柿野生雄性资源调查及其遗传多样性研究

柿（Diospyros kaki Thunb.）雄性种质资源非常稀缺，系统收集和梳理国内外现有雄性资源，阐明其遗传多样性并揭示其与主栽甜、涩柿品种及近缘种的亲缘关系，对雄性种质资源的高效利用有重要意义。在全国范围内开展柿野生种质资源调查，在湖北、广西、江苏等地发现野生雄性种质资源共计90余份。利用15对SSR引物对47份柿完全雄性资源、2份雄全同株资源和11份中国雌雄同株资源进行遗传多样性分析，以67份完全雌性资源、38份日本资源（其中包括25份雌雄同株资源）以及28份近缘种为对照。结果表明：15对引物共获得144个等位基因和316个基因型，平均每个引物获得9.60个等位基因和21.07个基因型，其中位点ssrDK11/DQ097479 遗传变异程度最高。从地理分布来看，柿雄性资源在适生的野外山区普遍存在，其中湖北木兰山来源的雄性资源遗传多样性最高，陕西省来源的雄性资源遗传多样性最低。UPGMA和ME聚类方法分别将193份资源分为16组和12组，两种聚类结果反映的总体规律类似：（1）各来源地间的野生雄性资源存在着一定程度的遗传隔离；（2）湖北木兰山野生雄性资源被分为遗传关系较远的两类，一类与江西、江苏、湖南野柿及‘火晶’等主栽涩柿品种和‘耀县五花柿’等雌雄同株涩柿品种亲缘关系较近，另一类与云架山雄性资源和罗田甜柿（包括‘甜宝盖’）亲缘关系较近；（3）大部分日本来源的柿资源可被单独聚为一类。

Survey of Wild Male Germplasm Resources of Diospyros kaki and Their Genetic Diversity Analysis

For the scarcity of persimmon male resources，the systematical collection and arrangement of the male resources as well as the clarity of genetic diversity of these resources and the genetic relationship among them together with other main cultivars and closely related species are essential for the efficient utilization of male resources. In this study，more than 90 wild male persimmon germplasm resources were found in Hubei，Guangxi，Jiangsu provinces，etc.，during the survey of wild persimmon germplasm resources nationwide. Subsequently，15 SSR primers were used to analyse the genetic diversity of 47 male persimmon resources，2 andromonoecy resources and 11 Chinese monoecious resources. Sixty-seven female resources，38 Japanese resources（including 25 monoecious resources）and 28 closely related species were used as reference out-group. The results indicated that a total of 144 alleles and 316 genotypes were obtained from 15 loci，with a mean value of 9.60 alleles and 21.07 genotypes per locus. ssrDK11/DQ097479 was the most informative locus. The geographical distribution of male persimmon resources indicated that they were common in wild comfortable remote area. The male resources originated in Mulan Mountain，Hubei Province and in Shaanxi Province showed the highest and the lowest genetic diversity，respectively. The 193 resources were divided into 16 and 12 clusters by UPGMA and ME cluster analysis，respectively. Similar general laws were found according to the two cluster analysis results：（1）a genetic isolation existed among wild male resources from different areas；（2）the wild male resources in Mulan Mountain，Hubei Province could be genetically divided into two clusters，one had a near relationship with Jiangxi，Jiangsu，Hunan wild persimmons as well as main astringent cultivars such as‘Huojing’and monoecious astringent cultivars such as‘Yaoxian Wuhuashi’，while the other was closer to Yunjia Mountain male resources and Luotian non-astringent persimmons（including‘Tianbaogai’）；（3）most Japanese resources could be classified into a same cluster exclusively.