zyxwvutsrqp Nord. J. Bot. - Section of tropical taxonomy zyxwvutsrq zyxwvuts Exotesta morphology of the Gardenieae - Gardeniinae (Rubiaceae) Claes Persson zyxwvu zyxwv Persson, C. 1995. Exotesta morphology of the Gardenieae-Gardeniinae (Rubiaceae). Nord. J. Bot. 15: 285-300. CGpenhagen. ISSN 0107-055X. 4. Exotesta morphology of'68 species in 59 genera of the Gardeniinae were examined with the aid of scanning and light microscopy. The shape of the exotesta cells in surface is either f isodiametrical or elongate and both shapes occur among New World and Old World genera. All genera, except for Monosalpinx, Posoqueria and Macrosphyra, are provided with secondary thickenings in the cell walls of the exotesta. The majority of the genera has a thickened radial wall. In the Asian genera Catunaregam, Deccania, Tamilnadia and the mainly Pacific genera, Atractocarpus and Trukia thickenings of the radial wall are very low or absent. In nearly half the genera the inner tangential wall is provided with thickenings, usually in the shape of a continuous plate. However, in four African genera, Atractogyne, Didymosalpinx, Mitriostigma and Sherbournia, the wall is provided with elongate anastomosing ribs. Atractocarpus differs markedly from the rest of-the Gardeniinae by its ring-like thickenings in the inner tangential wall. Thickenings, in the shape of a continuous plate, in the outer tangential wall is restricted to three neotropical genera, Alibertia, Amaioua and Borojoa. Presence of distinct knobs is common in palaeotropical genera, whereas in neotropical genera this feature is only present in Casasia and Kotchubaea. It is concluded that data on exotesta morphology alone does not support any of the previously proposed informal groups, but may still be an important character to deduce phylogenetic relationships, primarily on a level just above the genus. zyxwvut zyxwv C. Persson, Dept of Systematic Botany, University of Goteborg, Carl Skottsbergs Gaia 22, S-413 19 Goteborg, Sweden. Introduction in the Gardeniinae. Brachytome was placed by Tirvengadum & Sastre (1986) in a tribe of its own, Brachytomeae, and Amaioua was tentatively placed in the tribe Hypobathreae (Robbrecht 1980). Likewise, the inclusion of the aberrant neotropical genus Posoqueria is very doubtful. Posoqueria deviates markedly from other Gardeniinae genera by having a slightly zygomorphic corolla with an imbricate aestivation, and a basic chromosome number of x=17. Furthermore, the monophyly of Gardenieae was questioned in a preliminary cladistic analysis based on rbcL sequence data of the subfamily Ixoroideae (Andreasen & Bremer 1993). For a more detailed systematic and historical overview of Gardeniinae, see Robbrecht & Puff (1986). Bremekamp (1947; 1952) was the first to use exotesta (testa) morphology as a character in the taxonomy of the Rubiaceae. Observations on peels of exotesta in LM zyxwvut The present paper is concerned with the documentation, by scanning electron microscopy (SEM) and light microscopy (LM), of the exotesta morphology in the subtribe Gardeniinae, Gardenieae - Rubiaceae. The subtribe Gardeniinae consists of 64 genera (Persson 1993). The group has 2 a pantropical distribution, and consists mainly of trees, shrubs or lianas, very rarely geofrutices (Robbrecht 1988). It is characterized by nondehiscent few- to many-seeded, relatively large fruits, containing a juicy pulp formed from the septum andor the placenta (Robbrecht & Puff 1986). As is usual in other members of the Ixoroideae the aestivation of the corolla is normally contorted. The absence of a juicy pulp from several genera (e.g. Brachytome, Anomanthodia and Amaioua), has caused doubts concerning their inclusion zyxwvutsr zyxwvutsrqponm Accepted 13-10-1994 0 NORDIC JOURNAL OF BOTANY Nord. J. Bot. 15 (3) (1995) 285 zyxwvutsr zyxwvut zyxwvutsr revealed a large variation which played an important part in his classification. In his paper on Acranthera (1947) he used the minutely, densely pitted testa cells of Acranthera as one reason to separate the genus from Mussaenda, which has testa cells with few, large pits. Exotesta was important also when segregating several genera from Mussaendeae to form the tribes Pomazoteae and Urophylleae Bremekamp (1952). These tribes were with slight modifications, later (Bremekamp 1966), raised to subfamily level. Verdcourt (1958), however, rejected the importance of the testa character, mainly due to difficulties in distinguishing pits from knobs. Apart from Robbrecht & Puffs (1986) overview of the Gardenieae and related tribes, information on exotesta morphology is rather scanty, and is largely confined to palaeotropical genera. Robbrecht (1978) described the seed coats of Preussiodora and later (1986) discussed the exotesta of Didymosalpinx and Massularia. Tirvengadum (1978) provided LM-photos of exotesta peels in surface view of 16 South East Asian species, but a description is wanting. The same author published (1993) some SEMphotos of Kailarsenia and Larsenaikia, the latter genus comprising several Australian species previously included in Kailarsenia by Puttock (1989). Bridson & al. (1980) discussed the thin walled seed coat of the East African genus Phellocalyx. Bridson & Robbrecht (1985 a, b) described the exotesta of Hyperacanthus and provided a SEM-photo of the South East Asian genus Brachytome. In separating the African species of Porterandia (Aoranthe) Somers (1988) also provided SEM-photos for some of the Asian species. Lorence (1986) described the exotesta of the neotropical genus Glossostipula. Likewise has Rodriguez (1976) studied a few species of Gardeniinae from Venezuela. All these papers are mostly dealing with one genus or based entirely on LM observations. To obtain a proper understanding of the exotestal cells Robbrecht & Puff ( 1 986) considered both sections in LM and surface views in SEM to be necessary. Consequently in their inclusive study of the Gardenieae and related tribes, they used both LM and SEM. They dealt with 47 genera of the Gardeniinae and discerned 5 main types and 15 subtypes of exotesta cells in the group. They also distinguished several variants within the subtypes (smooth v. ornamented walls and elongated v. f isodiametrical cells). Unfortunately only photos of eight genera were provided and it was not always indicated which species that was studied. Tirvengadum & Sastre (1986). On the other hand, no genera are treated, which were described after 1988, e.g. Larsenaikia Tirvengadum (1993). Likewise, the transfer of several Borojoa species to Alibertia (Schultze-Motel 1986) is not accepted, pending enough evidence for this transfer to be presented. A few genera listed in Robbrecht (1988) have been synonymized and thus not treated in this study, see Persson (1993). No material was at hand of the following five genera: Dolichodelphys, Neofranciella, Melanopsidium, Phellocalyx and Sulitia. All seeds originated from herbarium material except those of Sherboumia bignoniiflora,’ which came from alcohol material. Specimens from the following herbaria were used: AAU, BISH, BM, BR, CAY, F, G, GB, K, L, NY, P, S, SING, US WAG and, WU (abbreviations according to Holmgren et al. 1990). Seeds were, in most cases, taken from specimens identified by specialists such as D. D. Tirvengadum (South East Asia), D. Bridson, E. Petit, E. Robbrecht, B. Verdcourt or N. Hall6 (Africa) and J. A. Steyermark, D. Lorence or J. Dwyer (Neotropics). For taxa examined in SEM the following methods were used in order to remove the often membranaceous outer tangential wall. The seeds were soaked for a few minutes in 65% HN03 or in a mixture of 10% CrO, and 50% HN03, boiled in saturated water-solution of NaOH or 10% HN03 for 10-30 min., then rinsed twice in distilled water. Thereafter followed a treatment with ultrasonicator for c. 15 s at 7-16 pm, before putting them on the stubs. In some cases the seed coat had a tendency to loosen from the endosperm when treated in acid or basic solutions. If air dried, loosened seed coats often shrink and cell shape changes drastically. Better results were then obtained when dehydrating in a critical point dryer. Sections of seeds for SEM were made by hand using a razor. Additional observations by LM were made both in surface view and in cross-section. In order to make crosssections, seeds were put in 70% alcohol. The percentage was increased by 5% every two hours until reached 99%. The last bout lasted c. 12 hours. The seeds were then treated in a infiltration medium for 24 h followed by embedding in plastic. Microtome cross-sections of the seeds were stained either with toluidine blue or safranin. For surface views of exotesta in LM, peels were bleached in chlorine, stained in safranin or toluidine blue, prior to embedding in Euparal. The terminology is mainly in accordance with Robbrecht & Puff (1986). zyxwvu zyxwvut Material and methods Sixty eight species, representing 59 genera of the Gardeniinae were investigated. The circumscription is that of Robbrecht (1988), and consequently Brachytome is included, although it was placed in a separate tribe by 1. The authors of the species are specified in the appendix. 286 Observations Exotesta cells in the Gardeniinae are normally provided with secondary thickenings, rarely parenchymatic. Thick zyx zyx Nord. J . Bat. 15 ( 3 ) (1995) zyxwv zyxwvutsrqpo zy Tab. 1. Summary of Gardeniinae exotesta characters. The following abbreviations are used: sh = shortly; anast. = anastomosing; OTW = Outer tangential wall; RW = Radial wall; ITW = inner tangential wall. X designates secondary thickenings present as a + continuous plate or a mesh work of longitudinal and transversal ribs. - designates absence of secondary thickenings in the cell walls. + designates presence of knobs. - designates absence of knobs. Taxon Group Aidia cochichinensis VI Aidiopsis forbesii 111 Alibertia concolor I Alibertia edulis I Alibertia macrantha I1 Amaioua guianmris I Anomanthodia aff. auriculata IX Atractocarpus chartacea VIII Atractogyne gabonii VII Benkura malabarica VI Borojoa patinoi I Borojoa stipularis I Brachytome wallichii VI Brenania brieyi VI Burchellia capensis IX Byrsophyllum ellipticum IX Calochone acuminata I1 Casasia clusiifolia VI Catunaregam spinosa IV Ceriscoides turgida 111 Coddia rudis IX Deccunia pubescens IV Didymosalpinx abbeokutae2 VII Dioecrescis etythroclada X Duroia hirsuta IX Duroia saccifera IX Euclinia longiflora I1 Fagerlindia fasciculata VI Gardenia coronaria I11 Gardenia mannii 111 Gardenia volkensii VI Genipa americana V Glossostipula concinna' IX Hyperacanthus amoenus VI Kailarsenia tentaculata 111 Kochummenia stenopetala IX Kotchubaea sp. X Macrosphyra longistyla XI Mantalania sambiranensis 111 Massularia acuminata IX Mitriostigma axillare VII Monosalpinx guillaumettii XI Morelia senegalensis 111 Oligocodon cunliffeae I1 Oxyanthus speciosus X Oxyceros horridus VI Pelagodendron vitiense 111 Pleiocoryne fernandensis I1 Porterandia anisophylla I11 Posoqueria sp. XI Preussiodora sulphurea I1 Pseudaidia speciosa VI Pseudogardenia kalbreyeri I1 Pseudornantalania macrophyylla IX Randia aculeata var. dasyphylla I1 Randia urmata 11, I11 Randia cf. calycina I1 Randia ruiziana I11 Rothmannia wittii IX Schumanniophyton magnijlcum X Sherbournia bignoniiflora VII Sphinctanthus microphyllus I11 Stachyarrhena heterochroa2 IX Sukunia longipes I1 IV Tamilnadia uliginosa Tarennoidea wallichii IX I1 Tocoyena formosu Trukia dyadum IV Cell shape elongate isodiam. - sh. elongate elongate elongate isodiam. elongate elongate isodiam. elongate sh. elongate elongate elongate sh. elongate elongate elongate elongate isodiam. isodiam. - sh. elongate sh. elongate sh. elongate - elongate elongate sh. elongate elongate sh. elongate elongate elongate isodiam. isodiam. sh. elongate sh. elongate sh. elongate elongate elongate elongate sh. elongate elongate elongate sh. elongate sh. elongate - elongate elongate elongate elongate sh. elongate - elongate isodiam. elongate sh. elongate elongate isodiam. sh. elongate isodiam. isodiam. sh. elongate isodiam. elongate isodiam. isodiam. isodiam. isodiam. elongate elongate elongate isodiam. elongate isodiam. sh. elongate elongate isodiam. sh. elongate - elongate OTW X X X - RW ITW X X X X X X X X X - - thickenings very X X X X X X X X X thickenings very X X thickenings very thickenings very X X X X X X X X transverse ribs X X X X X X Knobs low circular thickenings anast. ribs X simple ribs - X X - low X X X - low low X anast. ribs simple ribs - simple and anast. ribs - X X X X transverse ribs X X - simple ribs - - X X X X simple and anast. ribs anast. ribs - - X X X X X X X X - simple ribs X X - X zyxwvutsrq zyxwv 'surface of seeds deeeply folded; exotesta consists of many layers. - - X X X X X X X -/x X X X X simple and anast. ribs X .. thickenings very low anast. ribs X X X simple and anast. ribs low on ribs X thickenings very low' X - -_ x X - - low zyxwvut zyxwvuts zyxw zyxwvutsr enings occur often both in the inner tangential wall (ITW) and the radial wall (RW), sometimes only in the RW, rarely both in the radial and outer tangential (OTW) walls. The thickenings consist normally of a continuous plate, but is sometimes made up of several elongate, anastomosing or simple, free ribs in the ITW. Rarely, transverse ribs occur in the ITW and/or the OTW. The RW and ITW are usually provided with pits of various sizes and are sometimes ornamented with knobs. The shape of the cells is either elongate or isodiametrical. (Tab. 1). For simplicity of presentation I have divided the material into 11 groups. These groups are based on the occurrence, type and position of thickenings, presence or absence of knobs, and cell shape. * * seeds (Fig. 1G-H) in all species with this type of exotesta, with the exception of Randia species and Tocoyena formosa. In Calochone acuminata the cells are almost parenchymatic in the center of the flattened part of the the seed, but have distinct thickenings at the edge of the seed (Fig. 11). The RW of the neotropical species with this type of exotesta is generally thicker than it is in palaeotropical ones. In Pseudogardenia kalbreyeri the ITW appears to be thickened on SEM micrographs (Fig. 2A), but no thickenings could be observed on cross-sections in LM. Group I11 (Figs 2H-0; 3A-J) Group I (Fig. 1A-E) Occurrence: Alibertia concolor, A. edulis, Amaioua guianensis, Borojoa patinoi, B. stipularis. Characteristics: Cells elongate; OTW and RW thickened, ITW not thickened, or rarely with rib-like thickenings. Remarks: Scattered simple elongate ribs were observed in the ITW of Borojoapatinoi (Fig. 1E). The seed surface at the edge of the flattened seed is sometimes folded in Alibertia edulis (Fig. 1B) and Amaioua guianensis. Large differences in the height of the RW within the cells in Alibertia edulis result in a “wavy” seed surface. Group I1 (Figs 1F-0; 2A-G) Occurrence: Alibertia macrantha, Calochone acuminata, Euclinia longifora, Oligocodon cunlcffeae, Pleiocoryne fernandensis, Preussiodora sulphurea, Pseudogardenia kalbreyeri, Randia aculeata, R. armata, R. cf. calycina, Sukunia longipes, Tocoyena formosa. Characteristics: Cells ened. & isodiametrical; only RW thick- Remarks: All species of this type have & lenticular seeds. The exotesta cells are markedly higher and the secondary thickenings are heavier near hilum and the edge of the Occurrence: Aidiopsis forbesii, Ceriscoides turgida, Gardenia coronaria, G. mannii, Kailarsenia tentaculata, Mantalania sambiranensis, Morelia senegalensis, Pelagodendron vitiense, Porterandia anisophylla, Randia armata, R. ruiziana, Sphinctanthus microphyllus. * Characteristics: Cells isodiametrical to elongate; ITW and RW thickened, OTW not thickened; ITW consists of a continuous plate or of a meshwork. Remarks: In the ITW, transitions occur from a meshwork of irregular ribs and large pits to a continuous plate with small pits. Pelagodendron vitiense, Kailarsenia tentaculata and Porterandia anisophylla form the first extreme and Randia ruiziana and Sphinctanthus microphyllus form the other. The thickenings of exotesta cells in Pelagodendron vitiense are sometimes provided with low knobs (Fig. 3C). The OTW of Morelia senegalensis is provided with thin, transverse ribs (Fig. 3B). The seeds of Mantalania sambiranensis and Sphinctanthus microphyllus have markedly higher cells near the edge and hilum than those in the center of the flattened side. On SEM micrographs of Randia armata a thickened ITW was found (Fig. 3F), although non could be discerned on cross-sections in LM. In Randia ruiziana the secondary thickenings of the RW are separated into an inner and an outer layer, the former layer somewhat thinner than the latter (Fig. 3G). Both layers are provided with pits. The OTW of Mantalania sambiranensis is often provided with a thick cuticle (Fig. 3A). zyxwvutsrqp zyxwvutsrqp Fig. 1. (A-B) Alibertia edulis; (A) cross-section of normal cells, showing thickenings in domed OTW and RW; (B) cross-section of edge cells, showing folded seed. - (C) A. concolor, cross-section, showing plane OTW and RW. - (D) Amaioua guianensis, cross-section, showing thickenings in OTW and RW. - (E) Borojoa patinoi, cross-section, showing thickenings in OTW and RW, and single ribs in ITW (arrow). - (F-H) Aliberria macrantha; (F) cross-section, showing thickenings in RW and a thin, parenchymatic OTW; ( G ) surface view of normal cells; (H) surface view of thick walled cells close to the edge of the seed. - (I-J) Calochone acuminata;(I) surface of half seed, showing parenchymatic cells in the middle and thickened cells towards the edges; (J) surface, showing thickenings in RW of edge cells. - (K)Euclinia longiflora, surface, showing thickenings in RW. - (L) Oligocodon cunliffeae, surface view, showing thickenings in RW. - (M) Pleiocorynefernandensis, surface view, showing thickenings in RW. (N-0) Preussiodora sulphurea; (N) surface view, showing thickenings in RW. (0)cross-section, showing thickenings in RW. Scales: C, D-H, K. L and 0=10 pn; A, J and M=30 pm; B, C and N=100 pm; I=lmm. 288 Nard. I. Bot. 15 (3) (1995) zyxwvutsrqpon Nord. J. Bot. IS (3) (1995) 289 zyxwvutsr zyxwvutsrqp zyxwvu Group IV (Fig. 3K-N) and RW thickened, ornamented with distinct knobs. ITW normally a continuous plate, OTW not thickened. zyxwv zyxwvut zyxwvuts Occurrence: Catmaregam spinosa, Deccania pubescens, Tamilnadia uliginosa, Trukia dryadum. Characteristics: Cells isodiametrical to elongate; I T W with a thickened continuous perforated plate, RW very low, thickened, or RW without thickenings. OTW not thickened. Remarks: The RW is provided with very low thickenings in Catmaregam spinosa, Deccania pubescens and Tamilnadia uliginosa (Fig. 3K-M). The thickenings of the RW in Deccania pubescens and Tamilnadia uliginosa are sometimes lower than those of the ITW.The thickenings of the RW are absent from Trukia dryadum. The ITW is provided with small rounded to elongate pits in Catunaregam spinosa and Deccania pubescens, labyrinthiform perforations in Tarnilnadia uliginosa (Fig. 3M). The RW of the latter has a undulating outline in surface view. The exotesta of Trukia dryadum has normally elongate cells, but scattered f isodiametrical cells occur interspersed among the elongate ones. Both kinds of cells are rarely provided with low knobs on the thickenings in the ITW. * Group V (Fig. 30) Occurrence: Genipa americana. Characteristics: Cells elongate; RW thickened, ITW often provided with transverse ribs, OTW not thickened or rarely provided with thin transverse ribs. Remarks: The ITW is usually provided with large pits, which are also frequently present in the RW. The thickenings of the ITW in Hyperacanthus amoenus consist often of a meshwork, sometimes of anastomosing or free ribs (Fig. 45). Rarely, the thickenings of the ITW are absent from this species. In Gardenia volkensii ssp. spatulifolia the cells are narrower and have smaller pits close to the edge of the seeds. Group VII (Figs 4N-0; 5A-D) Occurrence: Atractogyne gabonii, Didymosalpinx abbeokutae, Mistriostigma axillare; Sherbournia bignoniiflora. Characteristics: Cells elongate; RW and ITW thickened, OTW not thickened. ITW provided with several elongate, anastomosing ribs. Remarks: Both the RW and the ITW are provided with knobs in Atractogyne gabonii and Mitriostigma axillare. In Didymosalpinx abbeokutae the exotesta is many-layered (Fig. 5B). The individual cells in this species are extremely long, and the ITW is normally provided with elongate, anastomosing ribs. Group VIII (Fig. 5E) Occurrence: Atractocarpus (Randia) chartacea. Remarks: The ribs of the ITW are normally transverse, rarely longitudinal. The OTW has often a thick cuticle. Group VI (Fig. 4A-M) Occurrence: Aidia cochichinensis, Benkara malabarica, Brachytome wallichii, Brenania brieyi, Casasia clusiifolia, Fagerlindia fasciculata, Gardenia volkensii ssp. spatulifolia, Hyperacanthus amoenus, Oxyceros horridus, Pseudaidia speciosa. Characteristics: Cells isodiametrical to elongate; ITW Characteristics: Cells f isodiametrical; ITW with circular thickenings, OTW and RW without thickenings. Remarks: The height of the exotesta cells is very low. Thickenings occur in the center of the I T W , shaped rf: like a spiral or in two concentric rings. Group M (Figs 5F-0; 6A-K) Occurrence: Anomanthodia aff. auriculata, Burchellia capensis, Byrsophyllum ellipticum, Coddia rudis, Duroia Fig. 2. (A) Pseudogardenia kalbreyeri, surface view, showing thickenings in RW and possible thickenings in ITW. - (B) Randia aculeata, surface view, showing thickenings in RW. - (C-E) R. calycina; ( C ) surface view of half seed; (D) cross-section, showing thickenings in RW; (E) surface view, showing thickenings in RW. - (F) Sukunia longipes, surface view, showing thickenings in RW. - (G) Tocoyenaformosa,surface view, showing thickenings in RW. - (H) Aidiopsisforbesii, surface view, showing thickenings in RW and ITW, and pits in the RW. - (I-J) Ceriscoides turgida, (I) surface view, showing thickenings and pits in ITW; (J) oblique surface view, showing thikenings and pits in RW. - (K)Gardenia coronaria, surface view, showing thickenings and pits in RW and ITW. - (L) G. mannii, surface view, showing thickenings and pits in RW and ITW.- (M-N) Kailarsenia tentaculata; (M) surface view showing thickenings and pits in ITW and RW; gU) cross-section, showing thickenings in RW and ITW. - (0)Mantalania sambiranensis; surface view of one cell, showing thickenings and pits in RW and ITW; - Scales: B, D, H and L=20 pm; E-F, I-J, and M-0=30 pm; G=50 pn; A and K=100 pm; C=lmm. 290 zyxwvu zyxwvutsrqp Nord. J. Bot. IS (3) (1995) zyxwvutsrqpon zy Nord. J. Bot. 15 (3) (1995) 29 1 zyxwvutsr zyxwvu zyxwvutsrq hirsuta, D. saccifera, Glossostipula concinna, Kochummenia stenopetala, Massularia acuminata, Pseudomantalania macrophylla, Rothmannia wittii, Stachyarrhena heterochroa, Tarennoidea wallichii. Characteristics: Cells elongate; RW with smooth thickenings, OTW and ITW without thickenings, or ITW with rib-like thickenings. Remarks: In Massularia acuminata, Duroia saccifera, Rothmannia wittii and Stachyarrhena heterochroa free or anastomosing elongate ribs frequently occur in the ITW (Figs 5N; 6D & H). The ribs are k circular in crosssection in the three latter species, whereas in Massularia acuminata the ribs appear more flattened, sometimes forming an almost continuous sheet. The ribs are frequently beset with low knobs in Stachyarrhena heterochroa, a feature which sometimes also occurs on the RW of Tarennoidea wallichii. The exotesta of Stachyarrhena heterochroa normally comprises four to five layers (Fig. 61), but at the edges of the lenticular seeds there may be as many as ten (Fig. 6J), resulting in a clearly thicker exotesta at the edges of the seed. The edge is also thickened in Duroia saccifera, but in this case due to a greater height of the RWs towards the edge of the seed. In Rothmannia wittii and Kochummenia stenopetala the thickenings of the RW are sometimes separated into two horizontal bands (Fig. 6F). The thickenings of the RW in Pseudomantalania macrophylla consist probably of several horizontal bands (Fig. 6E). The RWs in Coddia rudis are thickened at the base, with the result that the RWs of two neighbouring cells appear as an inverted T in crosssection (Fig. 5L). Surface views of this species reveal intrusions from the lumen in the RWs. In Byrsophyllum ellipticurn, on the other hand, the RWs are somewhat thickened in the uppermost part (Fig. 5K). The thickenings of the RW in Glossostipula concinna occur on deeply folded parts of the surface of the seed (Fig. 6B). The OTW of Burchellia capensis, Coddia rudis and Massularia acuminata is often provided with a thick cuticle (Fig. 51 & L). Group X (Fig. 6L-0) Occurrence: Dioecrescis erythroclada, Kotchubaea sp., Oxyanthus speciosus, Schumanniophyton magnificum. Characteristics: Cells shortly elongate to elongate; RW thickened, provided with knobs; ITW and OTW not thickened, or ITW sometimes provided with rib-like thickenings. Remarks: Simple elongate ribs often occur in Dioecrescis erythroclada, Kotchubaea sp. and Oxyanthus speciosus (Fig. 6L-N). These ribs may rarely be slightly branched in Dioecrescis erythroclada and Oxyanthus speciosus. In Dioecrescis erythroclada the ribs are often provided with knobs (Fig. 6L), more rarely so in Kotchubaea sp. Group XI (Fig. 6P-R) Occurrence: Macrosphyra longistyla, Monosalpinx guillaumettii, Posoqueria sp. Characteristics: Exotesta cells isodiametrical to elongate, parenchymatic. Remarks: The pulp as well as the endosperm in Macrosphyra longistyla are difficult to distinguish from the parenchymatic exotesta cells, resulting in difficulties to observe the latter cells. The exotesta of Monosalpinx guillaumettii consists probably of several, tightly compressed layers (Fig. 6Q). zyxwvut Discussion In their survey of the Gardenieae and related tribes, Robbrecht & Puff (1986) divided the exotesta into 5 main types (A, B, C, D and E) and the main types were subdivided into a total of 15 subtypes. The subtypes were further subdivided into several variants (smooth v. ornamented walls and elongated v. & isodiametrical cells). The types were mainly based on the occurrence, position and shape of thickenings along the walls. When using this zyxwvutsrqpo Fig. 3. (A) Muntuluniu sarnbirunensis; cross-section of edge cells, showing showing thickenings in RW and ITW. - (B) Morelin senegalensis, surface view of one cell , showing thickenings in RW and ITW, and remnants of transverse ribs in OTW. - (C) Pelugodendron vitiense, surface view of one cell, showing thickenings in ITW and RW. - (D-E) Porterundiu anisophylla; (D) surface view, showing thickenings in ITW and RW; (E) cross-section, showing thickenings ITW and RW. - (F) Rundiu umutu, surface view, showing thickenings in ITW and RW. - (G-H) R. ruiziuna; ( G ) surface view of one cell, showing thickened inner and outer layers of secondary wall, the latter has become detached from the inner layer; (H) surface view, showing thickenings in ITW and RW, the inner and outer layers of the secondary wall being coherent. - (I-J) Sphinctunthus rnicrophyllus; (I) surface view of one cell, showing thickenings and pits in ITW and RW; (J) cross-section, showing thickenings in ITW and RW. - (K) Cutunaregarn spinosa, surface view, showing thickenings in lTW and low RW. - (L) Deccuniu pubescens, oblique cross-section, showing thickenings in ITW. - (M) Turnilnudiu uliginosu, surface view, showing thickenings in ITW and low RW. - (N) Trukia dryadurn, surface view, showing thickenings in ITW and RW without thickenings. - (0)Genipu umericunu, surface view showing thickenings in RW and transverse ribs in ITW. The cells somewhat shortened and widened due to rough treatment. - Scales: F-G. I-J, L and M=10 p mi B-C H, K, N and 0=20 pm; A=30 pm; D and E=100 pm. 292 Nord. .I. Bot. 15 (3) (1995) zyxwvutsrqpon zyxwvutsrqponmlkjihgfedcb Nord. J. Bot. IS (3) (1995) 293 zyxwvu zyxwvut zyxwvut zyxwvut typology several different types may sometimes apply to a single seed. Moreover, the difference between the types D and E (the degree of development of thickenings along the radial wall) is not always easy to discern and transitions occur between these types. In my material I found that the degree of development of secondary thickenings along the RW and the occurrence of simple or slightly branched ribs along the ITW are not useful criteria for ordering of the observations. Instead, I have used, occurrence, position and type of thickenings, internal ornamentation cell shape in surface view to subdivide the studied material into groups. Since every single feature of the exotesta seems to vary independently, they should in phylogenetic analyses be treated as separate characters. Thus, the presented groups are to be considered entirely phenetic gropings and are not supposed to reflect phylogeny. Most genera of the Gardeniinae have secondary thickenings in cell walls of the exotesta. Thickenings in the shape of a continuous plate throughout the OTW are found exclusively in three neotropical genera of group I, Alibertia (some species), Amaioua and Borojoa (Fig. 1A-E). Within the subfamily Ixoroideae, this character occurs also in many genera of the Pavetteae, and in some genera of the Hypobathreae (Robbrecht & Puff 1986). Robbrecht & Puff also found thickenings in the OTW of the South East Asian genus Anomanthodia. However, A. aff. auriculata, examined by me, showed no sign of thickenings in the OTW (Fig. 5G). In the African genus Morelia (group 111) and the neotropical genus Genipa (group V) thin transverse ribs occur sometimes in the OTW (Fig. 30). Similar thickenings in the OTW were also found in Aoranthe Somers (1988), former members of African Porterandia, which were tentatively included in Isertieae by Robbrecht (1988). A thick cuticle is often present in several genera with elongate cells, e.g. Mantalania (group III), Genipa (group V), and in some genera of group IX,such as Coddia (Fig. 5L), Burchellia (Fig. 51) and Massularia. This cuticle could be taken for a thickened OTW if only using the SEM. In the majority of genera, the RW is provided with well developed thickenings. However, in three genera of group IV, Catunaregam (Fig. 3K), Deccania (Fig. 3L) and Tamilnadia (Fig. 3M), the thickened RW is very low. Thickenings are completely absent from the RW as well as the OTW and the ITW in the genera of group XI, (Fig. 6P), Monosalpim and Posoqueria, entirely parenchymatic exotesta. The East African genus Phellocalyx has also been reported to completely lack secondary thickenings (Bridson et al. 1980). Secondary thickenings are also absent from the RW of Trukia (Fig. 3N) of group IV, and of Atractocarpus (group VIII), which have thickenings only in the ITW. In the African genus Calochone, thickenings in the RW occur only at the edge of the seeds (Fig. 11), whereas the exotesta cells are parenchymatic in the center of the “flat” side of the lenticular seed. This condition was also reported from another African genus, Pleiocoiyne (Robbrecht & Puff 1986). In cross-section the RW has normally a straight to oval outline. In Coddia, however, the thickenings of the RW are markedly heavier in the lowermost part (Fig. 5L),and two neighbouring cells form an inverted T in cross-section. Another exception is Byrsophyllum which has a somewhat thicker RW in the uppermost part (Fig. 5K). In Rothmannia (Fig. 6E) and Kochummenia the thickenings of the RW are sometimes divided into two horizontal bands. A similar characteristic is found in Pseudomantalania (Fig. 6E) where the thickenings of the RW appear to be made up of a large number of horizontal bands. A thickened ITW shaped as a continuous pitted plate (e.g. in Figs 35;4B) occurs in group 111, IV and VI. Most representatives of these groups belong to 19 palaeotropical genera, but also the neotropical genera Randia and Sphinctanthus (group III), and Casasia (group VI) show this feature. The size of the pits in the ITW and RW varies greatly between the species and often varies within the same seed, where the pits may be smaller towards the edge (e.g. Gardenia volkensii ssp. spatulifolia). The exotesta of Didymosalpinx has a seemingly fibrous appearance. Robbrecht (1986) interpreted the individual cells of this genus to be provided with thickenings in the angle of the RW and the ITW, and furthermore to be provided with additional thickenings in the ITW. However, Robbrecht & Puff (1986) described the cells as their C1 type, with thickenings in both in the OTW, the RW, and the ITW.I interpret the individual, extremely elongate cells of Didymosalpinx to be provided with very low thickenings in the RW and with elongate, anastomosing ribs in the ITW. zyxw Fig. 4. (A) Aidiu cochichinensis, surface view, showing thickenings, knobs and pits in ITW and thickenings in RW. - (B-C) Benkuru rnulubaricu;(B) cross-section, showing thickenings in RW and ITW; (C) surface view, showing thickenings, distinct knobs and pits in ITW. - (D) Brachytome wallichii, surface view, showing thickenings, knobs and pits in ITW and thickenings in RW. - (E) Brenuniu brieyi, surface view of half cell, showing thickenings, distinct knobs and pits in ITW and RW. - (F) Casusiu clusiifoliu, surface view, showing thickenings, distinct knobs and pits in lTW, and thickenings in RW. - (G) Fagerlindia fusciculatu, surface view, showing thickenings, knobs and pits in RW and ITW. - (H-I) Gardenia volkensii ssp. spatulifoliu; (H) surface view of cells in the middle of the flattened side, showing thickenings and wide pits in ITW, and a thickened RW; (I) cross-section, showing thickenings, knobs and pits in ITW and RW. - (J) Hyperacanthus umoenus, surface view, showing thickenings, knobs and pits in RW and ITW. - (K) Oxyceros horridus, surface view, showing thickenings and knobs in RW and ITW and pits in the latter. - ( L M ) Pseuduidia speciosu; (L) cross-section, showing thickenings, knobs and pits in RW and ITW; (M) surface view - (N-0) Atructogyne gabonii; (N) oblique surface view of one cell, showing anastomosing ribs with knobs in ITW; (0)surface (photo in LM). - Scales: A-C, L N = 2 0 pm; D-E- G-I-and K=30 pm; J=50 pm; F and 0=100 pm. 294 Nord. J . Bot. IS ( 3 ) (1995) zyxwvutsrqpon zyxwvutsrqponmlkjihgfedcba zyxwvutsrqponm zyxwvutsrqpon Nord. J. Bot. 15 (3) (1995) 295 zy zyxwvutsr zyxwvutsrqp zyxwvu zyxwvutsrqp zyxwvutsrq Another similar feature of the ITW is the single, often slightly branched, elongate rib, which occur in some exotesta cells in certain species. These ribs occur scatteredly in several groups and were observed in the following taxa: Borojoa (group I), Duroia saccifera (Fig. 5N),Massularia, Rothmannia and Stachyarrhena in group IX, and in Dioecrescis (Fig. 6L), Oxyanthus, KotcFubaea in group X. In Stachyarrhena and Duroia saccifera the ribs are often sparsely branched, whereas they are often unbranched in the other genera. In Genipa (group V) the ITW of the exotesta is usually provided with transverse ribs (Fig. 30), more rarely with longitudinal ribs. Robbrecht & Puff (1986) also found branched elongas ribs in one or more species in the exotesta of Alibertia, and in some of the exotesta cells in Coddia and Rothmannia globosa. Unfortunately, Robbrecht & Puff did not distinguish between several anastomosing ribs and single, slightly branched ribs. Judging from the photos in their paper, the ribs in Rothmannia globosa and Oxyanthus latifolius are several in each cell. In the ITW of Rothmannia wittii and Oxyanthus speciosus, studied by me, only single ribs could be discerned. The f circular thickenings in ITW of the exotesta cells (Fig. 5E) in the Australian and New Caledonian genus Atructocarpus (group VIII) do not occur in any other member of Gardeniinae, and they seem to be rare in the Rubiaceae. However, to judge from a photo in Bremer (1989), Argostemma, in the tribe Argostemmatae seems to have similar thickenings in the ITW. Multi-layered exotesta occurs in the African genus Didymosalpinx (group IV, Fig. 5B) and the neotropical genus Stachyarrhena (group IX). Outside the Gardeniinae, this feature has only been found in the genus Paragenipa, tribe Hypobathreae (Tirvengadum & Robbrecht 1985). It is also suspected that Monosalpinx possess this feature, since it has an unexpectedly thick exotesta, considering its parenchymatic cells. In contrast to the exotesta of Didymosalpinx, the number of layers in Stachyarrhena is markedly higher at the edges of the seed, resulting in a clearly thicker exotesta in this part. The members of group VI and group X are characterized by having distinct knobs on the RW and ITW, but knobs occur also in two genera placed in group VII. It occurs mainly among palaeotropical genera, but Casasia (group VI) and Kotchubaea (group X), both neotropical, also share this feature. In Stachyarrhena and Tarennoidea (group IX, Fig. 6K) and Pelagodendron (group 111, Fig. 3C) the majority of the exotesta cells are smooth, but a minor part may be provided with low knobs. Robbrecht & Puff (1986) pointed out that size and shape of exotesta cells may vary greatly within the same seed, and this was confirmed in Kailarsenia by Tirvengadum (1993). In the present study I found pronounced within-seed differences among the African members in group 11, in which the cells near the edge were often found to be markedly higher than the cells in the middle of lenticular seeds, e.g. in Calochone, Oligocodon and Pseudogardenia. This is also the case in Kailarsenia, Mantalania, Porterandia (group 111) and Duroia saccifera (group IX). In Alibertia macrantha (Fig. 1G-H) and Calochone the thickenings are also considerably thicker towards the edge. However, in Sphinctanthus the thickenings become higher and thinner closer to the edge. In Gardenia volkensii ssp. spatulifolia the cells become narrower and the pits become smaller towards the edge of the seed. Rarely, the shape of the cells changes from very elongate in the middle of the flat side to shortly elongate at the edges of the seed (e.g. Mantalania). Cell shapes seem to be somewhat erratically distributed among neotropical and palaeotropical genera. In neotropical genera the shape is generally elongate, but isodiametrical cells occur in four genera. Likewise a minor part of the African genera has isodiametrical shape, whereas the majority has elongate cells. Asian and Pacific genera generally have shortly elongate to elongate exotesta cells, but the isodiametrical cells of Atractocarpus and Sukunia form two striking exceptions. As a result of the limited number of species examined in each genus, only a few conclusions can be drawn about the infrageneric variation in exotesta morphology. Interesting to note is the variable exotesta of the neotropical genus Alibertia. Alibertia macrantha (Fig. 1F-G) has thickenings only in the RW, whereas A. concolor (Fig. 1C) and A. edulis (Fig. 1A-B) also have thickenings in the OTW, a feature which also occurs in the two Borojoa species examined (Fig. 1E). Alibertia edulis and species of Borojoa share the same type of pollen (Persson 1993). This seems to support the transfer of three species of Borojoa to Alibertia, made without arguments by Schultze-Motel (1986). The peculiar “double” thickenings in the RW of Randia ruiziana (Figs 3G-H) makes this species clearly distinct from R. armata, R. aculeata * zyxw zyxwvut Fig. 5. (A-B) Didymosalpinx abbeokutae; (A) surface view, showing extremely elongated cells; (B) cross-section, showing inultilayered exotesta. - ( C ) Mitriostigma millare, surface view, showing thickenings in RW and anastomosing ribs in ITW. - (D) Sherbournia bignoniijbra, surface view, showing thickenings in RW and anastomosing ribs in ITW. - (E)Atractocarpuschurtaceu, surface view, showing ring like thickenings in ITW. - (F-G) Anomanthodiu aff. uuriculatu; (F) surface view, showing thickenings in RW; (G) surface view of entire seed. - (H-I) Burchelliu cupensis;(H) cross-section, showing thickenings in RW; (I) surface view (LM), showing thickenings in RW. - (J-K) Byrsophyllum ellipticum;(J) surface view, showing elongated cells, somewhat widened due to treatment; (K) oblique cross-section, showing somewhat thicker RW at the top. - (L) Coddia rudis, cross-section, showing thickenings in RW thickened at base. - (M) Duroia hirsuru, cross-section, showing thickenings in RW and thin OTW. - (N-0) D. sacc{fera;(N) cross-section, showing thickenings in RW and ribs in ITW; (0)cross-section, showing differences in height between edge cells and “normal cells”. - Scales: L and N=10 pm; B, E, G, I and M=20 pm; C and D=30 pm; J=50 pm; A, F, H, K and 0=100 Pm. 296 Nard. J . Bot. 15 ( 3 ) (1995) zyxwvutsrqponmlk zyxwvutsrqponm zyxwvutsrqponmlkji Nord. J. Bot 15 ( 3 ) (1995) 297 zyxwvutsrqp zyxwvutsr zyxwvu zyxwvutsr zyxwvut and R. cf. calycina. Randia ruiziana differs also from the other Randia species by having a thickened ITW, although the ITW may sometimes be thickened in R. armata. The great variation in exotesta morphology in Randia is congruent with its pollen morphology, which is also rather variable (Persson 1993). All species of Gardenia have thickenings in the RW and the ITW.Moreover, the exotesta is provided with knobs in G. volkensii ssp. spatulifolia (Fig. 41), a feature also found in G. augusta (Robbrecht 1988). The variation of exotesta morphology in the Gardeniinae is. like that in pollen morphology (Persson 1993), considerable, and makes the group one of the most diverse in the Rubiaceae. In 1986, Robbrecht & Puff tried to propose three informal subgroups of the Gardeniinae. No single exotestal character can support these groups, suggesting that either exotesta characters have been subjected to parallel evolution, or these groups are not monophyletic. Nevertheless, the amount of variation suggest that there is plenty of phylogenetic information in exotesta morphology, and therefore exotesta characters are potentially useful in cladistic analyses. It seems that they are primarily useful at a level just above that of the genus. K. Schumann. - Verh. Kon. Ned. Akad. Wet., Afd. Nat., ser 2, 18: 1-297. - 1966. Remarks on the position, the delimitation and subdivision of the Rubiaceae. - Acta Bot. Neerl. 15: 1-33. Bremer, B. 1989. The genus Argostemrnu (Rubiaceae-Argostemmateae) in Borneo. - Ann. Miss. Bot. Gard. 76: 7-49. Bridson, D., Gasson, P. & Robbrecht, E. 1980. Phellocalyx, a new tropical African genus in Rubiaceae (Gardenieae). Kew Bull. 35(2): 315-321. - & Robbrecht, E. 1985a. Further notes on the tribe Pavetteae (Rubiaceae). - Bull. Jard. Bot. Nat. Belg. 55: 83-115. - & Robbrecht, E. 1985b. Validation of the African genus Hyperacanthus E. Mey. (Rubiaceae tribe Gardenieae). Kew Bull. 40:273-286. Holmgren, P. K., Holmgren, N. H. & Barnett, L. C. 1990. Index Herbariorum I. The Herbaria of the World. 8th edn. - Regnum Veg. 120 1 4 9 3 . Lorence, D. H. 1986. Glossostipula (Rubiaceae), a new genus from Mexico and Guatemala. - Candollea 41: 453-461. Persson, C. 1993. Pollen morphology of Gardenieae-Gardeniinae (Rubiaceae). - Nord. J. Bot. 13: 561-582. Puttock, C. F. 1989. Kailarsenia Tirvengadum emend. Puttock (Rubiaceae: Gardenieae) in Australia. - Austrobaileya 3( 1): 5142. Robbrecht, E. 1978. Some observations in Preussiodoru Keay (African Rubiaceae, Gardenieae). - Bull. SOC.ROY. Bot. Belg. 111: 3-9. 1980. The Hypobathreae (Rubiaceae - Ixoroideae) 1. Delimitation and division of a new tribe. - Bull. Jard. Bot. Nat. Belg. 5 0 69-77. 1986. Studies in troDical African Rubiaceae. 10. Momhological observations oh the fruits and seeds of Didym&alpik (Gardenieae). - Bull. Jard. Bot. Nat. Belg. 56: 145-162. 1988. TroDical woodv Rubiaceae. Characteristic features and progrissions. Conkibutions to a new subfamilial classification. - Opera Bot. Belg. 1: 1-271. Robbrecht, E. & Puff, C. 1986. A survey of the Gardenieae and related tribes (Rubiaceae). - Bot. Jahrb. Syst. 108: 63-137. Rodriguez, P. 1976. Estudio sobre frutos camosos y sus semillas de las Rubiaceae de Venezuela. - Acta Bot. Venez. 11(1-4): 283-383. Schultze-Motel, J. 1986. Rubiaceae. - In: Mansfeld, R. (ed.), Verzeichnis landwirtschaftlicher und giirtnerischer Kulturpflanzen (ohne Zierpflanzen) Band 2. Springer, Berlin, Heidelberg, New York, Tokyo. Somers, C. 1988. Aoranthe (Rubiaceae) a new genus to accommodate the African species of Porterandia. -Bull. Jard. Bot. Nat. Belg. 58: 47-75. Tirvengadum, D. D. 1978. A synopsis of the Rubiaceae-Gardenieae of Ceylon (Sri Lanka). -Bull. Mus. natn. Hist. Paris 3e s&., no 521, Botanique 35: 3-33. - 1993. Larsenaikia, a new genus of the Rubiaceae from Australia. - Nord. J. Bot. 13: 175-184. - & Robbrecht, E. 1985. Remarks on three Hypobathreae, zyxwvutsr Acknowledgements - I wish to thank Lennart Andersson for reading the manuscript. I also express my gratitude to Mrs Annie Sloth for skilful technical assistance and Mr Christian Tange N0rgaard for encouraging me in the laboratory work. The major part of this study was done while holding a scholarship from NorFA (Nordisk forskerutdanningsakademi)at Botanisk Institut, Aarhus University, k h u s , Denmark. Financial support was also received from the Royal Swedish Academy of Sciences. References zyxwvutsrqp Andreasen, K. & Bremer, B. 1993. A phylogenetic analysis of the subfamily Ixoroideae (Rubiaceae), based on rbcL sequence data. - International Conference on the Systematics of the Rubiaceae. abstracts. Missouri Botanical Garden. mimeographed. Bremekamp, C. E. B. 1947. A Monograph of the genus Acranthera Am. ex Meisn. (Rubiaceae). - J. Arnold Arbor. 28(3): 261-308. - 1952. The African species of Oldenlandia L. sensu Hiem et Fig. 6. (A) Duroia sacciferu, surface view showing elongated cell with thickenings in RW. - (B) Glossosripula concinna, cross-section, showing deeply folded seed with thickenings in the RW (arrow). - (C) Kochummenia stenopetala, oblique surface view, showing the two-parted thickenings of the RW. - (D) Mamularia acuminata, surface view, showing thickenings in RW and ribs in ITW. - (E) Pseudomuntalania rnucrophylla, surface view, showing parallel band-like thickenings of the RW. (F-G) Rothmannia witrii; (F) cross-section, showing two-parted thickenings of the parted RWs. (G) surface view, showing thickenings in RW. - (H-J) Stachyarrhena heterochroa; (G) (H) surface view, showing thickenings in RW and branched rib in ITW; (I) cross-section at the flattened part of the seed, showing a 4-5 layered exotesta; (J) cross-section at the edge of the seed, showing an exotesta of c. 10 layers. - (K) Tarennoidea wallichii, surface view, showing thickenings in RW with very small knobs. - (L) Diocrescis erythroclada, surface view, showing thickenings in RW and simple ribs provided with knobs in ITW . - (M) Kotchubueu sp., surface view, showing thickenings in RW. - (N) Oxyanthus speciosus, surface view, showing thickenings provided with knobs in RW. - (0)Schurnanniophyton magnificum, surface view, showing thickenings provided with knobs in RW. - (P) Macrosphyra longiswla, surface view in LM, showing parenchymatic exotesta. (Q-R) Monosalpinx guilluumettii; (Q) cross-section of thick parenchymatic seed coat; (R) surface view in LM, showing parenchymatic exotesta. - Scales: F and G=10 pm; B, C, E, H and I=20 pm;D, K-M and 0=30 p;A, N, P and R=50 pm; J and Q=lOO p. 298 Nord. J. Bot. 15 (3) (1995) zy zyxwvutsrqponmlk zyxwvutsrqponm zyxwvutsrqpon Nord. J. Bot. 15 (3) (1995) 299 zyxwvu zyxwvutsrqp zyxwvuts zyxw zyxwvutsr zyxwvuts zyxwvut (K). Gardenia coronaria Buch.-Ham, collector unknown 30 April 1919 (US). G. mnnnii St. John & Kuykendall, Kuykendall 135 (BISH). G. volkensii ssp. spatulifolia Stapf & Hutch., Dahlstrand 2203 (GB). Genipa americana L., Nee 37622 (NY). Glossostipula concinna (Standl.) Lorence, Steyermark 369 13 (F). Hyperacanthus amoenus (Sims) Bridson, Lambinon & Reekmans 82/142 (BR). Kailarsenia tentaculata (Hook f.) Tirveng., Larsen & Larsen 32716 (AAU). Kochummenia stenopetala (King t Gamble) Wong, Curtis 1306 (K). Kotchubaea sp., Silva & Rosfio 3728 (NY). Macrosphyra longistyla (DC.) Hook. f. ex Hiem, Lykke 24 ( M U ) . Mantalania sambiranensis Capuron ex Leroy, Bora 26993 SF (P). Massularia acuminata Appendix (G. Don.) Bullock ex Hoyle, Louis & et al. 1028 (WAG). Mitriostigma axillare Hochst., Marloth 4186 (K). Monosalpinx Specimens examined guillaumettii N. Hall6, Guillaumet 1218 (P). Morelia senegalenAidia cochinchinensis Lour., Selling s.n. 2 June 1949 (S). Aisis A. Rich. ex DC., Thomas 98 (S). Oligocodon cunliffeae diopsis forbesii (King & Gamble) Tirveng., Comer 28954 (Wemham) Keay, 5 Jan. 1947 collector unreadable 348 (P). (SING). Alibertia concolor (Cham.) Schum., Irwin & et al. Oxyanthus speciosus DC., Back6 19 (L). Oxyceros horridus 30295 (AAU). A. edulis (L.C. Rich.) A. Rich. ex. DC., Contre- Lour., 8-2 (SING). Pelagodendron vitiense Seem., Qoro 14090 ras 9283 (S). A. mucranth Standl., Holm-Nielsen & et al. (K). Pleiocoryne fernandensis (Hiem) Rauschert, Fleury 26187 21539 (AAU). Amaioua guianensis Aubl., Klein 26 b (S). (P). Porterandia anisophylla (Jack. ex Roxb.) Ridl., Maxwell Anomanthodin aff. auriculata (Wall.) Hook f., Brooke 8854 (G). 76-728 (AAU). Posoqueria sp., S t a l et al. 694 (GB). PreusAtractocarpus chartacea (F. Muell.), comb. ined. [=Randia siodora sulphurea (Schum.) Keay, Martin & Duncan 71 (BR). Pseudaidia speciosa (Bedd.) Tirveng., Dalzell s.n. (K). Pseudochartncen (F. Muell.) F. Muell.], Clark et al. 1353 (K). Atracrgardenia kulbreyeri (Hiern) Keay, Bos 4580 (BR). Pseudomanogyne gabonii Pierre, Hall6 2573 (BR). Benkura mulabarica (Lam.) Tirveng., Meebold 13753 (S). Borojoa patinoi Cuatrec., talania mucrophylla Leroy, Capuron 962 (P). Randia aculeata Cuatrecasas 16525 (F). B. stipularis (Ducke) Cuatrec., Schunke L. var. dasyphylla Steyermark, Britton & Broadway 2459 (NY). 91 1 (F). Brachytome wallichii Hook f., Clarke 45422 (G). Bre- Randia a m t a (Swartz) DC., de Granville et al. 10688 (CAY). nania brieyi (De Wild.) Petit, Toussaint 2171 (BR). Burchellia R. cf. calycina Cham., Hahn 2762. R. ruiziana DC. Fleury 405 (CAY). Rothmannin wittii (Craib) Tirveng., Murata & et al. capensis R. Brown, Strey 8814 (AAU). Byrsophyllum ellipticum (Thw.) Hook f., Kostermans 26700 (G). Calochone acuminata 64-1 (AAU). Schumanniophyton magn$cum (Schum.) Harms, Keay, Hedin 1918 (P). Casasia clusiifolia (Jacq.) Urb., Small & Herb, G. Le Testu 8160 (BM). Sherbounia bignoniiflora (Welw.) Hua, Puff 900413-1/3 (WU). Sphinctanthus microphylMosier s.n. 2 Mar. 1915 (S). Catunaregam spinosa (Thunb.) lus Schum., A:n Malme 1872 (S). Stachyarrhena heterochroa Tirveng., Faurie 1089 (BM). Ceriscoides turgida (Roxb.) Tirveng., Poilane 14 134 (P). Coddia rudis (E. May ex Harv.) Standl., de Nevers 5800 (MO). Sukunia longipes A. C. Sm., Verdcourt, Balsinhas 228 (BM). Deccania pubescens (Roth.) Parks 20335 (BISH). Tamilnadia uliginosa (Retz.) Tirveng. & Sastre, Tirvengadum 2006 (AAU). Tarennoidea wallichii Tirveng., Fischer s.n. 29 June 1912 (K). Didymosalpinx abbeok(Hook. f.) Tirveng. & Sastre, Maxwell 88-1088 (AAU). Toutae (Hiem) Keay, Binuyo 41238 (BR). Dioecrescis erythrocoyem f o m s a (Cham. & Schlecht.) Schum., Regnell I11 93 clada (Kurz) Tirveng., Pierre 4240 (P). Duroia hirsuta (P. & E.) Schum., Harling 7120 (GB). D. saccifera (R. & S . ) Schum., (S). Trukia dryadum ( S . Moore) Fosberg, Mauriasi & et al. BSIP Ducke 228/8 (S). Euclinia longij7ora Salisb., J. Louis 8598 15082 (AAU). (BR). Fagerlindia fasciculata (Roxb.) Tirveng., J. D. H. s. n. Rubiaceae from Rodrigues, Seychelles and Sri Lanka. Nord. J. Bot. 5: 455-461. - & Sastre C. 1986. Etude taxonomique et systkmes de ramification chez Aidia et genre affins, et chez Brachytome (Rubiaceae). - Bull. Mus. natn. Hist. nat., Paris 4e skr., 8 section B, Adansonia, no 3: 257-296. Verdcourt, B. 1958. Remarks, on the classification of the Rubiaceae. - Bull. Jard. Bot. Etat. 28(3): 209-290. 300 N o d I. Bot. 15 (3) (1995)