Synonyms: Platysma citrinum (Taylor) Nyl., Mem. Soc. Sci. Nat. Cherbourg 5: 100, 1858 [1857]. - Cetrariopsis pallescens var. citrina (Taylor) Thell & Randlane, Crytog. Bryol. Lichenol. 16: 44,1995. Description and discussion in Randlane et al. (1995: 44). Distribution and habitat: Indonesia (Java, West Irian) and Taiwan (Fig. 7); corticolous in mountainous forests (alt. 1300 - 3000 m). This southern variety (with its main distribution area in Java) is recognized by its smooth and uniformly light or bright yellow thallus on both surfaces as well as minute flat pseudocyphellae on the lower side. Altogether 59 specimens examined. Selected specimens examined. Indonesia. Java. Pangerango, alt. 2820 m, Schiffner, 1894, no. 2987a (FH, M); Preanger, Mt. Gedeh, alt. 2300 m, Schiffner, 1894, no.3387 (FH, M); Kandang Badals, alt. 2600 m, Yates, 1927, no. 2827 (LD, US); East Java, Andjasmora-complex, alt. 2100 m, Groenhart, 1937, no.2538 (Herb. Aptroot); Res. Pasoeroean, Goenoeng Ardjoena, Tretes - Lalidjiwa track, Du Rietz, 1927, no. 105-lb (UPS). Taiwan. Mt. Arisan, Asahina (US). Nephromopsis rugosa Asahina, J. Jap. Bot. 11: 12,1935. Type: Japan, Prov. Musasi, Mt. Kobusi, Asahina, 22.07.1933 (DUKE, isotype). Synonym: Cetraria rugosa (Asahina) M. Sato, in Nakai & Honda, Nova Flora Japonica 5: 46,1939. Thallus foliose, up to 20 cm in diameter, with rounded lobes up to 2.5 cm wide; upper surface yellowish or glaucous olive, often with a significant green tinge, remarkably regularly reticulated; medulla white; lower surface light brown, yellowish or whitish, strongly reticulated; pseudocyphellae on the lower side in the form of minute flat white spots located mainly on ridges. Rhizines sparse. Pycnidia marginal and laminal, on the black emergent projections, which are often situated along the ridges of the upper surface. Pycnoconidia bifusiform, 5 x 1-1,5 jim. Apothecia marginal on the lower side of the thallus, rounded or reniform, up to 20 mm in diameter, disc brown, faced upwards. Exciple three-layered. Asci 35-40 x 10 fim, axial body 4 jim, ascospores oblong, 7-9 x 3-5 (im. Chemical constituents: usnic acid in the cortex; two chemotypes: I - olivetoric acid; II - physodic and oxyphysodic acids, additionally fatty acids (+/-) in the medulla. Medulla C+ red in chemotyp I, C-, KC+ red in chemotype II. Distribution and habitat: Russian Far East, Japan; Mongolia (Schubert & Klement, 1971) (Fig. 8); corticolous on coniferous (Abies, Larix, Picea ) or deciduous trees (Quercus) in forests between 700-1700 m elevation.. N. rugosa can be morphologically recognized by its significantly reticulated thallus which often has a characteristic greenish tinge. Spot tests with C and KC are also of great help. Altogether 21 specimens examined. Selected specimens examined. Russia. Far East, Primorye Terr., Kedrovaya Padj, Guriljova, 1951 (TU; chemotype I). Primorye Terr., Sikhote Alin, Dzigitovka River valley, 44°50’N 136°10’E, Skirina, 1982, no. 9280 (LD; chemotype II). Sikhote Alin, Kitovoye Rebro, alt. 800 m, 44°33’N 136°80’E, Skirina, 1982, no. 1449 (LD; 33 11 chemotype II). Japan. Berg Buko, Miyshi, 1891 (UPS; chemotype I). Prov. Shinano, Mt. Mikuniyama, Kurokawa, 1958, Lich. Jap. Exs. no. 254 (TAIM; chemotype II). Honshu, Prov. Musashi, Mikuni Pass., alt. 1750 m, Shibuichi, Lich. Rar. et Critici Exs. no. 154 (B, LD, US; II chemotype). Nephromopsis stracheyi (Bab.) Mull. Arg., Flora 74: 374,1891. Basionym: Cetraria stracheyi Bab., Hooker’s J. Bot. Kew Gard. Misc. 4: 245, 1852. Type: Himalaya, Kathi, 7200 fit., Strachey & Winterbottom (BM, holotype; H-NYL 36138, isotype!). Synonym: Platysma stracheyi (Bab.) Nyl., Flora 52: 443, 1869. Thallus foliose, thick, coriaceous, up to 20 cm in diameter, with rounded wide iobes up to 3 cm wide; upper surface greenish or yellowish grey, smooth or only slightly wrinkled; medulla white; lower surface light or yellowish brown, smooth or reticulated; pseudocyphellae conspicuous, medium to large, oval or rounded, flat to concave, situated directly on the surface. Rhizines sparse, short and simple. Pycnidia rare, marginal, immersed. Pycnoconidia bifusiform, 5 x 1.5 jam. Apothecia marginal on the lower side, sometimes extremely large (up to 20 mm in diameter), disc brown, rounded or somewhat irregular, faced upwards. Exciple three-layered. Asci 35 x 10 jum, axial body small (3 jim), ascospores ellipsoid, 7-8 x 2,5-3 jim. Chemical constituents: usnic acid in the cortex; olivetoric (chemotype I) or anziaic acid (chemotype II) in the medulla. Medulla C+ red in both chemotypes. Distribution and habitat: India, Nepal, Taiwan; China (Wei, 1991) (Fig. 9); corticolous on trees at low and medium altitudes (up to 2800 m). N. stracheyi is mostly similar to N. nephromoides in general habit of the thallus and form of pseudocyphellae; differences can be easily noticed in the size of apothecia (much bigger in N. stracheyi) and spot test with C in medulla (positive in N. stracheyi). Altogether 13 specimens examined. Selected specimens examined. India. NW Himalayas, Almora Distr., Dhakuri, alt. 9500 ft., D. Awasthi & A. Awasthi, 1950, no. 642 (UPS, US). Nepal. On ascent to Sandakhpoo, alt. 1100 ft., Awasthi, 1953, no. 2470A (FH, UPS). Langtang Area, Dunche, alt. 2800 m, G. Miehe & S. Miehe, 22.03.1986, no. 346 (GZU). Taiwan. Miaoli Co., Nankonchi, alt. 2000 m, Kao, 1959, no. 121 (US). Nephromopsis yunnanensis (Nyl.) Randlane & Saag, Mycotaxon 44: 488, 1992. Basionym: Platysma yunnanense (“yunnense”) Nyl., Lich. Nov. Zeland.: 150, 1888. Type: China, Yunnan, alt. 1800 m, Delavay, no. 1602 (H-NYL 36134, lectotype!). Synonym: Cetraria yunnanensis (Nyl.) Zahlbr., Trudy Troitskos.- Kyakhtinsk. Otd. Priamursk. Otd. Imp. Russk. Geogr. Obshch. 12: 89, 1911 [1909]. Description in Randlane et al. (1995: 39-40). Distribution and habitat: endemic to China (prov. Yunnan) (Fig. 10); corticolous on coniferous (Picea) or deciduous trees (Quercus) in mountainous forests (alt. 1800-2800 m). The most attractive character of N. yunnanensis is the abundance of laminal pycnidia on pale emergent projections (all the other Nephromopsis species bear black projections, if they have any); pseudocyphellae are typically developed on ridges and outgrowths of the very rugose lower surface. Specimens examined in Randlane et al (1995: 40). 12 Fig. 7. World distribution of N. pallescens var. pallescens ( • ) and N. p. var. citrina (★). N. rugosa. Fig. 8. World distribution of Fig. 9. World distribution oiN. stracheyi. Fig. 10. World distribution of N. yunnanensis. ACKNOWLEDGEMENTS. - The authors are grateful to the keepers of herbaria mentioned in the text for kindly sending the lichen specimens. Special thanks are due to our good friend and colleague Dr. Arne Thell, Lund, for the anatomical data. Dr. Teuvo Ahti, Helsinki, Dr. Ingvar Kamefelt and the late Prof. Josef Poelt, Graz, are thanked for valuable comments and discussions on the subject. The research described in this publication was made possible in part by Grant No. LLO 100 from the International Science Foundation and by Grant No. 1297 from the Estonian Science Foundation. 13 REFERENCES CULBERSON C.F., 1972 - Improved conditions and new data for the identification of lichen products by a standardized thin-layer chromatographic method. Journal of Chromatography 72: 113-115. CULBERSON C.F., CULBERSON W.L. & JOHNSON A., 1981 - A standardized TLC analysis of fi-orcinol depsidones. The Bryologist 84: 16-29. CULBERSON C.F. & KRISTINSSON H., 1970 - A standardized method for the identification of lichen products. Journal o f Chromatography 46: 85-93. HUNECK S., POELT J., AHTI T., VITIKAINEN O. & COGT U., 1984 - Zur Verbreitung und Chemie von Flechten der Mongolischen Volksrepublik. Erforsch. biol. Ress. der Mongolischen Volksrepublik 4: 51-62. KUROKAWA S., 1980 - Cetrariopsis, a new genus in Parmeliaceae, and its distribution. Memoirs o f the National Science Museum 13: 139-142. LAI M.-J., 1981 - Studies on the cetrarioid lichens in Parmeliaceae of East Asia. Quarterly Journal o f Taiwan Museum 33: 215-229. MULLER ARGOVIENSIS, 1891 - Lichenologische Beitrage 35. Flora 74: 371-382. NYLANDER W., 1869 - De reactionibus in Cetrarieis. Flora 52: 441-444. PARK Y. S., 1990 - The macrolichen flora of South Korea. The Bryologist 93:105-160. RANDLANE T., SAAG A. & THELL A., 1997 - A second updated world list of cetrarioid lichens. The Bryologist 100: 109-122. RANDLANE T., SAAG A., THELL A. & KARNEFELT I., 1994 - The lichen genus Tuckneraria Randlane & Thell - a new segregate in the Parmeliaceae. Acta Botanica Fennica 150: 143-151. RANDLANE T., THELL A. & SAAG A., 1995 - New data about the genera Cetrariopsis, Cetreliopsis and Nephromopsis (fam. Parmeliaceae. lichenized Ascomycotina). Cryptogamie, Bryologie Lichenologie 16(1): 35-60. SCHUBERT R. & KLEMENT O., 1971 - Beitrag zur Flechtenflora der Mongolischen Volksrepublik. Feddes Repertorium 82 (3-4): 187-262. STREIMANN H., 1986 - Catalogue of the lichens of Papua New Guinea and Irian Jaya. Bibliotheca Lichenologica 22: 1-145. THELL A., MATTSSON J.-E. & KARNEFELT I., 1995a - Lecanoralean ascus types in the lichenized families Alectoriaceae and Parmeliaceae. Cryptogamie Botany 5: 120-127. THELL A., RANDLANE T., KARNEFELT I., GAO X. & SAAG A., 1995b - The lichen genus Allocetraria (Ascomycotina, Parmeliaceae). In: Daniels F.J.A., Schultz M. & Peine J. (eds.), Flechten Follmann. Contributions to lichenology in honour of Gerhard Follmann. University of Cologne, pp. 353-370. WEI J., 1991 - An enumeration o f lichens in China. Beijing, International Academic Publishers, 278 p. 14 VI 34 Randlane, T., Saag, A., Thell, A. & Kamefelt, I. 1994. The lichen genus Tuck­ neraria Randlane & Thell — a new segregate in the Parmeliaceae. — Acta Botanica Fennica 150: 143-151. Acta Bot. Fennica 150:143-151, 1994 The lichen genus T uckneraria R andlane & Thell — a new segregate in the Parm eliaceae TIINA RANDLANE, ANDRES SAAG, ARNE THELL and INGVAR KARNEFELT Randlane, T., Saag, A., Thell, A. & Kamefelt, I. 1994: The lichen genus Tuckneraria Randlane & Thell — a new segregate in the Parmeliaceae. — Acta Bot. Fennica 150:143-151. Helsinki. ISSN 0001-5369 ISBN 951-9469-44-3 The new lichen genus Tuckneraria Randlane & Thell is described. The separation from Nephrom opsis is based mainly on anatomical characters in the reproductive structures, such as shape and size o f ascospores and structures o f exciple and ascus, but also on morphological characters — thallus surface features and the presence o f cilia. T he genus Tuckneraria includes the three species T. laureri (Kremp.) Randlane & Thell, T. laxa (Zahlbr.) Randlane & Thell, T. pseudocom plicata (Asah.) Randlane & Saag and the newly described T. ahtii Randlane & Saag. N ephrom opsis nipponensis (Asahina) M.J.Lai is considered synonymous with Tuckneraria pseudocom plicata. Key words: lichenized Ascomycotina, Nephromopsis, Parmeliaceae, Tuckermannopsis, Tuckneraria Tiina Randlane and Andres Saag, Institute o f Botany & Ecology, Tartu University, Lai Street 38, EE-2400 Tartu, Estonia; Ingvar Kamefelt and Arne Thell, Dept, o f Systematic liotany. University o f Lund, Oslra Vallgatan 18-20. S-223 61 Lund, Sweden INTRODUCTION The generic complex comprising Cetrariopsis -C e ir e lio p sis-N e p h ro m o p sis (A scom ycotina, Parmeliaceae) has so far not been monographed on a worldwide basis. The species within these rare Asiatic genera are poorly known and have been referred to during the last few decades only in some local florislic studies (Rassadina 1971, Golubkova 1981, Awasthi 1982, Park 1990, Kurokawa 1991), with the exception o f a sig­ nificant paper on cetrarioid lichens o f East Asia (Lai 1980). A team o f reseachers is now carrying out a detailed study o f this group and the present paper is the first report o f the project. U ntil recently, the genus N e p h ro m o p sis Miill.Arg. has been delimited mainly on the basis o f morphological and chemical characters (Lai 1980, Randlane & Saag 1991, 1992) such as loosely attached foliose thallus; the tendency o f marginal apothecia to be situated on the lower surface; the p resence o f lam inal p seu d o­ cyphellae over the lower cortex; the occurrence o f pycnidia, frequently on emergent projections, marginally and/or laminally; the com plex o f secondary com pounds including usnic acid, fatty acids, orcinol depsides and depsidones and anthraquinones. However, anatomical studies o f ascocarps clearly dem onstrate that sp ecies 144 Tiina Randlane et al. treated under this genus vary significantly in their shape of ascospores, ascus type and excipular structure. As the division of species into groups based on anatomical qualities is also correlated with morphological characters, the segregation of a new genus is fully justified. MATERIAL AND METHODS About 250 herbarium specimens from B, DUKE, FH, GZU, H. KW, LD, M, MB, S, TAIM, TNS, TU, UPS, US, the majority of them belonging to T. laureri (Kremp.) Randlane & Thell, were studied. Chemical analyses according to the standardized TLC methods (Culberson 1972, 1974) were carried out in Tartu University. The acetone extracts were run in solvent systems B, C and G (Culberson et al. 1981). After spraying with 10% sulphuric acid, the plates were air dried and then heated at about 10 0 - 120°C for up to 15 min. Anatomy of ascocarps and conidiomata was examined at the University of Lund. Sections were made with a Kryomat, Leitz freezing microtome and stained in lactophenol cottonblue. After pretreatment with 10% KOH solution, asci were squashed in a 0.3% Lugol’s solution. The characters were studied with a Zeiss Axioscope light microscope, and photomicrographs made with a Zeiss M 35 W camera. TAXONOMY Tuckneraria Randlane & Thell, gen. nova Thallus foliaceus, mediocris (ad 7 cm latus), pallide flavescens, virescens vel glaucus; laciniae oblongae aut suborbiculares; partim ascendentes; marginibus interdum ciliatis; margine sorediata aut soredia desunt. Superficies inferior pallida, fusca vel nigra; pseudocyphellata; rhizinata. Apothecia marginalia, orbicularia vel reniformia, ad 10 mm lata; sporae 8-nae, subglobosae, 5-7 x 4-5 fim; asci 30-50 x 10-14 p.m. Pycnidia marginalia, papillaria vel spinuliformia; conidia 4-5 x 1-1.5 |im, extremis nonnihil infiatis. Type spccics: Tuckneraria pxem loconiplicala (Asahina) Randlane & Saag ACTA BOT. FENNICA 150 (1994) Thallus foliose, medium (to 7 cm broad), smooth or only slightly rugose; light yellow, yellowish-green or yellowish-grey, +/- loosely attached to the substratum; lobes elongate or rounded, with ascending margins and numerous or occasional marginal cilia; with or without marginal soredia; lower surface whitish, light to dark brown or black; white or light brown, usually small and plain pseudocyphellae situated on lower cortex; rhizines simple, at times long and numerous; both cortices para­ plectenchymatous; cortical hyphae strongly ge­ latinized. Apothecia marginal, disc brown, rounded or reniform, to 10 mm in diameter, facing upor downwards; exciple 2-layered; ascospores simple, globose - subglobose, 5-7 x 4-5 um, 8 per ascus; asci 30-50 x 10-14 |im, clavate, spores arranged +/- uniseriately; asci Tuckerm a n n o p sis-typ e (Karnefelt et al. 1992) with rather small tholus, very broad ocular chamber and broad axial body (2.5-4 um). Pycnidia on marginal (occasionally laminal on both surfaces) emergent projections; pycnoconidia bifusiform, 4-5 x 1-1.5 ^m. Chemical constituents: usnic acid +/- in the cortex; lichesterinic- and protolichesterinic-type fatty acids, caperatic acid and orcinol de­ psidones (physodic, conphysodic, alectoronic, collatolic acids) in the medulla. The name for the new genus is compiled from the names of the related lichen genera Tuckermannopsis and Nephromopsis, com-bining them with C e tr a ria . The species within Tuckneraria are morphologically reminiscent of the species in N ephrom opsis but in ascocarp anatomy more like Tuckermannopsis. These two genera are probably the most closely related entities to the new genus. Still, there are certain differences even in the morphological characters between Nephromopsis and Tuckneraria (Figs. 1-5): the thallus of the species within N ephrom opsis is usually coriaceous, thick and often strongly Figs. 1-5. Morphology of the genus Tuckneraria. — I: T. aluii, part of the frequently ciliated thallus, China, Yunnan, Handel-Mazetli 660 (US). — 2: T. laureri, showing the sorediate margin and the marginal apothecium, Austria, Stubaier Alpen, 1958 Steiner (LD). — 3: T. pseudocomplicata, part of the ciliated thallus; Japan, Shikoku, Awa, Fujikawa 29560 (TNS). — 4: T. laxa, characterized by the same type of cilia; Taiwan, Miaoli Co., Lai 7770 (TAIM). — 5: Lobes of the same specimen with marginal pycnidia and cilia. Scale in Figs. 1, 2, 4 = 1 cm, in Figs. 3, 5 = 1 mm. p = pycnidia. s = soredia. ACTA BOT. FENNICA 150 (1994) 35 Tuckneraria a new segregate in the Parmeliaceae 145 146 Tiina Randlane et a l ACTA BOT. FENNICA 150 (1994) ACTA BOT. FENNICA 150 (1994) Tuckneraria a new segregate in the Parmeliaceae reticulated or rugose; the lobes are usually rounded, not elongate; the marginal cilia are absent; pseudocyphellae on the lower surface are large, well delimited, either on special outgrowths or concave. In Tuckneraria cortical hyphae are always strongly gelatinized (Figs. 7-9). The ascospores o f these two genera essentially differ, being ellipsoid in N ephrom opsis and subglobose in Tuckneraria, while the anatomical characters of exciple (Fig. 8) and asci (Figs. 6, 10, 11) present similarities as well as differences (Table 1). Pycnoconidia are bifusiform (5 x 1-1.5 pm) in both genera (Figs. 12, 13). Secondary chemistry o f Nephromopsis and Tuckneraria is similar: both have usnic acid in the cortex; both have lichesterinic- and protolichesterinic-type fatty acids and occasionally caperatic acid in the medulla together with the orcinol depsidones physodic and conphysodic acids; a few Nephromopsis species also contain orcinol de-pside olivetoric acid and the anthraquinone en-docrocin or secalonic acid, none of which are present in Tuckneraria. The best characters for the separation of Tuckneraria from Nephromopsis are the general habit o f the thallus and the ascospore shape. Species within Tuckermannopsis are easily se­ parated from both genera by their lack o f pseu­ docyphellae on the lower cortex. The genus Tuckneraria includes 4 species growing on deciduous and coniferous trees mainly in eastern and south-eastern Asia. Tuckneraria ahtii Randlane & Saag, spec, nova Thallus foliaceus, laciniae 4 -1 0 mm latae; marginibus interdum ciliatis; superficies superior virescens vel fuscesenti-flavens. Superficies infe­ 147 rior pallide fusca, in centra fere nigra, pseudocyphellata; rhizinae fuscae, ad 5 mm longae. Apothecia marginalia, ad 8 mm longa et 5 mm lata; ascosporae subglobosae, 5 -7 x 4 -5 pm; asci clavati, 30-50 x 10-14 pm. Pycnidia marginalia, papillaria vel spinuliformia; conidia 5 x 1-1.5 pm recta, extremis nonnihil inflatis. Acidum usnicum + /- in cortice superiore; acidum lichesterinicum et protolichesterinicum in medulla. Type: China. Prov. Yunnan, Lijiang County, Mt. Yulongshan, lower central E slope, Ganheba, 3 200-3 300 m, 27°06'N, I00°14'E, on Abies. 23 April 1987, TeuvoAhti, Jian-Bin Chen & Li-Song Wang, 46 649 (H, holotype; TU, isotype). Thallus foliose, upper surface pale glaucous or yellowish- brown, lower surface light to dark brown or almost black in central parts. Lobes rounded at the tips but usually elongate in ge-neral habit, 4 to 10 mm wide, bearing con-spicuous black m arginal pycnid ial projections and sometimes also numerous or occasional pale or brown cilia (Fig. 1). Pseudocyphellae on lower cortex plain, white or light brown, in-frequent on som e sp ecim en s. R h izines brown, sim ple, sometimes very long, to 5 mm, and numerous. Apothecia marginal, with oblong or re­ niform brown disc, to 8 x 5 mm; ascospores subglobose, 5 -7 x 4 -5 pm; asci 3 0 -5 0 x 1 014 pm, narrowly clavate; axial body 2 .5 -4 pm. Pycnidia on emergent projections, usually mar­ ginal and numerous, som e laminal pycnidial projections may be present on both surfaces (on the lower cortex growing out from rim o f pseu­ docyphellae); pycnoconidia bifusiform, 5 x 1 1.5 pm (Fig. 12). Chemistry: usnic acid present or absent in the cortex; lichesterinic- and protolichesterinic- Figs. 6—13. Anatomy of the genus Tuckneraria. — 6: Cross section of an apothecium of T. pseudocomplicata, showing the two layered exciple, Japan, Honshu, Tagawa 319 (US).—7: Upper cortex of T. laureri; Austria, Tirolia, Zahlbruckner 463 (LD). — 8: T. laxa, cross-section of thallus with strongly gelatinized hyphae; Taiwan, Miaoli Co., Lai 7770 (TAIM). — 9: Upper cortex of T. pseudocomplicata with strongly gelatinized cortical hyphae; Japan, Honshu, Tagawa 319 (US). — 10: Asci of T. laureri, with a small tholus and a rather broad axial body; Zahlbruckner 463 (LD). — 11: Asci of T. ahtii', China, Xizang, Zong Yu-chen & Liao Yin-shang 307 (LD). — 12: Pycnoconidia of T. ahtii', Nepal, Himalayas, Miehe 11836 (GZU). — 13:. Pycnoconidia of T. laureri, Austria, Stubaier Alpen, Steiner 1958 (LD). Scale = 10 Jim.; exc 1 = upper excipular layer, exc 2 = lower excipular layer, th = tholus, ab = axial body. 148 ACTA BOT. FENNICA 150 (1994) Tiina Randlane et al. Table 1. Comparison of characters of Tuckneraria with those of the related genera Nephromopsis and Tuckermannopsis. Nephromopsis Tuckneraria Tuckermannopsis s. str. Thallus foliose, coriaceous foliose, paper thin foliose, paper thin Lobe shape rounded rounded to elongate elongate Lower surface rugose or reticulated + /- smooth smooth Marginal cilia absent present present Pseudocyphellae on lower surface large, distinct small, indistinct absent Exciple 3-layered 2-layered 2-layered Asci 30-70 x 9—14 |Am 30-50 x 10-14 |im 25-40 x 8-15 jim Ascospores ellipsoid, 5-10 x 2.5-5 jam subglobose, 5-7 x 4-5 um globose, 3.5-5 x 3.5-5 jim Axial body 2-4 (im 2.5-4 |im 3-4 |xm Cortical substances usnic acid usnic acid atranorin Medullary substances a) fatty acids present present present b) secalonic acids present absent absent c) orcinol depsides & depsidones olivetoric or physodic acid alectoronic, collatolic, physodic acid alectoronic, collatolic, olivetoric or physodic acid type fatty acids always present in medulla, while caperatic acid is an accessory substance. Distribution: China, Nepal, Taiwan. Specimens of this lichen species have usual­ ly been erroneously identified as Nephromopsis delavayi Hue, even though several characters do not correspond with the original description of N. delavayi (Hue 1899-1900). The most im­ portant character is the shape and size of the ascospores: N. delavayi has ellipsoid ascospores (7-11 x 4-5 Jim) and therefore probably belongs to the genus Nephromopsis, while the ascospores of the species described here are subglobose (57 x 4-5 |im). Other characters such as the size and reticulation of the thallus, absence of cilia and the apothecial measurements highlight the essential differences between these two entities. According to Lai (1980), the type material of N. d e la v a yi contains secalonic acid and is morphologically identical with Nephromopsis ornata (Mull.Arg.) Hue. We are in agreement about the synonymy of N. delavayi with N. ornata proposed by Lai but propound here a new species, Tuckneraria ahtii, to include the specimens that in many herbaria have been wrongly determined as N. delavayi. Teuvo Ahti collected wonderful material from China, Yun­ nan, and it was in the Helsinki herbarium in 1992 that we first began to speculate about the new species. Specimens examined — China. Prov. Yunnan, Mt. Yulongshan nearLijiang, 3 450-3 500 m, Handel-Mazzetti 3563 (US, FH), Lijiang County, Mt. Ndaza Ko, 4 000 m, Rock (Zahlbruckner-Redinger: Lich. Rar. Exs. 31 S); Lijiang County, Yangtze watershed, eastern slopes of Lijiang Snow Range, Rock I I 773 (UPS); Prov. Xizang, 3 300 m, Zong Yu-chen & Liao Yin-shang 307. Nepal. Langtang area, Pamdang Karpo, 4 620 m, Miehe 13 056f (GZU), Langschisa Glacier, 4 090 m, 4 400 m, 4 480 m, 4 530 m Miehe 11 725b, 13 846, 12 424, 11 835 (GZU), Dupku, Helambu, 4 090 m, Miehe 7396e (GZU), Pangtang, 4 300 m, Miehe 2284 (GZU). Taiwan. Prov. Taichung, Mt. Armashan, Lai 6860 (TAIM). ACTA BOT. FENNICA 150 (1994) Tuckneraria a new segregate in the tarm eliaceae T u ck n eraria lau reri (Kremp.) Randlane & Thell, comb, nova Cetraria laureri Kremp., Flora 34: 673. I SSI. Nephromopsis laureri (Kremp.) Kurok.. J. Jap. Hoi. 66: 156. 1991. Cetraria complicata Laurcr in Fr., Lichenogr. Eur. Ref.: 459. 1831 (nomen nudum). Cetraria straminea Kremp. ex Schwend. in Niigeli, Beitr. Wiss. Bot. 2; 154. 1860; syn. nov. Thallus light yellow on upper surface, white to pale brown on lower surface. Lobes rounded, up to 5 mm broad, ascending in the margins, bearing marginal soredia (som etim es almost isidia- like structures) and scattered cilia (Fig. 2). Pseudocyphellae on lower cortex white, plain, rounded or irregular, often surrounded by a light brown line, at times absent from some specimens. Rhizines scattered. Apothecia very rare, marginal, with brown disc and sorediose thalline margin; ascospores subglobose, 5 -6 x 4 -4 .5 (im; asci clavate, 3 5 45 x 10-12 |im (Fig. 10); axial body 2.5 jim. Pycnidia on marginal emergent projections; pycnoconidia bifusiform, 5 x 1-1.5 tun (Fig. 13). Chemistry: usnic acid in the cortex; lichesterinic- and protolichesterinic-type fatty acids in the medulla. Medulla Pd-, K -, C -, K C-. Distribution: montane forests o f Central Europe (the Alps, the Carpathians); Asia (Rus­ sia, China, M ongolia, Japan, Nepal); SouthAmerica (Venezuela, Colombia). T. laureri is the only sorediate taxon within the C e tra rio p sis-N e p h ro m o p sis-T u c k n e ra ria complex, thus representing a ‘secondary’ spe­ cies. In accordance with the ‘sp ecies-pairs’ theory (Poelt 1970), it is the most widely distri­ buted o f all other — "primary' — species dis­ cussed here, growing widely in Eurasia and found also in the northern part of South America. Because o f its superficial morphological sim i­ larity (yellowish thallus, marginal soredia) to the North American and European lichen Tucker­ m a n n o p sis o a k e sia n a (T uck.) H ale, it has som etim es been confused with it. However, T uckneraria la u reri and T itckn ern n a n n o p sii oakesiana cannot be phylogenetically closely related be-cause o f essential differences in their rep ro -d u ctiv e structures and seco n d a ry chemistry. The present generic position o f the tormer species is not satisfactory either, but this problem will be discussed in a future paper. 36 149 More than 170 specim ens were examined from different parts o f the whole distribution area. Selected specimens examined — Austria. Stcierniark, See-Eben, I 400 m, Poelt (GZU); Tirol, Allgiiu, 1 720 m, Schauer (M). Germany. Salzburg, Radstadt, 1 320 m, Schauer (M); Oberbayern, Garmisch, 1 280 m, Schauer (M). Italy. Tirol, Bolzano, Hausmann (Erbar. Crittog. Ital., 464; M, S). Yugoslavia. Alpes Julia, Pokljuka, Rudno Polje, 1 800 m, Vezda, Lich. Sel. Exs. 847 (LD, S). Romania. Hunedoara, Retezat Mts., 45°23'N 22° 49'E, 1 450-1 550 m, Moherg 10 765 (UPS). Ukraine. Zakarputsku region, district Rahivska, Velikii Bichkiv, 800 m, Makarevich 8261 (KW). China. Sikang, Kangling, Chungo Valley, Yara, 4 050 m, Smith 14 011 (UPS). Japan. Honshu, Prov. Shinano, Mt. Takesbi-mine, 1 700 m, Kashiwadani 15 043 (TNS). Mongolia. Ara-Khangai region, Zenkhcr district, Suvraga ridge, 2 200 m, Biazrov 714 (LD). Nepal. Langtang area, Dubku Helambu, 4 090 m, Mielte 7396e (GZU). Russia. Irkutsk region, HamarDabau Mt. Range, Bolshaya Osinovka, Trass 1031 (TU); Habarovsk region, Badzhal Mt. Range, Urnii, Randlane 208 (TU). Colombia. Risaralda, volcano Santa Rosa, 4*49'N, 75°28'W, 4 130 m, Wolf 1172 (B). Venezuela. Merida, Apartaderos, 8°45'N, 70°45'W, 3 500 m, Kalb (LD). T uckneraria laxa (Zahlbr.) Randlane & Thell, comb, nova Nephromopsis ciliaris (Ach.) Hae var. laxa Zahlbr., Fedde, Repert. 33: 61. 1933. — Nephromopsis laxa (Zahlbr.) M.Sato, J. Jap. Bot. 14: 783. 1938. — Cetraria laxa (Zahlbr.) Sato, Parmeliales (I), in Nakai et Honda, Nova Flora Japonica 5: 51. 1939. Cetraria daihuensis Rasanen, J. Jap. Bot. 16: 85 1940. — Nephromopsis daihuensis (Rasanen) Rasanen, Kuopion Luonnon Yst. Yhd. Julk. B 2(6): 47. 1952. Thallus pale yellow on both surfaces, lobes elongate and narrow (up to 2 mm broad) with abundant marginal cilia, yellow ish-brow n or darker brown to black at the tips. P seudo­ cyphellae on lower cortex in the form o f tiny, white, plain spots. Rhizines scattered, simple or occasionally branched. Apothecia very rare, marginal, with a brown rounded disc; ascospores subglobose, 5 x 4 |im; asci clavate, 4 0 -4 5 x 12-13 pm; axial body 3 Jim. Pycnidia marginal, on + /- emergent pro­ jections. Pycnoconidia not seen. Chemistry: usnic acid in the cortex; lichesterinic- and protolichesterinic-type fatty acids in the medulla. Medulla Pd-, K -, C -, KC-. Distribution: endemic to Taiwan. 150 ACTA BOT. FENNICA 150 (1994) Tiina Randlane et al. This species stands somewhat alone in the genus due to its highly characteristic morpho­ logy (uniformly pale colour o f the thallus on both surfaces, narrow and elon gate lob es, abundant marginal cilia, extremely small pseu­ docyphellae) (Figs. 4, 5). Evidently its distri­ bution is restricted to the island o f Taiwan. However, the presence o f pseudocyphellae on the lower cortex and marginal position o f apot­ hecia in T. laxa, as well as the anatomical struc­ tures o f the asci and thallus (Fig. 6), clearly place it in Tuckneraria. Specimens examined — Taiwan. Nimandaira, Mt. Arisan, Asahina, (H). Miaoli County, Mt. Dapachienshan, Lai 7770 (TAIM). Hua-lien County, Shyu-lin village, 2 700 m, Koponen 18 024 (H). T u c k n e r a r ia p s e u d o c o m p lic a t a Randlane & Saag, comb, nova (A sa h .) Cetraria pseudocomplicata Asahina, J. Jap. Bot. 12: 804. 1936. — Nephromopsis pseudocomplicata (Asahina)M.J. Lai, Quart. J. Taiwan Mus. 33: 224. 1980. Cetraria rhytidocarpa Mont. & Bosch 1'. nipponensis Asah.. .1. Jap. Bol. 24: 228. 1954: syn. nov. Nephromopsis nipponensis (Asahina) M.J.Lai, Quail. J. Taiwan Mus. 33: 223. 1980. Thallus greenish on upper and white or light brown on lower surface. Lobes rounded, to 7 mm broad, bearing scattered marginal cilia (Fig.3). Pseudocyphellae on lower cortex not numerous; in the form o f rounded or irregular white, small, plain patches, often with light brown margins. Rhizines pale, simple, long (to 3 mm). Apothecia marginal, rounded or re­ niform, to 6 mm in diameter, with reddish-brown disc; exciple 2-layered (Fig. 6); ascospores subglobose, 5 -6 x 4 -5 |im; asci clavate, 3 0 35 x 12-14 [im; axial body 3.5 (im. Pycnidia numerous, situated on marginal emergent pro­ jections; pycnoconidia bifusiform, 5 x 1-1.5 Jim. Chemistry: usnic acid present or absent in the cortex; as for the medullary compounds, two different chemotypes can be distinguished. The first c h e m o ty p e, form erly N e p h r o m o p s is pseudocom plicata, contains alectoronic acids (a and 13 forms) as the major, and a-collatolic acid as a m inor su b sta n ce in the m edulla; lichesterinic- and protolichesterinic-type fatty acids may occur rarely. The second chemotype, formerly N ephrom opsis nipponensis, contains equally constantly orcinol depsidones physodic and conphysodic acids as well as lichesterinicand protolichesterinic-type fatty acids. Both chem o-types respond similarly to medullary colour tests: P d -, K -, C -, KC+ red and no morpho-logical or anatomical differences. Distribution: Eastern Asia (Sakhalin island, Japan, Taiwan). T. pseudocom plicata is chosen to be the type species o f the new genus because o f its supposed central position in this group o f taxa. It evidently has some affinities to all the species in Tuckneraria. Its chemical diversity can be interpreted in terms o f evolutionary potential. About 60 specimens were examined from Japan and Taiwan. Selected specimens examined — The first chcinolype with alectoronic acids. Japan. Prov. Suruga, Ml. Fuji, Culberson & Culberson 10805, 10 807 (US), Lake Saiko, Culberson & Culberson 10 803 (M); Prov. Yamanashi, Adzumazawa, Mitomimura, 2 300 m, Omura 395 (US); Prov. Shinano, Mt. Tadesina, Kurokawa 51 747 (M), Mt. Yalsugalake, Kurokawa 58 303 (TAIM); Honshu, Prov. Nam. Ml. Odaigakara. Tarawa 3 l (), Komagalakc. Finnic 0759. Taiwan. Tailung County, Yakou, 2 750 m, Lai 9484 (US) — besides alecloronic acids also lichesterinic- and protolichesterinic-type fatty acids. The second chemotype wilh physodic, conphysodic, lichesterinic and protolichesterinic acids. Japan. Prov. Kai, Mt. Yatsu-ga-lake, Asahina (lectotype of Cetraria rhytidocarpa f. nipponensis; DUKE); Prov. Musashi, Mt. Kumatori, 1 900 m, Shibuichi, 4533 (Kurokawa: Lich. Rar. Crit. Exs. 153; H, LD, M, TAIM, TU); Prov. Musashi, Titibu, Mt. Ryogami, Kurokawa 55 0573-b (M); Prov. Hida, Mt. Ontake, Asahina, Lich. Jap. Exs. 56, (H); Honshu, Koponen (H). ACKNOWLEDGEMENTS We dedicate this publication to Professor Teuvo Ahti who, in his modest way, has always played the role of urgently needed link betw een the E uropean and Asian lichenologists and Western and Eastern science. The authors are grateful lo the keepers of herbaria B, DUKE, FH, GZU, H, KW, M, MB, S, TAIM, TNS, UPS, US for kindly sending the lichen specimens. Professor Josef Poelt is thanked for encouraging us in this project and for giving his Nepal materials totally into our hands. We are also indebted lo Professor Rolf Saniesson for useful remarks on nomenclature. The project has received financial support from the Swedish Institute and the Finnish-Chinese Botanical Foundation. ACTA BOT. FENNICA 150 (1994) Tuckneraria a new segregate in the Parmeliaceae 151 REFERENCES Awasthi, D.D. 1982: Lichen genus Cetraria in India and Nepal. — Bull. Bot. Surv. India 24:1-27. Culberson, C.F. 1972: Improved conditions and new data for the identification of lichen products by a stan­ dardized thin-layer chromatographic method. — J. Chromatogr. 72:123-125. Culberson. C.F. 1974: Conditions for the use of Merck silica gel 60 F 254 plates in the standardized thinlayer chromatography technique for lichen pro­ ducts. — J. Chromatogr. 97:107-108. Culberson, C.F.. Culberson. W.L. & Johnson, A. 1981: A standardized TLC analysis of 13-orcinol de­ psidones. — Bryologist 84:16-29. Golubkova, N.S. 1981: Conspectus of lichen flora of Mon­ golian People’s Republic. — 200 pp. Nauka. Leningrad (in Russian). Hue. A.M. 1899-1900: Lichenes extra-Eumpaei a pluribus collectoribus ad Museum Parisiense missi. — Nouv. Arch. Mus. Hist. Nat. 4(1): 27-220. Kurokawa. S. 1991: Japanese species and genera of the Parmeliaceae. — J. Jap. Bot. 66:152-159. Karnefelt, I., Mattsson, J.-E. & Thell, A. 1992: Evolution and phylogeny of cetrarioid lichens. — PI. Syst. Evol. 183:113-160. Lai, M.-J. 1980: Studies on the cetrarioid lichens in Parme­ liaceae of East Asia (1). — Quail. J. Taiwan Mus. 33:215-229. Park, Y.S. 1990: The macrolichen flora of South Korea. — Bryologist 93:105-160. Poelt. J. 1970: Das Konzept der Arlenpaare bci der Flechten. — Deutsche Bot. Ges. Neue Folgc 4:187-198. Randlane, T. & Saag, A. 1991: Some chemosysteinatical data about the lichen genus Nephromopsis in the U.S.S.R. — Folia Crypt. Eston. 28:26-30. Randlane, T. & Saag, A. 1992: Additional data about the genus Nephromopsis (Lichenes, Parmeliaceae). — Mycotaxon 44:485-489. Rassadina, K. A. 1971: Fam. Parmeliaceae. — In: Handbook of the lichens of the U.S.S.R.: 282-386. Nauka, Leningrad (in Russian). Received 29 September 1993 VII Thell, A., Randlane, T., Kamefelt, T., Gao, X.-Q. & Saag, A. 1995. The lichen genus Allocetraria (Ascomycotina, Parmeliaceae). — In: Daniels, F. J., Schulz, M. & Peine, J. (eds.). Flechten Follmann. Contributions to lichenology in honour o f Gerhard Follmann. University o f Cologne, Germany, 353 - 370. FLECHTEN FOLLMANN. CONTRIBUTIONS TO LICHENOLOGY IN HONOUR OF GERHARD FOLLMANN. - EDITED BY FRED J. A. DANIELS, MARGOT SCHULZ, AND JORG PEINE. PUBLISHED BY THE GEOBOTANICAL AND PHYTOTAXONOMICAL STUDY GROUP, BOT­ ANICAL INSTITUTE, UNIVERSITY OF COLOGNE, COLOGNE, GERMANY: PP. 353 - 370; 1995. THE LICHEN GENUS ALLOCETRARIA (ASCOMYCOTINA, PARMELIACEAE) A rne T hell Department of Systematic Botany, University of Lund, Lund, Sweden T iin a R a n d l a n e Institute of Botany and Ecology, Tartu University, Tartu, Estonia In g v a r K a r n e f e l t * Department of Systematic Botany, University of Lund, Lund, Sweden X ia n g -Q u n G ao Botanical Museum, University of Uppsala, Uppsala, Sweden and A ndres S aag Institute of Botany and Ecology, Tartu University, Tartu, Estonia Key Words Cetrarioid lichens, Allocetraria, Parmeliaceae, axial body, filiform pycnoconidia Abstract The cetrarioid lichen genus Allocetraria (Parmeliaceae) includes eight species distin­ guished by a palisade plectenchymatous cortex, asci with a very broad axial body, globose or subglobose ascospores, and filiform pycnoconidia. Two new species, A. flavonigrescens T h e l l et R a n d l. and A. sinensis G a o , are described, and the new combinations A. denticulata (H u e) T h e l l et R a n d l., A. globulans ( N y l .) T h e l l et R a n d l., and A. oakesiana (T u c k .) R a n d l. et T h e l l are proposed. A. potaninii (OXN.) R a n d l. et S a a g is synonymized with A. stracheyi (B a b .) KUROK. et L a i. The anatomy of the genus is carefully described for the first time. Dedicated lo Prof. Dr. G h r u a r d F o l l m a n n on occasion o f his 65lh birthday and retirement from the University o f Cologne, Germany, with respect to his appreciated piona ;mg lichenological works. 354 A. Thell, T. Randlane, I. K arnefelt et al. Introduction The genus Allocetraria, originally including three species from high altitudes in South-east Asia, was introduced in the Parmeliaceae by KUROKAWA and L ai (1991). It was separated from Cetraria on the dichotomously or subdichotomously branched lobes, the special appearance of the pseudodyphellae, the palisade plectenchymatous cortex, and on the unique chemistry (fig. 7). Three species, A. ambigua (B a b .) K u r o k . et L a i , A. isidiigera KUROK. et L a i , and A. stracheyi (B a b .) K u r o k . et L ai were combined here. The authors did not pay attention to ascomatal and pycnidial characters consequently. However, our studies of these structures confirm the necessity of a separate genus for this species group also on the grounds of filiform pycnoconidia, Lecanora type of asci with an unusually broad axial body (T h e l l et al. 1994), and globose or subglobose ascospores (tab. 1). These are basic characters for the recognition of the genus Allocetraria. Three more species have earlier also been combined in Allocetraria by lane and Saag (1992). Two of them, however, A. cucullata Rand­ (B e l l a r d i ) R a n d l . et S a a g and A. nivalis (L.) R a n d l . et S a a g , have asci of the Cetraria type and bifusiform pycnoconidia and were recently separated into a new genus, Flavo­ cetraria K a r n . et T h e l l (K a r n e f e l t et al. 1994). After recent studies of the type material of A. potaninii (O x n .) R a n d l . et Sa a g it became clear that this species must be synonymized with A. stracheyi (B a b .) KUROK. et L a i . Three fur­ ther species, however, Nephromopsis globulans (N y l .) L a i , Tuckermannopsis oakesiana (TUCK.) H a l e , and Cetraria denticulata H u e , are transferred to Allo­ cetraria in this paper. The two first entities were examined carefully and both show the characteristic features of the genus. No material was seen of C. denticu­ lata but the Latin diagnosis clearly unveils its affinities to Allocetraria. Unfor­ tunately, we did not succeed to get any material of the newly described A. isi­ diigera, but the species is obviously a good member of this well delimited genus. Material and Methods About 120 herbarium specimens from FH, GZU, H, KW, LD, LE, PC, S, TU, and UPS were examined. Methods used for anatomical studies have been described in detail by K a r n e f e l t et al. (1992). The secondary chemistry was investigated with standardized methods (CULBERSON 1972). 355 Allocetraria (Parmeliaceae) Taxonomy Allocetraria K urok . et Lai B u ll. nat. Sci. Mus. Tokyo, Ser. B, 17: 60; 1991. Type species: Allocetraria stracheyi (B a b .) K u r o k . et L a i . Thallus foliose or fruticose, suberect to erect; upper surface yellow to yellowish green or brown; lower surface of the same colour as upper or light tan or brown or black; lobes prostrate, unbranched or dichotomously branched, in most of the spe­ cies quite narrow, elongated, and characteristically convex; soredia or isidia pre­ sent on one species each; rhizines sparse; pseudocyphellae on the lower surface either piunctiform and irregular or sublinear along the margins, absent in some species; cortical layers usually more or less palisade plectenchymatous; medulla white or coloured from light yellow to bright orange; medullary hyphae 3 - 6 |im in diameter; algae near the upper cortex. Apothecia lateral and marginal or submarginal to almost laminal, with brown disc and yellowish thalline margin, up to 7 - 8 mm in diameter; exciple 2-layered, 30 120 jim thick; asci usually rather narrowly clavate to cylindrical, 30 - 70 x 8 - 18 Um, tholus small, ocular chamber cylindrical and broad, axial body broad to very broad, 5 - 9 jim; ascospores globose to subglobose, rarely broadly ellipsoid, more or less uniseriately arranged, 5 - 10 x 5 - 8 |im; paraphyses usually straight, sparsely branched with swollen tips; pycnidia marginal to submarginal, rarely laminal, immersed or on emergent projections, dark pigmented; pycnoconidia fili­ form, usually slightly sublageniform, 1 0 -1 9 x 0 .5 - 2 |jm. Chemical substances: usnic acid in the cortex; different fatty acids (caperatic, li­ chesterinic, protolichesterinic) in the medulla together with secalonic acids and other related pigments. One species contains fumarprotocetraric acid. The genus Allocetraria , including eight species mainly distributed in South-east Asia, is characterized on both structural characters, characters in the reproductive structures, and characters in the secondary chemistry. All species appear to be only rarely fertile and mature asci have been found only in Allocetraria ambigua, A. globulans, and A. oakesiana. Therefore much attention must be payed to the structure of the lobes when recognizing different species. Of the structural characters is the palisade plectenchymatic arrangement of the cortical hyphal cells the most important. The asci are above all characterized by 38 356 A. T h e ll, T. R a n d la n e , I. K a r n e f e lt et al. an unusually large axial body in addition to globose or subglobose ascospores (tab. 1). The pycnidia furthermore produce a unique form of rather long, slightly sublageniform, filiform pycnoconidia. All species are also characterized by con­ tent of usnic acid in the cortex and several medually substances, lichesterinic-, protolichesterinic, and secalonic acids. Tab. 1: Allocetraria compared with some other cetrarioid genera Characters Allocetraria Flavocetraria Tuckerman­ nopsis s. str. * yellow brown 1-layered, pa­ raplectenchy­ matous 1-layered, pa­ raplectenchy­ matous 1-layered, pa­ raplectenchy­ matous present absent present present absent present 0.8 -1.6 jim oblong citriform 5 -7.5 x 1 - 1.5 um 0.3 - 1.5 um bi fusiform 3 -4 um bifusiform present 2.5 -4 um c. 6 x 1 jim c. 5 x 1 |om 4 -5 x 1 - 1.5 um usnic acid atranorin usnic acid alectoronic, collatolic, oli­ vetoric, or physodic acid alectoronic, collatolic, and physodic acid Cetraria upper surface yellow upper and lower 1-layered, pa­ usually lisade plecten- 2-layered chymatous pseudocy­ phellae cilia usually present absent axial body 5 -9 um conidi al shape filiform and size 10-19 x 0.5 -2 torn brown present cortical sub­ stances usnic acid medullary substances except fatty acids fumarprotocefumarprotoce- endocrocin traric and se­ and parietin traric acid calonic acid — Tuckneraria yellow to greenish bifusiform * The genus Tuckermannopsis is still not clearly delimited; T. americana, T. chlorophylla, T. ciliaris, and T. platyphylla are studied here The closest relatives are presumably to be looked for among the aggregate of ce­ trarioid genera which is presently recognized. Some characters, however, seem to be isolated such as the very broad axial body and the long filiform to slightly sub­ lageniform pycnoconidia (tab. 1). Furthermore there is no ring structure in the tholus, which is otherwise characteristic of the genera Arctocetraria, Cetraria, Flavocetraria, and partly Nephromopsis. The fatty acids lichesterinic and protoli­ chesterinic acids are in common with Cetraria, Flavocetraria, and Nephromopsis. In Nephromopsis in addition both usnic and secalonic acids occnr 357 Allocetraria (Parmeliaceae) A majority of the species included in the genus occur at rather high altitudes, preferably above 3000 m, in mountainous areas in South-east Asia. They have been collected in India, Nepal, China, Mongolia, and Taiwan, but most frequently in the Himalayas. One species, however, Allocetraria oakesiana is distributed in montane forests in North America and Europe. The species can be both corticolous and terricolous. Key to the Species la. lb. 2a. 2b. 3a. 3b. 4a. 4b. 5a. 5b. 6a. 6b. 7a. 7b. 8a. 8b. 1. Thallus sorediate or isidiate ......................................................................... 2 Thallus not sorediate or isidiatte .................................................................. 3 Marginal soredia always present..................................................... A. oakesiana Sparse isidia present.......................................................................A. isidiigera Lobes occasionally more or less radial symmetric................................ A. stracheyi Lofcies distinctly dorsiventral ........................................................................ 4 Lobes more or less convex on the upper side ................................................ 5 Lobes plane or concave on the upper side..................................................... 7 Lobes apically indented................................................................. A. denticulata Lobes not apically indented ......................................................................... 7 Lower surface pale yellow to brown, concave; medulla white or coloured, Pd- . ..................................................................................................... A. stracheyi Lower surface dark brown to black, plane to slighdy convex; medulla always white, Pd+ (orange) ............................................................ A. flavonigrescens Epiphytic; upper surface from pale yellow to brown, lobes slightly concave; medulla white or coloured............................................................ A. globulans Always on soil; upper surface pale yellow, lobes plain or concave; medulla always white ............................................................................................ 8 Lower surface brown, with marginal pseudocyphellae in the form of white li­ nes ................................................................................................ A. sinensis Lower surface pale yellow, without marginal white lines; lobes comparatively wide and distinctly concave........................................................... A. ambigua A llocetraria am bigu a (B ab .) K u r o k . et L a i Bull. nat. Sci. Mus. Tokyo, Ser. B, 17; 60; 1991. Bas. Cetraria ambigua Bab., H o o k e rs J. Bot. 4: 244; 1852. Type: Bompras, Garhwal, Himalaya, on wood and mosses, alt. 16 000 ft., R. S t r a c h e y and E. W in t e r b o t t o m 6 (lectotype, BM); seen. The type material of Allocetraria ambigua also contains a small branch of Allocetraria stracheyi. Thallus suberect to erect foliose, forming 2 - 3 cm high tussocks, upper surface pale yellow, smooth, lower surface pale yellow, smooth or slightly rugose, with sparse concolorous rhizines; lobes comparatively wide (up to 3 mm), lobe mar­ gins and tips concave: pseudocyphellae marginal; upper and lower cortex ca. 358 A. T h e l l , T. R a n d l a n e , I. KA r n e f e l t et al. 30 |itm, composed of 4 - 6 layers of cells, smaller near the surface; medulla always white, 120 - 160 jum. Apothecia rare, marginal, with brown disc, up to 5 mm in diameter; exciple 30 - 35 |im thick; ascospores broadly ellipsoid, 7 - 8 x ca. 5 jim; pycnidia marginal to submarginal, immersed or on emergent projections, dark pig­ mented; pycnoconidia 12 -14 x 0.5 - 2 |im. Chemical substances: usnic acid (+/-) in the cortex; fatty acids (lichesterinic and protolichesterinic) in the medulla; se­ calonic acid A and/or C may also be present. Allocetraria ambigua is easily recognized on the broad with rather narrow, con­ cave apices (fig. 1). Morphologically it is sometimes very similar to Flavocetraria nivalis, and these two species sometimes also grow together. The species is ende­ mic to the Himalaya. Specimens examined: India. Bompras, Garhwal, Himalaya, alt. 16000 ft., STRACHEY and WINTERBOTTOM, no. 6 (BM, lectotype). China. South-eastern Kansu, Min Shan range, S from Bashi-Denga, Tebbu, H um m el, 1930, no. 14 190 (S). Prov. Shaanxi, Mt. Taibai, Baxiantai, alt. 3550 m, GAO, 13.07.1988, no. 3184 (LD). Qinghai, Central Tibet, Northcentral Tangula Shan, SE of Geladandong Glacier, 33° 27' N, 91° 13' E, alt. 5360 m, DlCKORE, 31.08.1989, no. L-07 (GZU). Tibet, KOMARKOVA, no. 32, no. 369 (GZU). Nepal. Khumbakama-Himal, Dhankuta, Barun Valley, alt. 5000 m, WRABER, 10.1972 (GZU). Fig. 1: The typically concave thallus of Allocetraria ambigua, China, Shaanxi, Mt. Taibai, Baxiantai, on ground, 3650 m, GAO 13.07.1988 (LD). Bar = 1 mm Allocetraria (Parmeliaceae) 2. 359 A llocetraria den ticu lata (H ue) THELL et RANDL. comb. nov. Bas.: Cetraria denticulata Huh, Nouv. Arch. Mus. (Paris), I, 4: 85; 1899. Type: China, Yunnan, Yen-tze-hay, on ground, R. P. D e la v a y 08.08.1888 - not avail­ able at PC. Thallus greenish-yellowish, foliose, erect, 2 - 3 cm long, flattened at the bases and caespitose; lobes 0.8 - 1.5 mm broad, apically indented and with marginal projec­ tions along the margins; marginal projections at their tops either whitish or bear­ ing pycnidia or small spines; upper surface smooth, plane or somewhat convex; lower surface similar to the upper or slightly brownish and somewhat concave; upper cortex presumably palisade plectenchymatous, 25 - 60 jim, richly incrusted with crystals; lower cortex similar to the upper but more white, with fewer cells and crystals not as frequent; algal cells 5 - 6 jim broad, in clusters; medulla 25 60 |im thick with 3.5 - 5.5 (im broad hyphae. Apothecia unknown; pycnidia on marginal emergent projections, small, black; conidiophores 2- 3 celled; pycnoco­ nidia filiform of somewhat different appearances, 12-15x1 jim. Chemical com­ pounds: unknown, but obviously usnic acid in the cortex. The value of this taxon is still uncertain simply because it is known only from the type locality. C etraria denticulata was described from China by H u e (1899) and probably forgotten later, mentioned again only by Wei (1991). Unfortunately, we have not been able to see any material of this taxon. However, according to the detailed Latin diagnosis H ues species should definitely be included in the genus A llocetraria. The species is according to the description well separated from other taxa according to the indented lobe tips, the special appearance of the marginal projections and the thick upper cortex. We have asked for type material of Cetrara denticulata at PC, but without any result. 3. A llo cetra ria fla vo n ig re sce n s T h e l l et R a n d l. sp. nov. Type: Nepal, Langtang, Pemdang Karpo, SW exposed rocks, on Juniperus, 29.09.1986, G. M ie h e and S. M ie h e 13 056 (GZU); selected here. Thallus subfruticosus ad foliosus, suberectus ad prostratus, irregulariter ramosus, usque 4 cm longus, lobi concavi 1-3 mm lati, facies supera pallide lutea, infera fusca ad nigra valde rugosa, rhizinae et pseudocyphellae desunt; apothecia non vi­ sa; pycnidia marginalia ad laminalia immersa vel in prominentiis brevibus, pycno­ conidia non visa. 39 360 A. Thell , T. R a n dla n e , I. K arnefelt et al. Thallus subfruticose to foliose, suberect to prostrate, up to 4 cm, irregularly branched; lobes narrow, convex, 1 -3 mm broad, thick; upper surface pale yellow with black spots, lower surface brown to black, strongly wrinkled; rhizines and pseudocyphellae absent; upper cortex ca. 30 jim, composed of 4 - 6 layers of cells, smaller near the surface; medulla white, 170 - 280 pm; lower cortex 15 20 pm, composed of 3 -4 layers of equal sized cells, the outer cell-layer entirely black. Apothecia not seen; pycnidia marginal to laminal, immersed or on short projections, dark pigmented; pycnoconidia not seen. Chemical substances: usnic acid in the cortex, one unknown fatty acid and fumarprotocetraric acid in the me­ dulla; an unknown violet pigment is also noticed. Unfortunately we have not been able to detect either conidia nor ascospores in this species. However, the structural characters in the thallus and especially the cortical hyphal cells support that the new species should be accommodated here rather than in any other cetrarioid genus within the Parmeliaceae. The dark rugose lower surface and the black spotted upper side are the most characteristic and diagnostic features for Allocetraria flavonigrescens (fig. 2). A. flavonigrescens grows on the ground in the alpine tundra above 3000 m, occasionally at the base of shrubs. It is distributed in Nepal. Specimens examined: Nepal. Langtang area, Pemdang Karpo, alt. 4620 m, G. Miehe and S. Miehe, 29.09.1986, no. 13 056 (GZU - holotype). Himalaya, Khombu Glacier, alt. 16000 ft. above tree line, Izzard, 1954 (UPS). 4. Allocetraria globulans (N yl.) T h e ll et R an d l. comb. nov. Bas.: Platysma globulans Nyl., Flora (Regensburg) 70: 134; 1887. Syn.: Cetraria globulans (N yl.) Z a h lb r., Trav. Sous-sect. Troitzkossawsk-Khiakta, Sect. Pays d'Amour Soc. Imp. russe Geogr. 12: 89; 1911. Nephromopsis globulans (N y l.) Lai, Quart. J. Taiwan Mus. 13: 222; 1980. Type: China, Yunnan, R. P. D e lav ay , 1885, no. 1570 (holotype, H-NYL 36 135); seen. Thallus foliose; upper surface yellowish (from light yellow to almost brown), lower surface brown, with sparse concolorous rhizines; lobes plane to concave, prolonged, up to 8 mm wide, dichotomously branched, secondary lobes usually quite narrow, about 1-1.5 mm wide. Some pseudocyphellae-like structures may also be present on marginal warts. Upper cortex undistinctly palisade plectenchy­ matous, strongly crystallized, 15-30 pm, composed of 3 -4 layers of equal-sized cells. Lower cortex similar to the upper but somewhat thinner and less crystal­ Allocetraria (Parmeliaceae) 361 lized, the lower cells brown-pigmented. Medulla light yellow but specimens with white medulla also occur, and sometimes the yellowish layer is seen on the lower parts of die thallus only. Apothecia marginal and submarginal, up to 8 mm in dia­ meter, with a brown disc. Exciple 40 - 120 jum thick, asci narrowly to rather Figs. 2 - 6: Morphology in Allocetraria. (2) A. flavonigrescens, habit, showing the light, black spotted upper surface and the dark, strongly wrinkled lower surface, Nepal, Langtang, Pemdang Karpo, SW-exposed rocks, on Juniperus, 29.09.1986, G. MIEHE and S. MIEHE 13 056 (GZU). (3) Part of the type material of A. globulans, China, Yun­ nan, D e la v ay 1570, holotype (H-NYL 36 135). (4) A. sinensis, China, Shaanxi, Mt. Taibai, Baxiantai, on ground, 3650 m, 13.07,1988, GAO 3159 isotype (LD). (5) Detail structures of the same specimen showing the dark brown underside with pseu­ docyphellae (ps) along the margin. (6) A. stracheyi with typically convex lobes, In­ dia, Kumaon, God River, among mosses and dead leaves, alt. 4700 ft. (?), R. S traCHEY and J. E. WINTERBOTTOM, lectotype (H-NYL 36 055). Bar = 1 mm 362 A. T h e ll, T. R a n d la n e , I. K a r n e f e lt et al. broadly clavate, 45 - 70 x 15 - 20 jim, tholus small, axial body very broad, 5 - 9 (im, ascospores globose, 7 -10 |im in diameter or subglobose, 6.5 - 9 x 5 - 6.5 |im; pycnidia numerous marginally and a few laminally, located on black and somewhat emergent projections which often grow out from special warts of the thallus; pycnoconidia 10 - 19 x 0.5 - 2 [im. Chemical constituents: usnic acid in the cortex; lichesterinic, protolichesterinic, and secalonic acids A and C (+/-) in the medulla. Allocetraria globulans is sometimes difficult to separate from A. stracheyi but the upper surface of the latter is always pale yellow and the lobes more or less convex (fig. 6). Some specimens of A. globulans are rather broad-lobated (fig. 3). The structure of the ascus top and the pycnoconidial shape remove all doubts about the systematic position of the species (figs. 9 - 10). This epiphytic species is distributed in China (Yunnan County), Nepal (Langtang area and Topkegola). It grows on Potentilla fruticosa, Rhododendron sp., etc. in high altitudes, 3200 4700 m. Specimen examined: China. Yunnan, Delavay, 1885, no. 1570 (H-NYL 36 135, holotype; H-NYL 36 136, H-NYL 36 137). Nepal. Langtang area, Chisedang Lekh, above Palpa, alt. 3700 - 3900 m, POELT, 07.09.1986, no. 86-1195 (GZU). Langtang area, above Langshisa Karka, Shalbachun Glacier, alt. 4500 m, POELT, 16.09.1986, no. 86-2342 (GZU). Langtang area, Dupka, alt. 4000 m, G. MIEHE and S. M iehe, 30.07.1986, no. 7307a (GZU). Langtang area, Upper Langtang, Yala, alt. 4790 m, G. MIEHE and S. MIE­ HE, 10.07.1986, no. 5005 (GZU); alt. 4830 m, G. MIEHE and S. MEHE, 02.07.1986, no. 4403 (GZU). Langtang area, Skidscha Kunda, alt. 4670 m, G. MIEHE and S. MEHE, 11.08.1986, no. 8590 (GZU); alt. 4370 m, G. M iehe and S. MEHE, 11.08.1986, no. 8693 (GZU). Langtang area, Pemdang Karpo, alt. 4880 m, G. MEHE and S. MEHE, 05.10.1986, no. 13 482d (GZU). Eastern Nepal, Topkegola, between Rupa and Saju Pokhari, alt. 15000 ft., Awasthi, 30.05.1953, no. 2407 (UPS). 5. A llo cetra ria isidiigera K u r o k . et L a i Bull. nat. Sci. Mus. Tokyo, Ser. B, 17: 62; 1991. Type: China, Xisang (Tibet), Nylalam, on Rhododendron stem, alt. 3910 m, J.-C. Wei and J.-B. CHEN, no. 1857 (holotype HMAS, isotype TNS). This species is distinguished from Allocetraria stracheyi by the presence of sparse isidia ( K u r o k a w a and L a i 1991). We have not been able to get any material of this rare lichen, known only from the type locality. 363 Allocetraria (Parmeliaceae) 6. A llocetraria oakesiana (TUCK.) R a n d l. et THELL comb. nov. Bas.: Cetraria oakesiana TUCK., Boston J. nat. Hist. 3: 445; 1841. Syn.: Platysma oakesianum (T uck.) N y l., Mem. Soc. imp. Sci. nat. Cherbourg 3: 172; 1855. Tuckermannopsis oakesiana (TUCK.) H a le in EGAN, Bryologist 90: 164; 1987. Type: White Mountains, alpine regions, on small branches of dwarf firs, O a k e s, 25.06.1839 (FH, lectotype); seen. Cetraria oakesiana T u ck . var. spinulosa M er­ r i l l , Bryologist 13: 25; 1910. Cetraria bavarica TUCK. KREMPELH., Flora 34: 273; 1851. Type: Germany, Oberbayern, KREMPELHUBER, 06.1851 (isolectotype, UPS); seen. Thallus foliose to subfoliose, upper surface greenish yellow, lower surface brown to light tan, somewhat wrinkled, with sparse rhizines; lobes concave, moderately broad, ca. 1 -4 mm wide, weakly wrinkled, usually with abundant marginal soralia, soredia light yellow; rhizines and pseudocyphellae absent; upper and lower cortex paraplectenchymatous but the hyphae are sometimes anticlinally arranged. Both cortical layers ca. 20 pm, composed of ca. 3 rather gelatinized cell layers each; cells near the surface somewhat smaller than other cells. Medulla in many specimens yellowish in lower parts. Apothecia lateral, marginal to laminal, up to 6 mm in diameter, with brown disc and a thin thalline margin which may turn sorediate; exciple ca. 80 pm thick; asci narrowly clavate to cylindrical 30 - 40 x 7-12 pm, tholus small, ocular chamber cylindrical and broad, axial body broad, 6-7 |im, ascospores globose, ca. 5 x 5 pm; pycnidia marginal, immersed to raised (globose or spinulose; the latter type of variation has been described by M e r r i l l as var. spinulosa), pigmented or non-pigmented, sometimes with corti­ cal tissue beneath, pycnoconidia 11 x 12 x ca. 1 pm. Chemical compounds: usnic acid in the cortex; caperatic, lichesterinic, protolichesterinic, and secalonic acids in the medulla. The two unidentified fatty acids reported by (D ey 1978) are obviously lichesterinic and protolichesterinic acids. This species has since long been treated as a member of the genus Cetraria. How­ ever, detailed investigations of the reproductive structures clearly revealed that this species is better accommodated in the genus Allocetraria. Earlier Allocetraria oakesiana was usually also considered as a closely allied species to Tuckneraria laureri, due to the similar thallus colour and the sorediate margins. Later studies demonstrated that although the spores are subglobose in both taxa, the pycnoconi­ dia are totally different, i. e., bifusiform in T. laureri and, in addition, there are several minor morophological and chemical differences ( R a n d la n e et al. 1994). 40 364 A. Th e l l , T. R a n d l a n e , I. K a r n e f e l t et al. A relationship between Allocetraria oakesiana and Tuckermannopsis chlorophyl- la has also been presumed earlier by KARNEFELT et al. (1992). The investigations of the reproductive structures again clearly demonstrated that these two species are not closely allied. A. oakesiana differs in having a much broadar axial body (fig. 8), filiform conidia and presence of usnic acid, and seems in these important aspects to be a good member of Allocetraria. The cortex in A. oakesiana is not ty­ pically palisade plectenchymatous, like it is in most species within the genus. However, in many taxa the structure of the cortex seems to be variable and repre­ sents an uncertain character ( K A r n e f e l t et al. 1992,1993, R a n d l a n e and S a a g 1992, MATTSSON and LAI 1993, M a t t s s o n 1993). Allocetraria oakesiana differs from the other species included in the genus in being distributed also outside South-east Asia. It is present in China (W e i 1991), but also in North America (Canada, USA) and Europe (Austria, Germany, Italy, Slovakia, Russia, Ukraine). It grows on coniferous and deciduous trees in montane forests. Figs. 7 -10: Anatomy in Allocetraria. (7) Upper cortex of A. sinensis. Note the typically anticlinally arranged hyphae, most typical in the lower cortex (arrows indicating), Nepal, 02.07.1986, MIEHE 4403 (GZU). (8) Asci of A oakesiana, showing very broad axial bodies (ab), USA, T uck erm an n. (9) Pycnidium and pycnoconidia of A. globulans, Nepal, 02.07.1986, MIEHE 4403 (GZU). (10) Pycnoconidia of A. globulans, the same specimen Allocetraria (Parmeliaceae) 365 Selected specimens (74 specimens from Europae and North America examined): Austria. Steiermark, St. Radegrund, Graz, HOLZINGER, 1874 (LD). Untersteiermark, V r a n g , 1986 (S). Salzburg, alt. 1040 - 1080 m, H ertel, 17.07.1978, Lich. Alp. no. 317 (LD, S). Germany. Ostbayem, K rempelhuber , 06.1851 (UPS, isolectotype). Bavaria, Alpen, A r ­ n o ld , 1893 (LD). Wendelstein, Nuremberg, JAMES, 07.06.1956 (LD). Ammergauer Al­ pen, Schwaben, Fleckenau, FORSTER and SCHROPPEL, 1960, Lich. Alp. no. 195 (LD, S). Italia. Sudtirol, Predazzo, A r n o ld , 14.08.1879 (LD, S). Prov. d'Udine, Passo del Pura, alt. 1020 m, C lerc , 05.07.1982, no. 8608 - 8610 (UPS). Slovakia. Slovakia boreoorient., Nizke Poloniny, Montis Hrubky, alt. 1180 m, PlSUT, 06.09.1964, Lich. Slov. Exs. no. 68 (LD). Ukraine. Carpati Poloniny, Montis Pop Ivan, alt. 1300 - 1400 m, Su z a , 1928, Lich. Bohemoslovak. no. 87 (LD, S, TU). Canada. Algoma Distr., Lake Superior Provinical Park, 47° 40' N, 84° 50' W , Br o d o , 07.09.1965, no. 7063 (LD). Ontario, Al­ goma Distr., MacGregor Lake, 47° 17' N, 84° 35' W , alt. 1000 ft., BRODO, 09.09.1969, Lich. Canad. Exs. no. 97 (LD, TU). USA. White Mountains, alpine regions, O akes , 25.06.1839 (FH, lectotype). New York, St. Lawrence Co., Star Lake, KARNEFELT, 05.09.1981, no. 81-65-20 (LD). West Virginia, Randolph County, Cheat Bridge, HALE, 05.1956, Lich: Am. Exs. no. 64 (LD, S). 7. Allocetraria sinensis Gao sp. nov. Type: China. Shaanxi, Mount Taibai, alt. 3400 m, on ground, G ao 3052 (holo­ type HM AS, isotypes UPS, LD). Thallus erectus, foliosus, leviter adnatus, lobi concavi 1-3 mm lati, facies supera laevis nifidula viridi-lutea ad lutea, margine fusca ad nigra, facies infera fusca sat laevis et nitida, pseudocyphellae lineam secus marginem formantes; apothecia non visa; pycnidia marginalia, leviter prominentia, pycnoconidia filiformia, 12-16 pm longa. Thallus erect, foliose, 2 - 3 cm high, loosely attached to the substrate, forming tussocks, more or less dichotomously branched; lobes concave, 1-3 mm broad; upper surface greenish yellow to yellow, brown to black on the margin, smooth, slightly shiny; lower surface brown, rather smooth and shiny; pseudocyphellae marginal, forming a white, continuous line; projections present, short; epicortex 3-4 pm, non-cellular; upper cortex 28 -30 pm, composed of 1 -2 layers of more dark-stained and small cells in outer part, 2 - 3 layers of less stained and large cells in inner part; lower cortex 17-23 pm, 2-3 layers of cells in inner part; not double-structured, lowermost part pigmented; medulla 85 - 100 pm. Apothecia not seen; pycnidia marginal, somewhat raised; pycnoconidia 12-16 pm long. 366 A. Th e ll , T. R a n d l a n e , I. Ka r n e f e l t et al. Chemical compounds: usnic acid in the cortex; lichesterinic, protolichesterinic, and an unidentified fatty acid in the medulla. Allocetraria sinensis differs basically from A. ambigua and A. stracheyi by a dark brown underside with pseudecyphellae forming a white line along the margin (fig. 4 - 5). This species grows on the ground among mosses in the alpine tundra above 3000 m. It is found only in South-west China. Specimens examined: China. Prov. Shaanxi, Mount Taibai, alt. 3400 m, Gao, no. 3052 (UPS, LD, isotypes); alt. 3550 m, Gao, 13.07.1988, no. 3184 (LD); alt. 3650 m, Gao, 13.07.1988, no. 3159 (LD). Nepal. Langtang area, above Yala, alt. 4830 m, G. MEEHE and S. Miehe, 05.07.1986 (GZU). 8. Allocetraria stracheyi (B ab .) K u r o k . et L a i Bull. nat. Sci. Mus. Tokyo, Ser. B., 17: 62 - 63; 1991. Bas.: Evernia stracheyi B ab., H o o k e rs, J. Bot. 4: 244; 1852. Type: India, Kumaon, Gori River, among mosses and dead leaves, alt. 4700 ft. (?), R. S t r a c h e y and J. E. WlNTERBOTTOM (lectotype H-NYL 36 055, isolectotype BM); seen. Platysma everniellum NYL., Mem. Soc. Sci. nat. Cherbourg 5: 100; 1857 (based on Evernia stracheyi B ab.). Cetraria everniella (N y l.) K re m p e lh ., Verhandl. zool.-bot. Ges. Wien 18: 315; 1868. Cetraria potaninii OXN., J. Cycle Bot. Acad. Sci. Ukraine 7-8: 168; 1933. Type: China, Tibet, Montes Kamenses, inter stats. Tasso et Penczamu, Tassaschanj, POTANIN, 31.05.1893 (KW, holotype); seen. Allocetraria potaninii (O x n .) R a n d l. et S a a g , Mycotaxon 44: 492; 1992. Thallus prostrate to suberect foliose, up to 5 cm, upper surface pale yellow, smooth, lower surface pale yellow to brown, wrinkled, with sparse marginal rhi­ zines; lobes convex, comparatively thick; upper and lower cortex ca. 30 pm, com­ posed of 3 - 5 layers of cells, smaller cells near the surface; medulla white to orange, 200 - 230 pm thick. Apothecia rare, marginal, with brown disc, up to 5 mm in diameter, mature asci not found; pycnidia marginal, immersed or on emergent projections, dark pigmented; pycnoconidia 12 - 14 x 0.5 -2 pm. Chemi­ cal substances: usnic acid in the cortex; fatty acids (lichesterinic and protoli­ chesterinic) and secalonic acids (A and C) together with other related pigments (e. g., endocrocin) in the medulla. This species has mainly been known under the name of Cetraria everniella (N y l.) K re m p e lh . However, K u r o k a w a and L a i (1991) cleared out the nomen- 367 Allocetraria (Parmeliaceae) clatural and consequently taxonomical misinterpretation of this name. The taxon was described originally as Evernia stracheyi by B a b in g t o n (1852) on the basis of material collected in the Himalayas. When it was transferred to Platysma, NyLANDER (1857) gave it a new name Platysma everniellum, because of an older ho­ monym, Platysma stracheyi (B ab .) N yl. (= Nephromopsis stracheyi [Bab.] M uell.- A R G .; see further discussion in KUROKAWA and L a i 1991). Allocetraria stracheyi is similar to A. ambigua but differs in having more narrow, plane or convex lobes with a wrinkled lower surface (fig. 6). A further specific name described by O x n e r (1933), Cetraria potaninii, was based on material from Tibet. He indicated that the type was preserved in Leningrad (LE). It was there­ fore feared that the type material was destroyed during World War II, but fortu­ nately it was still present in Kiev (KW) and kindly sent to us. After having studied the material, we could only conclude this species being synonymous with A. stracheyi (B ab .) KUROK. et Lai. A. stracheyi has a rather wide distribution. It is terricolous in the alpine tundra (above 3000 m) in India, Nepal, China, Taiwan, and Mongolia. Selected specimens (49 specimens examined): India. Himalaya, Gori River, Kumaon, alt. 4700 ft. (?), STRACHEY and WlNTERBOTTOM (H-NYL 36 055, lectotype). Himalaya, Kumaon, Almora Distr., Phurkia-Pinderi Glacier, alt. 11500 ft., D. D. AWASTHI and A. M . A w a s th i, 23.05.1950, no. 781 (H, UPS). Nepal. Langtang area, from Kyangjin to Nubama Dhang, alt. 3750 - 3900 m, POELT, 11.09.1986, no. 86-1329 (GZU). Eastern Nepal. Topkegola, Thagalabhanjyang, alt. 14000 ft., D. D. A w a s th i, 29.05.1953, no. 2350 (UPS). Langtang area, Niang Tscha, alt. 4800 m, G. M iehe and S. MIEHE, 12.10.1986, no. 13 962 (GZU). Langtang area, Yala, alt. 4830 m, G. MIEHE and S. Miehe, 05.07.1986, no. 4603a (GZU). Langtang area, Donga, Tangsep, alt. 4730 m, G. MIEHE and S. Miehe, 04.09.1986, no. 10 683 (GZU). Langtang area, above Pemdang Karpo, alt. 4640 m, G. M iehe and S. MlEHE, 30.09.1986, no. 13 075 b. Upper Langtang, Kangsa, alt. 4300 m, G. M iehe and S. Miehe, 19.09.1986, no. 12 187 (GZU). Upper Langtang, Pemdang Karpo, alt. 4880 m, G. MlEHE and S. MlEHE, 26.09.1986, no. 12 901 (GZU). 33 km NW Pokhara, Machhapuchhare basecamp, 28° 30' N, 83° 52' E, alt. 3965 m, T h o r, 27.11.1979, no. 1415 (S). China. Tibet, inter stat. Tasso et Penczamu, Tassa-schanj, P o ta n in , 31.05.1893 (KW, holotype of Cetraria potaninii ). Sikang, Taofu Distr., Taining (Ngata), alt. 4500 -4600 m, Sm ith, 07.09.1934, no. 14 032 (UPS). Mon­ golia. Bayan-Khongor region, Gurvan-Bulak Distr., Lake Khuh-nur, alt. 2600 m, BiazROV, 18.08.1972, no. 700 (LD). Hangai, GOLUBKOVA (LE). 41 368 A. T h e ll, T. R a n d la n e , I. K a r n e f e lt et al. Acknowledgements The authors are grateful to the keepers of the herbaria FH, GZU, H, KW, LE, PC, S, and UPS for sending kindly the lichen specimens. We want to thank Prof. Dr. J. P o e lt , Graz, and our collegues at the University of Lund for numerous advices and valuable comments on the manuscript. Dr. P. L a ssen , Lund, is thanked for help with the Latin diagnoses. This project has been supported by the Swedish Institute, Stockholm, the Estonian Science Foundation, Tallinn, and the Royal Physiographic Society, Lund. References 1. Asahina, Y. Lichens. -In: K ih ara, H. (Ed.) Fauna and Flora of Nepal Himalaya 1: 43 -63; 1955. 2. B a b in g to n , C. 3. CULBERSON, C. F. Lichenes himalayenses. -H o o k e rs J. Bot. 4: 243 -252; 1852. Improved conditions and new data for the identification of li­ chen products by a standardized thin-layer chromatographic method. - J. Chroma­ togr. 97:107 - 108; 1972. 4. DEY, J. P. Fruticose and foliose lichens of the high-mountain areas of the southern Appalachians. -Biyologist 81: 1 -93; 1978. 5. Egan, R. S. A fifth checklist of the lichen-forming, lichenicolous, and allied fungi of the continental United States and Canada. -Bryologist 90: 77 - 173; 1987. 6. HUE, A.-M. Lichenes extra-europaei. - Nouv. Arch. Mus. (Paris) I, 4: 27 - 220; 1899. 7. K a rn e fe lt, I. M attsson , J.-E., and T h e ll, A. Evolution and phylogeny of cetrarioid lichens. -Plant. Syst. Evol. 183: 113 - 160; 1992. 8. K a rn e fe lt, I., M attsso n , J., E., and T h e ll, A. The lichen genera Arctocetraria, Cetraria, and Cetrariella (Parmeliaceae) and their presumed evolutionary affinities. - Bryologist 96: 394 -404; 1993. The genus Flavocetra­ ria KARNEF. et THELL (Parmeliaceae) and its affinities. - Acta bot. fenn. 150: 79 - 9. K a rn e fe lt, I., T h e ll, A., R a n d la n e , T., and Saag, A. 86; 1994. 10. KREMPELHUBER, A. VON. Cetraria bavarica, eine neue deutsche Flechtenart, ent- deckt und beschrieben von A. VON KREMPELHUBER, Konigl. Bayer. Salinen- u. ForstcommissSr in Miinchen. -Flora 34: 273 -275; 1851. 11. KREMPELHUBER, A. VON. Exotische Flechten aus dem Herbar des K. u. K. Botanischen Hofkabinettes in Wien. - Verhandl. zool.-bot. Gesellsch. Wien 18: 305 - 320; 1868. 369 Allocetraria (Parmeliaceae) 12. K u r o k a w a , S. and L ai, M.-J. Allocetraria, a new lichen genus in the Parmelia­ ceae. -Bull. nat. Sci. Mus. Tokyo, Ser. B, 17: 59 -65; 1991. 13. Lai, M.-J. 14. Studies on the cetrarioid lichens in Parmeliaceae of East Asia. I. - Quart. J. Taiwan Mus. 13: 139 -242; 1980. M a tts s o n , J.-E. A monograph of the genus Vulpicida (Parmeliaceae, Ascomycetes). -Op. bot. 119: 1-61; 1993. 15. M a tts s o n , J.-E. and L ai, M. J. Vulpicida, a new genus in Parmeliaceae (liche­ nized Ascomycetes). -Mycotaxon 46:425 -428; 1993. 16. M e r r ill, G. K. Two new Cetraria forms and three new combinations. - Bryolo­ gist 13: 24-30; 1910. 17. N y la n d e r , W. Essai d'une nouvelle classification des lichens. - Mem. Soc. imp. Sci. nat. Cherbourg 3: 161 -202; 1855. 18. N y la n d e r , W. Enumeration gen6rale des lichens, avec l'indication sommaire de leur distribution gdographique. - Mem. Soc. imp. Sci. nat. Cherbourg 5: 85 - 146; 1857. 19. N y la n d e r , W. Addenda nova ad lichenographiam europaeam. - Flora (Regens­ burg) 70: 129 - 136; 1887. 20. O x n e r, A. N. Species lichenum novae ex Asia. -J. Cycle Bot. Acad. Sci. Ukraine 7-8: 167- 172; 1933. 21. R a n d la n e , T. and S aag , A. New combinations of some cetrarioid lichens (Par­ meliaceae). -Mycotaxon 44: 491 -493; 1992. 22. R a n d la n e , T., S a a g , A., T h e ll, A., and K a r n e f e lt , I. The genus Tuckneraria et T h e ll, a new segregation in the family Parmeliaceae. - Acta bot. fenn. 150: 143-151; 1991. 23. T h e ll, A., M a tts s o n , J.-E., and K a r n e f e lt , I. Lecanoralean ascus types in the R a n d l. lichenized families Alectoriaceae and Parmeliaceae. -Crypt. Bot.: in press; 1994. 24. T u c k e rm a n n , E. Further notices of some New England Lichenes. - Boston J. nat. Hist. 3: 438-464; 1841. 25. WEI, J. • An enumeration of lichens in China. - International Academic Publishers, Beijing; 1991. 26. Z a h lb r u c k n e r , 73 -95; 1911. A. Transbaikalische Lichenes. -Trav. Soc. imp. russe Geogr. 12: 370 Received: October 10th, 1994 Accepted: December 16th, 1994 A. T h e ll , T. R a n d l a n e , I. K arnefelt et al. Addresses of the authors: Mr. A. T h e ll Prof. Dr. I. K a r n e fe lt Department of Systematic Botany University of Lund Ostra Vallgatan 18-20 S-22361 Lund Sweden Dr. T. R a n d la n e Mr. A. S a a g Institute of Botany and Ecology Tartu University Lai Street 38 EE-200 Estonia Mr. X.-Q. G a o Botanical Museum University of Uppsala Villavagen 6 S-74236 Uppsala Sweden VIII Thell, A., Goward, T., Randlane, T., Kamefelt, E. I. & Saag, A. 1995. A revision of the North American Lichen genus Ahtiana (Parmeliaceae). — Bryologist 98(4): 596-605. The Bryologist 98(4), 1995, pp. 596-605 Copyright © 1995 by the American Bryological and Lichenological Society, Inc. A Revision of the North American Lichen Genus Ahtiana (Parmeliaceae) A r n e T hell Department of Systematic Botany, University of Lund, Ostra Vallgatan 18-20, 223 61 Lund, Sweden T re v o r G o w a rd Herbarium, Department of Botany, University of British Columbia, Vancouver, British Columbia, VOE 2B1 Canada (Mailing address: Edgewood Blue, Box 131, Clearwater, B.C. VOE 1N0, Canada T iin a R a n d l a n e Institute of Botany and Ecology, Tartu University, Lai Street 38, EE-2400 Tartu, Estonia E. I. K a r n e f e l t Department of Systematic Botany, University of Lund, Ostra Vallgatan 18-20, 223 61 Lund, Sweden A n dre s Saag Institute of Botany and Ecology, Tartu University, Lai Street 38, EE-2400 Tartu, Estonia Abstract. The formerly monotypic genus Ahtiana (type species: A. sphaerosporella (Miill. Arg.) Goward is shown to include two additional species, A. aurescens (Tuck.) Thell & Randl. and A. pallidula (Riddle) Goward & Thell. All three species are endemic to North America, and are char­ acterized by their greenish yellow upper surface, globose ascospores, and caperatic acid in the medulla. The habitat ecology o f these lichens is very specialized. Ahtiana is presumably most closely related to Tuckneraria Randl. & Thell and other cetrarioid genera having globose ascospores. Goward (1985) originally segregated Ahtiana sphaerosporella from Parmelia s. lat. on the basis of the former’s emergent pycnidia, globose asco­ spores, leptodermatous cortex, and presence of medullary caperatic acid. Despite its parmelioid habit with laminal apothecia and pycnidia, Ahtiana sphaerosporella was shown to be closely allied to Cetraria pallidula Riddle (Goward 1985). More re­ cent studies on the morphology and anatomy of cetrarioid lichens, carried out in Lund and Tartu, clearly demonstrate that both Cetraria pallidula and Cetraria aurescens Tuck, should be placed in Ah­ tiana. The new combinations, Ahtiana aurescens (Tuck.) Thell & Randlane and Ahtiana pallidula (Riddle) Goward & Thell, are therefore presented here. As thus defined, Ahtiana is a morphologically well delimited group related to other cetrarioid gen­ era having globose or subglobose ascospores, i.e., Allocetraria Kurok. & Lai, Esslingeriana Hale & Lai, Tuckermannopsis Gyelnik, and Tuckneraria Randlane & Thell (Table 1). Kamefelt et al. (1992) showed by means of cladistic analysis that these five genera form a monophyletic group within the Par­ meliaceae. Esslingeriana, Tuckneraria, and Allo­ cetraria have already been examined by Esslinger (1971), Lai (1980), Randlane et al. (1994), Kuro­ kawa and Lai (1991), and Thell et al. (1995a,c). However, the largest genus in the group, Tucker­ m annopsis, still requires critical examination (Kamefelt et al. 1992, 1993). M a t e r ia l a n d M e t h o d s About 70 specimens have been consulted for this study from: a s u , b, bm, b r y , c a n l , c o lo , d u k e , f h , g z u , h , l d , m in n , n y , s fsu , u a c , u b c , uc, us, and was. For anatomical observations, the lichens were sectioned with a Leica Cryostat 1800 Cryocut freezing microtome and stained in lactophenol cotton-blue. The asci were squashed in a 0.3% Lugol’s solution after pretreatment with a 10% K O H solution. The secondary chemistry was investigated on HPTLC plates using standardized TLC methods (Culberson 1972). R esults A h t i a n a Gow ard, T h e B r y o l o g i s t 88: 370. 1985. T y p e species : Ahtiana sphaerosporella (Miill. Arg.) Go­ ward Thallus foliose, more or less closely appressed, up to 8 cm across; upper surface pale-yellow to pale yellow-green, wrinkled; lower surface usually light tan; pseudocyphellae absent; isidia and cilia present in one species; pycnidia laminal or marginal, black, conspicuous, usually abundant, up to 0.2 mm across; upper cortex paraplectenchymatous with crystals of usnic acid, 15-25 fim thick, composed of ca. 3 cell 0007-2745/95/596-60551.25/0 1995] T a b l e 1. Comparison of Ahtiana with other cetrarioid genera having globose or subglobose ascospores. The genus Tuckermannopsis is still not clearly delimited; results in this table are based on T. americana, T. chloropfiylla, T. ciliaris, and T. orbata. Species Ahtiana Allocetraria Tuckermannopsis Esslingeriana yellow tan or yellow yellow brown or yellow grey black brown light brown to blackish Pseudocyphellae Isidia Cilia Apothecial position Pycnidial position Conidial shape and size absent absent or sparse absent or sparse marginal to laminal laminal usually bifusiform, 5-7 x l fim usnic acid present absent absent marginal marginal filiform, 10-19 x 0.5-2 #im usnic acid absent absent absent marginal to laminal mostly laminal bifusiform, 5—7 x 1 fim atranorin absent absent present or absent mostly marginal marginal bifusiform, ca. 5 x 1 pm atranorin caperatic, lichesterinic, and protolichesterinic acids lichesterinic and protoli­ chesterinic acids one unidentified fatty acid lichesterinic and protoli­ chesterinic acids secalonic acid fumarprotocetraric acid in one species endocrocin corticolous or terricolous mainly in Southeast Asia corticolous in western North America Cortical substances Medullary substances a) fatty acids b) anthraquinones c) orcinol & (3-orcinol depsides & depsi­ dones Distribution and ecology — — — corticolous in eastern or western North America — — Tuckneraria yellow to greenish whitish, light to dark brown or black present sparse present marginal mostly marginal bifusiform, 4-5 x 1-1.5 fim usnic acid lichesterinic and protoli­ chesterinic type fatty ac­ ids — — alectoronic, collatolic, olivetoric or physodic acids corticolous' in northern Eurasia and North America alectoronic, collatolic and physodic acids corticolous mainly in Southeast Asia THELL ET AL.: AHTIANA Upper surface Lower surface [VOL. 98 THE B RYO LO G IST 598 T a b le 2. Comparison of the three species included in Ahtiana. Lobe width measured on the widest part o f fully developed lobes. Species Color of upper surface Color of lower surface Lobe width Rhizinae Structure of the cortex Isidia Cilia Apothecia Disc color Thalline margin Pycnidia Distribution Ecology A. pallidula A. aurescens A. sphaerosporella light yellow to pale greenish yellow pale to light tan light yellow to pale greenish yellow light yellow 0.5-2.0 (3.0) mm often abundant pachydermatous paraplectenchyma absent or sparse to rather frequent on central lobes absent or sparse, dark or black common, marginal mostly restricted to thallus cen­ ter usually dark brown wrinkled common, marginal or lami­ nal, usually somewhat raised eastern N. America corticolous, most common on Thuja 4-10 mm often abundant pachydermatous paraplectenchyma absent pale yellowish green, in part olivaceous whitish to pale tan, in part olivaceous 2-3 (4) mm often abundant leptodermatous paraplectenchyma absent absent absent less common, marginal to submarginal always light brown wrinkled common, marginal or lami­ nal, immersed common, laminal, mostly restricted to thallus cen­ ter always light brown smooth common, mainly laminal, immersed to raised western N. America corticolous, most common on Larix or Pseudotsuga western N. America corticolous, most common on Pimis albicaulis or Abies layers, lower cortex also paraplectenchymatous, 1520 um thick, composed of 2-3 layers of somewhat brownish cells; medulla white, medullary hyphae 3-4 Mm thick; algal cells up to 10 um . Apothecia lrequent, laminal or marginal, up to 13 mm across, disc brown, entire; hymenium including subhymenium 50-60 fim high; exciple 2-layered, upper layer 10-30 ^im thick, composed of periclinally ar­ ranged hyphae, lower layer 35-50 um thick, para­ plectenchymatous; asci clavate, 45-60 p m high, ax­ ial body 2-5 um; ascospores globose or subglobose, more or less uniseriately arranged, 8 per ascus, 46 x 4-6 n m \ paraphyses straight, somewhat branched, with thickened apices; pycnoconidia usu­ ally bifusiform or occasionally bacillariform, citriform or sublageniform, 5-7(-9) x ca. 1 fim. Chemical substances.—Usnic acid in the cortex; caperatic acid as major and lichesterinic-protolichesterinic type fatty acids as minor compounds in the medulla. The three species included in Ahtiana are char­ acterized mainly by their foliose habit, their more or less closely appressed, pale yellowish lobes, their lack of pseudocyphellae, and their narrowly clavate asci containing uniseriately arranged subglobose spores. The yellow-green colour of the lobes reflects the presence of usnic acid in the cortical layer. This substance also occurs in the related genera Alloce­ traria and Tuckneraria, in which the ascospores and ascus form are also similar. Ahtiana-, however, dif­ 43 fers from these and other cetrarioid genera in having caperatic acid instead of lichesterinic acid as a major medullary compound (Table 1), although it must be admitted that some species of Allocetraria and Tuckneraria contain caperatic acid as an accessory substance. The genus Allocetraria', however, is easily distin­ guished from Ahtiana by the former’s very broad axial body, filiform pycnoconidia, and palisade plectenchymatous cortex. Several cetrarioid species have recently been transferred to this genus, mainly distributed in eastern Asia (Thell et al. 1995c). Presumably Ahtiana is most closely allied to Tuckneraria, also an essentially eastern Asian genus consisting of five corticolous species. However, in that genus pseudocyphellae are present on the lower surface (Randlane et al. 1994; Thell et al. 1995a), the lobe margins are frequently ciliate, and the apo­ thecia are always marginal. Although these admit­ tedly morphological points of distinction appear not to be supported by corresponding differences in apothecial structure or thallus chemistry, we prefer for the moment to maintain Ahtiana and Tuckneraria as separate genera; however, further studies are in progress. Points of separation with other related genera are summarized in Table 1. The structure of the cortical hyphae is rather vari­ able in Ahtiana (Table 2). All species have a para­ plectenchymatous cortex, but the thickness of the walls in relation to the cell lumina differs between the species. In A. pallidula and especially A. aures- 1995] THELL ET AL.: AHTIANA 599 F ig u r e s 1-3. Morphology of Ahtiana. — 1. A. aurescens, Trana 8793 (m in n ). — 2. A. pallidula, Ryan 25308 (a su ). — 3. A. sphaerosporella, Bird 14348 (wis). (Bar in Figs. 1-3 = 1 cm). cens, the walls are obviously thicker than the lu- mina, and may be termed pachydermatous (Frey 1936; Hale 1973). In A. sphaerosporella, by contrast the walls are thinner than the lumina, and may be referred to as leptodermatous (Goward 1985). An identical range of variation was observed in the low­ er excipular layer, with A. aurescens and A. pallidula having smaller lumina than A. sphaerosporella. Ascus structures show almost no variation within Ahtiana. The asci are of the Tuckermannopsis-form, and are rather narrowly clavate with an amyloid tholus, a broad ocular chamber, and a broad axial body (Thell et al. 1995b). The ascus form also occurs in the related genera Allocetraria, Esslingeriana, Tuckerm annopsis, and T uckneraria. The asco­ spores are more or less uniseriately arranged in un­ squashed asci. Ahtiana has typically dumbbell­ shaped pycnoconidia, although slightly disc-bar­ shaped ones, in which the ends are narrow and the thickenings subapical, are also present (Fig. 15). The range of variation is especially large in A. pallidula (see below). In A. sphaerosporella, the pycnoconidia may be as much as 9 (im long, although in the other species they are usually not longer than 7 um. All species have a corticolous ecology and are appar­ ently restricted to North America. A h t ia n a a u re sce n s comb. nov. (Tuck.) Thell & Randlane (F ig . 1, 4-5) Cetraria aurescens Tuck., Proc. Amer. Acad. Arts Sci. I: 208.1847. T yp e : U.S.A., N e w H a m p s h ir e , White Mts., Tuckerman, 1848 (FH-Tuck)—Lectotype selected here. Platysma aurescens (Tuck.) Nyl., Synops. Lich. I: 313. 1860. Tuckermannopsis aurescens (Tuck.) Hale, T h e B r y o l o g is t 90: 164. 1987. Thallus foliose, closely adnate, up to 6 cm across, surface smooth, lobes 0.5-2(-3) mm wide, often with somewhat indented but rounded tips, lobe margins slightly raised; upper surface pale yellow to pale greenish yellow, somewhat ridged; lower sur­ face pale to light tan, strongly reticulate; isidia sparse to rather frequent in central parts of few specimens; cilia absent or sparse and black; rhizines abundant; pycnidia marginal or laminal, usually somewhat raised; upper and lower cortex composed of pach­ ydermatous hyphae. Apothecia marginal, rarely submarginal, usually numerous, especially in cen­ tral portions of thallus, up to 7 mm in diameter, thalline margin sometimes strongly wrinkled, disk usually dark brown; exciple 2-layered, upper layer composed of horizontally arranged hyphae, lower layer pachydermatous; asci narrowly clavate, 4560 x 13-17 Mm, axial body 2.0-3.5 /im; ascospores 4.0-5.5 x 4.0-5.5 um in diameter; pycnoconidia bifusiform (dumbbell shaped), ca. 5 x 1 fim. Chemical substances .—Usnic acid in the cortex; caperatic acid in the medulla. One unidentified fatty acid reported by Dey (1978) is, according to our investigations, allied to lichesterinic-protolichesterinic acids. Ahtiana aurescens is easily distinguished from A. pallidula and A. sphaerosporella by its narrow lobes, 600 THE BRYOLOGIST [VOL. 98 1995] THELL ET AL.: AHTIANA typically indented tips, and marginal apothecia (Fig. 1, 4-5). A. aurescens occurs at temperate latitudes in eastern North America, where it is restricted to the Appalachian Mountains and Great Lakes region between 34° and 48°N (Fig. 16). In some districts, it has been reported to be common (Degelius 1940; Dey 1978), although in at least the eastern and northern portions of its range it appears to be rather rare (Fink 1910; Wetmore 1981; Wong & Brodo 1992). A htiana aurescens has most frequently been reported on Pinus (Degelius 1942; Hale 1979), as well as on “old rails” (Fink 1935). In northern areas; however, it also colonizes Thuja and other conifers in swampy areas (Fink 1910, Harris 1977; S. Clayden, pers. com m .), whereas in the southern Appa­ lachians it has been reported from “hardwood trees” (Dey 1978). Hale (1979) commented that this spe­ cies is often associated with Im shaugia placorodia. Selected specimens examined. —CANADA. O n t a r i o . Lake Nipissing, Macoun, 1884 (FH-Tuck). Thunder Bay District, Tibell 5290 (ld) 12 km S of Smooth Rock Falls, N W o f Cochrane, Wetmore 44037 (m in n ) . N e w B r u n s w ic k . Carleton Co., Clayden 77-132 (ubc). U.S.A. A la b a m a . F ra n k lin Co., Harris 28482 (n y ). A l ­ a b a m a /G e o r g ia . L o o k o u t M ts., Calkins 354 (c o lo ). C o n n e c t i c u t . New London Co., N o r w ic h , Setchell, 02.15.1882 (us). M a s s a c h u s e tts . Bristol Co., New Bed­ ford, Willey 12930 (b). M ic h ig a n . Baraga Co., Harris 8095 (m in n ) . Keweenaw Co., Wetmore 49101 (m in n ). M in n e s o t a . Carlton Co., Kettle falls, Fink 1423 (m inn). Cass Co., Snowball Lake, Fink 869 (m in n ). St. Louis Co., Wetmore 28037 (m inn). N e w H a m p sh ire . Grafton Co., N o rth Woodstock, Cummings, 07.1884 (ny). W h ite M ts., Tuckerman, 1848 (le c to ty p e , FH-Tuck). N e w Y o r k . Essex Co., Harris, 1901 (m in n ). N o r t h C a r o l i n a . Jackson Co., Trana 7949 (m in n ). P e n n s y lv a n ia . Pike Co., Wetmore 55833 (m in n ). Tennessee. Greene Co., Harris 27228 (ny). V ir g in ia . Bath Co., Hale 12412 (c o lo ) . V e r m o n t . R u t ­ la n d Co., Brandon, Dutton 2698 (m in n ). V ir g in ia . H ig h ­ la n d Co., Anderson, 09.06.1936 (ny). W is c o n s in . V ilas Co., Newberry 2279 (wis). A h tia n a P a llid u la nov. (Riddle) Goward & Thell comb, (Fig. 2, 6-7, 9, 11, 13-15) Cetraria pallidula Riddle, T h e B r y o lo g i s t 18: 27. 1915. Type: U.S.A. Washington Territory, near Mt. Adams, 1881, Pringle 218 (FH-Tuck, holotype). Nephromopsis pallidula (Tuck.) Riddle, The B r y o lo g i s t 18: 27. 1915.-invalid publ. Tuckermannopsispallidula (Riddle) Hale, T h e B r y o lo g is t 90: 164. 1987. 601 Thallus foliose with ascending lobes, rather loose­ ly adnate; lobes 4-10 mm wide, more or less linearelongate, apically rounded; upper surface pale yel­ low to pale greenish yellow, occasionally partly cov­ ered with a white pruina, strongly ridged; lower sur­ face pale yellow, strikingly reticulate; cilia absent; rhizines often abundant; pycnidia common, mar­ ginal or laminal, immersed; upper and lower cortex composed of pachydermatous hyphae. Apothecia rather frequent, marginal to submarginal, situated on ascending lobes, thalline margin wrinkled, disc entire, predominantly pale brown; exciple 2-layered, upper layer composed of horizontally arranged hy­ phae, lower layer pachydermatous; asci narrowly clavate, 45-60 x 13-17 /im, axial body 3.0-4.5 pm broad; ascospores 4-6 x 4-6 pm ; pycnoconidia usu­ ally bifusiform (dumbbell-shaped, rarely disc-barshaped) but several other pycnoconidial types (bacillariform, citriform, and sublageniform) are also found, 5-7 x ca. 1 pm . C h em ical substances .—Usnic acid in the cortex; caperatic acid as major and lichesterinic-protolichesterinic type fatty acids as minor compounds in the medulla. The pycnoconidia of A. pallidu la are highly vari­ able, with bacillariform, bifusiform, citriform, and sublageniform pycnoconidia having all repeatedly been observed in a single pycnidium (Fig. 15). Such an astonishing array of pycnoconidial types has not previously been reported in the cetrarioid lichens. The pycnoconidia are invariably of the same length, measuring 5-7 pm . In conformity with/4, aurescens, the apothecia and pycnidia have a marginal or sub­ marginal position (Fig. 2, 6-7). A htiana pallidu la is endemic to western North America, where it occurs between about 37° and 59°N (Fig. 16). Throughout most of this latitudinal range it is restricted to the Coast Mountains and Cascades, but is absent from the outer coast. Be­ tween about 46° and 52°N, its range extends inland to the Columbia and Rocky Mountains—presum­ ably in response to a corresponding inland extension of relatively oceanic climatic conditions (Goward & Ahti 1992; McCune 1984). In British Columbia, A. pallidu la is generally confined to Pseudotsuga m en ziesii (Goward & Ahti 1992), though farther south it occurs also over Pinus ponderosa (Tucker 1973) and especially L a rix occidentalis (R. Rosen- F ig u r e s 4-8. Morphology o f Ahtiana. — 4. Close-up of the type material of A. aurescens, showing the narrow lobes with the typically indented tips (white arrow), Tuckerman 1848 (lectotype, FH-Tuck). — 5. Ciliate portion (black arrows) of the same material. — 6. Close-up of A. pallidula, showing an apothecium and numerous, mostly marginal pycnidia, Goward 91-663 (ubc). — 7. An ascending lobe and apothecium from the type collection of A. pallidula, Pringle 218 (holotype, FH-Tuck). — 8. Close-up of A. sphaerosporella showing laminal apothecia and pycnidia. Note the smooth edge of the apothccia, Irnshaug 6037 (ld ) . (Bar in Figs. 4-8 = 1 mm). 602 THE BRYOLOGIST [VOL. 98 FI o u r e s 9-15. Anatomy of Ahtiana. — 9. Pachydermatous lower cortex, A. pallidula, Toren 2823 (sfsu ), — 10. Leptodermatous upper cortex, A. sphaerosporella, Imshaug 6037 (l d ). — 11. Transverse section of an apothecium o f A. pallidula. Note the strongly gelatinized paraplectenchymatous hyphae in the lower excipular layer (= e2), Thiers 23274 ( sfsu ). — 12. Transverse section of an apothecium of A. sphaerosporella with rather large lumina in the lower excipular layer (= e2), Imshaug 6037 (l d ). — 13. Ascus of A. pallidula is typical of the genus, although the umseriate arrangement has been lost here, due to the squash technique (ab = axial body), Thiers 23274 (sfsu ). — 14. Laminal pycnidhim of A. pallidula, Toren 2823 (sfsu ). — 15. Pycnoconidia o f different types present in the same pycnidium of A. pallidula, Thoren 2823 (sfsu ). (Bar in Figs. 9-15 = 10 uni.) treter pers. com .). This species is most typical of lowland conifer forests, but in drier regions it is also found in middle elevation forests. Ahtiana pallidula seems to be of rather sparse, often localized, occur­ rence throughout its range. 44 Selected specimens examined. —CANADA. B r it is h C o l u m b ia . Vancouver Isl.: Goward 91-663 ( u b c ). U.S.A. C a l if o r n ia . Plumas Co., Toren 2823 ( sfsu ); Shasta Co., Hale51865 (us); Siskiyou Co., Edge of Marble Mountain Wilderness, Ryan 25308 ( a su ); Tehama Co., Thiers 39J68 (sfsu ). I d a h o . Latah Co., Schroeder L568 1995] THELL ET AL.: AHTIANA s] des. • Ahtiana aurescens; ■ A. pallidula-, ★ A. sphaeros- (ms). O re g o n . Bur Springs, Toren 23274 (sfsu ); Klamath Co., Brown 848 (u s). Id a h o . Bonner Co., Thiers 17316 (sfsu ). W a s h in g to n . Klicktat Co., Goldendale, Foster S I2,325 (c o lo ). Mt. Adams, Pringle 218 (FH-Tuck); Ya­ kima Co., Lodge Pole Pine Camp,, Howard 3310 (c o l o ). Skamania Co., Ice Cave, Suksdorf, 09.26.1900 (n y ). Thallus foliose, closely adnate; lobes 2-3(-4) mm wide, more or less linear-elongate, apically rounded; upper surface pale yellowish green (occasionally be­ coming dark olivaceous green), often with blackish margins, strongly wrinkled to more often becoming folded; lower surface whitish to pale tan or in part olivaceous, reticulate; cilia absent; rhizines often abundant; pycnidia common, laminal, immersed to emergent; upper and lower cortex composed of leptodermatous paraplectenchyma. Apothecia numer­ ous, laminal, mostly restricted to thallus centre, thalline margin continuous, smooth, disc entire, light F ig u r e 16. W o rld distribution o f the three Ahtiana 603 porella. A h t ia n a s p h a e r o s p o r e lla (Mull. Arg.) Goward, T h e B r y o lo g is t 8 8 : 3 7 0 . 1 98 5. (Fig . 3, 8, 10, 12) Parmelia sphaerosporella Mull. Arg., Flora 74: 378. 1891. T ype: Canada. British Columbia, Galton Mountains, Lyall, 1861 (b m , holotype)—verified by S. Clayden, 1985. 604 [VOL. 98 THE BRYOLOGIST brown; exciple 2-layered, upper layer composed of horizontally arranged hyphae, lower layer leptodermatous; asci rather broadly clavate, 40-50 x 1318 nm \ axial body 3-5 /mi; ascospores 4-6 x 4-6 jim in diameter; pycnoconidia bifusiform (dumb­ bell shaped or slightly disc-bar shaped), 5-7(-9) x ca. 1 p m . quoia National Park, Wetmore 50453, 51065 (a su , m in n). M o n t a n a . Columbia Falls, Williams 10.1895 (m in n , us). Glacier Co., Glacier National Park, Imshaug 6037 (ld ), O r e g o n . Hood River Co., M t. Hood, Shushan sl-2506 (b ry ). W a s h in g t o n . Olympic National Park, Sharpe 9933 Chem ical substances .—Usnic acid in the cortex; caperatic acid as major compound in the medulla together with other fatty acids of lichesterinic-protolichesterinic type as minor compounds. Ahtiana sphaerosporella is endemic to western North America, where it occurs in open, well illu­ minated, conifer forests at temperate and boreal latitudes (Fig. 16). In the Canadian portion of its range, it is restricted primarily to Pinus albicaulis, on which it may often be abundant. However, it also occurs although usually as scraps, on Abies la- This research was made possible in part by Grant No. LCZ 000 from the International Science Foundation. The Swedish authors are grateful for financial support from the Swedish National Research Council (NFR). The first au­ thor wishes to thank the Knut and Alice Wallenbergs Foundation for supporting a visit to British Columbia in 1994. We all wish to thank Roger Rosentreter (Idaho) for notes on the ecology of the western species. The keepers of the herbaria are also thanked for kindly sending spec­ imens. siocarpa, L a rix lyallii, Picea engelm annii, Pinus contorta, P .flexilis, and Pseudotsuga m en ziesii (Go­ B ir d , C. D . & A. H . M a r s h . ward 1985; Goward & Ahti 1992; Kalgutar & Bird 1969). As noted by Goward & Ahti (1992), the zonal distribution of A. sphaerosporella is determined largely by that of Pinus albicaulis, which is itself primarily confined to subalpine and alpine eleva­ tions. In the Canadian Rockies, for example, Bird and Marsh (1973) reported A. sphaeroporella over an elevational range from 1,300 m to 2,350 m, with a peak occurrence between 2,000 and 2,275 m. Out­ lying stands of Pinus albicaulis as low as 1,000 m have also been found to support healthy populations of this species (Goward 1985). In the American portion of A. sphaerosporella 's range, Pinus albicaulis appears to be much less im­ portant as a host tree. Hale (1979), for example, reported this lichen to be “widespread on conifers, especially Abies and Picea, at higher elevations.” In California, Hale and Cole (1988) stressed that it is “almost always” found over Abies. In the Pacific Northwest, A. sphaerosporella has been observed over A bies lasiocarpa, A. concolor, Pinus m onticola, and Pseudotsuga m enziesii. The phenomenon of substrate switches in relation to this species has al­ ready been briefly discussed by Goward (1985). A h ­ tiana sphaerosporella is easily separated from the two other species in the genus in having laminal apothecia with a smooth rim (Fig. 3, 8). Selected specimens examined.—CANADA. A lb e r t a . Livingstone Range, Bird 14348 (wis). Oldman River Wa­ tershed, Bird & Lakusta 16164,14823,14921 (b ry , c o lo , ld ) . B r it is h C o lu m b ia . Bella Coola valley, Ohlsson 2341 (wis). Chilliwack, Goward 78-1221 (ubc). Crowsnest Pass, Goward81—1710 (ubc). Mt. Robson Provincial Park, Marsh 1112 (NY) U.S.A. C a l if o r n ia . Amador Co., Weier 980 (uc); El Dorado Co., Stone 1964 (d u k e ). Fresno Co., Thiers 13431 (a s u ); Siskiyou Co., Cooke 30039E (us). Tulare Co., Se­ (asu). A cknow ledgm ents L it e r a t u r e C it e d 1973. Phytogeography and ecology of the lichen family Parmeliaceae in south­ western Alberta. Canadian Journal of Botany 51:261— 288. C u lb e r s o n , C. F. 1972. Improved conditions and new data for the identification of lichen products by a stan­ dardized thin-layer chromatographic method. Journal of Chromatography. 97: 107-108. D e g e u u s , G. 1942. Contributions to the lichen flora of North America. I. Lichens from Maine. Arkiv for Botanik 30: 1-62. D ey , J. P. 1978. Fruticose and foliose lichens of the highmountain areas of the southern Appalachians. T he B r y o lo g i s t 81: 1-94. E s s lin g e r , T. L. 1971. Cetraria idahoensis, a new species of lichen endemic to western North America. T h e B r y o lo g i s t 74: 364-369. F in k , B. 1910. Lichens of Minnesota. Contributions U.S. National Herbarium 14. Washington Gov. Print. Of­ fice. 269 pp. -----. 1935. The Lichen Flora of the United States. University of Michigan Press, Ann Arbor, M I. 426 pp. F re y , E. 1936. Vorarbeiten zu einer Monographic der Umbilicariaceen. Berichte der Schweizerischen Botanischen Gesellschaft 45: 198-230. G o w a r d , T. 1985. Ahtiana, a new lichen genus in the Parmeliaceae. T he B r y o lo g is t 88: 367-371. -----& T. A h t i. 1992. Macrolichens and their zonal distribution in Wells Gray Provincial Park and its vi­ cinity, British Columbia, Canada. Acta Botanica Fennica 147: 1-60. H a le , M. E. 1973. Fine structure of the cortex in the lichen family Parmeliaceae viewed with the scanningelectron microscope. Smithsonian Contributions to Botany 10: 1-92. -----. 1979. How to Know the Lichens. W m .C . Brown Co., Dubuque, Iowa. 246 pp. -----& M. C o le . 1988. Lichens of California. Cali­ fornia Natural History Guides 54: 1-253. H a r r is , R. C. 1977. Lichens of the Straits Counties, Michigan. University of Michigan Herbarium. Ann Asbor, MI. 150 pp. K a l g u t k a r , R. M. & C. D. B ir d . 1968. Lichens found on Larix lyallii and Pinus albicaulis in southwestern Alberta, Canada. Canadian Journal of Botany 47:627648. THELL ET AL.: AHTIANA J.-E. M a tts s o n & A. T h e ll. 1992. Evo­ lution and phylogeny of cetrarioid lichens. Plant Systematics and Evolution 183: 113-160. ---- , ---- & ---- . 1993. The lichen genera ArcK a r n e fe lt, I., tocetraria, Cetraria and Cetrariella (P arm eliaceae) an d th e ir presum ed evolutionary affinities. T he B r y o lo g is t 96: 394-404. S. & M.-J. L a i. 1991. Allocetraria, a new lichen genus in the Parmeliaceae. Bulletin of the Na­ tional Scienece Museum ser. B 17: 59-65. L a i, M.-J. 1980. Studies on the cetrarioid lichens in Par­ meliaceae of east Asia (1). Quarterly Journal of the Taiwan Museum 33: 215-229. M c C u n e ,B . 1984. Lichens with oceanic affinities in the Bitterroot Mountains of Montana and Idaho. T he B r y o lo g is t 87: 44-50. R a n d la n e , T., A. S a a g , A. T h e ll & I . K a r n e fe lt. 1994. The lichen genus Tuckneraria Randlane & Thell—a new segregatein the Parmeliaceae. Acta Botanica Fen­ nica 150: 143-151. K u ro k a w a , 60S A., I. K A r n e fe lt & T . R a n d la n e . 1995a. Tuck­ neraria togashii, a new combination of a cetrarioid lichen in the Parmeliaceae from Japan. Journal of the Hattori Botanical Laboratory (in press). --- -, J.-E. M a tts s o n & I. K a r n e fe lt. 19956. Lecanoralean ascus types in the lichenized families Alectoriaceae and Parmeliaceae. Cryptogamic Botany (in press). — — , T. R a n d la n e , I. K a r n e fe lt, X. G a o & A. S a a g . 1995c. The lichen genus Allocetraria (Ascomycotina, Parmeliaceae). Bibliotheca Lichenologica (in press). T u c k e r, S. 1973. New records and comments on lichens in California. T he B r y o lo g is t 76: 209-211. W e tm o re , C. M. 1981. Lichens of Voyageurs National Park, Minnesota. The B r y o lo g is t 84: 482-491. W o n g , P . Y . & I. M. B ro d o . 1992. The lichens of south­ ern Ontario, Canada. Syllogeus 69: 1-79. Canadian Museum of Nature, Ottawa. T h e ll, IX 45 Saag, A., Randlane, T. & Thell, A. Phylogenetic analysis o f cetrarioid lichens with globose ascospores. (Submitted.) PHYLOGENETIC ANALYSIS OF CETRARIOID LICHENS WITH GLOBOSE ASCOSPORES Andres SAAG*, Tiina RANDLANE* & Ame THELL** institute of Botany & Ecology, University of Tartu, Lai Street 38, EE-2400 Tartu, Estonia **Department of Systematic Botany, University of Lund, Ostra Vallgatan 18-20, S-223 61 Lund, Sweden Abstract. The group of cetrarioid lichens with globose ascospores in narrowly clavate asci includes 30 species from six genera. Phylogenetic analyses were carried out, using programs PAUP 3.1.1, MacClade 3.04, and the Parsimony Jackknifer 4.22, to check the correspondence of the present taxonomy to the probable evolution of the taxa involved. A separate clade, comprising three Ahtiana species, is formed in most of the trees. The genus Allocetraria is paraphyletic with respect to Dactylina. The taxa belonging to the genera Esslingeriana, Tuckneraria, and Tuckermannopsis today, do not form separate groups and evidently need further studies. Key words: cetrarioid lichens, Parmeliaceae, Ahtiana, Allocetraria, Dactylina, Esslingeriana, Tuckneraria, Tuckermannopsis, cladistic analysis. Introduction The group of cetrarioid lichens, comprising 22 genera and more than 130 species today (Randlane et al. 1997), is definitely polypheletic. Still, phylogenetic affinities between several cetrarioid genera are rather obscure. The first review about the phylogeny of the whole group of cetrarioid lichens was presented by Kamefelt et al (1992) where cladistic analyses were carried out on 50 species as terminal taxa. This analysis was based mainly on anatomical characters that had been originally revised. As a result, the analysed taxa were grouped in three separate aggregates, described by important anatomical characters: taxa with an “apical ring structure”; taxa with “uniseriate asci”; taxa with "‘broadly clavate asci”. Independent cladistic analyses on the same group of lichens were also carried out by us (Saag & Randlane 1995). Of about 120 cetrarioid lichen species known at that time, 83 were chosen for the analysis to evaluate the systematic arrangement of the taxa. Morphological and chemical characters (character states) were identified or verified originally, while descriptions of anatomical characters were based mainly on literature data. Evolution for considerable number of species (from genera Nephromopsis, Cetreliopsis, Cetrelia) remained unresolved, showing several polytomies on the respective part of the consensus tree. The rest of the species were grouped in a way that shows considerable similarity with the main three assemblages pointed out by Kamefelt et al. (1992). Similar results are particularly remarkable because of the fact that the two analyses were based on different data matrices, with emphasis on anatomical characters in the Swedish study and morphological and chemical characters in our analysis. Today the polyphyletic origin of cetrarioid lichens is generally acknowledged. Three evolutionary lines referred to above, based on reproductive structures and structural 1 characters mainly, have been recognized for the group (Thell 1996). The first line, considered monophyletic, includes species from seven genera: Arctocetraria, Cetraria s. str., Cetreliopsis, Coelopogon, Flavocetraria, Masonhalea, and Nephromopsis. They are all characterized by the ellipsoid ascospores in narrowly clavate asci, with a small axial body. An amyloid ring structure in tholus is a significant character that is present in Cetraria, Flavocetraria and occasionally in Nephromopsis. The second evolutionary line comprises six genera: Ahtiana, Allocetraria, Dactylina, Esslingeriana, Tuckneraria, and Tuckermannopsis. These species are anatomically characterized by subglobose to globose ascospores borne in narrowly clavate asci (earlier called “uniseriate asci”). Some other cetrarioid lichens, which also have almost subglobose ascospores but quite different type of asci (e.g. species of Vulpicida and Platismatia), are not included. The assumption about monophyletic origin of this line is supported by two earlier cladistic analyses of various species of cetrarioid lichens (Kamefelt et al. 1992, Saag & Randlane 1995). In both analyses the clade comprising species with globose ascospores in narrowly clavate asci was clearly separated. The third line is represented by several genera and informal groups: Asahinea, Cetraria fendleri group, Cetrariella, Cetrelia, Cornicularia, Kaernefeltia, Melanelia commixta group, Nimisia, Parmelaria, Platismatia, and Vulpicida. Ellipsoid ascospores in broadly clavate asci, with a broad axial body are characteristic to these species. This heterogenous gouping is evidently paraphyletic in the previous treatments of cetrarioid taxa. A number of parmelioid lichens probably also belong here. The cetrarioid species of the second evolutionary line are treated here in more detail. Four genera of the six, Ahtiana, Allocetraria, Dactylina and Tuckneraria, have been thoroughly revised lately (Thell et al. 1995a, b, Kamefelt & Thell 1996, Randlane et al. 1994). The sole species of the monotypic genus Esslingeriana was described completely by Esslinger (1971). Tuckermannopsis is the only entity that still needs to be revised. Nevertheless, the data characterizing these taxa are scattered in many papers and evolutionary affinities between all the species included have not been evaluated yet. Material and methods Herbarium material from B, DUKE, FH, GZU, H, KW, LD, LE, M, MB, PC, S, TAIM, TNS, TU, UPS, US has been used for this study. Morphological characters were examined using a stereomicroscope Technival 2; anatomical methods and equipment used are described in detail in Thell et al. (1995a); secondary chemistry was investigated according to the standardized methods of TLC (Culberson 1972, Culberson et al. 1981). Computer programs for phylogenetic analysis PAUP 3.1.1 (Swofford 1993) and MacClade 3.04 (Maddison & Maddison 1992) were used. Cladistic analyses were carried out by the first author. Both programs were run on a Macintosh Color Classic. Using the program PAUP 3.1.1, the follwing heuristic search settings were applied: character-state optimization - ACCTRAN; MULPARS; MAXTREE = 1000, 3000, 6000; addition sequence - simple; 1 tree held at each step during stepwise addition; treebisection-recollection (TBR) branch-swapping performed; multi-state taxa interpreted as uncertainty; characters weighted equally. The same settings were used in the successive approximations character weighting method. Tree support was investigated using Bremer support and bootstrap in PAUP 3.1.1 and the program Parsimony Jackknifer 4.22 (Farris 1995). 2 Data Taxa analysed. The following 30 cetrarioid species from the six above-listed genera were applied as terminal taxa of the ingroup: Ahtiana aurescens (Tuck.) Randlane & Thell, A. pallidula (Tuck, ex Riddle) Goward & Thell, A. sphaerosporella (Mull. Arg.) Goward, Allocetraria ambigua (Bab.) Kurok. & M. J. Lai, A. endochrysea (Lynge) Kamefelt & Thell, A. flavonigrescens Thell & Randlane, A. globulans (Nyl.) Thell & Randlane, A. madreporiformis (Ach.) Kamefelt & Thell, A. oakesiana (Tuck.) Randlane & Thell, A. sinensis X. Q. Gao, A. stracheyii (Bab.) Kurok. & M. J. Lai, Dactylina arctica (Richardson) Tuck., D. ramulosa (Hook.) Tuck., Esslingeriana idahoensis (Essl.) Hale & M. J. Lai, Tuckermannopsis americana (Spreng.) Hale, T. chlorophylla (Willd.) Hale, T. ciliaris (Ach.) Gyeln., T. gilva (Asahina) M. J. Lai, T. inermis (Nyl.) Karnefelt, T. microphyllica (W. L. Culb. & C. F. Culb.) M. J. Lai, T. orbata (Nyl.) M. J. Lai, T. platyphylla (Tuck.) Hale, T. platyphylloides (Asahina) M. J. Lai, T. subalpina (Imshaug) Kamefelt, T. ulophylloides (Asahina) M. J. Lai, Tuckneraria ahtii Randlane & Saag, T. laureri (Kremp.) Randlane & Thell, T. laxa (Zahlbr.) Randlane & Thell, T. pseudocomplicata (Asahina) Randlane & Saag and T. togashii (Asahina) Randlane & Thell. For choosing the outgroup we had, in principle, two alternatives: a taxon either from the group close to Cetraria (the first evolutionary line of cetrarioid lichens) or from the group related to the so-called “parmelioid Cetrariae” (the third evolutionary line). The form of asci - narrowly clavate - is the same in the ingroup and in the group close to Cetraria, while the third group has typically broadly clavate asci. Therefore, the parmelioid and allied genera were discarded at first. Two genera of the seven included in the first evolutionary line - Cetraria s. str. and Flavocetraria - were seriously considered as possible outgroups. Ascocarps of Flavocetraria are characterized by ellipsoid ascospores, narrowly clavate asci with a small axial body, and presence of an amyloid ring structure. These characters are similar to Cetraria s. str. and differ from those of the ingroup. Still, the stucture of upper and lower cortices and secondary chemistry of Flavocetraria is more similar to the studied taxa. Finally, Flavocetraria cucullata (Bellardi) Kamefelt & Thell was selected as outgroup for one series of analyses. Later the species from the third evolutionary line were also evaluated as possible samples for outgroup. In earlier analyses (Kamefelt et al. 1992, Saag & Randlane 1995) C. fendleri appeared as one representative of the sister group to the assemblage which is treated as ingroup here. Anatomically, ascocarps of C. fendleri are characterized by ellipsoid ascospores, broadly clavate asci, a large axial body, and absence of an amyloid ring structure. The ingroup differs from the species under discussion in two former and is similar in two latter characters. Therefore, C. fendleri was additionally chosen as outgroup. Both selected species - Flacocetraria cucullata and Cetraria fendleri - were used to root the trees in separate series of analyses. Characters. All in all, 36 morphological, anatomical, and chemical characters were used for the analysis. Lichen substances were not treated independently from each other but grouped into biochemically related sets, as suggested in our previous study (Saag & Randlane 1995). 46 3 Character states were coded as 0, 1, 2, 3 and 4; all multistate characters were treated as unordered. The characters and character states were the following. 1. Form of thallus: adnate (0), ascending (1) 2. Symmetry of thallus: dorsiventral (0), radial-symmetrical (1) 3. Interior of thallus: of densely arranged hyphae (1), of loosely arranged hyphae (1), becoming hollow (3) 4. Form of lobes: length of lobes ~ width of lobes (0), lobes longer than wide (1) 5. Width of lobes: up to 3 mm (0), up to 6 mm (1), up to 12 mm (2) 6. Colour of upper surface: yellow (0), brown (1), grey (2) 7. Colour of lower surface: whitish (0), yellow (1), brown (2), black (3) 8. Pseudocyphellae on upper side: absent (0), present (1) 9. Pseudocyphellae on lower side: absent (0), present (1) 10. Form of pseudocyphellae on lower side: spots (0), lines (1) 11. Cilia: absent (0), present (1) 12. Rhizines: absent (0), present (1) 13. Soredia: absent (0), present (1) 14. Isidia: absent (0), present (1) 15. Structure of the cortex (orientation of hyphae): both corteces paraplectenchymatous, i.e. hyphae randomly oriented (0), upper cortex palisade plectenchymatous, i.e. hyphae anticlinally oriented (1), both corteces palisade (2), both corteces prosoplectenchymatous, i.e. hyphae parallel to cortex (3) 16. Cell wall thickness (compared to cell lumina): leptodermatous (0), pachydermatous (1) 17. Position of apothecia: marginal only (0), marginal to submarginal (1), laminal (2), terminal (3) 18. Ascus shape: broadly clavate (0), narrowly clavate (1) 19. Ascus form: melanelia type (0), tuckermannopsis type (1), cetraria type (2) 20. Tholus: small (0), large (1) 21. Axial body: medium [3-5 jim] (0), broad [>5 |im] (1), narrow [<3 pm] (2), very narrow [<1 pm] (3) 22. Shape of ascospores: globose (0), subglobose (1), broadly ellipsoid (2), ellipsoid (3) 23. Length or diameter of ascospores: short [<6 pm] (0), long [>6 pm] (1) 24. Position of pycnidia: marginal only (0), marginal and laminal (1), laminal only (2) 25. Emergency of pycnidia: emergent (0), immersed (1) 26. Shape of pycnoconidia: bacillariform (0), oblong citriform (1), dumb-bell shaped incl. disc-bar shaped (2), filiform (3), sublageniform (4) 27. Length of pycnoconidia: short [<7 pm] (0), medium [7-10 pm] (1), long [>10 pm] (2) 28. Usnic acid: absent (0), present (1) 29. Atranorin: absent (0), present (1) 30. Fatty acids: absent (0), present (1) 31. Substance of fatty acids: lichesterinic-protolichesterinic type acids (0), caperatic acid (1), rangiformic acid (2), unidentified (3) 32. Secalonic acids: absent (0), present (1) 33. Orcinol depsides and depsidones: absent (0), present (1) 34. Substance of orcinol depsides and depsidones: alectoronic and collatolic acids (0), physodic acid (1), olivetoric acid (2), microphyllinic acid (3), gyrophoric acid (4) 35. P-orcinol depsidones: absent (0), present (1) 36. Substance of p-orcinol depsidones: fumarprotocetraric acid (0), physodalic acid (1) The data matrix and terminal taxa are presented in Table 1. 4 Table 1. Data matrix and terminal taxa. Character numbers and coding of character states correspond to the text. & = and (polymorphism), / = or (uncertainty), - = gap, ? = character state unknown. Names\Characters Ahtiana aurescens Ahtiana pallidu la Ahtiana sph aerosporella A llocetraria am bigua A llocetraria endochrysea A llocetraria Jlavonigrescens A llocetraria globulans A llocetraria m adreporiform is A llocetraria oakesiana A llocetraria sinensis A llocetraria strach eyi D actylina arctica D actylina ram ulosa E sslingeriana idahoensis Tuckermannopsis am ericana Tuckermannopsis chlorophylla Tuckermannopsis ciliaris Tuckermannopsis g ilva Tuckermannopsis inerm is Tuckermannopsis m icrophyllica Tuckermannopsis orbata Tuckermannopsis pla typ h ylla Tuckermannopsis p latyph ylloides Tuckermannopsis subalpin a Tuckermannopsis u lophylloides Tuckneraria ahtii Tuckneraria laureri Tuckneraria laxa Tuckneraria pseu docom plicata Tuckneraria togashii F lavocetraria cucullata C etraria fen d leri 0 0 I 2 0 0 0 0 3 4 5 6 0 7 0 0 11111 8 9 0 12 3 4 1 5 1 1 6 7 112 2 8 9 0 1 2 2 2 2 2 3 4 5 2 2 6 7 2 2 3 3 8 9 0 1 3 3 3 2 34 3 3 5 6 0 0 10 0 0 10 1 1 10 0 0 1 0&1 0 0 10 1 0&1 1 1 11 10 10 10 10 0 0 10 10 0 0 10 10 10 10 10 10 10 10 0 0 10 0 0 0 0 0 0 0 0 2 0 0 0 0 0 0 110 110 0 0 10 0 0 0 10/1 110 0 0 0 10 0 10 0 0 110 2 1 2 0/1 2 10 1 0 112 0 0 11 0 0 11 0 0 11 0 0 11 0 0 0 1 0 0 11 0 0 11 0 0 2 1 0 0 2 1 0 111 0 0 11 0 12 0 0 0 10 0 10 0 0 0 2 0 0 0 10 0 110 0 0 0 1 0 1 0 1 1 2/3 2 1 2 2 1/2 1/2 1 3 2 2 2 2 0 2 2 2 2 2 1/2 2/3 0/2 1 0/2 1/2 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 2 2 2 2 2 0/2 2 2 2 2&3 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0/2 0 1 1 1 1 0 2 10 0 ? 0 ? 0 1 0 3 0 1 0 ? 10 0 3 13 0 1 0 1 0 0 0 1 0 ? 1 1 1 1 0 0 1 1 0 0 0 0 0 ? 10 10 10 10 10 13 1 1&2 1 10 110 110 110 0 0 0 0 ? 0 10 0 11 0 1 0&1 1 1 0&1 ? 2 1 ? ? 1 0&1 0 110 112 1 0 0 0 0&1 ? ? 00 0 0 0 0&1 0 0 2 1 0 0 2 1 0 111 0 0 10 0 0 0 1 0 0 10 2 2 2 3 3 ? ? ? ? ? ? ? ? ? ? 110 0 110? 110 0 110 0 0 0 0 0 0 3 2 2 1 0 1 0&1 1 0 1 0&1 1 0 1 0&1 10 10 10 10 1 0 1 0&2 10 10 10 10 1 0 1 0&1 10 10 10 10 10 0 10 00 112 0 100 110 0 100100 0 10 0 100 0 10 0 113 0 0 10 0 0 10 0 0 10 1 0 1 O&l 10 10 10 10 10 10 1 0 1 O&l 10 10 0 0 10 0 00 00 010101 0100 0100 0100 1 1&4 0 1 1 100 10 0 00 1 1&2 0 10 0 00 13 0 0100 00 00 00 00 00 00 1 O&l 0 00 00 0- 0 00 0010 0 00 0 01 1 1 1 0 000 0 00 0 0 0 0 0 00 0 000 0 - 0 - 1 1 0 0&1 0 - 0 1 00 0-0100 10 0 10 0 10 0 0 0 0 0-0000 0-0100 0-0000 0 - 0 1 10 1 10 0 0 0 10 0 10 0 0-0000 0-0000 0-0100 0-1 100 0 - 0 1 10 0-1100 0 - 1 1 10 1 1 0 10 0 0-0100 0-1 100 0-0101 0-0100 1 10 0 0 0 0-01 1 1 10 1 1 0 0 10 1 1 1 0 10 1 1 0 0 10 1 1 0 0 10 1 1 0 1 10 0 0 0 0 0-0100 0/2 0 0 0 ? ? ? ? ? ? ? ? 110 1 110 0 110 1 ? ? ? ? 110 110 110 110 110 110 110 1 0 0 0 0 0 0 0 0 1 10 0 0 10 0 11 ? ? ? ? ? ? 10 110 0 0 0 0 0 ? ? ? ? ? ? 0 0 1 1 0 0/2 0 0 1 0 110 0 0 0 0 0 110 0 0 0 0 0 110 0 0 0 0 0 1 1 0 0 0 0 10 110 0 1 2 13 0 0 0 0 1111 0 0 0/1 2 2 ? ? 3 2 3 2 3 2 3 2 3 2 10 10 2 0 2 0 2 0 2 0 1 2 0 0 ? 0 0 1 ? ? 2 0 2. 0 ? ? 2 2 2 2 0 0 0 0 Results Two series of separate analyses were carried out using different outgroups (Cetraria fendleri and Flavocetraria cucullata). With Cetraria fendleri as outgroup, 36 equally parsimonous trees were obtained (heuristic search, all characters equally weighted, length of trees =121 steps, Fig. 1). The strict consensus tree and the 50% majority-rule consensus of 36 trees were also retained (not shown). The successive approximations character weighting method produced 108 equally parsimonous trees and, after the second reweightening of characters, 81 equally parsimonous trees were obtained (strict consensus tree on Fig. 2). Successive weighting is particularly useful when applied to data sets with much homoplasy (Tehler & Egea 1997). Character reweighting by maximum value of rescaled consistency indices generated remarkably low weights to some conspicuous morphological characters often used in key-books, such as presence of isidia and soredia (Table 2). This is quite acceptable, in our opinion, as these, asexual structures are certainly derived, but they cannot be treated as shared characters. In other words, we suppose that sorediate and isidiate species did not share sorediate or isidiate ancestors but came from sexual species instead. The most highly weighted characters are either connected with the anatomy of the thallus (characters 2, 3, 15) or medullary secondary compounds (31, 34). Anatomical characters of the ascocarps, presumably highly evaluated, are uninformative in this analysis as the ascus form is the same in the whole ingroup. Cetrariafendleri Ahtiana pallidida Ahtiana aurescens Ahtiana sphaerosporella Tuckermannopsis inermis Tuckermannopsis platyphylla Esslingeriana idahoensis Tuckermannopsis microphyllica Tuckermannopsis ciliaris Tuckermannopsis americana Tuckermannopsis gilva Tuckermannopsis orbata Tuckermannopsis platyphylloides Tuckermannopsis chiorophylla Tuckermannopsis ulophylloides Tuckermannopsis subalpina Tuckneraria laureri Tuckneraria togashii Tuckneraria pseudocomplicata Tuckneraria ahtii Tuckneraria laxa Allocetraria globulans Allocetraria oakesiana Allocetraria ambigua Allocetraria stracheyi Allocetrariaflavonigrescens Allocetraria sinensis Allocetraria endochrysea Allocetraria madreporiformis Dactylina arctica Dactylina ramulosa Fig. 1. One of 36 equally parsimonous cladograms (based on characters with equal weights; Cetraria fendleri used to root trees). Length 121, consistency index 0.405, retention index 0.654. 6 Table 2. Weights of all characters after the second reweighting according to maximum value of RC indices. 1. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17. 18. 19. 20. 21. 22. 23. 24. 25. 26. 27. 28. 29. 30. 31. 32. 33. 34. 35. 36. Character ______ Form of thallus Symmetry of thallus Interior of thallus Form of lobes Width of lobes Colour of upper surface Colour of lower surface Pseudocyphellae on upper side Pseudocyphellae on lower side Form of pseudocyphellae on lower side Cilia Rhizines Soredia Isidia Structure of cortex Cell wall thickness Position of apothecia Ascus shape Ascus form Tholus Axial body Shape of ascospores Length (diameter) of ascospores Position of pycnidia Emergency of pycnidia Shape of pycnoconidia Length of pycnoconidia Usnic acid Atranorin Fatty acids Substance of fatty acids Secalonic acids Orcinol depsides & depsidones Substance of orcinol depsides & depsidones p-orcinol depsidones Substance of P-orcinol depsidones Inform. Y Y Y Y Y Y Y N Y Y Y Y Y Y Y Y Y N N N Y Y Y Y Y Y Y Y Y Y Y Y Y Y Y N Weight____________ States 01 86 01 1000 012 1000 01 182 012 184 012 417 0123 313 01 1000 01 120 01 250 01 167 01 417 01 63 01 0 0123 1000 01 103 0123 688 01 1000 01 1000 0 1000 012 250 01 0 01 0 012 103 01 167 0123 467 012 250 01 458 01 417 01 400 1000 0123 01 143 222 01 1000 01234 01 250 01 1000 Some clades, which can be seen on one of the equally parsimonous cladograms of the initial analysis (Fig. 1), are also distinct in all following analyses (Fig. 2). The biggest clade consists of eight A llocetraria and two D actylina species. Next clade includes five T uckneraria species. Four species of Tuckerm annopsis (the so-called T. ciliaris group) also form a separate clade, while the other members of that genus are solved differently in different analyses. The fourth clade, including three Ahtiana species, is supported by 92% of trees (characters with equal weights); trees produced by the successive approximations character weighting method always show the same Ahtiana clade. 47 7 E= -d= Cetrariafendleri Tuckermannopsis inermis Ahtiana aurescens Ahtiana pallidula Ahtiana sphaerosporella Esslingeriana idahoensis Tuckermannopsis platyphylla Tuckermannopsis microphyllica Tuckermannopsis ciliaris Tuckermannopsis americana Tuckermannopsis gilva Tuckermannopsis chlorophylla Tuckermannopsis orbata Tuckermannopsis platyphylloides Tuckermannopsis ulophylloides Tuckermannopsis subalpina Tuckneraria laureri Tuckneraria togashii Tuckneraria pseudocomplicata Tuckneraria ahtii Tuckneraria laxa Allocetraria globulans Allocetraria oakesiana Allocetraria ambigua Allocetraria stracheyi Allocetrariaflavonigrescens Allocetraria sinensis Allocetraria endochrysea Allocetraria madreporiformis Dactylina arctica Dactylina ramulosa Fig. 2 Strict consensus tree of 108 equally parsimonous cladograms (based on characters reweighted; Cetraria fendleri used to root trees). Lenght of shortest trees 35 723, consistency index 0.644, retention index 0.800. The Bremer support test was carried out next. In this test all the trees are kept successively one step longer than the shortest tree, until all the groups are lost in consensus. Bremer’s length difference has also been referred to as the decay test or decay index (Parmasto 1996, Tehler & Egea 1997). In our analysis, inclusion of trees one step longer than the shortest tree, causes the majority of the tree to collapse into a polytomy, and only the A llocetraria - D actylina clade remains separate. The latter begins to decay by the Bremer support value of three. With F lavocetraria cucullata as outgroup, 1557 equally parsimonous trees (heuristic search, all characters equally weighted, length of trees = 123 steps) were obtained. Both strict consensus tree and 50% majority-rule consensus (Fig. 3) were saved. The clade of Ahtiana , consisting of three species, is supported by 78% of trees. The clade of A llocetraria (including D actylina), which was strongly supported in the first analysis, collapses into a polytomy. Still, in the strict consensus tree, two species of D actylina form a separate clade with two A llocetraria species (A. endochrysea and A. m adre­ poriform is). The clade including Tuckneraria species is not supported by this analysis either. One new clade, which consists of 11 species of Tuckermannopsis and one species of Esslingeriana, is composed in all cladograms. When characters were reweighted by maximum value of rescaled consistency indices, five equally parsimonous trees were obtained. Strict consensus tree (Fig. 4) of them is essentially different from that achieved by the analysis where characters were with equal weights (Fig. 3); besides, some groups, similar to the clades mentioned in the first series of analysis (with Cetraria fen d leri as an outgroup), are formed. These are: the clade of three species of A htiana ; the clade of eight species of A llocetraria and two species of Dactylina-, the clade of four species of Tuckermannopsis (T. ciliaris group). Tuckneraria species do not form a clade in this analysis. Flavocetraria cucullata Ahtiana aurescens Ahtiana pallidula Ahtiana sphaerosporella Allocetraria ambigua Allocetrariaflavonigrescens Allocetraria globulans Allocetraria oakesiana Allocetraria sinensis Allocetraria stracheyi Tuckneraria ahtii Tuckneraria laureri Tuckneraria laxa Tuckneraria pseudocomplicata Tuckneraria togashii Allocetraria endochrysea Allocetraria madreporiformis Dactylina arctica Dactylina ramulosa Tuckermannopsis orbata Tuckermannopsisplatyphylloides Tuckermannopsis chlorophylla Tuckermannopsis ulophylloides Tuckermannopsis inermis Tuckermannopsis subalpina Esslingeriana idahoensis Tuckermannopsis platyphylla Tuckermannopsis microphyllica Tuckermannopsis ciliaris Tuckermannopsis americana Tuckermannopsis gilva 98 68 100 90 100 73 58 78 63 64 65 100 100 100 100 75 100 100 100 100 100 Flavocetraria cucullata Allocetraria ambigua Allocetraria stracheyi AUocetraria globulans Allocetraria oakesiana Allocetrariaflavonigrescens Allocetraria sinensis Allocetraria endochrysea Allocetraria madreporiformis Dactylina arctica Dactylina ramulosa Tuckneraria laxa Tuckneraria ahtii Tuckneraria laureri Tuckneraria pseudocomplicata Tuckneraria togashii Ahtiana aurescens Ahtiana pallidula Ahtiana sphaerosporella Tuckermannopsis orbata Tuckermannopsis platyphylloides Tuckermannopsis chlorophylla Tuckermannopsis ulophylloides Tuckermannopsis inermis Tuckermannopsis subalpina Esslingeriana idahoensis Tuckermannopsis platyphylla Tuckermannopsis microphyllica Tuckermannopsis ciliaris Tuckermannopsis americana Tuckermannopsis gilva Fig. 3. Strict consensus tree and 50% majority rule consensus tree of 1557 equally parsimonous cladograms (based on characters with equal weights; Flavocetraria cucullata used to root trees). Length of shortest trees 123, consistency index 0.415, retention index 0.650. 9 Flavocetraria cucullata Allocetraria endochrysea Allocetraria madreporiformis Dactylina arctica Dactylina ramulosa Allocetraria sinensis Allocetrariaflavonigrescens Allocetraria ambigua Allocetraria stracheyi Allocetraria globulans Allocetraria oakesiana Tuckneraria laxa Tuckneraria ahtii Tuckneraria pseudocomplicata Tuckneraria laureri Tuckneraria togashii Ahtiana aurescens Ahtiana pallidula Ahtiana sphaerosporella Tuckermannopsis orbata Tuckermannopsis platyphylloides Tuckermannopsis ulophylloides Tuckermannopsis inermis Tuckermannopsis subalpina Tuckermannopsis chlorophylla Esslingeriana idahoensis Tuckermannopsis platyphylla Tuckermannopsis microphyllica Tuckermannopsis ciliaris Tuckermannopsis americana Tuckermannopsis gilva Fig. 4. Strict consensus tree of 5 equally parsimonous cladograms (based on characters reweighted; Flavocetaria cucullata used to root trees). Length of shortest trees 43 164, consistency index 0.658, retention index 0.808. In addition, analyses with both outgroups were carried out using the parsimony jackknifer programme Jac (Farris 1995), to identify well-supported monophyletic groups. In Jac, the data are resampled with a jackknifing technique, i. e. in every replicate c. 66% of the characters are chosen at random, without replacement for parsimony analysis. The resampling procedure can be repeated up to 10 000 times, as it was done in the present study. The objective of the jackknife method in Jac is the same as that of bootstrapping in PAUP (Tehler & Egea 1997), but considered much faster than other available techniques. The resulting tree (not presented) shows that all the clades described above are not supported by this very efficient procedure, except for a group consisting of two Dactylina species, and another group comprising Allocetraria globulans and A. oakesiana. Bootstrapping (500 replicates) was used as well - on the same purpose. Three small groups were supported by this method; Allocetraria globulans - A. oakesiana; Tucker­ mannopsis americana - T. ciliaris - T. gilva; Allocetraria endochrysea - A. madre­ poriformis together with the two Dactylinas as a subclade. Discussion The aim of the present phylogenetic analysis was to search for the monophyletic groups inside the ingroup and check the correspondence of the present taxonomy to the probable evolution of taxa involved. Originally monotypic genus Ahtiana was segregated from Parmelia s. lat. on the basis of emergent pycnidia, globose ascospores, leptodermatous cortex, and presence of medullary caperatic acid (Goward 1985). Despite its parmelioid habit with laminal apothecia and pycnidia, Ahtiana sphaerosporella was shown to be closely allied to 10 Cetraria pallidula. Today the genus includes three species. In our present analysis it appears as a separate clade in most of the cladograms of equally weighted characters and also in the successive weighting strict consensus trees of both series of analyses (with different outgroups). This fact supports the latest changes in the systematics of the genus transferring C. pallidula and C. aurescens to the originally monotypic Ahtiana (Thell et al. 1995a). The genus Allocetraria, at first including only three species from high altitudes in south-east Asia, was introduced by Kurokawa & Lai (1991). It was separated from Cetraria because of the dichotomously branched lobes, the special appearance of pseudocyphellae, the palisade plectenchymatous cortex, and the unique chemistry. The authors did not pay attention to ascomatal and pycnidial characters. Later studies of these structures confirmed the necessity of a separate genus (Thell et al. 1995b). Today, ten species are combined in the genus; eight of them were included in our analyses. We have not seen the herbarium material of two taxa (A. denticulata and A. isidiigera), and their descriptions (Hue 1899; Kurokawa & Lai 1991) are too poor to present them properly in the data matrix. The value of these taxa is rather uncertain - both species are known from the type localities only; in addition, they are sterile according to literature. The paraphyletic nature of the genus with respect to two species of Dactylina is obvious. The closeness of these two genera was noticed only recently, when two former Dactylinas - A. madreporiformis and A. endochrysea - were transferred to Allocetraria (Kamefelt & Thell 1996). Presence of filiform pycnoconidia and asci with extremely broad axial body are the essential characters for separating Allocetraria. Dactylina is distinguished by the radialsymmetrical thallus becoming hollow, by the terminal position of apothecia, and also by the secondary chemistry. The palisade plectenchymatous arrangement of cortical hyphae, which is rather unusual in the group of cetrarioid lichens, is characteristic of both Allocetraria and Dactylina. The splitting of the species involved into two separate genera is not supported by our present study. At the same time, the analyses based on morphological, anatomical, and chemical characters only, do not show enough confidence when using more severe methods such as jackknifmg or bootstrapping. We share the opinion recently approved by the symposium on taxonomy, evolution and classification of lichens and related fungi (January 9-11, 1998, London) that on such occasions quick changes in nomenclature are not advisable. At the present stage of lichenological studies, additional phylogenetic analyses using the modem molecular data should be carried out, before proposing extensive nomenclatural changes. Tuckneraria includes five species. Most of them were transferred from Nephro­ mopsis to the newly described genus because of important anatomical characters (globose ascospores, Tuckermannopsis-type asci, small axial body etc.) (Randlane et al. 1994). Today the genera Nephromopsis and Tuckneraria are considered even to represent different evolutionary lines (Thell 1996) of cetrarioid lichens. The idea of close affinities of Tuckneraria and Ahtiana has also been proposed (Thell et al. 1995a). The monophyletic origin of genus Tuckneraria is supported in one series of our analyses only (Cetraria fendleri used to root trees). In the other series {Flavocetraria cucullata used to root trees), the species of Tuckneraria do not form a separate clade but are branched out successively. Genus Tuckermannopsis was described by Gyelnik (1933) in a very short manner: “Affmis generi Nephromopsi sed thallus subtus pseudocyphellis deficientibus”. Today 48 11 much more is known about the genus but the correct description is still not presented. According to different authors (Lai 1981, Hale in Egan 1987, Kurokawa 1991, Weber in Egan 1991), various species have been transferred to Tuckermannopsis’, many of them have later been combined again into other genera (Ahtiana, Allocetraria, Melanelia, Vulpicida). At present it is generally accepted that globose ascospores in narrowly clavate asci, large axial body, dumb-bell shaped pycnoconidia, and moderately small foliose brown to greenish thallus (absence of usnic acid in the cortex) are the important characters in delimiting the genus. Eleven species are now recognized in Tuckermannopsis. Our phylogenetic analyses reveal further problems with this taxon. Monophyletic origin can be declared only for the so-called Tuckermannopsis ciliaris group (T. ciliaris is also the type species of the genus). The clade consisting of four close species is strongly supported in all parsimonous trees. On the whole, the genus Tuckermannopsis, in its generally accepted treatment, should be considered paraphyletic. For instance, E. idahoensis, the sole species of the, genus Esslingeriana, is predominately connected with Tuckermannopsis platyphylla and this pair of species always branches out next to Tuckermannopsis ciliaris group. Other members of the genus do not form a distinguished group. In our opinion, any further taxonomical rearrangements in this genus are not justified before additional - preferably molecular research has been carried out. Acknowledgements This research was made possible by Grant No. LLO 100 from the Joint Estonian International Science Fund and by Grant 401 from the Estonian Science foundation. The third author was supported by the Swedish Royal Science Academy, Per Erik Lindahls Foundation. Triin Randlane is thanked for revising the English text. References Culberson, C. F. 1972. Improved conditions and new data for the identification of lichen products by a standardized thin-layer chromatographic method, - Journal of Chromatography 72: 123-125. Culberson, C. F., Culberson, W. L. and Johnson, A. 1981. A standardized TLC analysis of fi-orcinol depsidones. - Bryologist 84: 16-29. Egan, R. S. 1987. A fifth checklist of the lichen-forming, lichenicolous and allied fungi of the continental United States and Canada. - Bryologist 90: 77-173. Egan, R. S. 1991. Changes to the “Fifth checklist of the lichen-forming, lichenicolous and allied fungi of the continental United States and Canada”. - Bryologist 94: 396-400. Esslinger, T. 1971. Cetraria idahoensis, a new species of lichen endemic to western North America. - Bryologist 74: 364-369. Farris, J. S. 1995. Guide to the parsimony Jackknifer, version 4.22. Computer program distributed by the Natural History Museum, Stockholm. Gyelnik, V. 1933. Lichenes varii novi critique. - Acta pro Fauna et Flora Universalis, Ser. 2, 1:3-10. Hue, A.-M. 1899. Lichenes extra-europaei. - Nouvelles Archives du Museum (Paris) I, 4: 27-220. 12 Karnefelt, I., Mattsson, J.-E. & Thell, A. 1993. The lichen genera Arctocetraria, Cetraria and Cetrariella (Parmeliaceae) and their presumed evolutionary affinities. - Bryologist 96: 394-404. Karnefelt, I. & Thell, A. 1996. A new classification for the Dactylina/Dufourea complex. - Nova Hedwigia 91: 595-605. Kurokawa., S. 1991. Japanese species and genera of the Parmeliaceae. - Journal of Japanese Botany 66:152-159. Kurokawa, S. & Lai, M.-J. 1991. Allocetraria, a new genus in the Parmeliaceae. Bulletin of the National Science Museum (Tokyo), Ser. B, 17: 59-65. Lai, M-J. 1981 [1980]. Studies on the cetrarioid lichens in Parmeliaceae of east Asia. I. Quarterly Journal of the Taivan Museum 33: 215-229. Maddison, W. P. & Maddison, D. R. 1992. MacClade: Analysis of Phylogeny and Character Evolution, version 3. Sinauer Associates, Sunderland. Randlane, T., Saag, A., Thell, A. & Karnefelt, I. 1994. The lichen genus Tuckneraria Randlane & Thell - a new segregate in the Parmeliaceae. - Acta Botanica Fennica 150: 143-151. Randlane, T., Saag, A. & Thell, A. 1997. A second updated world list of cetrarioid lichens. - Bryologist 100(1): 109-122. Saag, A. & Randlane, T. 1995. Phylogenetic affinities of cetrarioid lichens. Cryptogamie Botany 5(2): 128-136. Swofford, D. L. 1993. PAUP: Phylogenetic analysis using parsimony, version 3.1.1. Computer program distributed by the Illinois Natural History Survey. Tehler, A. & Egea, J. M. 1997. The phylogeny of Lecanactis (Opegraphaceae). Lichenologist 29: 397-414. Thell, A. 1996. Anatomy and taxonomy of cetrarioid lichens. Summary of doctoral dissertation. Department of Systematic Botany, Lund University. Thell, A., Goward, T., Randlane, T., Karnefelt, E. I. & Saag, A. 1995a. A revision of the North American Lichen genus Ahtiana (Parmeliaceae). - Bryologist 98(4): 596-605. Thell, A., Randlane, T., Karnefelt, T., Gao, X.-Q. & Saag, A. 1995b. The lichen genus Allocetraria (Ascomycotina, Parmeliaceae). - In: Daniels, F. J., Schulz, M. & Peine, J. (eds.). Flechten Follmann. Contributions to lichenology in honour of Gerhard Follmann. University of Cologne, Germany, 353 - 370. 13 CURRICULUM VITAE Andres Saag Bom in Tartu, Estonia, November 21, 1955. Married. Estonian citizen. Address: Institute of Botany & Ecology, University o f Tartu, Lai St. 38, Tartu, EE2400 Estonia. Tel.: +372 7 431 252, fax: + 372 7 441 272, e-mail: asaag@ut.ee Education 1974 graduated fromTartu Secondary School No. 5. 1974-1979 University of Tartu, Faculty o f Geography & Biology, speciality — ecology. 1993-1998 Ph. D. student in the Institute of Botany & Ecology, University of Tartu. Professional employment 1979-1981 Institute of Forestry. 1981-1991 Laboratory of Bioindication, University o f Tartu. 1991-1998 Institute of Botany & Ecology, University of Tartu. Fields of research 1. Systematics and chemistry of cetrarioid lichens (fam. Parmeliaceae). 2. Lichen flora of Estonia, mapping, and databasing of lichenological herbarium. Number of scientific publications: 25. Results have been presented at international symposia in Latvia, Lithuania, Russia, Ukraine, Sweden, France, Austria. Other scientific activities 1. Supervisor o f 2 undergraduate and 1 postgraduate students. 2. Co-editor of journal Folia Cryptogamica Estonica since 1997. 3. Member o f International Association for Lichenology since 1992. 49 193 CURRICULUM VITAE Andres Saag Siindinud 21. novembril 1955. a. Tartus; abielus; eestlane. Aadress: Tartu Ulikooli botaanika ja okoloogia instituut, Lai t. 38, Tartu, EE2400 Eesti. Tel.: (27) 431 252, faks: (27) 441 272, e-mail: asaag@ut.ee Haridus 1974 Tartu V Keskkool. 1974-1979 Tartu Ulikooli bioloogia-geograafiateaduskond, okoloogia eriala. 1993-1998 Tartu Ulikooli botaanika ja okoloogia instituudi doktorant. Teenistuskaik 1979-1981 Eesti Metsainstituudi looduskaitse labor. 1981-1991 Tartu Ulikooli bioindikatsiooni labor. 1991-1998 Tartu Ulikooli botaanika ja okoloogia instituut. Uurimisvaldkonnad 1. Tsetrarioidsete samblike (sug. Parmeliaceae) taksonoomia ja samblikuainete keemia. 2. Eesti lihhenofloora, samblike kaardistamine, lihhenoloogilise herbaariumi andmebaasi loomine. Teaduspublikatsioonide iildarv: 25. Uurimistoo tulemusi olen esitanud rahvusvahelistel konverentsidel Latis, Leedus, Venemaal, Ukrainas, Rootsis, Prantsusmaal, Austrias. Muu erialane tegevus 1. Kahe uliopilase ja iihe magistrandi juhendaja. 2. Ajakirja Folia Cryptogamica Estonica kaastoimetaja alates 1997. 3. Rahvusvahelise Lihhenoloogia Assotsiatsiooni liige alates 1992. 194 DISSERTATIONES BIOLOGICAE UNIVERSITATIS TARTUENSIS 1. Toivo Maimets. Studies of human oncoprotein p53. Tartu, 1991, 96 p. 2. Enn K. Seppet. Thyroid state control over energy metabolism, ion transport and contractile functions in rat heart. Tartu, 1991, 135 p. 3. Kristjjan Zobel. Epifiiiitsete makrosamblike vaartus ohu saastuse indikaatoritena Hamar-Dobani boreaalsetes magimetsades. Tartu, 1992,131 lk. 4. Andres Mae. Conjugal mobilization of catabolic plasmids by transposable elements in helper plasmids. Tartu, 1992, 91 p. 5. Maia Kivisaar. Studies on phenol degradation genes of Pseudomonas sp. strain EST 1001. Tartu, 1992, 61 p. 6. Allan Nurk. Nucleotide sequences of phenol degradative genes from Pseu­ domonas sp. strain EST 1001 and their transcriptional activation in Pseu­ domonas putida. Tartu, 1992, 72 p. 7. UIo T'amm. The genus Populus L. in Estonia: variation of the species biology and introduction. 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