Available online at www.notulaebotanicae.ro Print ISSN 0255-965X; Electronic ISSN 1842-4309 Not. Bot. Hort. Agrobot. Cluj 37 (1) 2009, 45-50 Notulae Botanicae Horti Agrobotanici Cluj-Napoca Application of Numerical Taxonomy to Lip Morphology in the Genus Polystachya Hook (Orchidaceae) in Nigeria Abayomi Ezekiel FALORUNSO, Adeniyi Akanni JAYEOLA 1) Department of Botany, Obafemi Awolowo University, Ile-Ife, Nigeria: E-mail: afolorun@oauife.edu.ng Abstract SCLA, a numerical taxonomic method was applied to lip morphology in the genus Polystachya Hook. (Orchidaceae) in Nigeria. he basic data matrix was prepared by coding for the presence or absence of the attributes of characters involved. he data were standardized so that the values of a particular character were transformed into values ranging from zero to one. he SCLA showed that reproductive characters are much better than the vegetative characters earlier used in the distribution of Polystachya species into their sections. In the case of reproductive characters, more clusters were reported; this may be an indication of more sections in Polystachya than those earlier reported. Keywords: Polystachya species, cluster analysis, reproductive character, vegetative character, sections Introduction Polystachya is a genus of about 200 species, mostly African but also occurring in Madagascar, Central and South America and Asia (Cribb and Rasmussen, 2004). Epiphytic, occasionally litophytic or terrestrial perennial herbs (Croix, 1997). Hutchinson and Dalziel (1968) listed ity seven species for west Tropical Africa, with Nigeria having 24 species. New species from Tropical west and South-Central Africa were described by Cribb and Croix (1996) among which is P. anthoceros from Nigeria. Segerback (1983) described 104 species of orchids for Nigeria, but Jayeola (1991) otherwise encountered over 400 species for Nigeria. he greatest challenge for the utilization of orchids in Nigeria for both scientiic and aesthetic purposes is the absence of simple identiication keys devoid of the usual highly technical details of taxonomy ( Jayeola and folorunso, 2002). he existing keys are too technical to be used by non-taxonomists such as foresters and horticulturists. Orchids are good lagship species and possible good environmental indicators (Cribb and Croix, 1996). hey contribute immensely to the national economy of Hawaii, Malaysia, Singapore and hailand, whereas in Nigeria there is little information about orchids. here is a great deal of variation in the loral structure of Polystachya to the extent that most species can be recognized on its basis ( Jayeola and folorunso, 2002). he lowers are known to vary with respect to colour, size and shape. Previous literature on the use of lip show that they are of diferent sizes and shape (Victoria, 1994; Luer, 1995; Catling, 1997). he lip otherwise known as the labellum is the relatively morphologically complex median inner perianth segment (petal) that, in most orchids, is the primary attractant and landing stage for pollinators, most commonly insects (Dafni, 1992; Nilsson, 1992; van der Cingel, 1995; Rudall and Bateman, 2002). Numerical taxonomy makes use of quantitative methods for the classiication of organisms into groups (Sokal and Sneath, 1963). Most of the techniques used in numerical taxonomy to measure overall similarity between taxa, as well as classifying them into groups are described by Sokal and Sneath (1963). Morphometric techniques have long been established as valuable tools for exploring the development, population diferentiation and systematics of plants (Bookstein et al., 1985; wiens, 2000; forey and MacLeod, 2002; Jensen, 2003; Bateman and Rudall, 2006). hese morphometric studies have typically employed between 20 and 50 quantiied characters, generally consisting of a heterogenous mixture of metric (continuous), meristic, scalar and presence / absence characters (Bateman and Rudall, 2006). Earnest study of the species of Polystachya has become imperative, as some species are being threatened by habitat loss, as for example Polystachya cooperi (Pollard and Darbyshire, 2004). he purpose of the present study is to describe the results of the use of single linkage cluster analysis on the lip of the genus Polystachya in Nigeria and to delimit the species into their traditional sections. Materials and methods Orchid lowers were obtained from two sources: fresh lowers were obtained from the orchidarium, in situ, while dried lowers were collected, ex situ, from the Botany Herbarium (IfE), Obafemi Awolowo University, Ile-Ife (Tab. 1). fresh lowers were dissected to expose the loral segments and dorsal petals or lips were excised. Preparations of dried herbarium specimens for study required boiling Folorunso, A. E. et al./ Not. Bot. Hort. Agrobot. Cluj 37 (1) 2009, 45-50 46 P_paniculata P_paniculata_var_n P_o_var_odorata P_o_var_trilepidis P_sp_novum P_supfiana P_albescens P_subulata P_ramulosa P_laxiflora P_stricta P_calluniflora P_golugensis P_saccata P_bifida P_tesselata P_modesta P_fusiformis P_orosun_var_novum P_polychaete P_o_var_orosun P_adansoniae P_cooperi P_galeata P_elegans P_mukandaensis P_dolichophylla P_affinis he phylogenetic relationships of the species of Polystachya is as shown in fig. 1. Six main clusters are observable at 0.5 similarity level. Cluster A, which is the largest P_rhodoptera P_alpina Results and discussion cluster encompasses many species of Polystachya covering sections like Cultriformes, Aines, Grandilorae, Callunilorae, Isochiloidae and Dendrobiantae. he species are P. dolichophylla, P. ainis, P. elegans, P. mukandaensis, P. cooperi, P. galeata, P. odorata var. orosun, P. adansoniae, P. orosun var. novum, P. polychaete, P. tessellata, P. modesta, P. fusiformis, P. saccata, P. biida, P. callunilora, P. golugensis, P. coriscensis P. camaridiodes, P. rhodoptera, P. alpina, P. ramulosa, P. laxilora, and P. stricta. Cluster B, comprised of P. albescens and P. subulata, included in the section Caulescent. Clusters C and D belong to Polystachya section and thus include two varieties of P. paniculata in addition to P. odorata var. odorata and P. odorata var. trilepidis. he characters used for the cluster analysis are representatives of both vegetative and reproductive characters of the species Polystachya. he species of Polystachya here listed are those reported for Nigeria (Hutchinson and Dalziel, 1968) and the 8 species representing entirely new records for the orchid lora of Nigeria ( Jayeola and folorunso, 2002). he result of the cluster analysis revealed a lot of intraspeciic relationships among the species of Polystachya P_coriscensis P_camaridiodes the lowers in ammonium chloride (NH4Cl) solution for thirty minutes to revive the tissues of the lips for detailed study under the microscope. he lips were then let to cool for two to three hours ater which they were dissected in the laboratory for examination. All data were scored from a minimum of ten accessions for each species. he mean values of these data were then coded (Tab. 2). A total of twenty six characters were recorded for each OTU. he basic data matrix of 26 x 32 was prepared by coding for the presence or absence of the attributes of characters involved. Data were standardized, so that the values of a particular character were transformed into values ranging from zero to one, using the method of Sokal and Sneath (1963). Single linkage cluster analysis (SCLA) was performed to study the phylogenetic relationships of members of the genus. 0.96 0.84 0.72 Similarity 0.6 0.48 0.36 0.24 0.12 0 0 4 8 12 16 OTU fig. 1. Phylogenetic relationships of the species of Polystachya studied 20 24 28 32 Folorunso, A. E. et al./ Not. Bot. Hort. Agrobot. Cluj 37 (1) 2009, 45-50 47 studied. Cluster A, which is the biggest of the clusters is a conglomerate of ive sub-clusters. for instance P. dolichophylla and P. ainis which have yellow lowers and whose mid lobes are short, form the irst sub – cluster comprised of P. elegans, P. mukandaensis, P. cooperi and P. galeata. All these species have green lowers with the apex of their lateral lobes acute. he third sub-cluster includes P. odorata var. orosun, P. andansoniae, P. orosun var. orosun, P. polychaete, P. tesselata, P. modesta, P. fusiformis, P. saccata and P. biida. hese are the species with white lowers together with purple markings. he fourth sub-cluster includes species like P. callunilora, P. golugensis, P. coriscensis, P. camaridiodes and P. rhodoptera with pale green lowers. he ith sub-cluster comprises of P. alpina, P. ramulosa, P. laxilora, P. stricta and they are largely white with yellow colour that is tinged with pink. Cluster B comprises P. albescens and P. subulata. hey have greenish white lowers and belong to the Caulescent section. he species in cluster C are with whitish yellow lowers tinged with rose. hose in cluster D have orange lowers with red markings. Conclusions his work has shown that reproductive characters are much better than the vegetative characters earlier used by Kraenzlin (1926) in the distribution of Polystachya species into their sections. with the use of reproductive characters more clusters have been reported which also indicate the occurrence of many more sections in Polystachya than those earlier reported. References Bateman, R. M. and P. J. Rudal (2006). Evolutionary and Morphometric Implications of Morphological Variation Among Flowers Within an Inlorescence: A case-study Using European Orchids. Annals of Botany. 98:975-993. Bookstein, F. L., B. Chernof, R. Elder, J. Humphries, G. Smith and R. Strauss (1985). Morphometrics in evolutionary biology. Philadelphia, PA: Philadelphia Academy of Natural Sciences. Catling, P. M. (1997). he taxonomic status of Fernald’s ragged Orchid of Newfoundland, Plantanthera lacera var. terraenovae. Lindleyana. 12(2):79-88. Van, der Cingel (1995). An atlas of orchid pollination: European orchids. Rotterdam: Balkema. Cribb, P. J. and I. Croix (1996). New Polystachya species (Orchidaceae) from tropical west and south-central Africa. Kew Bulletin. 51(3):571-577. Cribb, P. and J. Rasmussen (2004). Field guide to the Ethiopian Orchids. Croix, I. (1997). African Orchids in the wild and cultivation.W3 Tropicos, Kew Monocot list, IPNI. Dafni, A. (1992). Pollination ecology. Oxford: Oxford University Press. Forey, P. and N. MacLeod (2002). Morphology, shape and phylogenetics. In: Taylor and Francis (Eds.). Systematics Association Special Vol. 64. London. Hutchinson, J., J. M. Dalziel (1968). Flora of West Tropical Africa. 3(1):215-225. Jayeola, A. A. (1991). Computer-aided taxonomic study of the angraecoid orchids of Nigeria and Cameroon. Obafemi Awolowo University. Ph.D. hesis. 16-91. Jayeola, A. A. and A. E. Folorunso (2002). Floral morphology of the Polystachya Hook. (Orchidaceae) in Nigeria. Nigerian Journal of Horticultural Science. 6:70-75. Jensen, R. J. (2003). he conundrum of morphometrics. Taxon. 52:663-671. Kraenzlin, F. (1926). A monograph of the genus Polystachya. Feddes Repertorium; Beih. Vol.39. Luer, C. A. (1995). A new species of Pleurothallis (Orchidaceae) from French Guyana Bulletin du Museum. National d’ Histoire Naturelle section B. Adansonia Botanique. Phytochimie.16(24):231-233. Nilsson, A. (1992). Orchid pollination biology. Trends in ecology and Evolution. 7:255-259. Pollard, B. J. and I. Darbyshire (2004). Polystachya cooperi. 2006 IUCN Red List of hreatened species. Rudall, P. J. and R. M. Bateman (2002). Roles of synorganisation, zygomorphy and heterotopy in loral evolution: the gynostemium and labellum of orchids and other lilioid monocots. Biological Reviews. 77:403-441. Segerback, L. B. (1983). Orchids of Nigeria. A. A. Balkema Publishers. A member of Swets and Zeitlinger Publishers. Netherlands. Sokal, R. R. and P. H. A. Sneath (1963). Principles of Numerical Taxonomy. W.H. Freeman and Co. San Francisco, Publishers. Victoria, S. (1994). Bletia greenwoodiana (Orchidaceae), a new species from Durango, Mexico. Brittonia, 46(3):208-210. Wiens, J. J. (2000). Phylogenetic analysis of morphological data. Washington, DC: Smithsonian Institution. .OTU coded values of the species of Polystachya Lipcelled P_galeata P_stricta P_elegans P_golugensis P_o_var_odorata P_o_var_orosun P_o_var_trilepidis P_mukandaensis P_adansoniae P_orosun_var_novum P_alpina P_paniculata P_callunilora P_dolichophylla P_polychaete P_coriscensis P_camaridiodes P_albescens P_paniculata_var_novum P_cooperi P_supiana P_sp_novum P_saccata P_biida P_rhodoptera P_afinis P_tesselata P_laxilora P_subulata P_fusiformis P_modesta P_ramulosa 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 1 0 0 0 48 Lip-pubes- Lip-apex- Lip-apexcent acute depre-ssed 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 0 0 1 0 1 0 1 0 1 0 0 0 0 1 1 1 0 0 0 0 0 0 0 1 1 1 0 0 1 1 1 1 1 0 0 0 0 0 0 0 0 1 0 0 1 Lipmarginentire 0 0 0 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 Apexmidlobeacute 1 1 1 1 0 1 0 1 1 1 1 1 1 0 1 1 1 1 1 1 1 0 0 1 1 1 1 1 1 1 0 1 Marginincurvedbelow 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Venationnetwork Liplarge Lipsmall Lipmoderate Lipleshy Lipwithoutcallus 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 0 0 0 0 1 0 0 1 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 0 0 1 0 0 0 0 0 0 0 0 0 1 0 0 1 0 1 1 0 0 1 1 0 1 0 0 0 0 0 1 0 0 0 0 1 1 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 1 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 1 1 0 1 0 0 0 0 1 0 0 1 .OTU coded values of the species of Polystachya Lipclawed P-galeata P-stricta P-elegans P-golugensis P-o-var-odorata P_o_var_orosun P_o_var_trilepidis P_mukandaensis P_adansoniae P_orosun_var_novum P_alpina P_paniculata P_callunilora P_dolichophylla P_polychaete P_coriscensis P_camaridiodes P_albescens P_paniculata_var_novum P_cooperi P_supiana P_sp_novum P_saccata P_biida P_rhodoptera P_ainis P_tesselata P_laxilora P_subulata P_fusiformis P_modesta P_ramulosa 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 1 1 0 1 1 0 0 0 0 0 0 0 0 0 0 Lip-notLip-transwell deveparent loped 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 1 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 Lippunctate 0 0 0 0 0 0 0 0 0 0 1 0 0 1 0 0 1 1 0 0 0 1 0 0 1 0 0 1 1 0 0 0 LipLip-lobed- lobed-at atmiddle lower part 0 0 0 0 1 0 0 0 0 0 0 1 0 1 0 1 1 1 1 0 0 1 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 1 0 0 0 0 0 0 1 0 0 0 0 0 0 0 Lipaslongaswide Flowerswhite Flowers-red Flowersgreen Flowersyellow Flowerstinged-pink 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 1 1 0 0 0 1 0 1 0 1 0 0 1 0 0 1 0 1 0 1 0 0 1 0 1 0 0 1 1 0 0 0 1 1 0 1 1 1 0 0 0 0 0 0 0 0 0 0 1 0 0 1 0 0 1 0 0 0 0 0 0 0 1 0 0 0 0 0 0 1 0 1 1 0 0 0 1 0 0 0 0 0 0 1 1 1 1 0 1 0 0 0 0 0 0 1 0 1 1 1 0 0 1 1 1 1 0 1 1 0 0 0 0 0 1 1 1 1 0 0 0 1 0 0 0 1 1 1 1 0 0 1 1 0 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 Flowers-49 greenishyellow 0 0 0 1 0 0 0 0 0 0 0 0 0 0 1 0 1 0 0 0 0 0 0 0 0 0 1 0 0 0 1 0 Tab. 1. Representative Specimens of the Polystachya species in Nigeria. 1. 50 2. 3. P. dolichophylla Schltr. Keay 39, Nindam F.R. Zaria. P. ainis Lindl. Burtt B27, Abeokuta to Ijebu Ode FHI 24666 Keay Usenigbe F.R. Sapoba FHE 22714 Keay, Owo, Ondo Prov. Talbot 3776, Ohbe, Degema, Randole Barter 1863, Onitsha P. elegans FHI 25376 Keay Sankwala, Ogoja prov. P. mukandaensis De Willd. FHI 22410 Keay, Olokemeji F.R. Abeokuta Tab. 1 P. golungensis Rchb.f. King 22, Kogin Delli, Zaria Prov. 17. Olokemeji F.R. Keay. FHI 22712 Abeokuta, Akure F.R. Keay 28294 Ondo 18. P. coriscensis Rchb.f. Sanford WS/185/66, Benin Prov. Sapoba P. camaridiodes Summerh. Wright 101, Calabar 4. a. homas 1902 Onika Ohene FHI 6128 Janes, Onitsha Talbot 861, Oban 19. 5. Cooper 80, Obudu Scarp, Ogoja P. cooperi Summerh. FHI 15190; WS 5458 IFE 20. P. rhodoptera Rchb.f Talbot 903 Oban, Sapoba Kennedy 1917 21. P. alpina Lindl. Talbot 835, Oban Niagi 22. P. ramulosa Lindl. Sanford WS/184/66 Benin Prov. Sapoba Talbot 3647, Oban Degema P. laxilora Summerh. FHI 1366, WS/1521/65 Idanre Hills P. galeata Brenan 9116, Okomu F.R. Benin 6. 7. 8. 9. 10. 11. 12. 13. a. FHI 227738a Keay Wright 128, Ajagbodudu, Sapele b. Talbot 3725 Degema Cooper 82, Ogoja Plain FHI 25304 Keay Coombe 151 FHI 22429 Keay, Eleyele, Ibadan P. odorata var. orosun Sanford 4135, Orosun peak Idanre P. adansoniae Rchb. F. FHI 40169 Onochie, Niger Prov. Gwari, Bornu a. Sanford WS 216a, 221/66 Boyogbe, 5 Benin Province; FHI 25317 Ejiofor b. Ogoja Province FHI 22438 Keay Okomu F.R., Benin Prov. Talbot 867,927 Kundene Obari P orosun var. novum. Sanford 4123, Orosun Peak, Idanre P. polychaete Kraenzl. Talbot 927, 928 Oban P. tesselata Lindl. King 54, Kushanka, zaria Prov. Elliot 80, FHI 33190 Onochie, Stubbs Creek F.R. Eket FHI 13348 Keay, Calabar P. modesta Rchb. F. Cole 13, Zaria king 31, Mongu F.R. Plateau Province FHI 22443 Keay, Sanga River F.R., Zaria, WS/1848765 Agodi Gate, Ibadan. P. fusiformis ( hou.) Lindl. IFE WS/5668 23. 24. 25. 26. Daramola FHI 29809, Talbot 918 Oban P. stricta Rolfe Winbush 611; King 350 Manbia Plateau P. albescens Ridl. WS 423 IFE P. subulata Finet FHI 22898 Keay Ribaka F.R. Zaria Prov. FHI 37667 Keay, Mangu F.R. Plateau Prov. Summerhayes 91, Kaliy Zankwa Zaria, Rchb.f. 27. P. sp. novum WS/443/66 sapoba F.R. 28. P. supiana Schltr. Talbot 440, Kwa F. Oban, FHI 3306, Eket; IFE WS/491/66. Sapoba P. saccata V.S.Summerh. 90 Kagarko, Zari FHI 37620 Keay and Janes Gimi River F.R. Zaria 29. P. odorata var. odorata Barter 1483, Onitsha 0.5cm Kennedy 1686 Sapoba Richards 3409, Shasha F.R. Ijebu FHI 33176 Onochie Orukini Unyene, Eket FHI 33209 Onochie Lower Enyong F.R. Itu FHI 37639 Keay and Janes Sanga River F.R. Zaria 30. 14. P. odorata var. trilepidis Summerh. FHI 20729bJanes, Mt. Orosun, Idanre, Ondo. FHI 21559 Keay and Onochie FHI 3374 Symington. 15. P. biida Summerh. IFE WS/5990, WS/199/66 sapoba F.R. Obudu Plateau Ogoja 31. 16. P. callunilora Kraenzl. WS/3541 IFE 32. P. paniculata (SW.) Rolfe; Keay, FHI 25371, Akure F.R.; Ejiofor FHI 24653 homson FHI 1509, Amedzofe; Ngogi, FHI 15341 P. paniculata (SW.) Rolfe subsp. novum