P1. Syst. Evol. 183: 113- 160 (1992) --Plam Systematics and Evolution © Springer-Verlag 1992 Printed i~ Austria Evolution and phylogeny of cetrarioid lichens I. KARNEFELT, J.-E. MATTSSON, and A. THELL Received February 14, 1992; in revised version April 3, 1992 Key words: Ascomycot&a, Lecanorales, Parmeliaceae, cetrarioid genera.- Cladistics, asci, conidiomata. Abstract: The ACHARIANgenus Cetraria has not been scrutinised in the same way as the genus Parmelia in terms of circumscription of segregate genera. A few generic names have been introduced, but mainly in checklists without any real indication of what these names stand for. After a detailed investigation of characters in the apothecia, conidiomata, anatomical structures, as well as morphology and secondary chemistry, it is clear that several, more or less distinctive, phylogenetic lines exist among taxa which earlier were accommodated in Cetraria. Some of these distinctive phylogenetic lines are distinguished by ascus characters: (1) clavate asci with a small axial body, an apical ring structure in the tholus and ellipsoid ascospores; (2) uniseriate asci with a large axial body and globose ascospores; and (3) taxa with broadly clavate asci with a large axial body and ellipsoid ascospores. Sixty-three character states were studied and results from the character analyses were evaluated by cladistic analysis against 43 examined terminal taxa. Seven further taxa were included in one analysis. Results from the cladistic analyses give rather strong support for a new systematic treatment of at least certain groups in the cetrarioid lichens. A formal systematic arrangement is not made here but will be presented elsewhere. The characters investigated are illustrated in 78 half tone pictures and one line drawing. Five cladograms are presented. Cetraria sensu ACHARIUS The generic name Cetraria was originally introduced by ACHARIUS (1803) and separated from Parmelia on the marginal position o f apothecia and pycnidia. This type o f systematic judgement, based on very simple diagnostic characters, was c o m m o n at this important period o f botanical history, and most o f the older, wellknown lichen genera were recognized and described in this simple manner. A problem with m a n y o f these older genera is that they are paraphyletic groups and do not correspond to natural monophyletic groups. The original eight taxa which ACHARIUS (1803) included in Cetraria, i.e., C. islandica, C. cucullata, C. nivalis, C. lacunosa, C. fallax, C. glauca, C. sepincola, and C. juniperina, almost all have very weak affinities to each other. The segregation of parmelioid foliose and other unrelated groups F r o m 1803 until the present, some 100 taxa were described in Cetraria. Some o f the most diverse forms, altogether 24 species, are now recognized as distinct genera, 114 I. KXRNEFELT8~ al.: i.e., Asahinea (CULBERSON• CULBERSON1965), Cetrelia (CULBERSON& CULBERSON 1968), and Platismatia (CULBERSONt~ CULBERSON 1968). These taxa differed in the foliose habit, types of cortical layers, and the secondary chemistry. Earlier, the position of the apothecia was often used as a character to identify cetrarioid taxa, however, we have found this character to be very variable and of less importance. In the parmelioid Cetrariae discussed here, apothecia are developed close to the margins or submarginally of the upper surface; occasionally some are lamina1. In addition to these well-known segregates of the genus Cetraria, several other species groups also have been separated at generic level, a position which, however, has not been widely accepted, for example, Cetrariopsis (KUROKAWA 1980), Cetreliopsis (LgI 1980), Esslingeriana (LAI 1980), Nephromopsis (MOLLER 1891), and Tuckermannopsis (GYELNIK1933). The latter two widely misunderstood genera, Nephromopsis and Tuckermannopsis, are based on very weak definitions. In addition, anatomical characteristics have generally been misinterpreted in the most recent accounts of generic segregates. The name Nephromopsis has been used occasionally for the separation of a large group of foliose corticolous species richly developed in SE Asia (LAI 1980, PARK 1990). However, in other accounts a similar group of species was maintained in Cetraria sensu lato (AWASTHI1982, 1988; YOSHIMURA 1982). The genus Nephromopsis was characterized, inter alia on the nephromoid apothecia, i.e. the apothecia develop marginally on the lower surface. Pseudocyphellae, in addition, appear to be developed on the lower surface as they are in the true Cetrariae. The rather weakly defined Asiatic genus Cetreliopsis (LAI 1980) characterized by a blackish under surface with small punctae, presumably also belongs in this aggregate. Tuckermannopsis was recently used in several works and checklists (EGAN 1987, HALE & COLE 1988), being applied to species separated from Nephromopsis which lack pseudocyphellae on the lower surface by GYELNIK (1933). Several presumably unrelated taxa have now been combined in Tuckermannopsis without any additional evidence for close affinities between them. Closely allied to these taxa is presumably also the monospecific Esslingeriana (LAI 1980), recognized only on such characters as a blackish lower surface, which lacks rhizines. Erect, brown species of Cetraria K~RNEFELT (1979) outlined a possible basis for the future circumscription of Cetraria in particular, centred around 14 species characterized by an erect foliose habit. Apothecia and conidiomata are developed marginally on the lobes of this group of species. In addition a number of other important characters; the generally canaliculate shape of the lobes; the presence ofpseudocyphellae on the lower surface; marginal projections which in most cases bear pycnidia; brown pigmentation; and the generally pachydermatous, paraplectenchymatous outer cortical layers overlying an inner layer of periclinally arranged hyphae, suggest that these species form a rather natural group (K,~RNEFELT1979). Earlier, Cetraria richardsonii was also generally associated with this group. However, it showed a number of rather unique characteristics such as an extremely thick upper cortical layer largely composed of periclinally arranged prosoplectenchymatous hyphae and large ecorticate areas on the lower surface, characters serving to separate it as a distinct genus, Masonhalea (K,~RNEFELT1977). Phylogeny of cetrarioid lichens 115 Evolutionary change and traditional taxonomy in Cetraria After separation of the major foliose species groups, i.e. Cetrelia, Platismatia, and the nephromoid entities, in addition to the typical erect foliose species group related to Cetraria islandica, the genus Cetraria still includes a number of species groups which are at present not clearly defined. This portion of Cetraria, which appears to have only few characters in c o m m o n with the species related to C. islandica, shows many important structural and chemical features, possibly the result of an ancient development in other lines and directions in response to changing environmental conditions. Three major groups are described in detail below and referred to briefly here as: (1) Taxa with an "apical ring structure", including mainly the C. islandica group; (2) taxa with "uniseriate asci", including mainly Tuckermannopsis and related entities; and (3) taxa with broadly "clavate asci", including, e.g., the C. commixta- and C. juniperina groups. Some of these aggregates also show stronger affinities to quite different taxa such as Bryoria sect. Subdivergentes and Melanelia rather than with Cetraria. The problems we must address therefore, are those inherent in forging a new systematic framework, different from the traditional view followed in floristic treatments and arrangements in many herbaria worldwide, and the impact that this changed framework will have on conventional lichen taxonomy. Material and methods Results are based on herbarium material from ALA, BM, BP, CANL, COLO, H, FH, G, GZU, L, LAM, LE, LD, M, MB, NY, O, S, SFSU, TUR, UBC, UME, UPS, US, and WIS in addition to private herbaria kept for the present time in UPS by GAO. During a visit to the Soviet Union during the fall of 1991 the senior author also had the possibility to study a large herbarium kept in the university of Vladivostok and a large private collection kept by IRINA SKIRINAat an associated department. A further large private collection was also studied in Lazo, Primorsky Kray, kept by SVETLANACHABANENKO.During the same visit the senior author had the possibility to become acquainted in the field with some of the large lobed foliose entities studied in this work. Anatomical studies are based on our own work, and literature references have only been used for comparison. For anatomical observations, fragments of lichens were sectioned with a Kryomat, Leitz freezing microtome and sections in lactophenol cottonblue. After pretreatment with 10% KOH-solution, asci were squashed in a 0.3% Lugol's solution. Hymenial characters were studied with a Zeiss Axioscope light microscope, and photomicrographs made with a Zeiss M 35 W camera. Conidiomata were also examined in detail, as well as anatomical sections of thalli for the specific structure of cortical layers. Approximately 500 microscopical slides were prepared. Chemical data are also based mainly on our own results from standardized TLC methods. Cladistic analyses were carried out according to Hennigian principles using methods discussed by DUNCAN& STUESSY (1984), ELDREDGE• CRACRAFT(1980), and HUMPHRIES & FUNK (1984). Apomorphies are discussed under the chapter Character states below. Synapomorphies were established through out-group comparison, and analysed in the Hennig 86 and PAUP package systems, run on a COPAM computer with MS-DOS and on a Macintosh SE. Taxa analysed Almost 60 taxa were examined using methods described above of which 50 listed here in alphabetical order were included in the cladistic analyses as terminal taxa" Bryoria ab- 116 I. K~RNEFELT• al.: breviata, Cetraria alvarensis, C. andrejevii, C. arenaria, C. aurescens, C. australiensis, C. californica, C. canadensis, C. chlorophylla, C. commixta I, C. commixta II, C. commixta III, C. cucullata, C. delisei, C. everniella, C. ericetorum, C.fastigiata, C.fendleri, C. hepatizon, C. inermis, C. islandica, C. juniperina, C. kamczatica, C. laevigata, C. laureri, C. merrillii, C. nepalensis, C. nigricans, C. nigricascens, C. nivalis, C. oakesiana, C. odontella, C. orbata, C. pallidula, C. pinastri, C. platyphylla, C. sepincola, C. tilesii, C. viridis, Cetrariopsis wallichiana, Coelocaulon aculeatum, C. crespoae, C. muricatum, C. steppae, Cornicularia normoerica, Esslingeriana idahoensis, Masonhalea richardsonii, Nephromopsis strachei, Tuckermannopsis americana, and T. cilaris. Additional material examined included: Bryocaulon divergens, Bryoria trichodes, Cetraria subalpina, Cetrelia cetrarioides, Nephromopsis endocrocea, N. ornata, N. rugosa, Parmelia saxatilis, and Platismatia glauca. Further work The results presented here are introductory data only and will be followed by other papers on the systematics and evolution of the groups studied. Formal systematic changes of generic names and/or new combinations of taxa therefore have not been made here. Manuscripts on the systematics of Cetraria sensu str. (KARNEF~LT& al., unpubl.) and on the vulpinic acid-containing species of Cetraria (MATTSSON, unpubl.) are in preparation. A survey is to be published of the genus Tuekermannopsis (MATTSSON,unpubl.). Morphology, anatomy, and chemistry Character states Sixty-three different character states were analyzed in this survey of the cetrarioid groups for evaluation of phylogenetic affinities among various taxa. The characters are listed below with plesiomorphic states vs. apomorphic states, including brief comments on their occurrence (Table 1). Structural thalline characters 1. Thallus dorsiventral, with an upper and lower cortex, and an algal layer mainly concentrated on one side/vs, more or less terete, i.e. with only one type of cortex and algal layer on all sides. The apomorphic state is characteristic of Bryoria, Coelocaulon, and Cornicularia. 2. Thallus more or less rosette-forming, with at least the central portions adnate/ vs. thallus with erect lobe portions, i.e. all parts mainly vertical. The apomorphic state is characteristic for species of Cetraria sensu str. and for Coelocaulon. 3. Lobes more or less broad, the length about the same as the width/vs, lobes much longer than wide, often with long branches. The apomorphic state is characteristic for most examined entities apart from C. pallidula, C. aurescens, Cetrariopsis, and Nephromopsis. 4. Lobes adnate, lobe margins very close to the substrate/vs, at least one third of the lobe tips raised from the substrate. The apomorphic state is characteristic for all groups apart from the C. commixta group, C. pinastri, and C. oakesiana. 5. Pseudocyphellae present on the lower surface/vs, pseudocyphellae absent on the lower surface. The apomorphic state is characteristic for taxa with uniseriate and broadly clavate asci. 6. Pseudocyphellae absent from the upper cortex/vs, pseudocyphellae present on the upper cortex. The apomorphic state is characteristic for terete entities (by definition). Phylogeny of cetrarioid lichens 117 7. Pseudocyphellae present along margins of the lower surface/vs, pseudocyphellae absent from margins of the lower surface. The apomorphic state is characteristic for taxa with uniseriate and broadly clavate asci. 8. Marginal projections absent/vs, marginal projections present. The apomorphic state is characteristic for taxa with an apical ring structure in the tholus, apart from C. nivalis. 9. Soredia absent/vs, soredia present. Anatomical characters 10. Upper cortex one-layered/vs, upper cortex composed of two distinctive layers. The apomorphic state is characteristic for species of Cetraria sensu str., Coelocaulon, Cornicularia, and Masonhalea. 11. Isodiametric hyphal cells absent in external cortical layer/vs, external cortical layer composed of isodiametric hyphal cells. The apomorphic state is characteristic for C. merrillii and Bryoria abbreviata. 12. At least some hyphal cells in the upper cortex with large lumina/vs, all hyphal cells in the upper cortex with very small lumina. The apomorphic state is characteristic for C. delisei, C. fastigiata, C. kamczatica, and C. tilesii. 13. All cells in the upper cortex of about the same size/vs, external cortical hyphal cells with very small lumina. The apomorphic state is characteristic for the shikimic acid derivative-containing taxa and for C. sepincola and C. oakesiana. 14. Algal layer thin/vs, algal layer very thick. The apomorphic state is only characteristic for the Cetraria eommixta group. 15. Algal layer absent beneath the excipulum/vs, algal layer present beneath the excipulum. 16. Algae beneath or surrounding the pycnidia/vs, no algae present close to the pycnidia. The apomorphic state is mainly characteristic for taxa with uniseriate asci except for the C. aurescens group. The state is also characteristic for taxa with broadly clavate asci and for the main part of Cetraria sensu str. Apothecial position 17. Apothecia otherwise placed/vs, apothecia lateral, placed at the margins far from the tip. The apomorphic state is characteristic for C. everniella and for terete taxa. 18. Apothecia otherwise placed/vs, apothecia frequently laminal, far from the lobe tips. The apomorphic state is characteristic for Cetrariopsis and for the C. commixta group. 19. Apothecia otherwise placed/vs, apothecia terminal at the lobe tips. The apomorphic state is characteristic for C. californica, Coelocaulon, and Cornicularia. 20. Exciple 3-layered/vs. exciple 1-2-layered. The apomorphic state is characteristic of all taxa except for Cetraria ericetorurn, C. islandica, Coelocaulon, Masonhalea, and Nephromopsis. 21. Lowest layer in exciple gelatinized to the same degree as other portions of the thallus/vs, lowest layer in exciple strongly gelatinized. The apomorphic state is characteristic for the C. juniperina group, Bryoria abbreviata, and C. californica. 118 I. KAiRN~FELT& al.: Anatomical and chemical characters of ascus and excipulum 22. Crystals absent or whitish crystals present in the exciple/vs, yellow crystals present in the exciple. The apomorphic state is characteristic for the entities with secondary products of shikimic acid type. 23. Crystals absent or yellow crystals present in the exciple/vs, whitish crystals present in the exciple. The apomorphic state is characteristic for Esslingeriana, Platismatia glauca, and C. sepincola. 24. Asci cylindrical or slenderly clavate/vs, asci broadly clavate. The apomorphic state is characteristic for a large group of species. 25. Asci of other type/vs, asci very broadly clavate. The apomorphic state is characteristic for C. fendleri, C. sepincola, and C. tilesii. 26. Asci of other type/vs, asci strictly uniseriate. The apomorphic character is characteristic for a large group of species and for C. andrejevii. 27. Tholus with normal amyloid reaction/vs, tholus with very strong amyloid reaction (IKI dark blue). The apomorphic state is characteristic for C. alvarensis, C. juniperina, C. pinastri, and C. tilesii. 28. No special amyloid structure in tholus/vs, amyloid ring structure present in tholus. The apomorphic state is characteristic for species of Cetraria sensu str. 29. Axial body of small to intermediate type/vs, axial body large. The apomorphic state is characteristic for taxa with uniseriate and broadly clavate asci. 30. Axial body of intermediate size to large/vs, axial body very small. The apomorphic state is mainly characteristic for species of Cetraria sensu str. 31. Ascospores oblong-ovate to ellipsoidal/vs, ascospores globose. The apomorphic state is characteristic for the entities with uniseriate asci. 32. Spores oblong-ovate/vs, spores globose to ellipsoid. Apomorphic state characteristic in all examined entities apart from those in Cetrariopsis, Cornicularia, and Nephromopsis. Conidiomatal characters 33. Pycnidia exclusively marginal/vs, at least some pycnidia laminal. The apomorphic state is common among the taxa characterized by broadly clavate asci, e.g., the C. commixta group, the group with mainly terete lobes and in C. canadensis and C. viridis. Pycnidia in more or less apical position and on lichens with terete lobes have also been interpreted as laminal. 34. Pycnidia raised above the cortex or immersed/vs, pycnidia on projections. The apomorphic state occurs in taxa with narrowly clavate asci, except for Masonhalea. 35. Pycnidia raised on projections/vs, pycnidia immersed i.e. the main tissue is located beneath the cortex. The apomorphic state is characteristic for all terete groups except Cornicularia. 36. Pycnidial wall l-layered/vs, pycnidial wall 2-layered. The apomorphic state is characteristic for the C. islandica- and for the C. commixta groups. 37. Pycnidial wall I-layered or 2-layered with a thin outer layer, c. less than 5 gm thick/vs, outer layer much thicker, 10-60 gm. 38. Pycnidial wall without pigmentation/vs, brown pigments in the gelatinous matrix of the pycnidial wall. The apomorphic state is characteristic for groups with uniseriate and broadly clavate asci. Phylogeny of cetrarioid lichens 119 39. Pycnidial wall without pigmentation or brown/vs, black pigments at least in some other parts of the pycnidial wall than around the ostiole. The apomorphic state occurs among taxa with shikimic acid pathway metabolites. 40. Pycnidial wall without yellowish crystals/vs, yellow crystals in pycnidial wall. The apomorphic state occurs in taxa with shikimic pathway metabolites. 41. Medullary layer attached directly beneath the pycnidial wall/vs, cortical tissue beneath the pycnidial wall. The apomorphic state is characteristic for groups with narrow clavate asci, and for C. inermis, C. chlorophylla, C. oakesiana, C. tilesii, and C. viridis. 42. Conidia of other shape/vs, conidia filiform. The apomorphic state is characteristic for the C. everniella group. 43. Conidia of other shape/vs, bottle-shaped. The apomorphic state is characteristic for C. alvarensis, C. juniperina, C. pinastri, and C. tilesii. 44. Conidia of other shape/vs, citriform. The apomorphic state is characteristic for C. canadensis, C. viridis, C. commixta, and C. andrejevii. 45. Conidia of other shape/vs, bacillariform. The apomorphic state is characteristic for Cetraria sensu str. It should be noted in characters 41-44 that more than 75% of the conidia should be of a shape other than the normal bifusiform type if a species is to be coded for the apomorphic state. 46. Conidia longer than 5 gm/vs, conidia up to 4 gm long. The apomorphic state is characteristic for C. eanadensis, C. viridis, C. commixta, and Masonhalea. Characters in the secondary chemistry 47. Aliphatic substances absent/vs, aliphatic substances present. The apomorphic state is characteristic for taxa with narrow clavate asci and for Masonhalea. 48. Caperatic acid absent/vs, caperatic acid present. 49. Protolichesterinic- and lichesterinic acids absent/vs, protolichesterinic or lichesterinic acids present. 50. Norrangiformic- and rangiformic acids absent/vs, norrangiformic or rangiformic acids present. 51. Orcinol depsides present/vs, orcinol depsides absent. The plesiomorphic state is characteristic for C. delisei group, C. everniella, Nephromopsis stracheyi, and Tuckermannopsis ciliaris. 52. Gyrophoric and hiascic acids absent/vs, gyrophoric and hiascic acids present. The apomorphic state is characteristic for the C. delisei group. 53. Olivetoric acid absent/vs, olivetoric acid present. 54. Orcinol depsidones absent/vs, orcinol depsidones present. 55. Alectoronic acid absent/vs, alectoronic acid present. 56. a-collatolic acid absent/vs, ct-collatolic acid present. 57. J3-orcinol depsides absent/vs. J3-orcinol depsides (i.e. chloratranorin or atranorin) present. 58. J3-orcinol depsidones absent/vs. J3-orcinol depsidones present. The apomorphic state is characteristic for C. hepatizon, C. islandica, and C. laevigata. 59. Fumarprotocetraric acid absent/vs, fumarprotocetraric acid present. The apomorphic state is characteristic for C. islandica and C. laevigata. 60. Protocetraric acid absent/vs, protocetraric acid present. The apomorphic state is characteristic for C. islandica. 120 I. KARNEVELT& al.: 61. Stictic acid absent/vs, stictic acid present. 62. Usnic acid present/vs, usnic acid absent. 63. Shikimic acid derivates absent/vs, shikimic acid derivates present. The apomorphic state is characteristic for the C. juniperina group. C o m m e n t s on s o m e c h a r a c t e r s Characters of the reproductive structures, i.e. true ascomycete characters such as ascus, hamathecial and conidiomatal structures, must be considered of major importance in the evaluation of higher systematic categories (KAI~NEVELT& Tr~ELL 1992). However, we must also emphasize that structural thalline characters cannot be entirely neglected in the circumscription of generic entities since distinctive structural changes must be connected to basic genetical differences in the course of the evolution. Thallus morphology The observed morphological variation in thalli of Cetraria is very wide. Two main types of thallus are discerned; firstly a generally more foliose type and secondly a more erect type. There are also many intermediate types of thallus organization, but basically material can be sorted into these two groups. 1. Foliose types seem to be more common, comprising taxa with more or less typically foliose, dorsiventral, horizontally spreading, somewhat rosette-forming, with more or less broad lobes, which may project a little from the substrate but which are otherwise adpressed and fastened to the substrate with scattered rhizinae. Pseudocyphellae may be present in the group both on the upper or on the lower surface, or along the margins. Soredia and cilia also occasionally occur on the margins in certain taxa (Figs. 2-3, 10-13, 15). 2. In eret types the thallus is basically organized into erect to suberect rather longish dorsiventral lobes (Fig. 6). Dorsiventral lobes in addition differentiate in upper and lower surfaces, where the upper surface may be partly hidden through development of a more or less canaliculate or subtubular structure (Figs. 7, 9). Rarely lobes are more terete in structure (Figs. 5, 14). These forms are not attached to the substrate by any means, but lobes continue to grow progressively upwards, dying off basally at the same time. Pseudocyphellae generally occur on the lower surface or on the margins of the lower surface but rarely on the upper surface (Figs. 4, 8). Cilia occur along the margins of a few species, but soredia are only very occasional (Figs. 10, 15). Lobe colour in both groups is partly the result of secondary metabolites, such as yellowish taxa having usnic acid deposited in the cortical layers (Fig. 7). A majority of the entities examined, however, are characterized by more brownish, greyish or blackish pigments of unknown chemical origin (K~;RNEFELT 1986). A n a t o m y o f the c o r t e x Anatomical structure is more or less correlated with the two basic structural types of thallus, i.e. two different anatomical types are seen. 1. In foliose types lobes are organized into an upper cortical layer, a photobiont layer, a medullary layer, and a lower cortical layer; both cortical layers are composed Phylogeny of cetrarioid lichens 121 of pachydermatous paraplectenchymatous hyphal cells (Figs. 16-17, 22-29). This type of cortical layer is, with very few exceptions, characteristic for all groups of foliose lichenized ascomycetes, i.e. also outside the Parmeliaceae. 2. In species characterized by erect to suberect lobes the anatomical organization is mainly of the same type, with upper and lower cortical layers, photobiont layer and medullary layer. A difference, however, is seen in the cortical layer where occasionally the cortices are organized in one external layer of pachydermatous hyphal cells overlying a layer of periclinally arranged prosoplectenchymatous hyphae (Figs. 18-21). In true fruticose groups, such as Alectoria, Bryoria, and Teloschistes, the cortical layers are formed by periclinally arranged prosoplectenchymatous hyphae. Thus, in the intermediately organized erect foliose thallus structure of Cetraria the anatomical organization could represent an intermediate form between true foliose and fruticose groups. Apothecia The importance of the position of the apothecia has previously been emphasized, particularly in the separation of genera in this group of lichens (AwAsTHI 1982, KUROKAWA 1980, LAX 1980, R~S/~NEN 1933). We have, however, found this character to be too variable in the material studied and of minor importance. Most commonly, apothecia appear to be placed in a type of marginal position and then arranged towards the upper surface in this group of species, i.e. in the erect foliose group of species and in many foliose groups (Figs. 10-12). The so-called nephromoid stage is more rare, i.e. apothecia develop on the margin but the disc is turned downwards towards the lower surface. This stage particularly has been recognized as of major importance in separation of the genus Nephromopsis. In addition to marginally oriented apothecia, laminal apothecia also occur in a few entities such as in Cetrariopsis (Fig. 3), but also in some species of Cetraria, i.e. in C. canadensis, C. viridis, and the C. commixta group. Asei The investigation of asci in this group of lichens revealed more astonishing data than we would have expected, in a way similar to the variation we discovered in Fig. 1. Schematic drawings of the three main types of ascus tips in cetrarioid genera. A Cetraria islandica (Cetraria-type) with a moderately large tholus (th), a conical ocular chamber, (oc), a very small axial body (ab), and a distinct ring structure (rs). B Tuckermannopsis ciliaris (Tuckermannopsis-type) showing a rather small tholus, a very broad ocular chamber and a very broad axial body. The broad axial body is the most common type in the lichen family Parmeliaceae. C Nephromopsis stracheyi (Nephromopsis-type) with a large tholus, a conical ocular chamber, and an axial body of intermediate type. Bars: 10 ~tm 122 I. KARNEFELTt~¢ al.: Phylogeny of cetrarioid lichens the alectorioid taxa (K.~RNEFELT• THELL 1992). Asci varied not only in general structure and shape, but also in the apical portions and the amyloidity to a degree which can hardly be accepted within the variation o f a genus. Concerning the general shape of asci, three different types were seen. The structure of the axial b o d y and the general amyloidity in the tholus are variable in i m p o r t a n t characters (Figs. 1, 42-61). In addition the ocular chamber showed a certain variation, however, this variation was more indistinct to be estimated in terms of character states. 5 Figs. 2-7. Morphology in cetrarioid genera. - Fig. 2. Nephromopsis rugosa with a nephromoid apothecium, Japan 1958, KUROKAWA 58549 (M). - Fig. 3. Cetrariopsis wallichiana, showing numerous, small, laminal apothecia, Nepal, C. Himalaya, MIEHE 15615 (GZU). - Fig. 4. Masonhalea richardsonii. Note whitish, pseudocyphellae-like structures on the lower surface, Alaska, VIERECK & JONES 5634 (LD). - Fig. 5. Fertile specimen of Coelocaulon acuIeaturn, Sweden, Blekinge 187t, SVANLUND(LD). - Fig. 6. Inland form of Cetraria arenaria, Michigan, IMSHAUG 20321 (WIS). - Fig. 7. A yellow usnic acidcontaining species of Cetraria, Cetraria cucullata, Alaska, MATTSSON 16t8a (LD). Bars: 10ram 9 13 1, 15 Figs. 8-15. Morphology in cetrarioid genera. - Fig. 8. Cetraria islandica, showing pseudocyphellae on lower surface, Finland, Tuusiniemi 1924, RASXNEN (H). - Fig. 9. Erect, canaliculate lobes of Cetraria fastigiata, Canada, Franklin Distr., 1985, EDLUND 95967 (LD). - Fig. 10. Cetraria chlorophylla, with typically sorediate margins, V/irmland 20/9 1954, SUNDELL (S). - Fig. 11. Tuckermannopsis ciliaris, Oregon 1961, HENSSEN 13585C (MB). - Fig. 12. Esslingeriana idahoensis, showing the extremely black and rugose lower surface, Idaho 1978, ANDERE~a 10489 (LD). - Fig. 13. Cetraria hepatizon, a brown, flatfish species of Cetraria, Austria, Ostalpen, HAFELLNER 3118 (GZU). - Fig. 14. Cetraria californica, frequently with terminal apothecia, Washington, Baylaity 1952, BECK~N~ 5789 (LAM). - Fig. 15. Sorediate, vulpinic acid-containing species, Cetraria pinastri, V~irmland 13/4 1950, SUNDELL (LD). Bars: 10mm 124 I. K~RNEFELT~¢ al.: Ascus shape 1. Asci rather slender, narrowly clavate to cylindrical. This type occurs in Cetraria sensu str. (Figs. 45-53). 2. Ascospores arranged more or less uniseriately in the group of subfoliose entities related to Tuckermannopsis (Figs. 34-36, 54-58). 3. Asci with a more broadly clavate form, occurring in the Cetraria commixta group, the Cetraria juniperina group, and in Bryoria sect. Subdivergentes (Figs. 38-41, 60-61). Asci which are broadly clavate, or almost pear-shaped, were found in the C. fendleri group and C. tilesii (Fig. 59). Axial body 1. Axial body very small as in Cetraria sensu str. (Figs. 1, 45-53). 2. Axial body very large, occurring in most taxa having cylindrical and broadly clavate asci (Figs. 1, 54-61). 3. Axial body of intermediate type occurring in Cetrariopsis, Masonhalea, Nephromopsis, and the Cetraria delisei group (Figs. 1, 42-44). Amyloidity of tholus 1. A distinctive ring structure occurs in Cetraria sensu str. (Figs. 1, 45-53). 2. Tholus strongly amyloid, in C. juniperina group (Fig. 61). Ascospores The structure of the ascospores is partly correlated with the structure of the asci and three main types were discerned: 1. Entities with cylindrical and uniseriate asci are basically characterized by globose to subglobose ascospores (Figs. 34-36). 2. Entities with broadly clavate asci have ellipsoid ascospores (Figs. 39-40). 3. A third group, however, occurs, in the genera Cetrariopsis, Cornicularia, and the outgroup Nephromopsis which are characterized by oblong-ovate ascospores (Figs. 42-43). Hamathecium The structure of the hamathecium was not found to be very variable in comparison with results obtained from the alectorioid genera (KARNEVELT& THEI.L 1992). The hamathecium comprises a type of straight, weakly branched and septate paraphyses with rather swollen tips (Figs. 30-41). A slight variation was seen in the investigated material, however, this could not be evaluated in the cladistic analysis. Conidiomata The structure and position of pycnidia, including the shape of conidia, showed an extremely broad and also partly unexpected variation in the material examined. Position The position of pycnidia varies from strictly laminal to marginal, and they may be either immersed, raised or on projections. Laminal pycnidia appear to be most rare, occurring only in C. canadensis, C. viridis and the C. commixta group. Phylogeny of cetrarioid lichens 125 Position in relation to cortex The position of pycnidia in relation to the cortex could be divided in three different grouPs: 1. Pycnidia on projections are frequent in entities with clavate asci. 2. Pycnidia immersed, i.e. most of the pycnidium below the cortical layer, is mainly characteristic for taxa with terete thalli. 3. Pycnidia on the cortex, i.e. the early stages of pycnidial formation take place in the uppermost part of the cortex, found in C. viridis. Structure of the wall Other characters of importance in the material examined proved to be related to the structure and pigmentation of the pycnidial wall: 1. The wall is made up of one layer of isodiametric cells. 2. Two, more or less distinctive layers can be discerned, i.e. in the C. islandica sensu str. group and in the C. commixta group (Figs. 66, 71). Pigmentation of the wall 1. Most entities examined have pycnidia with, in some cases very black pigmented walls (Figs. 62-63, 70-75, 79). 2. In the C. juniperina group, in addition, yellow crystals are also found in the pycnidial wall. Conidia The shape of the conidia varies in material examined (Figs. 62-81): 1. Bifusiform conidia are the most common and widespread type (Fig. 73). 2. Rod-shaped (bacillariform) conidia occur in the C. islandica group. 3. Bottle-shaped (sublageniform) conidia occur in the C. juniperina sensu str. group and in C. oakesiana (Figs. 79-80). 4. Filiform conidia are one of the most isolated forms and they occur in the C. everniella group (Fig. 69). 5. Lemon-shaped (citriform) conidia are also rare, being found in C. canadensis, C. viridis, C. andrejevii, C. commixta, C. pallidula, and occasionally Coelocaulon aeuleatum (Figs. 63-64, 70-71, 78, 81). Secondary chemistry Secondary metabolites can be of major importance in the classification of higher categories, such as the abundance of anthraquinones in the Teloschistales (K~;RNEFELT 1989) and also in lichens in general (CULBERSON& CULBERSON1968), while in other groups such as in the alectorioid genera, secondary metabolites do not correlate so well with structural features (I~RNEFELT & THELL 1992). In the material investigated, characters from secondary chemistry appear to be better correlated with characters in the asci mainly from two different groups of species. Three major groups of substances were found: 1. Higher aliphatic substances or fatty acids. These substances such as lichesterinic, protolichesterinic, rangiformic, and norrangiformic acids are mainly characteristic of species of Cetraria sensu str., but these substances are also found in species characterized by cylindrical and uniseriate asci. 126 I. KARNEVZLT& al.: Phylogeny of cetrarioid lichens Figs. 16-21. Thallus anatomy in cetrarioid genera. - Fig. 16. Longitudinal section of Nephromopsis stracheyi, China 2428 (Herb. GAO). -- Fig. 17. Longitudinal section of Cetrariopsis wallichiana, Nepal, C. Himalaya, M~E~IE 15615 (GZU). - Fig. 18. Longitudinal section of the two-layered cortex of Coelocaulon aculeatum, Sweden, Blekinge 1871, SVANLUND (LD). - Fig. 19. Masonhalea richardsonii, longitudinal section. Note the very thick prosoplectenchymatous lower layer of the upper cortex, Alaska, VIER~CK & JONES 5634 (LD). - Fig. 20. Cetraria arenaria, longitudinal section, showing two distinct layers of the upper cortex, New York 1987, DIRIG L 2136 (LD). - Fig. 21. Cross section of the slightly two-layered cortex of Cetraria islandica, Kola Peninsula, MATTSSON, 2187a (LD). Bars: 10 gin Figs. 22-29. Thallus anatomy in cetrarioid genera. - Fig. 22. Longitudinal section of Cetraria cucullata, Alaska, MATTSSON 1618a (LD). - Fig. 23. Cetraria kamczatica, longitudinal section. Note the very strongly gelatinized cortical hyphae of the cortex, Alaska 1973, KJ~RNEFELT2223 (LD). -- Fig. 24. Cetraria nigricaseens, longitudinal section, Canada 21.7. 1972, KARNEFELT 84 1423 (LD). - Fig. 25. Gelatinized, thin, large-celled cortex of Cetrariafastigiata, Canada, Franklin Distr., 1985, EDLUND95967 (LD). - Fig. 26. Cetraria laureri, longitudinal section, Austria, Stubaier Alpen 1958, STHNER (LD). - Fig. 27. Thin cortex of Tuckermannopsis ciliaris, longitudinal section, Newfoundland, BRODO22940 (LD). Fig. 28. Esslingeriana idahoensis, longitudinal section showing the pigmented lower cortex, Idaho 1971, SC~ROZOER L 1754 (LD). - Fig. 29. Longitudinal section of Cetraria hepatizon, showing the very dark pigmented lower cortex and the thick algal layer, Austria, Ostalpen, HAFELLNER3118 (GZU). Bars in Figs. 22-27, 29:10 gm, in Fig. 28:100 gm - Figs. 30-35. Apothecial anatomy in cetrarioid genera. - Fig. 30. Cross section of apothecium of Cetrariopsis wallichiana. Note the straight, sparsely branched paraphyses and narrowly clavate asci, Nepal, C. Himalaya, MIEttE 15615 (GZU). - Fig. 31. Hymenium and three-layered excipulum of Masonhalea richardsonii, Alaska, VmP,ECK & JON~S (LD). Fig. 32. Cetraria andrejevii, cross section of apothecium, showing the strictly uniseriate arrangement of ascospores in the asci, Canada, Manitoba, M~LLER 1969 (LD). - Fig. 33. Cross section of apothecium of Cetraria islandica with a three-layered excipulum, Kola Peninsula, MATTSSON 2187a (LD). - Fig. 34. Two-layered excipulum of Cetraria platyphylla, also showing uniseriate arrangement of ascospores in the asci, British Columbia, BRODO 21865 (LD). -- Fig. 35. Cross section of apothecium of Cetraria subalpina. Note the uniseriate globose-spored asci, Michigan, IMSHAUG1876 isotype (LD). Bars in Figs. 3031, 34-35:10 gm, in Figs. 32-33:100 gm - Figs. 36.41. Hymenial and apothecial structures in cetrarioid genera. - Fig. 36. Esslingeriana idahoensis, cross section of apothecium, showing uniseriate arrangement of ascospores in the asci and the white crystals present in the two-layered excipuhim, Idaho t978, ANDZRE~6 10489 (LD). - Fig. 37. Cetraria sepincola, cross section of apothecium, showing gelatinized hyphae of the lower excipulum, Poland, ZIELINSKA 56 (LD). - Figs. 38-39. Cross section of apothecium and hymenium of Cetraria hepatizon showing rather broadly clavate asci, Austria, Ostalpen, HAFELLNER 3118 (GZU). - Fig. 40. Hymenium and exciputum of Bryoria abbreviata with strongly gelatinized lower excipulum layer and rather broadly clavate asci, California, NASR 26435 (LD). - Fig. 41. Cetraria juniperina, with a similar type of hymenium and excipulum as Bryoria abbreviata, J/imtland 1933, SANTESSON (LD). Bars in Figs. 36-37, 39-41: 10pro, in Fig. 38: 100gin Figs. 42-49. Asci in cetrarioid genera. - Fig. 42. Small asci but broad paraphyses of Nephromopsis stracheyi. Note the oblong ellipsoid ascospores, China, 2428 (Herb. GAO). Fig. 43. Asci of Cetrariopsis wallichiana, also with oblong-ellipsoid ascospores, Nepal, C. Himalaya, MIEHE 15615 (GZU). - Fig. 44. Masonhalea richardsonii, asci with the same type of axial body seen in Nephromopsis and Cetrariopsis, Alaska, VIERECK & JONES 5634 (LD). - Fig. 45. Asci with an apical ring structure, Coelocaulon aculeaturn, Sweden, Blekinge 1871, SVANLUND (LD). - Fig. 46. The same type of asci are found in Cetraria kamczatica, Alaska 1973, KARNEFELT 2223 (LD). - Figs. 47-48. Cetraria andrejevii, uniseriate ar- - rangement of ascospores in the asci with a very small tholus but a distinct ring structure and paraphyses with very broad bases, Canada, Manitoba, MILLER 1969 (LD). -- Fig. 49. Asci of Cetraria islandica, Kola Peninsula, MATTSSON 2187a (LD). Bars: 10 ~tm Figs. 50-55. Asci in cetrarioid genera. - Fig. 50. Asci of Cetraria ericetorum subsp, reticulata, typical for Cetraria sensu str. with an apical ring structure and a very small axial body, British Columbia 1977, BRODO 22073 (LD). - Fig. 51. A similar ascus type, Cetraria nigrieans, Himalaya, RAT3p2092C (GZU). - Fig. 52. Asci of Cetraria cucullata are somewhat smaller, but the apical apparatus shows the same features, Sweden, ()land 1914, ANDERSSON (LD). - Fig. 53. The same type of asci are found in Cetraria nivalis, Norway, SANT~SSON 24533 (S). - Fig. 54. Asci and paraphyses of Cetraria inermis, showing several characters not seen in Cetraria sensu str., for example globose ascospores and a large, broad axial body, Canada, Mackenzie 1972, K~RNEVZLT 2370 (LD). - Fig. 55. A similar ascus type, seen in Cetraria chlorophylla, Vfirmland 20/9 1954, SUND~LL (S). Bars: 10 ~m Figs. 56-61. Asci in cetrarioid genera. - Fig. 56. Small cylindrical asci of Tuckermannopsis ciliaris, Oregon 1961, HENSSEN 13585C (MB). - Fig. 57. Ascus of Cetrariaplatyphylla with rather large, globose spores, British Columbia, Revelstoke National Park 1970, Orro 237 (GZU). - Fig. 58. Narrowly clavate asci ofEsstingeriana idahoensis, Idaho 1978, AND~IU~GG 10489 (LD). - Fig. 59. Cetraria fendleri, ascus very broad, clavate and with ellipsoid ascospores, but structures in the apical apparatus are of Tuckermannopsis-type, CAIN 28405 (SFSU). - Fig. 60. Rather broadly clavate asci of Cetraria merilli with a very distinct axial body, California 1896, CUMMINGS2013, type (FH). - Fig. 61. Asci of Cetrariajuniperina, showing the very strong amyloid-reaction of the tholus, Sweden, J/imtland 1933, SAm~ESSON (LD). Bars: 10 ~tm Figs. 62-69. Pycnidial characters in cetrarioid genera. - Fig. 62. Part of pycnidium of Nephromopsis stracheyi with short, thick pycnoconidia, China, 2428 (Herb. GAO). -- Fig. 63. Pycnidium of Cetraria andrejevii. Note the oblong citriform pycnoconidia, Canada, Manitoba, MILLER 1969 (LD). - Fig. 64. Some pycnoconidia of Coelocaulon aculeatum are citriform, Sweden, Blekinge 1871, SVANI~UND(LD). - Fig. 65. Part of pycnidial chamber of Cetraria nivalis with bifusiform pycnoconidia, Sweden, Dalarna 22 VII 1930, HASSELROT (S). -- Fig. 66. Pycnidium of Cetraria cucullata, also showing the bifusiform pycnoconidia, Sweden, 01and 1914, ANDERSSON (LD). - Fig. 67. Short and broad pycnoconidia of Cetraria laureri, Austria, Stubaier Alpen 1958, STEINER (LD). - Fig. 68. Conidiophores and bifusiform pycnoconidia of Tuckermannopsis ciliaris, Oregon 1961, HENSSEN 13585C (MB). - Fig. 69. Cetraria everniella, pycnidium and filiform pycnoconidia, Nepal, 1986 Po~I.T 86 1773 (GZU). Bars: 10 ~tm 134 I. K~RNEFELT & al.: Figs. 70-75. Pycinidial characters in cetrarioid genera. - Figs. 70-71. Cetraria commixta, pycnidium, and citriform pycnoconidia, Czechoslovakia, Bohemia septen. 1957, VEZDA Lich. Boh. exs. (LD). - Figs. 72-73. Pycnidial characters of Cetrariahepatizon with typically bifusiforrn pycnoconidia, Ontario State Islands 1977. W~TMORE29004 (GZU). - Fig. 74. Marginal pycnidium of Esslingeriana idahoensis with typically bifusiform pycnoconidia, Idaho 1978, ANOEREO~ 10489 (LD). -- Fig. 75. Pycnidium of Bryoria abbreviata, California, NAsa 26, 435 (LD). Bars in Figs. 70-71, 73-75: 10gin, Fig. 72: 100~tm Phylogeny of cetrarioid lichens 135 Figs. 76-81. Pycnidial characters in cetrarioid genera. - Fig. 75. "Parmelia-shaped" bifusiform pycnoconidia of Bryoria abbreviata, California, NASH 26, 435 (LD). - Fig. 77. Cornicularia normoeriea, bifusiform pycnoconidia, Spain, FOLLIVIANN& CRESPOXI 1974 (LD). - Fig. 78. Citriform pycnoconidia of Cetraria viridis, Arkansas, JoHNsoN 9-27 1937 (NY). - Fig. 79. Pycnidium of Cetraria alvarensis, showing the very dark pigmented wall and the bottle-shaped pycnoconidia, Estonia, Loo alvar, MATTSSON1943a (LD). - Fig. 80. Bottle-shaped pycnoconidia of Cetraria tilesii, U.S.S.R., Irkutsk, SMIRNOF23. VIII 1926 (LE). - Fig. 81. Cetraria eanadensis, citriform pycnoconidia, California, Lake Co, TmERS 28757 (SFSU). Bars: 10~tm Secondary products originating from the shikimic acid pathway. stances are characteristic for the C. jumperina group. 3. A m o n g the aromatic substances several different types occur in the entities: a. [3-orcinol depsidones are represented by fumarprotocetraric acids. b. Orcinol depsidones are represented by alectoronic and by acids. , These subinvestigated and stictic a-collatolic 136 I. K~RNEFELT~¢ al.: Phylogeny of cetrarioid lichens c. 13-orcinol para-depsides are represented by atranorin. d. Orcinol tridepsides are represented by gyrophoric and hiascic acids. Other substances represented are usnic acids and rarely also anthraquinones. Usnic acid ~ppears to be more widespread in the entities investigated and does not correlate with any particular structural character. Polysaccharide chemistry Polysaccharide chemistry has not been investigated in this material. See also K~RNEFELT & THELL (1992). Cladistic analysis Outgroup selection One of the main principles in phylogenetic analysis apart from the definition of monophyletic groups includes the establishment of the closest relatives to a studied ingroup (DUNCAN • STUESSY 1984, ELDREDGE & CRACRAFT 1980, HUMPHRIES 8L FUNK 1984). The accepted method of outgroup comparison for assessing homologies and synapomorphies was used. Seven different cases were considered of which we found Nephromopsis stracheyi to be the most appropriate outgroup for an analysis of all taxa. Naturally, we also aimed at finding an outgroup which could be considered most closely related to the group studied. Other entities considered were Bryocaulon divergens, Bryoria trichodes, Cetrelia cetrarioides, Melanelia stygia, Parmelia saxatilis, and Platismatia glauca. 1. Parmelia saxatilis and 2. Cetrelia cetrarioides Parmelia saxatilis and Cetrelia cetrarioides were excluded as possible closest relatives to the studied ingroup mainly because of too distinct differences in the structure of asci and hamathecium. Both these entities are characterized by rather large, strongly amyloid asci and rather large, thick-walled ascospores (HALZ 1987, KARNEFECT & Tr~ELC 1992). In both Cetrelia cetrarioides and in P. saxatilis, the hamathecium consists of narrow, sparsely branched paraphyses. 3. Platismatia glauca and 4. Bryoria trichodes Platismatia glauca and Bryoria trichodes were also excluded from the discussion of a possible outgroup basically since they had no pycnidial characters. Conidiomata appear to be rare as a whole in Platismatia (CuLBERSON & CULBERSON 1968). Conidiomata are apparently also extremely rare in Bryoria (BRODO& HAWKSWORTn 1977). Asci and hamathecial structures are otherwise reminicent of those in the cetrarioid group (K~RNF.FELT & TnELL 1992). 5. Bryocaulon divergens In Bryocaulon divergens we had material with both apothecia and pycnidia. The structure of asci and conidia also recall the same structure in some cetrarioid groups. However, the major structural differences in addition to the absence of higher afiphatic substances nevertheless speak against Bryocaulon being used as the closest possible related group. Table 1. Data matrix and terminal taxa corresponding with text. 0 = plesiomorphy, 1 = apomorphy 000000000111111111122222222223333333333444444444455555555556666 123456789012345678901234567890123456789012345678901234567890123 llll?l?O0110000111011001000010011010010000000000001000000000010 001110110000100000011101001010011011011100t0000000100000000000t 011100010000001100010000010100010100000010010010011000000000010 011100010100000000000000000101010101000010001010101000000000010 C. aurescens 000110100000000000000000010010110000011000000011001000000000000 C. australiensis 01111001010000?00???????????????0101000010001010101000000000010 C. californica 1111?1?00110000100111001000010011010000000000000001000000000010 C. canadens& 001110100000100100011001000010011010011000010100001000000000001 C. chlorophylla 001110101000101100010000010010110100000010000010101000000000010 C. commixta I 001010110000011101010001000010011101100000010100001001010000010 C. commixta II 001010110000011101010001000010011101100000010100001001100000010 C. commixta III 001010110000011101010001000010011101100000010100001000000000010 C. cucullata 011100010000101100010000000101010100000010000010101000000000000 C. delisei 011100010001101100010001000000010100000000000000000100000000010 C. ericetorum 011100010100000000000000000t0101010t000010001010101000000000010 C. evernietla 001110110000000110010000010010110100010001000010100000000000010 C. fastigiata 011100010001101100010001000000010101100010000000000100000000010 C. fendteri 001010100000000100010001100010010000010000000010101000000000010 C. hepatizon 001010110000011101010001000010010101010000000000001000000100110 C. inermis 011110000000000110010000010010110000000010001010101000000000010 C. islandica 0111001101000000000000000001000t0101100010001010101000000110010 C. juniperina 001110110000100100011101001010010100011100100000001000000000001 C. kamczatica 011100010001101?00010000000101010?????????????10101000000000010 C. laevigata 011100010000000000010000000101010101000010001010101000000110010 C. laureri 0011101110001001??0100000?00?00101011000100000?0?01000000000000 C. merrilli 001110100110001100010001000010011010010000000010001000000000010 C. nepaIensis 01111001000000?00?????????????'??0101000010001010101000000000010 C. nigricans 011100010000000?0001000000010101010??????000t010101000000000010 C. nigricascens 011100010000001100010000000100010100000010000010011000000000010 C. nivalis 011100100000100100010000000101010100000010000010101000000000000 C. oakesiana 00101010100010?1????????????????0000000011000011001000000000000 C. odontella 011100010000000?0001000000010101010??????0000010101000000000010 C. orbata 001110110000000100010000010010110001010000000010101000000000010 C. pallidula 000110101000000000010000010010111000000000010111001000000000000 C. pinastri 001010111000100100011101001010010100011100t00000001000000000001 C. platyphylla 001110100000001100010000010010110010010000000010001000000000010 C. sepincola 001110100000100000010011100010010000010010000010101000000000010 C. tilesii 0011101100010000000t1101101010010100011110100000001000000000001 C. viridis 001110100000100100011001000010011000011010010100001000000000001 Cetrariopsis wallich. 000100000000001?0101000000000001?????????????710101111000000000 Coelocauton aculeat. 1111?1?1010000010010000000010101110000001000101010t000000000010 C. erespoe 1111?1?t0100000100100000000101011100000010001010101000000000010 C. muricatum 1111?1710100000100100000000101011100000010001010101000000100010 C. steppae 11t1?1?1010000?1001?????????????1t00000010001010101000000000010 Cornicularia normoer. 1111?0?001000001001100010000t0000001010000000000001000000000010 Esslingeriana idahoen. 001110100000000100010010010010100011010000000010001000001000010 Masonhalea richards. 001100000100000110000000000000010000000010001100001001100000010 Nephromopsis straeh. 000000000000000000000000000000000000000000000000000000000000000 Tuckermannopsis am. 001110110000001100010000010010110000010000000000001001001000010 T. ciliaris 0011101t0000001100010000010010110000010000000000000010001000010 Bryoria abbreviata Cetraria alvarensis C. andrejevii C. arenaria ~ II L' v " II ~' II II " V ,, II 5', II . ., GI1 . I~ v A X II Nephromopsis stracheyi Cetrariopsis wallichiana Masonhalea richardsonii C. delisei II II C. fastigiata il i I (~'1 i q C. andrejevii ! ,I II II i ~ a~ a~ C. nigricascens C. cucuUata ,i~ I~ III II II C. nivalis A C. kamczatica II I II II I ~ i, ti I! II II II II ,.. ¢~ ~ ~1 ii C. nigricans Coelocaulon aculeatum C. ericetomm l v ei " " In I " " II ~ l ul N N v v V !I l~' 1 =- II x,~ ~o % ~II I=]~=.X~ ~ " --.1--r%%" "* II ~ ~ C. islandiea C. l a e v i g a t a C. aurescens C. chlorophylla C. laureri C. inermis C. evemiella C. orbala *' " 1 ~ - Esslingeriana u , II P_.. '~ ~ I:: :: ::I ~ ,.. : :III ~-~ idahoensis ~m~c~aT"ck~°'~*~ ~ eiliaris ~i~"~_ ~ II II v ,, ,,I ~ ~ ,i ¢~I C. hepatizon Comicularia nonnogl'ica nu ~nn nu~ ~ . . . . . Il ~ ~ ill " I "I II " . I II X II ....g i ~ ' ~ ~ B~yon~ fl ,, ,, ,, " X abbmviala C. califomica ~ C.m~-~u~ C. csnsd~nsis ~ . ,,,, C. viridis II '~ I~ commixtalI C. commixtaHI i o, ,.. nn " C. commixtaI t--~C, ~; ~ c~ vn C. alvarensis . v ~ ~ ' ~ " ~~ II II , . ,, . c~ Iv iv v ,, U C. tilesii C. j uniperina C. pinastri Fig. 82. One of 16 equally parsimonious cladograms derived from data in Table 1. Length 192, consistency index 32, retention index 68 I. K~RNEFELT& al.: Phylogeny of cetrarioid lichens 139 6. Melanelia stygia Melanelia stygia presumably belongs to the ingroup based on too many similar character states when it was tested as outgroup in the cladistic analysis. It branched out with the C. commixta group. 7. Nephromopsis stracheyi Nephromopsis stracheyi appears to be the best choice for the selected outgroup. Characters of the asci and pycnidia recall those in the other cetrarioid groups. In N. endocrocea there is also a tendency towards development of a ring structure in the tholus. Characters in the secondary chemistry, especially in the presence of higher aliphatic substances, suggest close affinities between the genus Nephromopsis and other cetrarioid groups. Cladistic analysis The cladistic analysis was carried out in two different steps: A. First, 42 investigated entities were run against the selected outgroup, in the first place to identify characters of importance for the definition of presumed monophyletic groups (Fig. 82). B. Second, the assumed monophyletic groups in the total analysis were selected and analysed alone against the same defined outgroup. A few basic character states, although in some cases, present only as parallelisms in the first analysis could be " ~'~ z~ • . ~ d d d o d 71 26 lll3 d d _ ~ -~o~'~ ~ ~ ~ ~ ~ © '3 d d - . ~ d I I[ +58 d - d ~0 112 +15 I//14s 10 Fig. 83. One of 6 equally parsimonious cladograms derived from data in Table 1. Length 45, consistency index 73, retention index 80. For legend see Fig. 82 140 I. K)kRNEFELT& al.: discerned as being of major importance in the definition of these branching systems or subgroups, i.e. 24. (asci broadly clavate also characteristic for a number of other foliose entities); (Figs. 83, 85, 86), 26. (asci uniseriate characteristic for a number of foliose entities), 28. (tholus with an amyloid ring structure characteristic for the C. islandica group), 31. (globose ascospores), and 34. (pycnidia on projections). Three rather distinctive groups were discerned (Fig. 82). 1. One of the best defined of these assumed monophyletic groups, i.e. the species related to Cetraria islandica, was supported by 12 apomorphic character states of which, however, character 28 (tholus with amyloid ring structure) must be considered of major importance. This larger branching system was then subdivided into smaller groups characterized by presence of different aliphatic substances. Basically this whole group comprising 17 terminal taxa, of which 7 further entities C. arenaria, C. australiensis, C. nepalensis, C. odontella, Coelocaulon crespoae, C. muricatum, and also C. steppae were added, could be accepted as representing a genus, which interestingly not only' includes erect foliose species of Cetraria, but also species presently included in Coelocaulon (Fig. 83). The same group of terminal taxa, apart from the seven added (Fig. 83), were also run along with some previously associated taxa, i.e. Masonhalea and Cetrariopsis, which were supported as representing distinctive branches in the three equally parsimonious cladograms generated (Fig. 84). 2. A second rather naturally delimited branch and assumed monophyletic group, named here as the "uniseriate group", comprising Tuckermannopsis ciliaris and related entities, was supported by 12 apomorphic character states of which 26 ~.~ "~ .~ ~ ~ °° o - ~ "~- "2 g -~ ~ .- _ J 62 l !46 - " 49 :45 - : 47 ;10 ~ C 3 37 41 36 ~ 5 0 51 24 " 20 J36 ]~ T . . ~~ !5 26 =L 12 + C20 59 58 16 13 12 51 41 32 20 16 15 4 3 Fig. 84. One of 3 equally parsimonious cladograms derived from data in Table 1. Length 64, consistency index 62, retention index 65. For legend see Fig. 82 Phylogeny of cetrarioid lichens 141 r~ 0 46 17 32 36 44 3~ ~ 9 35 ~ 62 =¢ ~ : : ~ 38 47 32 31 29 26 20 7 ii Fig. 85. One of 25 equally parsimonious cladograms derived from data in Table 1. Length 62, consistency index 59, retention index 51. For legend see Fig. 82 ~-~ 8 ~ -~ 36 " 18 38 41 16 I g" 13 12 135 -.141 I 161 I 158 m4 ~ 16 "~.13 :36 c32 62 39 I ~ 1 1 1 0 1 3 4~ 15 .q 62 1151 ~38 [I32 129 124 1120 ~ 7 5 13 Fig. 86. One of 85 equally parsimonious cladograms derived from data in Table 1. Length 82, consistency index 60, retention index 73. For legend see Fig. 82 142 I.K~RNEFELT&al.: (uniseriate asci), 29 (axial body large), and 31 (globose ascospores), are of major importance in the definition of an assumed genus. This group oftaxa was subdivided into two less distinctive branching systems defined on fewer character states. The whole group, comprising 11 terminal taxa, presumably, represents a rather welldefined genus (Fig. 85). 3. A third group, referred to as the "clavate group", comprising 16 terminal taxa was also supported by 12 different apomorphic character states (Fig. 86). Several smaller branching systems, however, could be discerned, e.g., the C. commixta group, the C. fendleri group, the Bryoria abbreviata group, and the C. juniperina group, which probably are more related to parmelioid genera. These smaller subgroups were all supported by several apomorphic character states and could presumably be interpreted as representing generic groupings. Evolution and phylogeny of cetrarioid lichens From the present material, it is obvious that three more or less distinctive groups can be discerned which are interpreted as representing different phylogenetic lines: 1. The first line comprises taxa characterized by a mainly erect foliose type of thallus, narrow clavate asci, ellipsoidal ascospores, and an apical ring structure in the tholus. To this group belong, basically, C. islandica and allied species, plus the erect, foliose, usnic acid-containing species in addition to a majority of species presently referred to Coelocaulon. 2. The second line comprises entities characterized by a more adpressed, foliose to subfoliose thallus, uniseriate asci, and globose to subglobose ascospores. To this group belong basically taxa which have been recognized as Tuckermannopsis and allied species, in addition to the more suberect forms in the C. everniella and the C. inermis groups. 3. The third line comprises taxa characterized by both more adpressed types of foliose to subfoliose thalli, but also more erect and terete forms, broadly clavate asci, ellipsoid ascospores and a large axial body. This main group also includes several more or less distinct subgroups, such as the Cetraria commixta, C. fendleri, and C. juniperina groups, in addition to some entities of more uncertain affinity. These three different phylogenetic lines must have differentiated very early in the evolutionary history of this group of lichenized fungi. The results of this differentiation, as we see it represented in modern species, in different types of structures in the asci must be recognized as being of fundamental importance in the systematic treatment of this lichen group. The outgroup Nephromopsis MCTLL. ARG., MOLLER (1891: 374). Type species: Nephromopsis stracheyi (CHURCH. BAB.) Mt3LL. ARG. Thallus foliose, upper surface brown, greyish, greenish to yellowish, often strongly wrinkled to rugose, lower surface very dark to blackish, with rhizines, lobes rather broad, c. 0.5-1.5 cm wide, pseudocyphellae developed mainly on the lower or rarely on the upper surface; cortex composed of paraplectenchymatous hyphae. Apothecia developed on the lower surface, marginal, asci rather small, thin- Phylogeny of cetrarioid lichens 143 walled, narrowly clavate, c. 25-40 x 6-10 gm, tholus rather large, ocular chamber conical, small, axial body small, distinct, sometimes with an indistinct ring structure, spores oblong ovate, 6-9 x 3-6 gin, paraphyses usually straight, sparsely branched with swollen tips; conidiomata marginal, raised, wall one-layered, non-pigmented, no cortical tissue beneath, conidia bifusiform. Chemistry: fumarprotocetraric, protocetraric, protolichesterinic, lichesterinic, usnic, and olivetoric acids, and endocrocin. Nephromopsis stracheyi was included in the cladistic analysis. The genus Nephromopsis was described on material from India characterized briefly on a foliose thallus fastened to the substrate from a central portion (M~LLER 1891). The apothecia were noted as being turned from the margin. Furthermore, M~LLER concluded that the thallus, apothecia, ascospores, and photobiont reminded him of Cetraria but the developmental type of apothecia reminded of Nephroma. Most species so far described are corticolous and occur in subtropical mountains. The circumscription of Nephromopsis is far from clear and different concepts have been used by various authors (LAI 1980, PARK 1990, RAS,~NEN 1952). LAI (1980) listed 10 species from E. Asia, i.e.N, asahinae, N. endocrocea, N. globulans, N. laxa, N. morrisonicola, N. nipponensis, N. ornata, N. pseudoeomplicata, N. rugosa, and N. stracheyi. AWASTHI(1982, 1988) chose to include all species earlier combined under Nephromopsis within Cetraria sensu lato. One species or group of species, according to LAI (1980), which presumably is most closely related to Nephromopsis is Cetraria rhytidiocarpa MONT. & V. D. BOSCH, separated as the genus Cetreliopsis by LAI (1980), and characterized by a blackish and punctate lower surface and with pseudocyphellae on the upper surface. We have not seen any authentic material of this species and it was therefore not included in the analysis. The ingroup T a x a c h a r a c t e r i z e d m a i n l y by narrow c l a v a t e to c y l i n d r i c a l asci, an a p i c a l ring s t r u c t u r e in the t h o l u s , and e l l i p s o i d a s c o s p o r e s Cetraria ACHARIUS (1803: 292). Type species: Cetraria islandica (L.) ACH. Thallus erect foliose, dark brown to paler brown, or yellowish green, occasionally reddish at the basal portions; lobes more or less canaliculate or almost subtubular, occasionally becoming markedly expanded towards the apical portions in a few species, occasionally also markedly foveolate, pseudocyphellae marginal or laminal on the lower surface, the margins with scattered, branched or unbranched cilia; cortex composed of an external layer of paraplectenchymatous hyphae overlying a more or less thin layer of periclinally arranged prosoplectenchymatous hyphae. Apothecia marginal on the upper surface, asci clavate to cylindrical, 40-60 × 713 ~tm, more or less uniseriate in a few species, tholus moderately large with an apical ring structure, ocular chamber conical with narrow beak, axial body small to very small, ascospores ellipsoid, c. 5-10 × 2.5-5 Ixm, paraphyses usually straight, sparsely branched with swollen tips; conidiomata on marginal projections, wall two-layered, non-pigmented, the outer layer usually thin, c. 5 I.tm, cortical tissue beneath, conidia bacillariform to bifusiform. 144 I. K~RNEFELT• al.: Chemistry: protolichesterinic, lichesterinic, and fumarprotocetraric acids. Two species contain usnic acid in the cortex. Thirteen species were included in the cladistic analysis, C. andrejevii, C. arenaria, C. australiensis, C. cucullata, C. ericetorum, C. islandica, C. karnczatica, C. laevigata, C. nepalensis, C. nigricans, C. nigricascens, C. nivalis, and C. odontella. Brown-pigmented species This group, characterized largely by erect, slightly canaliculate brownish pigmented lobes was originally defined for 14 mainly Northern Hemisphere species in terricolous habitats (K,~RNEFELT1979). As pointed out then, however, the group was not considered as forming a natural entity, but nervertheless was presented as a species group which appeared to have several common characters such as the structure of the lobes, the structure of cortical layers, position of apothecia and pycnidia and secondary chemistry. The term fruticose was used only as a way of expressing a certain type of thallus habit as indicated above, since true fruticose groups are mainly strap-like or thread-like with cortical layers composed of prosoplectenchymatous, periclinal hyphae as in Bryoria, Cladonia, Teloschistes, and Usnea. Among the species which clearly differed in several characters were C. inermis and C. subalpina which were characterized by lateral apothecia and globose to subglobose spores whereas a majority of species are characterized by marginal apothecia and ellipsoid ascospores (Figs. 35, 54; KARNEFELT 1979). Further characteristics such as the uniseriate arrangement of globose ascospores, a large axial body and a small tholus without an amyloid ring structure indicates a closer affinity to the uniseriate group. Two species dearly differed from other species in containing gyrophoric and hiascic acids, C. delisei and C.fastigiata. These species also entirely lacked higher aliphatic substances, which are otherwise characteristic of erect, brown species of Cetraria. The amyloid ring structure The present detailed anatomical investigations of asci and conidiomata confirmed the earlier observations of the senior author. The upper part of the rather large tholus in the erect brown Cetrariae is characterized by a distinctively amyloid ring structure in the apical portion of the asci (Figs. 46-51). This ring structure is lacking in the C. inermis group and also in the C. delisei group (Fig. 54). We consider this amyloid ring structure in the tholus to be fundamental for the definition of the species group as a whole. Among other supporting characters, apart from the large tholus with the ring structure, are a conical ocular chamber with a narrow beak and a small axial body (Fig. 1). One of the species, C. andrejevii, differs slightly in having a relatively small tholus (Fig. 47). Furthermore in C. nigricascens the ring structure is only very weakly developed. C. andrejevii is in addition characterized by a more uniseriate arrangement of the ascospores (Fig. 48). From earlier data these two species also differ by the contents of rangiformic and norrangiformic acids (K~,RNEFELT1979). Usnic acid pigmented species with amyloid ring structure In K.~RNEFELT'S (1979) attempt to define a central group of Cetrariae, the usnic acid-containing species, C. cucullata and C. nivalis, were not included. Anatomical Phylogeny of cetrarioid lichens 145 investigation of their asci, however, strongly confirms that these species are also very closely related to the brown species. They differ only slightly in having rather small asci, but all other characters in the apical portion of the asci including the ring structure are also found in the brown species discussed here (Figs. 52-53). KARNEVELT (1979) suggested that C. cucullata particularly appeared to be very closely related to C. kamczatica except for the loss of the brown cortical pigmentation. Cladistic analysis The cladistic analysis strongly supported that the brown species, including the usnic-acid containing species C. cucullata and C. nivalis, characterized by a ring structure in the tholus formed a monophyletic group (Fig. 83). As a whole, the monophyletic origin of the group was strongly supported by a number of additional apomorphic character states: 2 (thallus with erect lobes), 3 (lobes narrow and much longer than wide), 4 (lobes raised from the substrate), 28 (amyloid ring structure in tholus), 32 (ascospores globose to ellipsoidal), 34 (pycnjdia on projections), 41 (cortical tissue beneath pycnidial wall), 47 (aliphatic substances present), and 51 (orcinol depsides absent). An additional three apomorphic characters, however, with reversals on certain branches, also supported the separation of this group (Fig. 83). This group definitely should be maintained as a distinct genus. Since the type species of Cetraria, Cetraria islandica, belongs to this group of erect, foliose Cetrariae, the group naturally encompasses the genus Cetraria sensu str. Cetraria odontella and the genus Coelocaulon In connection with the systemic overview of the species previously accommodated in the genus Cornicularia, one species was suggested to be more appropriately treated as Cetraria odontella (KXRN~FELT 1986). Fertile material examined confirms this view. Interestingly the ring structure in the tholus is also distinctly developed in the fertile species of Coeloeaulon examined (see below). Coelocaulon LINK (1833: 165). Type species: Coelocaulon aculeatum (SCHREBZR) LINK Thallus erect foliose, dark brown to paler brown, occasionally reddish at the base; lobes more or less terete, occasionally becoming markedly broader towards the apical portions of lobes bearing apothecia, pseudocyphellae scattered, the surface furnished with scattered, branched or unbranched cilia; cortex comprising an external layer of paraplectenchymatous hyphae generally overlying a thicker layer of periclinal prosoplectenchymatous hyphae. Apothecia terminal on lobe tips, ascospores ellipsoid, asci clavate to cylindrical, 30-60 x 7-13 gm, tholus moderately large with a distinctive apical ring structure, ocular chamber conical with a narrow beak, axial body very small, ascospores ellipsoidal, c. 5-10 x 2.5--5 gm, paraphyses usually straight, sparsely branched with swollen tips; conidiomata developed on the tips of projections, wall one-layered, non-pigmented, cortical tissue beneath, conidia bacillariform. Chemistry: protolichesterinic, lichesterinic and, in one species, norstictic acids. One species, Coelocaulon aculeatum was at first included in the cladistic analysis. 146 I. K~RNEFELT tY¢ al.: The apical portion of the asci was also examined in C. crespoae and in C. muricatum. Material of C. steppae was also evaluated in one analysis (Fig. 83). The name Coelocaulon was reinstated during the late seventies for the C. aculeata group (HAWKSWORTHtY¢al. 1980). Five species were accommodated in the genus by K~RNEFELT (1986), i.e.C, aculeatum, C. crespoae, C. epiphorellum, C. muricatum, and C. steppae. The accommodation of C. epiphorellum in this genus, however, was only tentative, since this species differed from the others in several characters (K.~RNEFELT1986). This species has now been removed to the new genus Coelopogon (BRusSE 86 K~RNEFELT 1991). Of the remaining species, C. aculeatum, C. crespoae, and C. muricatum are frequently fertile and the apical portions of their asci interestingly are almost identical with those in the erect foliose Cetrariae discussed above (Fig. 45). Only C. steppae has so far never been found fertile. However, this species is structurally very similar to C. aculeatum and there is no doubt that these entities are closely related. C. aculeatum and C. muricatum are widespread terricolous species mainly in heaths and similar habitats both in Northern and Southern Hemispheres. The corticolous C. crespoae and terricolous C. steppae have more limited distributional ranges. This group of species is similar to Cetraria in several other characters apart from the structure of the apical portion of the asci, i.e. in the structure of the cortical layer (Fig. 18), pseudocyphellae, the position and structure of pycnidia as well as secondary chemistry. The major difference between the genera Coelocaulon and Cetraria is basically the organization of the thallus into terete vs. dorsiventral lobes (Figs. 5-8). To maintain Coelocaulon over Cetraria seems then only to express one simple structural character while a majority of other characters suggest that the species presently referred to Coelocaulon should be included in Cetraria, a view also strongly supported by the cladistic analysis (Figs. 83-84). T a x a w i t h c l a v a t e a s c i , e l l i p s o i d a l a s c o s p o r e s , and w i t h o u t a ring s t r u c t u r e in the t h o l u s , w i t h m o r e u n c e r t a i n a f f i n i t i e s Cetrariopsis KUROKAWA(1980: 140). Type species: Cetrariopsis wallichiana (TAYL.)KUROK. Thallus foliose, generally abundantly fertile, upper surface yellowish to olivegreen, lower surface paler or whitish; lobes rather broad, c. 0.5-2.0 cm wide, often strongly lacunose, pseudocyphellae more or less distinct, often a little raised from the surface; upper cortex composed of pachydermatous paraplectenchymatous hyphae, overlying a less distinct layer composed of strongly gelatinized hyphae, lower cortex composed of pachydermatous paraplectenchymatous hyphae. Apothecia lamina1 to submarginal, generally numerous, asci clavate, 4055 x 10-13 gm, tholus rather large, ocular chamber small, conical, axial body rather small, ascospores long, ellipsoid, c. 8-10 x 3.5-5 gm, paraphyses usually straight, sparsely branched with swollen tips; conidiomata not observed. Chemistry: alectoronic, u-collatolic, usnic, protolichesterinic and lichesterinic acids. Cetrariopsis wallichiana was included in the cladistic analysis. This distinctive foliose corticolous species, which occurs mainly in mountainous habitats in E. Asia, was separated almost at the same time by KUROKAWA(1980) Phylogeny of cetrarioid lichens 147 under the generic name Cetrariopsis, and only a month later by LAI (1980) under the generic name Ahtia. Both KUROg_AWAand LAI pointed out the unique structure of the generally numerous rather small, laminal apothecia (Fig. 3). Both KUROKAWA (1980) and LAI (1980), however, misinterpreted the structure of the cortical layer as composed of prosoplectenchymatous hyphal cells and therefore assumed a close relation to the genera Asahinea, Cetrelia, and Platismatia. True periclinal prosoplectenchymatous hyphae do not occur at all in those foliose groups. The genus was recognized as monospecific, but LAI (1980), however, in addition draw attention to a few other taxa which might belong in the same group, i.e. Cetraria citrina, C. pallescens, and C. teysmannii, all mainly known from E. Asia. Cetraria komarovii, known mainly from NE Asia presumably also might belong here. We have not had access to type material of these taxa, which is why these entities were not evaluated. They all seem to be characterized by a similar, rather broad lobed type of thallus according to the descriptions, as in Cetrariopsis. The apothecia are described as more marginal. Masonhalea K~RNEFELT (1977: 101). Type species: Masonhalea richardsonii (HooK.) KARNEF. Thallus foliose, dichotomously to irregularly branched, upper surface brown to greenish brown, lower surface partly whitish or brown; lobes moderately broad, c. 0.5-20 mm wide, slightly channeled, with rather conspicuous pseudocyphellae-like structures on the lower surface; upper cortical layer composed of a rather thin layer of strongly pachydermatous paraplectenchymatous hyphae overlying a massive prosoplectenchymatous layer, lower cortical layer composed of a relatively thin layer of anticlinal hyphae. Apothecia rather rare, lateral and marginal, asci clavate to cylindrical, c. 40 x 10 ~tm, more or less narrow clavate, tholus rather large, ocular chamber rather narrow and conical, axial body rather small, ascospores ellipsoid to subglobose, c. 6-6.5 × 3-6 ~tm, paraphyses usually straight, sparsely branched with swollen tips; conidiomata marginal, raised, wall one-layered, non-pigmented, cortical tissue beneath, conidia bacillariform, 4 x 0.75 ~tm. Chemistry: alectorortic acid. Masonhalea richardsonii was included in the cladistic analysis. This monospecific genus was separated from the erect, foliose Cetraria on a combination of characters such as the branching pattern of the lobes, especially the structure of the extremely thick upper cortical layer of periclinally arranged prosoplectenchymatous hyphae, the large ecorticate portions of the lower surface and the presence of alectoronic acid (K~.RNEFELT 1977). The single species is confined to arctic-alpine tundra in North America and Asia (Figs. 4, 19). The ascus apex showed a larger axial body than in the Cetraria islandica group (Fig. 44). Furthermore, there was no ring structure developed in the tholus. Altogether the structure of the ascus apex confirms that Masonhalea and Cetraria are not too closely related which was also supported in the cladistic analysis based on both the larger data matrix and from the smaller data matrix of the entities with an apical ring structure (Figs. 82, 84). The branch with Masonhalea was supported by the autapomorphic state 46 small conidia in addition to a number of parallel character states. Nothing suggests an alternative systematic treatment of this unique species and it appears to belong to a genus of its own. 148 I. K~RNEFELT • al." The Cetraria delisei group Cetraria delisei (BoRY ex SCrlAERER) NYLANDER (1866:114). Basionym: Cetraria islandica "~ delisei SCI~AERER(1850: 16) Cetrariafastigiata (DEL. ex NYL.) K~ZR~EFELT(1979: 84). Basionym: Cetraria delisei subsp.fastigiata NYL. (NORRLIN 1873: 325). Thallus erect foliose, dark brown to pale brown; lobes more or less canaliculate or almost subtubular, occasionally becoming markedly expanded towards the apical portions in a few specimens, pseudocyphellae marginal or laminal on the lower surface; cortex of one layer of paraplectenchymatous hyphae. Apothecia terminal on lobe tips, asci clavate, c. 40 x 15 gm, tholus moderately large, ocular chamber short and broad, axial body short and rather broad, ascospores ellipsoid, c. 5-10 x 2.5-5 gm, paraphyses usually straight, sparsely branched with swollen tips; conidiomata developed on the tips of projections, wall one-layered or two-layered, non-pigmented, sometimes with cortical tissue beneath, conidia bifusiform. Chemistry: gyrophoric and hiascic acids. Two species, C. delisei and C. fastigiata, were included in the original cladistic analysis to trace the presumed monophyletic groups (Fig. 82). Cetraria delisei and C. fastigiata were included in the group of brown erect foliose Cetrariae resulting from a number of common structural characters. Both terricolous species occur mainly in arctic-alpine habitats of the Northern Hemisphere. The presence of gyrophoric and hiascic acids in combination with the absence of higher aliphatic substances, however, suggests that these species have an isolated position in the group (KARNEFEI~T 1979). The investigation of ascus apices in C. delisei and C. fastigiata showed that they differed from other erect foliose species of Cetraria. A ring structure is never developed in the tholus. The erect foliose Cetrariae and the C. delisei group, are therefore not considered as closely related. In the light of these two important characters, i.e. ascus apex and secondary metabolites, it is strongly contended that C. delisei and C. fastigiata should be separated from species related to C. islandica. T a x a c h a r a c t e r i z e d by u n i s e r i a t e a r r a n g e m e n t of the a s c o s p o r e s , a l a r g e a x i a l body, and g l o b o s e a s c o s p o r e s The Cetraria aurescens group Cetraria aurescens TUCKERMAN (1847: 208). Cetraria pallidula TUCK. ex RIDDLE (1915: 28). Thallus foliose to subfoliose, upper surface pale yellowish green, lower surface pale, to light tan, with sparse rhizines; lobes moderately broad, c. 1-4 mm wide, slightly canaliculate, weakly wrinkled, occasionally with marginal soralia, with scattered pseudocyphellae on the upper surface; cortex of paraplectenchymatous hyphae. Apothecia lateral, marginal to laminal, asci uniseriate, cylindrical, 30-40 x 810 ttm, tholus small, ocular chamber cylindrical and broad, axial body rather distinctive, broad, ascospores globose, c. 6 × 6ttm, paraphyses usually straight, sparsely branched with swollen tips; conidiomata marginal, immersed to raised, wall one-layered pigmented or non-pigmented, cortical tissue beneath absent or present, different types of conidia. Phylogeny of cetrarioid lichens 149 Chemistry: caperatic, lichesterinic, protolichesterinic, and usnic acids. Cetraria aureseens and C. pallidula were included in the cladistic analysis. Cetraria aurescens and C. pallidula presumably represent related entities, mainly confined to corticolous habitats in mountainous regions in eastern, and western North America respectively. In the cladistic analysis based upon the data matrix from the entities with uniseriate asci and globose ascospores, C. aureseens and C. pallidula branched out separately on one side of a dichotomy, whereas the other branch comprised 9 other terminal taxa. Only one parallel character state, however, supported the separation of C. aurescens and C. pallidula from other taxa (Fig. 85). Cetraria laureri KREMPELHUBER(1851: 673) Thallus foliose to subfoliose, upper surface yellowish, greyish to whitish green, lower surface pale or brownish, with sparse rhizines; lobes moderately broad, c. 1-4 mm wide, slightly canaliculate, with ascending tips, lower surface with scattered, small pseudocyphellae; cortex of paraplectenchymatous hyphae. Apothecial position unknown, asci narrowly clavate, c. 35 x 10 ~tm, ascospores ellipsoid; conidiomata marginal, on projections, wall two-layered, non-pigmented, cortical tissue beneath, conidia bifusiform, c. 4 x 0.5 ~tm. Chemistry: protolichesterinic and usnic acids. The position of this corticolous species from mountainous habitats is rather uncertain. Many characters have not been properly evaluated because of the lack of richly fertile material. In the cladistic analysis C. laureri branched out in a dichotomy along with C. oakesiana on the other side. However, this branch was supported only by presence of usnic acid (Fig. 85). Cetraria oakesiana TUCKERMAN (1841: 445). Thallus foliose to subfoliose, upper surface yellowish green, lower surface pale tan to white, with sparse rhizines; lobes moderately broad, c. 1--4mm wide, slightly canaliculate, weakly wrinkled, occasionally with marginal soralia; cortex of paraplectenchymatous hyphae where the cells of the uppermost layer generally has much smaller lumina and are more strongly gelatinized. Apothecia lateral, marginal to laminal, asci uniseriate, cylindrical, 30-40 x 712 gm, tholus small, ocular chamber cylindrical and broad, axial body rather distinct and broad, ascospores globose, c. 5 x 5 gm, paraphyses usually straight, sparsely branched with swollen tips; conidiomata marginal, immersed to raised, wall onelayered, pigmented or non-pigmented, cortical tissue beneath absent or present, different types of conidia. Chemistry: caperatic, lichesterinic, protolichesterinic, and usnic acids. For discussion on possible affinities, see C. chlorophylla below. Cetraria chlorophylla (HuMB.) VAINIO (1896: 7). Basionym: Lichen chlorophyllus HUMBOLDT(1793: 20). Thallus foliose to subfoliose, irregularly branched, upper surface brown to pale brown, or grayish brown, lower surface paler brown to whitish, with sparse pale rhizines; lobes moderately broad, c. 1-4 mm wide, weakly canaliculate, with marginal soralia, scattered marginal cilia; cortex of paraplectenchymatous hyphae where those of uppermost layer generally have much smaller lumina and are more strongly gelatinized. 150 I. K~RNEFELT& al.: Apothecia extremely rare, lateral and marginal, asci uniseriate, cylindrical, 3040 x 7-10 ~tm, tholus small, ocular chamber cylindrical and broad, axial body rather distinct and broad, ascospores globose, c. 5 x 5 gm, paraphyses usually straight, sparsely branched with swollen tips; conidi6mata marginal, on projections, wall one-layered, non-pigmented, cortical tissue beneath, conidia bifusiform, 1.5 x 56 Ixm. Chemistry: protolichesterinic acid. Cetraria chlorophylla, widespread in the Northern Hemisphere (KARNEFELT 1987), and C. oakesiana, locally abundant mainly in eastern North America are both sorediate, structurally similar corticolous lichens (Fig. 10). Their presumed relation was also supported in the cladistic analysis (Fig. 84). Both entities seem to differ mainly in pigmentation, i.e. presence of usnic acid in C. oakesiana. Two taxa not examined here, C. togashii and C. ulophylloides, described from Japan by ASAmNA (1953), mainly on chemical characters, are presumably related to C. chlorophylla. The Cetraria inermis group Cetraria inermis (NYL.) KROG (1973: 299). Basionym: Cetraria crispa f. inermis NYL. (1887: 214). Cetraria subalpina IMSnAU~ (1951: 746). Thallus foliose to subfoliose, weakly branched to unbranched, upper surface brown, pale brown to olive green, lower surface pale to whitish, with sparse rhizines; lobes moderately broad, c. 1-5 mm wide, weakly canaliculate, rather smooth and glossy, pseudcyphellae on the margins of the lower surface; cortex of paraplectenchymatous hyphae. Apothecia lateral and marginal, asci uniseriate, cylindrical, 30-50 x 6-10 ~tm, tholus small, ocular chamber cylindrical and broad, axial body rather distinct and broad, ascospores globose to subglobose, c. 3.5-5 x 3.5-6 ~tm, paraphyses usually straight, sparsely branched with swollen tips; conidiomata marginal, raised, wall one-layered, non-pigmented, cortical tissue beneath, conidia bacillariform. Chemistry: lichesterinic and protolichesterinic acids in addition to two unidentified aliphatic substances. Cetraria inermis was included in the cladistic analysis. C. subalpina was examined anatomically. Cetraria inermis and C. subalpina were included in the group of erect, brown species of Cetraria for several reasons. However, the lateral position of the apothecia and the globose to subglobose ascospores suggests that these two species have a more isolated position (KARNEVELT1979). C. inermis occurs mainly on small shrubs in the Bering region, while C. subalpina has a more local range in NW North America. It should also be noted here that both taxa lack the characteristic amyloid ring structure in the tholus, which supports their separation from the true species of Cetraria sensu str. In this analysis C. inermis branched out in a dichotomy along with C. chlorophylla, C. laureri, and C. oakesiana on the other side as described above under C. orbata, united by the apomorphic character 41 (cortical tissue beneath the pycnidial wall; Fig. 84). Phylogeny of cetrarioid lichens 151 Cetraria platyphylla TUCKERMAN (1882: 34). Thallus foliose to subfoliose, upper surface brown to olive brown, lower surface pale brown, with sparse rhizines; lobes c. 3-6 mm wide, slightly canaliculate, wrinkled, papillose to coarsely isidiate, margins becoming dissected; cortex of paraplectenchymatous hyphae. Apothecia lateral, marginal to laminal, asci uniseriate, cylindrical, 30-40 x 810 pm, tholus small, ocular chamber cylindrical and broad, axial body rather distinct and broad, ascospores globose, c. 6 x 6 pm, paraphyses usually straight, sparsely branched with swollen tips; conidiomata marginal, immersed to raised, wall onelayered, pigmented or non-pigmented, cortical tissue beneath absent or present, different types of conidia. Chemistry: lichesterinic and protolichesterinic acids. Cetraria platyphylla is one of the most common corticolous species in the northwestern part of the Rocky Mountain system and is reminiscent in certain traits of both Tuckermannopsis ciliaris and of C. merrillii. There are, however, major differences between these entities in the secondary chemistry as well as in the ascus structure (Fig. 57). In the cladistic analysis, C. platyphylla branched out alone on one side of a dichotomy, with Esslingeriana idahoensis on the other side of the branch (Fig. 85). The C. platyphylla branch was supported by one parallel character state, the algal layer present beneath the exciple. Esslingeriana HALE & LAI, LAI (1980: 220). Type species: Esslingeriana idahoensis (EssL.) HALE & LAI Thallus foliose, upper surface gray, occasionally at lobe tips brownish, rather strongly rugose, lower surface blackish and strongly rugose, sparsely rhizinate; lobes not very broad, c. 0.5-7mm wide, pseudocyphellae and soredia lacking; cortical layer of pachydermatous paraplectenchymatous hyphae, the lower cortical layer strongly pigmented. Apothecia common, marginal, terminal, becoming laminal, asci rather small, uniseriate, clavate to cylindrical, 30-40 × 8-10 pm, tholus rather small, ocular chamber short, broad, axial body rather distinctive, short and broad, spores globose to subglobose, 5.5-6 × 6-7 pm, paraphyses usually straight, sparsely branched with swollen tips; conidiomata rather frequent, laminal or marginal, immersed, wall twolayered, pigmented, no cortical tissue beneath, conidia bifusiform, 5-7 × 1 pm. Chemistry: atranorin, endocrocin and one unidentified fatty acid. Esslingeriana idahoensis was included in the cladistic analysis. This monospecific genus, occurring in corticolous habitats in western North America, was described briefly by LAI (1980) based on the more detailed description of Cetraria idahoensis of ESSLINGER(1971). BRODO(1984) did not accept this genus, which in his opinion was atypical based only on slightly features in gross morphology. LAI (1980) emphasized particularly the occasionally laminal apothecia compared to those in Cetrariopsis. Other characters in his description were the mainly laminal pycnidia and the blackish lower surface (Fig. 12). In the cladistic analysis based on the smaller data matrix, Esslingeriana idahoensis branched out in a dichotomy along with C. platyphylla.One coded character seems to be unique for E. idahoensis, i.e. 23 (presence of white crystals in the exciple; Fig. 36) and furthermore the extremely blackish lower surface, composed of strongly pigmented cortical hyphae appears as a rather unique character (Figs. 28, 74). 152 I. K~RNEFELT8¢ al.: The Cetraria everniella group Cetraria ambigua CHURCH. BAB. (1852: 244). Cetraria everniella (NYL.) KREMPELHUBER (1868:315). Basionym: Platysma everniellum NYL. (1857: 100). Cetraria potaninii OXNER (1933: 168). Thallus erect, foliose, greenish yellow, dark brown to paler brown, or yellowish green on the upper surface, the lower surface concolorous or darker, occasionally darker at base and apices; lobes more or less dorsiventral or more convex, becoming slightly reticulate or ridged to faveolate, pseudocyphellae marginal or absent; cortex paraplectenchymatous, comprising an external layer of rather small hyphae overlying a layer of generally much larger hyphal cells. Apothecia lateral and marginal, asci uniseriate, cylindrical, 30-40 x 8-10 ~tm, tholus small, ocular chamber cylindrical and broad, axial body rather distinct and broad, ascospores globose to subglobose, c. 5-7 x 5-6 ktm, paraphyses usually straight, sparsely branched with swollen tips; conidiomata marginal, immersed, wall one-layered, pigmented, no cortical tissue beneath, conidia filiform. Chemistry: lichesterinic, protolichesterinic, secalonic, and usnic acids. Cetraria everniella was included in the cladistic analysis. C. ambigua and C. potaninii were also examined for anatomical structures. This group of three terricolous species occurs at rather high altitudes in the Himalayas, and is above all characterized on the autapomorphic character state 42 (conidia filiform; Fig. 69). In the cladistic analysis C. everniella branched out on one side of a dichotomy together with C. orbata supported only by one chemical character state, whereas the other branch included the genus Tuckermannopsis sensu str. (Fig. 85). Recently, this species group was separated as Allocetraria by KUROKAWA& LAI (1991). They did not recognize C. potaninii as belonging to the same genus but described a new species A. isidiigera. The genus was described mainly on morphological characters in addition to recognition of the palisadeplectenchymatous cortical layer. Cetraria orbata (NYL.)FINK (1919: 298). Basionym: Platysma orbatum NYLANDER (1869: 442). Thallus foliose to subfoliose, upper surface brown to pale brown, lower surface blackish, with sparse rhizines; lobes moderately broad, c. 1-4mm wide, slightly canaliculate, weakly wrinkled, with scattered pseudocyphellae on the upper surface; cortex of paraplectenchymatous hyphae. Apothecia lateral, marginal to laminal, asci uniseriate, cylindrical, 30-40 x 810 ~tm, tholus small, ocular chamber cylindrical and broad, axial body rather distinct and broad, ascospores globose, c. 6 x 6 gm, paraphyses usually straight, sparsely branched with swollen tips; conidiomata marginal, wall two-layered, pigmented, no cortical tissue beneath, conidia bifusiform. Chemistry: protolichesterinic acid. No clear elucidation of the systematic status of this entity, which occurs on conifers in western North America, was obtained from the cladistic analysis. In the analysis based on the smaller data matrix, C. orbata branched out nearest to C. everniella, as mentioned above (Fig. 85). We would have expected it to come Phylogeny of cetrarioid lichens 153 closer to C. platyphylla which C. orbata slightly resembles both in structural details and also in the secondary chemistry. Only material from western North America was used in this analysis. Tuckermannopsis GYELNIK (1933: 6). Type species: Tuckermannopsis ciliaris (AcH.) GYELNIK. Thallus foliose to subfoliose, upper surface pale brown to greenish, lower surface blackish, with sparse rhizines; lobes moderately broad, c. l ~ m m wide, slightly canaliculate, with scattered marginal cilia, weakly wrinkled, with scattered pseudocyphellae on the upper surface; cortex of paraplectenchymatous hyphae. Apothecia developed on the lower surface at the margins, asci small, uniseriate, cylindrical, c. 30 x 8 p-m, tholus small, ocular chamber cylindrical and broad, axial body rather distinctive and broad, spores globose to subglobose, c. 4-5 x 4-5 p,m; conidiomata marginal, raised, wall one-layered, pigmented, no cortical tissue beneath, conidia bifusiform, c. 5 x 1 p-m. Chemistry: alectoronic, a-collatolic, and olivetoric acids. Two species, T. ciliaris and T. americana, were included in the cladistic analysis. This genus was very briefly described by GYZLNIK (1933) as closely related to Nephromopsis and characterized only on the absence of pseudocyphellae from the lower surface. During the eighties the generic name Tuckermannopsis was mainly used for a number of corticolous North American taxa. E~AN (1987) lists 15 species, i.e.T, americana, T. aurescens, T. canadensis, T. chlorophylla, T. ciliaris, T. fendleri, T. juniperina, T. merrillii, T. oakesiana, T. orbata, T. pallidula, T. pinastri, T. platyphylla, T. sepincola, and T. viridis. HAI,Z & CoLE (1988) listed six of these species from California also as Tuckermannopsis. In Eastern Asia LAI (t980) earlier treated T. gilva, T. ham (= T. americana), and T. microphyllica. LAI (1980) in addition made the combinations T. orbata, T. platyphylloides, and T. ulophylloides. KUROKAWA (1991) recently listed 7 species from Japan of which T. hepatizon was combined here for the first time. Even though LAI (1980), briefly and partly also HALE & COLZ (1988), tried to define the genus, it is still far from clear how the generic name Tuckermannopsis should be used. HALE & COLE (1988) pointed out the marginal apothecia and the erect or immersed marginal pycnidia as opposed to laminal apothecia and pycnidia in related parmelioid genera. These characters, however, could rather include the definition of Cetraria in the older sense. In our analysis we attempted to find out the basis for a possible definition of Tuckermannopsis. Two important character states, i.e. 31 (globose ascospores) and 26 (asci strictly uniseriate) indicated a monophyletic group based on the larger data matrix comprising 42 entities. In the following analysis based on the smaller data matrix altogether 10 apomorphic character states supported this presumed monophyletic group (Fig. 85). Taxa characterized by broadly clavate asci, a large axial body, and e l l i p s o i d a l ascospores The Cetraria commixta group Cetraria commixta (NYL.) T. FRIEs (1871: 109). Basionym: Platysma commixtum NYL. (1860: 310). Cetraria hepatizon (AcH.) VA~NIO (1899: 278). Basionym: Lichen hepatizon Acn. (1799: 110). 154 I. K~RNEFELT• al.: Thallus foliose, upper surface brown to dark brown, rather glossy, lower surface blackish, with sparse rhizines; lobes moderately broad, c. 1-4mm wide, slightly canaliculate, weakly wrinkled; cortex of paraplectenchymatous hyphae. Apothecia marginal or laminal, asci clavate, 35-45 x 12-15 ~tm, tholus rather large, ocular chamber cylindrical and rather broad, axial body very distinct and rather broad, ascospores broadly ellipsoid, c. 6-8 × 3-4 Ixm, paraphyses straight, sparsely branched with swollen tips; conidiomata marginal or laminal, raised or on short projections, wall two-layered, outer layer sometimes thick, pigmented or not, no cortical tissue beneath, conidia citriform or bifusiform. Chemistry: a-collatolic, alectoronic, stictic, and norstictic acids. Three chemical types: C. commixta I with t~-collatolic acid, C. commixta II with alectoronic acid, and C. commixta III without any secondary substances, as well as C. hepatizon were included in the cladistic analysis. This saxicolous, arctic-alpine species group is characterized above all on its parmelioid habit reminiscent to a certain degree of, e.g., Melanelia stygia (L.) ESSL. Superficially, Cetraria commixta and C. hepatizon are very similar and they basically differ in anatomical and chemical characters, i.e.C, commixta, with citriform conidia, containing alectoronic- and a-collatolic acids and C. hepatizon with bifusiform conidia, containing stictic- and norstictic acids (Figs. 13, 29, 38-39, 7073). Cladistic analysis gave some evidence that these taxa form a distinct group. In the analysis comprising terminal taxa characterized by broadly clavate asci the group branched out alone on one side of a main dichotomy (Fig. 86). The Cetraria fendleri group Cetrariafendleri (NYL.) TUCKERMAN (1872: 35). Basionym: Platysmafendleri NYLANDER (1857: 105). Cetraria sepincola (EHRH.)ACHARIUS(1803: 297). Basionym: Lichen sepincola EHRHARDT (1783:203 = 1788: 95). Thallus foliose, upper surface brown to dark brown, lower surface tan, with sparse rhizines; lobes not very broad, c. 0.5-2 mm wide, slightly canaliculate, partly finely dissected at tips, sparsely ciliate; cortex of paraplectenchymatous hyphae. Apothecia numerous, common, marginal on lobe tips, asci broadly clavate, 3060 × 15-20 ~tm, tholus moderately large, ocular chamber short and broad, axial body short and broad, ascospores ellipsoid, c. 6-10 × 3-6 ~tm, paraphyses usually straight, sparsely branched with swollen tips; conidiomata marginal, raised, wall one-layered, pigmented, cortical tissue beneath absent or present, conidia bifusiform. Chemistry: protolichesterinic acid. Cetraria fendleri and C. sepincola were included in the cladistic analysis. Cetrariafendleri, occurs mainly in corticolous habitats in western North America and C. sepincola, a widespread corticolous species in the Northern Hemisphere, appear to be closely related. In the cladistic analysis the entities branched out on one side of a main dichotomy supported by character 25 (very broadly clavate asci; Fig. 59), 47 (presence of aliphatic substances), and 49 (lichesterinic and protolichesterinic acids present; Fig. 85). The C. fendleri group might presumably be separated at the generic level in view of the other branches in the cladogram, i.e. the nearby branches Cornicularia, Phylogeny of cetrarioid lichens 155 the Bryoria abbreviata group sensu lato, and the vulpinic acid-containing species of Cetraria (Fig. 86). To this group presumably also belongs three other species related to C. fendleri and found mainly in western North America, i.e.C, coralligera (W. WEB.) HALE, C. subfendleri ESSL. and C. weberi ESSL. (ESSLINGER1973). Material of these entities, however, was not examined here. Cormcularia HOFFMANN (1794: 36). Type species: Cornicularia normoerica (GuNN.) Du RIETZ Thallus caespitose, erect, stiffly attached, arising from a foliose-squamulose basal part; lobes mainly regularly dichotomously branched, more or less flattened to terete, stiff and cartilaginous, blackish to dark brown; cortical layer composed of a thin external layer of heavily pigmented anticlinal hyphae, overlying an extremely thick layer of periclinal prosoplectenchymatous hyphae. Apothecia terminal or subterminal, asci broadly clavate, 30-60 x 8-13 gm, thohis moderately large, ocular chamber conical, axial body rather large, ascospores ellipsoid to oblong-ovate, c. 5-10 x 2.5-5 gm, paraphyses usually straight, sparsely branched with swollen tips; conidiomata immersed, wall two-layered, pigmented, the outer layer usually thin, c. 5 gin, no cortical tissue beneath, conidia bifusiform. Secondary chemical products absent. Cornicularia normoerica was included in the cladistic analysis. The structure of this rather distinctive saxicolous species which occurs mainly in oceanic regions of the Northern Hemisphere was described in detail by KgRNEFELT (1986). Cornicularia branched out alone in the cladistic analysis based on the data matrix comprising entities with broadly clavate asci (Fig. 86). Cornicularia should, in our opinion, henceforth be treated as a monospecific genus. Bryoria sect. Subeh'vergentes (MOT.) BRODO & D. HAWKSW. BRODO & HAWKSWORTH(1977: 132). Type species: Bryoria subdivergens (DAHL) BRODO & D. HAWKSW. Thallus fruticose, caespitose, erect, decumbent or pendent, attached by a basal holdfast; branching variable; colour greenish grey to brown, dark brown or black; isidia, soredia and pseudocyphellae absent to abundant; cortical layer composed of a thin layer of paraplectenchymatous hyphae overlying a much thicker layer of periclinal, prosoplectenchymatous hyphae. Apothecia subterminal or lateral, common in most species, margins ciliate in a few species; asci clavate, c. 25-40gm long, 10-15 gm wide, tholus rather large, ocular chamber rather broad, axial body broad; ascospores hyaline, ellipsoid, c. 6.5 x 4.5gm; paraphyses usually straight, sparsely branched with swollen tips; conidiomata immersed, wall one-layered, pigmented, no cortical tissue beneath, conidia bifusiform. Secondary chemical products absent. Bryoria abbreviata was included in the cladistic analysis. This rather distinctive group, at present comprising four species distributed mainly in boreal and cool temperate regions, is described in detail by BRODO & HAWKSWORTH (1977). The other species B. divergescens, B. oregana, and B. subdivergens were not examined and evaluated here. B. abbreviata is discussed in this connection basically because of certain affinities with one particular entity which 156 I. K~RNEFELTt~¢al.: has been referred to Cetraria. In addition, characters of the asci and hamathecium strongly support that Bryoria should remain within the Parmeliaceae as it is presently circumscribed (K~,RNEFELT& THELL 1992). Section Subdivergentes differs above all from other species in Bryoria in the external cellular cortical layer in addition to the absence of secondary chemical compounds, and also in the two-layered exciple with the lower layer composed of strongly gelatinized hyphae (Fig. 40). Bryoria, sensu str., seems to have a threelayered exciple. COMMON(1991) also recently demonstrated that the cell wall chemistry is entirely different in sect. Subdivergentes compared with other species of Bryoria examined. The senior author earlier assumed that sect. Subdivergentes shared a number of common characters, such as, general morphology, the structure of cortical layer in addition to the secondary chemistry especially with Cetraria californica, which was only tentatively included in Cetraria (I~RNEFELT1986). The cladistic analysis basically supported this assumption that these entities are closely related (Fig. 86). Bryoria sect. Subdivergentesmight presumably be separated at the generic level, which is already in preparation COMMON • BRODO (pers. comm.). C. californica might also belong here. Cetraria cnlifornicn TUCKERMAN (1859: 203). Thallus caespitose, erect, attached by a basal holdfast; lobes dichotomously or irregularly branched, grey to brownish grey, central portions generally paler grey, pseudocyphellae occasionally distinct, cilia more or less frequent; cortical layer composed of a thin layer of paraplectenchymatous hyphae overlying a thicker layer of periclinal prosoplectenchymatous hyphae. Apothecia subterminal or lateral, rather frequent, margins ciliate; asci broadly clavate, c. 25-40 Ixm long, 10-15 ~tm wide, tholus rather large, ocular chamber rather broad, axial body broad; ascospores hyaline, ellipsoidal, 6-8.5 x 2.5-3.5 ~tm; paraphyses usually straight, sparsely branched with swollen tips; conidiomata immersed, wall one-layered, non-pigmented, no cortical tissue beneath, conidia bifusiform. Secondary chemical products absent. This remarkable species, is basically confined to a rather narrow coastal near range along the North American west coast. For discussion of the presumed relation between Cetraria californica and Bryoria sect. Subdivergentes see above. In addition C. californica has occasionally been misinterpreted and also partly confused with C. merrillii over a long period of time (K~RNEFELT1986). The secondary chemistry, in addition to characters in the structure and colour of the lobes supports that also these entities are presumably related. The ascus apex has no amyloid ring structure in this species and it should be pointed out that this entity is not related at all to the C. islandica group. In the cladisfic analysis this branch group was supported by 3 character states (Fig. 86). Cetrarin merrillii Du RIETZ (1926: 42). Thallus foliose to suberect, upper surface greenish to blackish brown, lower surface faintly brownish; lobes moderately broad, c. 1-5 mm wide, slightly canaliculate, weakly wrinkled, with sparse rhizines; cortex of paraplectenchymatous hyphae. Phylogeny of cetrarioid lichens 157 Apothecia subterminal or lateral, rather frequent, margins ciliate; asci clavate, c. 25-40 ~tm long, 10-15 l.tm wide, tholus rather large, ocular chamber broad, axial body broad; ascospores hyaline, ellipsoid, 6-8 × 3-6~tm; paraphyses usually straight, sparsely branched with swollen tips; conidiomata immersed, wall onelayered, pigmented, no cortical tissue, conidia bifusiform. Secondary chemical products absent. Cetraria merrillii belongs to one of the most variable groups discussed in this connection. The most extremely broad-lobed and blackish green individuals therefore can appear as entirely different from members of Bryoria sect. Subdivergentes. We find it difficult to decide here whether C. merrillii should be included in Bryoria sect. Subdivergentes or not. It should, however, be noted that this species also has no apical ring structure in the tholus and nothing to do with the genus Cetraria sensu str. The Cetraria juniperina group C. alvarensis (WAHLENB.)VAINIO(in LYNOE 1910: 76). Basionym: Lichenjuniperinus var. alvarensis WAHLENB. (1826: 827). C. canadensis (RAs.)RAS~NEN (1952: 36). Basionym: C.juniperina var. canadensis R~,s. (1933: 12). C. juniperina (L.) AcI-IA~it:s (1803: 298). Basionym: Lichen juniperinus L. (1753: 1147). C. pinastri (StoP.) S. F. GRAY (1821: 432). Basionym: Lichen pinastri StoP. (1772: 382). C. tilesii ACHARIt:S (1814: 228). C. viridis ScHWmN. (in HALSEY 1824: 16). Thallus foliose to subfoliose, deeply yellow to greenish yellow, loosely adnate to suberect, lower surface sparsely rhizinate; lobes moderately broad, c. 1-4 mm wide, slightly canaliculate; weakly wrinkled to faveolate; cortex paraplectenchymatous, comprising an external layer of rather small hyphae overlying a layer of generally much larger hyphal cells. Apothecia lateral, marginal to laminal, asci clavate, 30-50 × 10-15 ~m, tholus large, strongly amyloid in most species, ocular chamber cylindrical and broad, axial body very distinct and broad, ascospores broadly ellipsoid, c. 6-8 × 4-6 pm, paraphyses usually straight, somewhat branched with swollen tips; conidiomata marginal or laminal, wall one-layered, pigmented, no cortical tissue beneath, bottle shaped or citriform conidia. Chemistry: vulpinic, pinastric, and usnic acids. C. alvarensis, C. canadensis, C. juniperina, C. pinastri, C. tilesii, and C. viridis were included in the cladistic analysis. This group of six taxa will be discussed further in a new revision (J.-E. MATTSSON, unpubl.). Our original hypothesis was that this group formed a monophyletic group especially well separated from the C. islandica group. As seen in the cladistic analysis, several apomorphic character states also support this theory (Figs. 82, 86). Similarly with other species earlier accommodated in Cetraria the vulpinic acid-containing species also lack the characteristic amyloid ring structure in the tholus (Fig. 61). Taxa of uncertain affinities Authentic material was not studied of several additional species, i.e. Cetraria delavayi (HUE.) SATO, C. endoxanthoides AwAs., C. hypotrachyna M~LL. ARG., C. 158 I. K~RNEFELT• al.: isidioidea (RAs.) AWAS., C. laeteflava ZAHLBR., C. leucostigma L~.v. in JACQUEM., C. melaloma (NYL.) KREMPELH.,C. nephromoides (NYL.) AWAS. M o s t of these taxa appear to develop rather large foliose thalli and presumably belong close to genera such as Nephromopsis or Tuckermannopsis. Two of these entities, C. leucostigma and C. melaloma, however, are reminiscent o f erect foliose species o f Cetraria. It is necessary to study both type material and fertile material o f these entities for a correct evaluation o f their systematic position. We thank Drs IRWINM. BRODO and DAVID J. GALLOWAYfor their personal comments and remarks on the text as well as GALLOWAY'Simprovement of the English style. Professor DAVID L. HAWKSWORTHis also thanked for his general comments on the text. The study has been financially supported by the Royal Swedish Academy of Science (HIERTA-RETZIUS' Stipendiefond) and the Royal Physiographic Society in Lund. References ACHARIUS,E., 1799 ["1798"]: Lichenographiae suecicae prodromus. - Link6ping. - 1803: Methodus lichenum. - Stockholm. 1814: Synopsis methodica lichenum. - Lund. 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Fauna Flora Fennica 13.6: [I] - 20. 1899: Lichenes in Caucaso et in Peninsula Taurica annis 1884--1885 ab H. LOJKA et M. A. DECI-IY collectors enumeravit WAI~IO. -- Termrsz Ffizetek 22: 269-343. WAHLENBER~, G., 1826: Flora suecica. - Uppsala. YOSHIMURA, I., 1982: Lichen flora of Japan in color. - Osaka. - - - - - - - - - Address of the authors: I. KARNEVELT,J.-E., MATTSSON, and A. THELL, Department of Systematic Botany, University of Lund, Ostra Vallgatan 18-20, S-223 61 Lund, Sweden. Accepted April 3, 1992 by D. L. HAWKSWORTn