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136-019-2558-ตี orlly6S NRC Genetic diversity and population structure of pencil yarlr (Vigna lanceolata) (Phaseoleae, Fabaceae), a wild herbaceous legume endemic to Australia, revealed by microsatellite markers Phakchana Nubank6h, sarocha Pimtong, Prakit somta, Sujinna Dachapak, and Peerasak Srinives Abstrac! pencil yaDr (yi8no lan(eoldto Benth.) (Phaseoteae, Fabaceae) is a herbaceous legume endemic to Austmlia. A Pr-evious DorpholoSical study sufLested that pencil yam is a complex species of two or more related t:xa with seven distinct nrorPho genetic diversity and deter' logiial typrs lmorphorypes) and, thus, taxonomic levision is nec€ssary. ID this study, we assessed accessions fiom seveD morphotyPes using 18 microsateUite (simple o mined tire geneti. structure ofa pencilyam collection of62 sequence ipeatl markers with the aim to provide information for taxonomic study. In total. 138 alleles were d€tected with a ofo.61, mean of7.6i alleles per locus. polymorphism infornation content per marker vaded betwe€n 0.06 and 0.90 with a mean while the overall gene diversity was 0.62. Bayesian clustering, principal coordilate, and neighborjoining analyses consistently revealed that these accessions are grouped into two subpopulations with differenc€ in number ofllleles, allelic richness, and genc diversity. The population structure was not related to either morphotype or geographical origin. Gene diversity of yi8ro compaEble i. Ianceolata was higherih.r. that ofwild vigaa rodiata (1.)Wilczek and wild utnbell@ta (Thunb.)Ohwi & Ohashi, withthatofwild yiSno nungo (1.)Hepper, yiSna erilis Tateishi & Ma-xted, andvigna gfandifiora lPIain)Tateishi & Maxted and lowe! than that of wild rigra o[Arhrir (Willd.]Ohwi & Ohashi. These results indicated that the taxonomy ofy. lonceolato should be v reyised and that its gene diversitywas moderate compared wirh the other viild Visno sPecies (d, wordsj yi8no, microsatellite marker, simPle sequence rePeat, SSR. R6sum6: yiSnd laflceolotd Benth. (Phaseoleae, Fabaceae)est une l€gumineuse fourragCre end6mique de l'Australie. Une 6tude tyPes nrorphologique antErieure a sugg€rd que y, drreoloto est une espace conPlexe comPrcnant au moins deux taxons et sept auteurs ont morphologiques distincts {morphotypes), ce qui rend nEcessaire sa r€vision taxonomique. Dans cette dtude, les y. t5 6val;6 la diversit6 g€n6rique et ddtermin6 la siructure g6n6tique d une collecion de lanceoiaia comPortant 62 accessions de sept morphotypes il'aide de 18 rnarqueurs micrositellites (dp6titionde s€queoce simple), darls Ie but d obtenir de l'information *" du t.frua" t"*onomique. Au total, 13E allales ont 4t6 d6tect6s, avec une moyenne de 7.67 alliles Par locus. Le conteDu informationnel"n du polymorphi5me par marqueur variait de 0,06 i 0.9o, avec une moyenne de 0,61, alorc que la dive$it€ g€n6tique globale 6tait ae o,eZ.I agrigation b;y6sienne,les analrses erl coordonn6es Principales et la m6thode de NJ ont rev6l6 iyrte,notiq-u"-"rrr qr. ces accessions sont groupEes en deux sous-populations qui difftr€nt sur le Plan du nombre d allales d€ I origine ti richesse attetque et de la dive6it6 g6n€iique. La structure de la population n'6tait Pas reli6e au morphotyPe ou i plus grande que celle de yigno rodidto (1.)wilczek et ylSno umbellota E g6ographique. l-adiversit6 gdndtique de v. Ior.eolats Ctait o yigad m!r8o (1. Hepper sauvage, yi8na exilisTateishi & Maxted, et Vigna & irnunu_.1Oirwi a otrastri sauvages, comparable a celle de ) de viSlo an$laris (willd.) ohwi & ohashi sauvage. ces r6sultats 87an.ilrlora lPrain)Tateishi & Maxt€d, et plus faible que celle ! indiqu"i"ni qr" i" t*onomie de y. lanse;lrro devrait €tre rCvis6e et que sa diversitd 86n6tique dtait moddrde comPlrativement o e celle d'autres esptces de viSnd sauvages. lTraduit par la Rddactionl Moli-clis j yi8xo, marque!r microsatellite, reP6tition de s6quence simPle SSR. o lntroduction (Thunb.) ohwi and Ohashi), and cftole bean (Vi$lo relexo'piloso (Hayata)) (Smartt 1990;Tomooka et al. 2006). Some ofthese crops The genus yi8,m (Phaseoleae, Fabaceae) is a Iarge leguminous o such as cor,pea, mungbeat!, black gram and azuki bean are im_ taxon comprising seven subgenera of1o4 herbaceous sPecies dis' portant fronr an agro-economic ln addition. several ffibuted in topical and subtropical rcgions ofAfrica, Asia' America, Perspective. wild yigno species are cultivated as cover or hawested as and Australia (Lcwis et al. 2oos). Nine yiSra species are domesti- Sxound foods iIr the fol.in ofyoung leaves, flowers young cated as food crops in Asia, Afiica, and America. They include supplementary pods, seeds, and fresh tubers (Lawn and Cottrell 1988; Bambara groundnut (yignd nlbterrdnea (L.) Verdc.)' cowpea (Vigno mature Tomooka et al. 2006). These Iegumes are important sources of unSuiculara (L) Walp.). zombi Pea (vi8no vexillata (L.) A.Rich)' mung- (Ohwi) dietary protein, carbohydmte, vitamins, and minerals for people bean lvigr,a radiatn lL.) wilczek), azuki bean {yi8na an8tllorij and America. Therefore, yiSno is an important Ohwi & Ohashi), black gram {yigno murSo 1L.) Heppe!)' moth bean in Asia, Africa, \Vigno acoflitifolia (Jacq.) Mar6chal), rice bean {Vigno &rnDellota plant taxon that is ofinterest. Reccived 3 November2014. Accepted E January2015. University, p. Nubankoh and S. pimtong. progam in Agriculrural Biotechnolory, faculty of Agriculture at Xamphaeng Saen, Kasetsart Nakhon Pathom 73140, Thailand. p. Xamphaeng Saen, Kasetsart University, r. io-t , S. n .fr.p"fc anil Srinives. Department ofAgonomy, fac'lrlty ofAgriculture at Nakhon Pathom 731,10, Thailand. corEspondlng author: Prakit Somta {e_mail: [email protected]). Botany 93: 1E3-191 (201s) dx.doi.orS/10.1139/.jb'201'I{222 .l Prblished it M.nrcrescar.hpress.conlcjb on 31Jrnuary 201s. Botany Vol. 93, 2015 Five yignd species inctuding yryfla radiata lL.l Wilczek var. highly transferable to odrer related species ffangphatsonuuang iublobdrrl (Roxb.) Verdc. (wild mungbean), \4 vcxillou, \\gnd lafteolatLl et al. 2009). SSR markers can be detected by standard polymerase gelus Benrh. (pcncil yam or bush carrot), yiSra hrteola Uacq.) Benth. chain reaction (PCR) analysis and are easy to score. ln the (hairy pod cowpea), and yigra tnarino (Burm.) Merrill (beach cow' yi8!d, SSRS have been develoPed for azuki bean (warg et al. pea) are found in Australia. The first two species belong to the 2004; Chankaew et al. 2014), colvpea (Li et al. 2001; GuPta and subgeneE Cera[orroPis and Plectotropis, respectively, while the lat' GopalakrishrF 2O1O; Xu ct aI.2010; Kongraimun et a1.2012)' and ter thrce species belong to subSenus Vitna. Among these species, mungbean (Somta et al.2008: Seehalak et aI.2009i Somta et al. o]nly V.lanceolaLa is endemic to Australia, while tbe other four 2009i Tangphatsornruang et al. 2009). SSR markers from azuki species are indigeDous (Lawn and Watkinsolr 2002).Vignlan eolata is bean and mungbea[ showed a high rate of amplification in sev_ an important "bush tucker" food for Aborigi[es in Austmlia' eral other Vigra species (Somta et al. 2009; Tangphatsoralruang where the root is commonly eaten in the desert at present, and it et al. 2oo9; Chankaew et al. 2014). was evaluated a set of is a palatable and useful species in Pastures. Vigrra lanceolora is the ln this study, SsR polymorphism io most widely distributed Vlsna species fould throughout the 62 accessions ofV- lan.eolata. The objectives were (i) to assess the y. Australian tropics and subtropics encompassins a divemity of transfembility of SSR markers in I@nceoldta, (ii) to estimate the habitats from the foreshore to the ce[tral desert (Lawn and Ievel of genetic diversity of this species, and (iii) to determine Cottrell 1988; Lawn and Watkioson 2002). An analysis of more population structure within V. lonceolota species comPlex for fur_ thalr 400 accessions of the flve Australian wild vi8n4 species ther taxonomic study. showed that v. lanceolata is the most morphologically and ge" netically diverse species (Lawn and Watkinson 2002). SamPles Materials and methods collected frout fretd/natural habitats showed high variation in Plant materials and DNA extraction morphology and reproductive systems Annual or perennial Sixty-two accessions ofy.lotlceolotd from various Prove[ances in plants showing trailing, twining, and bush-t]?e habits with Australia were used in this study. They represented the seven rhizomes and (or) tuberous roots were found. Although many morphoo'pes of the species suggested by Lawn and Watkinson ) accessions Produced flowers only on the aboveground stems. (2002), including (i) northem pubescent, (ii) geotropic inflores' rnany othcrs are amphicarPic, and some have aerial inflores_ cence, (iii) var.fliJbrmis, (iv)central Austlalia, (v)erectbush' (vi)silver cences on plostrate stems, becominS geottopic with geocarPic leaflet, and (vii) var. lomsolata (SuPPlementaryTable SIi. su(h pods. Vigna laficeolota also Possesses useFul ldaptive traits Seeds ofeach accession were sown in an exPerimental field of IZ as early flowering and frost tolerance (Lawn alrd Holland 2003). Kasetsart University, KamPhaeng Saen Campus, Nakhon Pathom, which may be useful for crop improvement in the future. Thailand. Young leaves from a si gle plant of each accession were Three botanical vaieties have long been desdibed for V lanaedata' method described 66 collected and exEacted for DNA following the viz. vat. fliiorrnis,lanceolatd, and lotlfolia (Bentham 1854 and white by Lodhi et al. (1994)with the exceptionthat absolute ethanolwas a 1918: cited in Lawn and Watkinsolr 2002). However, results frorn used instead of95% ethanol for DNA precipitation. The DNA con- study based on morphological tlaits bv Law[ aDd Watkinson centration was determined by compariso[ with a knowD concen' having 46 (2002) suggested thal V. lanceolatu is a sPecies complex tration of tr DNA and adjusted to 2 n8 L-1 for SSR malker analysis. seven different morphoq?es (var. trtiJonnis northem pubescent ii9 form, geotropic inflorescence, silver leaflet type, centralAustnlia ssR marker analysis 9: type, robust erect type, and var. 14nceoloto). VarietyrliJomlis was Eighty SSR markers in total from azuki bean (Wang et aI 2004; reitricted to wet areas of coastal regions in the far north of Chankaew et al.
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