【Objective】 It is very important to make studies on the genetic diversity of wild, semi-wild and cultivated azuki beans for further detecting the origin, evolution and diffusion of azuki bean and effectively utilizing these germplasms and serving for azuki bean improvement. 【Method】 Eleven higher polymorphic SSR primers screened from 69 SSR primer pairs of azuki bean and black gram were used in amplification of 558 wild, semi-wild and cultivated azuki bean accessions from China, Japan, South Korea, Bhutan and Burma. The analysis of genetic diversity and trait association was done.【Result】 Eighty-six polymorphic bands were generated. An average of 7.28 polymorphic bands were detected from 1 pair SSR primers. The number of alleles changed from 6 to 10. There were specific bands were observed in wild, semi-wild and cultivated azuki beans. Specific bands of cultivated azuki bean were predominantly produced by Chinese cultivars and landraces. Specific bands of wild azuki bean were detected in accessions from China, Bhutan and South Japan. The order of genetic dispersion was wild＞semi-wild＞cultivated azuki beans. Semi-wild accessions has closer genetic relationship with wild azuki bean. The 558 accessions were divided into five main groups by cluster analysis according to genetic distance. The result of cluster appeared that the genetic types of azuki beans have obviously associated with different geographical regions. Japanese cultivated azuki beans have closer genetic relationship with wild and semi-wild accessions from Japan and South Korea. Wild accessions from Southwest China are closely related to wild accessions from Southeast Asia and landraces of Jiangsu Province. Trait association analysis showed that black gram BG111 marker located on linkage group 7 of azuki bean explained 49%, 44%, 31%, and 18% traits of number of nods on main stem, diameter of stem, top stem twining and branching numbers in wild and weedy types. BG48 of linkage group 1 and BG20 markers of linkage group 5 from black gram and azuki bean AZ24 marker of linkage group 9 accounted for 9%, 7%, 5%, and 6% traits of maturing days, plant height and seed number per pod, number of nods on main stem in cultivated azuki bean respectively.【Conclusion】 A higher genetic variation was detected in wild types than in cultivated accessions. Chinese cultivated azuki bean is originated from China and has the most diversely genetic variation. Number of nods, diameter of stem, top stem twining and branching numbers in wild and semi-wild types are related with black gram BG111 marker. Maturing days, plant height and the number of seeds per pod, number of nods on main stem are associated with black gram BG48 and BG20, azuki bean AZ24 markers in cultivated azuli bean, respectively.

【目的】研究野生、半野生、栽培型小豆的遗传多样性,进一步阐明小豆的起源进化与传播,提高小豆种质的利用效率。【方法】从69对小豆和黑吉豆SSR引物中,筛选出11对多态性好的SSR标记,并结合植株形态性状特征鉴定,对558份来自中国、日本、韩国、不丹、缅甸的野生、半野生和栽培小豆种质资源进行遗传多样性分析和性状关联分析。【结果】共检测到86个等位变异,平均每个位点等位变异数为7.82个,变幅6—10个。野生、半野生、栽培型小豆都有其特征带,栽培小豆的特征带绝大多数来源于中国;野生小豆的特征带仅出现在中国西南、不丹和日本南部地区的种质中。遗传离散度是野生型＞半野生型＞栽培型,半野生型小豆更接近野生型小豆。聚类分析把558个小豆种质分为5大类,归类有较明显的地理相关性和遗传类型的趋同性。日本栽培小豆与韩国和日本野生及半野生小豆亲缘关系近,中国西南野生小豆与东南亚野生材料遗传关系近,与江苏地方品种遗传关系较近。关联分析表明,位于小豆第7连锁群的黑吉豆BG111标记分别解释野生和半野生小豆的主茎节数、茎粗、顶蔓、主茎分枝数性状的49%、44%、31%和18%;栽培小豆中,第1连锁群的黑吉豆BG48、第5连锁群的BG20和第9连锁群的小豆AZ24标记分别解释生育期、株高和单荚粒数、主茎分枝数性状的9%、7%、5%和6%。【结论】野生小豆遗传多样性丰富、变异背景广泛;中国栽培小豆起源于中国,具有丰富的遗传变异。黑吉豆BG111标记与野生和半野生小豆的茎粗、顶蔓、主茎分枝数、主茎节数性状相关联;黑吉豆BG48和BG20、小豆AZ24标记分别与栽培小豆的生育期、株高和单荚粒数、主茎分枝数相关联。

Adzuki bean[Vigna angularis (Willd) Ohwi & Ohashi] is a majorfood legume in China, with special functions in people’ life and agricultural production. In order to evaluate thegenetic diversity of adzuki bean germplasm and provide information for gene mining and plant breeding, a total of 375 accessions of adzuki bean core collection were analyzed with 13 pairs of microsatellite (SSR) markers screened from SSR primers of adzuki bean and its relatives. A wide genetic variation was observed in all collections. Four to nineteen alleles per SSR primer pair were detected with an average of 10.23 alleles. The mean of polymorphism information content (PIC) and the percentage of polymorphic loci (P) for the collections in China were 0.561 and 93.523%, respectively. The dendrogram by UPGMA cluster analysis among populations showed that there were correlations between the groups clustered and their putative geographic origins, and the collections from northeast had a closer genetic relationship with that from middle-south region of China. SSR data showed that PIC in Hubei, Shannxi, and Anhui provinces was the first three high, which almost located in the central part of the three dimensions of PCO (principal coordinate analysis). We concluded that Hubei, Shannxi, and Anhui provinces were the origin or diversity center of cultivated adzuki bean in China. The results could be available for collection, conservation, and utilization of adzuki bean germplasm resources.

以375份小豆核心种质为试验材料,利用从小豆及其近缘种SSR引物中筛选出的13对引物进行遗传多样性分析。检测结果显示,小豆种质资源具有丰富的遗传多样性,共检测到133个等位变异,每对SSR引物检测到等位变异4~19个,平均10.23个,国内各省多态信息含量(PIC)平均为0.561,多态位点比例(P)平均为93.523%。聚类结果表明,小豆资源遗传关系与生态分区间有明显的联系,且东北地区资源与中南部资源遗传关系较近。湖北、安徽、陕西3省资源的PIC较高,且基本位于主坐标三维图的中心区域,推断湖北、安徽、陕西是中国栽培小豆的起源地或多样性中心。该结果有助于更好地对小豆种质资源进行收集、保护和利用。

Adzuki bean, originated from China, is either one of traditional exports or an important crop for agricultural structure adjustment. As the traditional varieties can not meet the demands of modern markets and the genetic bases of the cultivars have been narrow, it is necessary to develop new breeding materials or novel germplasm, to improve varieties of adzuki bean. China has a collection of over 5 000 adzuki bean resources, which is lacking in study and use in breeding. In the paper, genetic diversity of 158 accessions of cultivated and 18 wild types of adzuki beans were analyzed by using 24 pairs of SSR primers. The results showed that 18 pairs of SSR primers were detected to be polymorphic in cultivated adzuki beans and 21 in wild adzuki beans. The average number of allele (NA) per SSR locus for cultivated and wild adzuki beans was 3.0 and 2.6, respectively. The number of accessions used for cultivated and wild adzuki beans differed much, and the NA and genetic diversity index (polymorphic information content, PIC value) showed a higher genetic variation in wild types than in cultivated ones. Both cluster and principal coordinate analysis (PCO analysis) suggested that there was a high genetic differentiation between cultivated and wild adzuki beans, while cultivated adzuki beans from different provinces of China could not be distinguished from each other clearly, suggesting that there is a high rate of the gene exchange among local adzuki beans. The genetic backgrounds of the breeding cultivars revealed by SSRs are almost agreeable with their putative pedigree, indicating that these SSRs can be used to identify the genetic relationships of them, and used in marker-assist-selection (MAS) in breeding. The results also revealed that the adzuki bean germplasms occurred more than once with the same name in the National Germplasm Conservation Centre were different from each other in genetic background and so valuable to be conserved and used individually. The present study not only provides some informations in adzuki bean breeding, but can accelerate the application of SSR markers in diversity analysis, gene tagging and identification of new cultivars.

利用24对SSR引物对158份栽培小豆及18份野生小豆资源进行遗传多样性分析。结果表明，栽培小豆的遗传变异水平显著低于野生小豆，其中18对引物在栽培小豆中能检测到多态性，平均等位变异数为3.0个，21对引物在野生小豆中能检测到多态性，平均等位变异数为2.6个。栽培小豆种质间平均遗传相似性系数为0.724，野生小豆间为0.605。基于类平均法的聚类分析可以将栽培小豆和野生小豆区分开，这与主坐标分析的结果基本吻合。不同来源的栽培小豆群体间也有一定的遗传分化。SSR分析不仅验证了小豆品种间的遗传背景与其系谱来源相吻合，而且揭示了同名种质天津红小豆之间的遗传差异。本研究为我国小豆种质资源育种及SSR标记在小豆多样性分析、基因标记、品种鉴定等工作提供了信息。

【Objective】 Adzuki bean originated from China, and there are abundant germplasm in China. Evaluation of the genetic diversity can accelerate the application of them in breeding program. 【Method】 In this study, 51 pairs of SSR primers were used to assess the genetic diversity and structure among 137 Chinese and eight alien adzuki bean germplasm. 【Results】 A total of 222 alleles were detected and the alleles with frequency lower than 5% accounted for about 35.9%. The number of alleles changed from 2 to 13 with an average of 4.35 per locus. The PIC value varied from 0.014 to 0.838, and the mean PIC value for SSRs on each linkage group was 0.440. The similarity coefficient between accessions varied from 0.227 to 0.951 and averaged at 0.487. Adzuki bean from Hubei and Shaanxi provinces had much higher genetic variations, and their genetic backgrounds are much different from that of other provinces. Cluster analysis divided the 145 accessions into five groups, which is much agreeable with that obtained in analysis by using STRUCTURE package, and the accessions within five groups almost related with their origins of geographical distribution. 【Conclusion】 There are abundant genetic variations in Chinese adzuki bean. The genetic differentiation among adzuki bean from different origins is important in dividing ecological region for Chinese adzuki bean.

【目的】分析小豆起源国中国丰富的小豆种质资源的遗传多样性及群体结构,提高这些种质在育种中的利用效率。【方法】选用51对SSR引物对国内外145份小豆种质进行多样性评价,并分析了中国小豆种质资源间的遗传关系和遗传结构。【结果】共检测出222个等位变异,每SSR位点的等位变异数为2～13不等,平均为4.35个,其中分布频率低于5%的等位变异数占35.9%。多态性信息含量（PIC值）为0.014～0.838,平均为0.472。不同种质间遗传相似性系数为0.227～0.951,平均为0.482。比较分析发现,湖北、陕西等省小豆资源的遗传变异最丰富,且遗传背景与中国主产区小豆存在较大差异。基于NTSYS的聚类可以将145份小豆种质划分为5组,根据组内种质的地理来源,可分别命名为东北组、华北Ⅰ组、华北Ⅱ组、华东组和混合组,其中混合组主要由湖北、陕西及国外种质组成。利用STRUCTURE对小豆种质资源的遗传结构分析与NTSYS聚类结果基本一致,即种质的遗传背景与地理来源有关。【结论】中国小豆种质资源遗传变异丰富,不同地理来源小豆间存在遗传分化,可以作为小豆生态区划的重要参考依据。