Iran J Biotech. 2014 June; 12(2): e10315. DOI: 10.5812/ijb.10315 Research Article Published online 2014 June 15. Lathyrus Fabaceae-Fabeae Molecular Phylogeny of the Genus ( ) Based on matK cpDNA Sequence in Iran 1,* 2 3 Roghayeh Oskoueiyan ; Shahrokh Kazempour Osaloo ; Atefeh Amirahmadi 1Department of Biology, Ayatollah Amoli Branch, Islamic Azad University, Amol, I.R. IRAN 2Department of Plant Biology, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, I.R. IRAN 3Department of Biology, Damghan University, Damghan, I.R. IRAN *Corresponding author : Roghayeh Oskoueiyan, Department of Biology, Ayatollah Amoli Branch, Islamic Azad University, Amol, I.R. IRAN. Shahrokh Kazempour Osaloo, Department of Plant Biology, Faculty of Biological Science, Tarbiat Modares University, Tehran, I.R. IRAN, Tel: +98-9125180684, Fax: +98-88249642, E-mail: [email protected]@modares. ac.ir Received: ; Revised: ; Accepted: January 16, 2013 June 8, 2013 January 7, 2014 Background: Lathyrus More than 60 species of the genus are distributed in Southwest Asia. It is the second largest genus of the Vicia , tribe Fabeae, after , in the region (and in Iran with 23 species). In the regional Flora (Flora of Turkey Flora Iranica and flora of Lathyrus Pisum Iran), the genus has been divided into 9-10 sections. Here we analyzed the phylogeny of and its relationship with matK based on plastid gene sequences. Objectives: The present study utilized several approaches including maximum parsimony, Bayesian and maximum likelihood Lathyrus methods to evaluate the monophyly and relationship within the genus , both at the sectional level and species level, mainly based on the taxa growing in Iran. Materials and Methods: Lathyrus Pisum Vicia A total of 52 accessions, representing 38 species of , three species of and four species of Lens matK and as out-groups, were analyzed for reconstructing the phylogenetic relationship using chloroplast gene sequences. Maximum parsimony, Bayesian and maximum likelihood methods were used to construct phylogenetic trees. Results: Pisum Lathyrus Lathyrus Clymenum The present study indicated that was nested among species. Two members of the section, Lathyrus ochrus L. Clymenum Pisum, ( and ) with formed a weakly supported clade as sister to the larger polytomy comprising the Lathyrus Lathyrus Lathyrostylis Lathyrus Clymenum remainder of the species. Several sections of including , and were monophyletic. Lathyrus roseus Orobon Lathyrus (of the monotypic section ) were nested among the members of section . The newly taxon described L. alamutensis Lathyrostylis Linearicarpus Orobus Pratensis species , endemic to Iran, were nested among other species of . , and were Pratensis Aphaca L. Pratensis not monophyletic sections. and the monotypic section were the closest taxa. In our analysis, formed a Aphaca sister group relationship with the clade, not its own section. Conclusions: matK Lathyrus Shimodaira-Hasegawa (SH) test of the data set showed that all of the analyzed species formed their Pisum Lathyrus own clade and was sister to them. Furthermore, when we removed the two above-mentioned species, the analysis Pisum, Lathyrus retrieved as a well-supported clade being sister to the calde. Keywords: Fabaceae; Lathyrus; Tribe Fabeae 1. Background smaller centers in North and South America (2, 6-8). Many Lathyrus species of are used extensively as cover crops, for L. cicera L. hirsutus L. sativus The tribe Fabeae (= Vicieae) contains four genera, of fodder ( L., L. and L.), as orna- Lathyrus Vicia L. odoratus L. latifolius which L. and L. with ca 160 species are the mentals ( L. [sweet pea], L. [everlast- Pisum L. sylvestris L. sativus largest (1, 2). Lens Mill. with 4-6 species and L. with 4 ing pea] and L.) and as human food ( Vavilovia formosa L. ochrus species ( (Stev.) Fed. of the monotypic ge- [grass pea, Indian pea, chuckling vetch], (L.) DC. Vavilovia Pisum P. formo- L. montanus nus Fed. was treated as a part of as and Bernh. (with edible root tubers) and also sum Vicia (Stev.) Alef. (3) are small genera. is paraphyletic for erosion control, as green manure and for medicine; Lens Lathyrism and along with , as a monophyletic taxon, formed toxins are present in some species, causing (2). Lathyrus a sister group relationship with a clade of and More than 60 species of the genus have been distrib- Pisum Lathrys (2-5). Species of are distributed through- uted in the Southwest Asia (2). In the regional Flora in- out the Northern Hemisphere, tropical East Africa and cluding Flora of Turkey (9), Flora Iranica (10) and Flora of temperate South America. Its main center of diversity is Iran (11), the genus was divided into 9-10 sections. Almost in the Mediterranean and Irano-Turanian regions, with three decades ago, Kupicha (6) based on morphological Implication for health policy/practice/research/medical education: Lathyrus The present study aimed to evaluate the monophyly and relationship within the genus. Copyright © 2014, National Institute of Genetic Engineering and Biotechnology; Published by Kowsar Corp. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. Oskoueiyan R et al. characters, by assessing the older classifications of the ge- (AFLP) data to examine the systematic relationship in sec- Lathyrus nus, proposed her sectional system encompassing 13 sec- tion . Ben Brahim et al. (16) studied isozyme varia- Orobus tions, of which three were newly established: (L.) tion and phylogenetic relationships between ten species Lathyrostylis Pratensis Gordon, (Griseb). Bassler, Bassler, of the genus. Kenicer et al. (8) studied 53 species of the Orobon Orobastrum trn trn Neurolobus Bassler, Tamamshjan, genus based on nrDNA ITS, cpDNA L-F and S-G se- Linearicarpus Lathyrus Boiss, Viciopsis Kupicha, Kupicha, , quence data (except members of two Kupicha’s sections, Clyme- Orobon Viciopsis Aphaca (Miller) Dumort. Nissolia (Miller) Dumort, and ). Their results generally supported num Lathyrus Lathyrus Lathyrostylis (Miller) DC. and Noto Kupicha. She divided the monophyly of sections and , but Orobastrum Oro- Orobus section into three monotypic sections questioned monophyly of the larger section . bastrum Linearicarpus , and Viciopsis. She included section Cicercula Lathyrus (Medic.) Gren. and Godr. in section and 2. Objectives Notho- added a new section of South American species, Lathyrus In this study, we examined the phylogenetic relation- . We followed her system of classification in the Lathyrus ships among 38 species of the genus based on present study. matK chloroplast gene . All species of the genus growing Dogan et al. (12) conducted a numerical taxonomic Lathyrus in Iran and some species from Turkey were included. study on 52 species of Turkish using 40 external Unlike some previous studies (7, 8) the present study vegetative and floral morphological characters. They di- Gor- employed several approaches including maximum par- vided the genus into nine sections, of which section gonia simony, Bayesian and maximum likelihood methods to Dogan was described as a new taxon. Leht (13) based Lath- evaluate the monophyly and relationship within on cladistic analysis of 210 morphological characters yrus Pratensis , both at the sectional level and species level. suggest that all species of section Bassler should Aphaca belong to section . He placed the monotypic sec- 3. Materials and Methods Orobon L. roseus Steven tion (with species ) within section Orobus (13). Hitherto, several works based on molecular Lathyrus 3.1. Taxon Sampling data have been conducted on the phylogeny of . Lathyrus Asmussen and Liston (7) sampled 42 species of Fifty-two accessions for matK were analyzed. A total of Lathyrus based on cpDNA restriction site characters and their 38 species representing eleven sections of (6) Vici- Pisum Vavilovia formosa study included representatives of all sections but and three species of (including , opsis . They generally agreed with the sectional classifica- see 3) were included in the phylogenetic analyses using Orobon tion of Kupicha (6), yet they suggested that and each of the separate datasets as well as the combined da- Orobastrum Lathyrus, should be transferred to the section taset. The two species of Lens and Vicia were chosen as NothoLathyrus Orobus and to . Croft et al. (14) used Random outgroups according to previous studies (4, 5, 7, 8). List of Amplified Polymorphic DNA (RAPD) analysis on eight ac- the analyzed species along with their voucher informa- L. sativus cessions of and some allies species. Badr et al. tion and GenBank accession numbers are presented in (15) utilized Amplified-Fragment Length Polymorphism (Table 1). Table 1. mat Sampled Taxa Used in This Study With Their GenBank Accession Number ( K) o Species DNA Source (Location, Voucher) GenBank Accession No Section Aphaca - - Lathyrusaphaca L b Iran: B. Alizadeh 1005 (WARCNH) AB935057 Lathyrusaphaca L. a USDA 2865272 AF522084 Lathyrusaphaca L. a Portugal: Schaefer, H HM851115 Section Clymenum - - LathyrusClymenum L. a Portugal: Schaefer, H HM851116 Lathyrusochrus (L.) DC. b Turkey: H. Sagban 3137 (GAZI) AB935075 Lathyrusochrus (L.) DC. a Portugal: Schaefer, H HM851120 SectionLathyrostylis - - Lathyrusalamutensis Mozaffarian et al. b Iran: Mozaffarian, Ahvazi & Charkhchian 88388 AB935055 (TARI) Lathyrusarmenus (Boiss. & Huet) elak. C b Turkey: Aytaç 8317 (GAZI) AB935058