CONTRIBUTION OF PHYLOGENETIC CLASSIFICATION OF THE FAMILY APIACEAE IN THE MONGOLIAN FLORA Urgamal M. Institute of Botany, Mongolian Academy of Sciences urgamal@botany.mas.ac.mn Abstract Family Apiaceae consists of 74 species and 37 genera, 6 sub-tribes, 12 tribes (4 clades) and 2 sub-families (Saniculoideae and Apioideae) disjunctly distributed in the Mongolian flora. As determined by extensive molecular studies and critical evaluation of morphology, the 32 genera of the family Mongolian Apiaceae. A phylogenetic analysis was conducted to clarify inter and intraspecific relationships based on sequences of the ITS and 5.8S regions of nrDNA and cpDNA (by Downie et al., 1998, 2001). We updated classification system (by Downie et al., 2010) of the family Apiaceae in Mongolia and changed its system orders, phylogenetic placement tribes and some genus. Our analyses of these molecular data, in conjunction with results of identified 12 tribes (4 clades) in Mongolian Apiaceae at the tribal rank (Urgamal, 2013). First time phylogenetic classification of Apiaceae inferred from DNA sequence data in the Mongolian flora. Keywords: Apiaceae, tribe, nrDNA & cpDNA ITS, phylogenetic placement Recently research works for phylogenetic classification of the family Apiaceae was based on taxonomic congruence among the results of phylogenetic analyses of different molecular data sets, including chloroplast DNA (cpDNA) gene (rbcL, matK) and intron (rpl16, rps16, rpoC1) sequences, cpDNA restriction sites, and nuclear ribosomal DNA (nrDNA) internal transcribed spacer ITS region (ITS 1, 5.8S, and ITS 2) sequences. Sequences of the internal transcribed spacer region (ITS 1, 5.8S, and ITS 2) of nuclear ribosomal DNA (nrDNA) have been a staple source of data for the study of lower level phylogenetic relationships among plant taxa for more than ten years (e.g., Baldwin, 1992; Baldwin et al., 1995). We investigated the utility of nuclear ribosomal DNA (nrDNA) internal transcribed spacer (ITS) region and compare the results to those obtained from the chloroplast DNA (cpDNA). We report that the a higher percentage of parsimony informative characters than that of ITS and chloroplast DNA (cpDNA) loci examined to date. Materials and Methods Our materials including nrDNA ITS region markers is following accessions: Aegopodium kashmiricum (AF077872), Anethum graveolens (GQ148794), Angelica dahurica (JN603214), Anthriscus sylvestris (EU236159), Apium graveolens (GQ148789), Aralia chinensis (AF242256), Aulacospermum simplex (GQ379338), Bunium capillifolium (GQ379304), Bupleurum scorzonerifolium (EU001347), Carum carvi (AF077878), Cenolophium denudatum (AF077879), Cicuta virosa (AY524767), Cnidium monnieri (AY863065), Conioselinum vaginatum (FJ385041), Coriandrum sativum (HQ377194), Daucus carota (FJ415158), Eryngium planum (EU169002), Ferula dissecta (DQ379394), Ferulopsis hystrix (EU169271), Foeniculum vulgare (GQ148793), Hansenia mongholica (JF694084), Heracleum dissectum (EU185691), Oenanthe aquatica (AY691928), Paraligusticum discolor (AF077902), Pastinaca sativa (EU169305), Petroselinum crispum (GQ148797), Peucedanum terebinthaceum (JN603232), Pimpinella saxifraga (AY581801), Pleurospermum prattii (DQ270204), Prangos pabularia (AF077906), Saposhnikovia divaricata (EU592001), Schulzia albiflora (FJ385059), Seseli praecox (AY179024), Sium suave (DQ005693) and following accessions of cpDNA C1 (rpoC1): Aegopodium alpestre (U72434), Angelica dahurica (GQ248886), Aralia californica (U72480), Bupleurum chinense (U72455), Carum carvi (U72435), Cicuta virosa (U72447), Conioselinum chinense (U72452), Daucus carota (HQ593968), Ferula assafoetida (U72421), Foeniculum vulgare (GQ436171), Oenanthe pimpinelloides (U72442), Peucedanum morisonii (U72414), Pimpinella rhodantha (U72429), Prangos pabularia (U72431), Sanicula canadensis (U72461), Seseli krylovii (U72419), Sium suave (HQ594137). The molecular analysis included 32 species of nuclear and 29 species chloroplast DNA sequences, representing Results We updated classification system (by Downie et al., 2010) of the family Apiaceae in Mongolia and changed its system orders, tribes and phylogenetic genus placement of tribes and genus. Our analyses of these molecular data, in conjunction with results of identified 12 tribes (4 clades) in Mongolian Apiaceae at the tribal rank (Figure 1). Conclussion Aside from providing a framework for taxonomic revisions, the phylogenetic structure recovered in this study for subfamily Apioideae will lay the foundation for future investigations of evolutionary patterns of morphological characters and biogeography. An nrDNA 32 genera from the 12 tribes and 2 subfamilies of the family Mongolian Apiaceae and Araliaceae species is used as a outgroup taxa. Sequences of nuclear (nrDNA ITS region) and chloroplast (cpDNA rpoC1) from the published study (Downie et al., 1998, 2001) are analyzed using MEGA 5.05 software. The methodology for the alignment of nrDNA ITS region and cpDNA study data, either complete ITS 1 and ITS 2 nuclear ribosomal DNA region sequences were used. Phylogenetic analyses were carried out using Maximum Parsimony in MEGA version 5.05 software of partitioned ITS data sets resulted in highly consistent phylogenies, whereas analyses of combined molecular data resulted in trees of greatest resolution and overall branch support. Bootstrap values, however, were calculated for 1000 replicate analyses; decay values were also obtained. The resultant maximally parsimonious trees, were rooted with Aralia (Araliaceae - outgroup), having all of it`s states scored as “0”. Up to date, we have officially registered 74 species belonging to 37 genera, 6 sub-tribes, 12 tribes (4 clades) and 2 sub-families (Saniculoideae and Apioideae) belonging to the family Apiaceae that occur in the Mongolian flora (Figure 2). ITS-based classification of the subfamily is proposed that enables direct and precise references to specific groups and facilitates sampling and hypothesis testing in future systematic studies of family Mongolian Apiaceae. References Downie, S.R., Ramanath, S., Katzdownie, D.S. & Llanas, E. 1998. Molecular systematics of Apiaceae subfamily Apioideae: Phylogenetic analyses of nuclear ribosomal DNA internal transcribed spacer and plastid rpoC1 intron sequences. – Amer. J. of Bot. 85(4): 563–591 Downie, S.R., Katz-Downie, D.S. & Watson, M.F. 2000. A phylogeny of the flowering plant family Apiaceae based on morphological and chloroplast rpl16 and rpoC1 sequences: Towards a generic classification of subfamily Apioideae. – Amer. J. Bot. 87: 273-292. Downie, S.R., Plunkett, G.M., Watson, M.F., Spalik, K., Katz-Downie, D.S., ValiejoRoman, S.M., Terentieva, E.I., Troitsky, A.V., Lee, A.-Y., Lahham, J. & El-Oqlah, A. 2001. Tribes and clades within Apiaceae subfamily Apioideae: the contribution of molecular data. Edinburgh J. Bot. 58(2): 301-330. Downie, S.R., Spalik, K., Katz-Downie D.S. & J.-P. Reduron. 2010. Major clades within Apiaceae subfamily Apioideae as inferred by phylogenetic analysis of nrDNA ITS sequences. – Pl. Diversity and Evol. (formerly Botanische Jahrb.) 128: 111-136. Pimenov, M.G. & Leonov, M.V. 1993. The genera of the Umbelliferae. Royal Botanic Gardens, Kew, UK. Plunkett, G.M. & Downie, S.R. 1999. Major lineages within Apiaceae subfamily Apioideae: a comparison of chloroplast restriction site and DNA sequence data. – Amer. J. Bot. 86: 1014-1026. Plunkett, G.M. 2001. Relationship of order Apiales to subclass Asteridae: A re-evaluation of morphological characters based on from molecular data. - Edinburgh J. Bot. 8: 183-200. She, Menglan, Fading, Pu., Zehui, Pan., Watson, M.F., Cannon, F.M., Ingrid Holmes-Smith, Kljukov, E.V., Phillippe, L.R. & Pimenov, M.G. 2005. Apiaceae (Umbelliferae) Flora of China, tomus 14. Urgamal, M. 2009. Flora of Mongolia. vol. 10, (Apiaceae-Cornaceae). – Ulaanbaatar, "Bembi san", 130pp. (in Mongolian). Urgamal, M. 2012. Taxonomy of the family Carrot (Apiaceae Lindley 1836) in Mongolia. Erforschung Biologischer Ressourcen Der Mongolei. – Absracts Int.l Symp. ”Biodiversity of Research in Mongolia” Halle (Saale), Germany; 25-29, March 2012: 39. Urgamal, M. 2013. Plants of the family Umbelliferae Juss. of Mongolia. – Thesis of Ph.D. – Ulaanbaatar, Mongolia. (in Mongolian). Urgamal, M. 2013. Results of taxonomic investigation of the family Apiaceae in the Mongolian flora. – Proc. of Int. Conf. Plant Biodiversity and Ecosystem Services in Continental Asia, Ulaanbaatar, Mongolia, 26-27, June: 69-71. Vinogradova, V.M. 1994. Plantae Asie Centralis. (Araliaceae, Umbelliferae, Cornaceae) – Sankt.-Petersburg, Т. 10: 41-42. (in Russian). МОНГОЛ ОРНЫ ШҮХЭРТЭНИЙ ОВГИЙН ФИЛОГЕНЕТИК АНГИЛАЛЗҮЙН АСУУДАЛД Ургамал М. ШУА-ийн Ботаникийн хүрээлэн urgamal@botany.mas.ac.mn Хураангуй Өнөөдөр Монгол орны Шүхэртэний овогт 2 дэд овгийн 12 триба (4 clade), 6 дэд трибад хамаарах 37 төрлийн 74 зүйл ургана. Монгол орны Шүхэртэн овгийн триба ба бүлэглэлүүдийг төлөөлүүлж 15 төрөлд хамаарах 15 зүйлийн; харин төрлүүдийг төлөөлүүлж нийт 32 зүйл ургамлын эсийн бөөмийн (ITS and 5.8S regions of nrDNA) болон хлоропластын (ITS and 5.8S regions of cpDNA) ДНХ-н нуклеотидын дарааллыг (Downie et al., 1998, 2001) тогтоосон дүнгээр задлаг хийсний дүнд (Ургамал, 2013) овгийн ангиллыг шинэ систем (Downie et al., 2010)–ийн дагуу боловсруулж, триб болон зарим төрлүүдийн филогенетик дэс дараалалд өөрчлөлтийг хийв. Keywords: Apiaceae, триб, бөөмийн ДНХ, хлоропластын ДНХ, филогенетик ангилал View publication stats