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花生是我国重要的食用油和蛋白质来源作物,鉴定其DNA分子多态性对品种改良和资源评价具有重要的意义。从已公布的花生Genomic-SSR和EST-SSR引物中筛选出34对引物,用来分别鉴定花生4大类型各24份共96份品种资源的分子变异,其中龙生型资源全部来自广西,普通型资源中有11份从国外引进,有13份来自广西和国内其他省市,多粒型资源只有两份来自中国,其他22份分别来自印度、美国和非洲等地,珍珠豆型资源中有22份是来自中国各地的育成品种或农家品种,有2份来自国外。研究结果为:分别有10～16对SSR引物能在4大类型花生资源中扩增出多态性DNA片段;这些多态性SSR引物都具有多位点特性;首次为SSR分子标记设立了一个新的评价指标——区别指数,多态性SSR引物的区别指数最高达0.992;资源间的平均遗传距离,多粒型为0.59,普通型为0.48,珍珠豆型为0.38,龙生型为0.17。根据遗传距离采用最长距离法对4大类型花生资源分别进行了聚类分析,构建了资源间的遗传关系图,花生4大类型可进一步分成不同类群,资源间的亲缘关系与其来源相关。观察到PM15和PMc297的扩增产物具有类型特异性,PM15能在龙生型、普通型和多粒型花生资源中扩增出多态性条带,而在珍珠豆型花生中扩增条带完全相同,PMc297也有相似的扩增结果。由于在多粒型花生资源中检测出的遗传多样性最丰富,研究结果支持西班牙专家Krapovickas 1994年公布的花生栽培种分类系统。总之在花生4大类型内资源中能检测出丰富的SSR分子标记,开发出更多的SSR分子标记将能充分揭示花生分子水平的变异,从而使花生遗传图谱构建、分子标记辅助育种成为可能。
Peanut is one of the most important edible oil and protein source crops in our country. Identification of its DNA molecular polymorphism is of great significance to variety improvement and resource evaluation. Thirty-four pairs of primers were screened from published Genomic-SSR and EST-SSR primers to identify the molecular variation of 96 varieties of 24 varieties of peanut, 24 species each. All of them were from Guangxi, 11 were imported from abroad, 13 were from Guangxi and other provinces in China, only two were from multi-grain resources, and the other 22 were from India, the United States and Africa respectively. Among the pearl-bean resources 22 were bred varieties or farmer varieties from all over China, and 2 were from abroad. The results showed that 10 ~ 16 pairs of SSR primers could amplify polymorphic DNA fragments from four types of peanut resources respectively. These polymorphic SSR primers all had multi-locus characteristics. For the first time, a SSR marker The new index - difference index, the difference index of polymorphic SSR primers up to 0.992; the average genetic distance between resources, multi-grain type is 0.59, common type is 0.48, pearl bean type is 0.38, dragon type is 0.17 . According to the genetic distance, the longest distance method was used to cluster the four peanut resources. The genetic relationships among the resources were constructed. The four peanut types could be further divided into different groups. The genetic relationship among the sources was related to their origin. The amplification products of PM15 and PMc297 were observed to be type-specific. PM15 amplified the polymorphic bands in the native, common and multigrain peanut sources, whereas the amplified bands in peanut peanut Exactly the same, PMc297 has similar amplification results. Due to the most abundant genetic diversity detected in the multi-grain peanut resource, the results support the peanut cultivar classification system published by the Spanish expert Krapovickas in 1994. In conclusion, abundant SSR markers can be detected in four types of peanut resources, and more SSR markers will be able to fully reveal the variation of peanut molecular levels, thus making it possible to construct the genetic map of peanut and make molecular marker-assisted breeding.