豌豆属(Pisum)SSR标记遗传多样性结构鉴别与分析  

Identification and Analysis of Genetic Diversity Structure Within Pisum Genus Based on Microsatellite Markers

作　　者：宗绪晓[1] Rebecca Ford Robert R Redden 关建平[1] 王述民[1] 

机构地区：[1]中国农业科学院作物科学研究所/国家农作物基因资源与基因改良重大科学工程,中国北京100081 [2]BioMarka,Faculty of Land and Food Resources,The University of Melbourne,Melbourne,Australia3010 [3]Australian Temperate Field Crops Collection,Grains Innovation Park,The Department of Primary Industries,Private Bag260,Horsham,Australia3401

出　　处：《中国农业科学》2009年第1期36-46,共11页Scientia Agricultura Sinica

基　　金：国家"十一五"科技支撑计划项目(2006BAD13B05;2006BAD02B08);农业部作物种质资源保护项目[NB07-2130135-(25-30)-13];科技部植物种质资源共享平台建设项目(2005DKA21001-06);中澳政府间大型双边国际合作项目(ACIAR:CS1/2000/035)

摘　　要：【目的】评价豌豆属(Pisum L.)2个种5个亚种下,共8个资源类群的遗传多样性水平,揭示豌豆属下资源群体结构及其遗传关系远近,验证传统植物学分类的可靠程度,为充分发掘、利用豌豆野生种质提供必要信息。【方法】利用21对豌豆多态性SSR引物,对来自世界5大洲62个国家的豌豆属94份栽培种质(P.sativum ssp.sativum var.sativum)及其1个近缘野生种(P.fulvum),3个野生亚种(P.sativum ssp.abyssinicum、P.sativum ssp.asiaticum、P.sativum ssp.transcaucasicum)和3个野生变种(P.sativum ssp.elatius var.elatius、P.sativum ssp.elatius var.pumilio、P.sativum ssp.sativumvar.arvense)的103份野生种质进行SSR标记遗传多样性分析;利用NTSYSpc2.2d软件估算其遗传距离,进行主成分分析(PCA)并绘制三维空间聚类图;利用Popgene V1.32估算种质群间的Nei78遗传距离等参数并进行UPGMA聚类分析,采用MEGA3.1绘制种质群间聚类图;采用Popgene V1.32估算种质群的等位位点分布等参数,利用Fstat V2.9.3.2进行种质群间遗传多样性差异显著性测验。【结果】21对豌豆多态性SSR引物共扩增出104条多态性带,每对引物平均扩增出4.95个等位变异,其中有效等位变异占65.56%;PSAD270,PSAC58,PSAA18,PSAC75,PSAA175和PSAB72等SSR引物最为有效。SSR等位变异在豌豆属植物学分类单位中分布均匀,但分类单位种质群间的遗传多样性在多数情况下差异显著。豌豆属野生种P.fulvum的遗传多样性远低于栽培种P.sativum;豌豆栽培种下,P.sativum ssp.sativum var.sativum和P.sativum ssp.asiaticum的遗传多样性最高,P.sativum ssp.elatius var.elatius和P.sativum ssp.transcaucasicum次之,P.sativum ssp.elatius var.pumilio、P.sativum ssp.sativum var.arvense和P.sativumssp.abyssinicum最低。PCA分析发现,豌豆属种质资源由4个差异明显的基因库构成。"fulvum"基因库主要由野生种Pisum fulvum资源构成,"abyssinicum"基因库主要由栽培种下的P.sativum ssp.abyssinicum亚种资[ Objective ] Assessing the genetic diversity between wild and cultivated accessions of eight taxonomic groups in two species, five subspecies under Pisum genus, and analyzing the population structure and their genetic relationships among various groups of taxonomy, the study try to verify the fitness of traditionally botanical taxonomic system under Pisum genus and to provide essential information for the exploration and utilization of wild relatives of pea genetic resources. [Method] One hundred and ninety-seven Pisum accessions from 62 counties of five continents were employed for SSR analysis using 21 polymorphic primer pairs in this study. Except for cultivated field pea Pisum sativum subsp, sativum var. sativum （94 genotypes）, also included were wild relative genotypes that were classified as P. fulvum, P. sativum subsp, abyssinicum, P sativum subsp, asiaticum, P sativum subsp. transcaucasicum, P. sativum subsp, elatius var. elatius, P. sativum subsp, elatius var. pumilio and P sativum subsp, sativum vat. arvense （103 genotypes）. The PCA analyses and three-dimensional PCA graphs were conducted and drawn by NTSYSpc 2.2d statistical package. Nei78 genetic distances among groups of genetic resources were calculated, and cluster analysis using UPGMA method was carried out by using Popgene V1.32 statistical package, the dendrogram were drawn by MEGA3.1 statistical package. Allelic statistics were carried out by Popgene V1.32. The significance test between groups of genotypes was carried out by Fstat V2.9.3.2 statistical package. [Result] One hundred and four polymorphic bands were amplified using 21 SSR primer pairs with unambiguous unique polymorphic bands. 4.95 alleles were detected by each SSR primer pair in average, of which 65.56% were effective alleles for diversity. PSAD270, PSAC58, PSAAI8, PSAC75, PSAA175 and PSAB72 were the most effective SSR pairs. SSR alleles were uniformly distributed among botanical taxon units under pisum genus, but significant difference appeared in most pairwise compariso

关 键 词：豌豆属(Pisum L.) SSR 遗传多样性 植物学分类体系 基因库 

分 类 号：S643.3[农业科学—蔬菜学]