作 者 ：梁玉琴, 张嘉嘉, 梁晋军, 李芳东, 韩卫娟, 傅建敏
期 刊 ：林业科学 2015年 51卷 6期 页码：71-80
Keywords:Diospyros kaki, genetic diversity, SSR markers, genetic relationship, germplasm identification,
摘 要 ：[目的] 对河南省柿种质资源的遗传多样性进行分析,揭示河南省柿种质资源的遗传多样性程度,分析不同资源之间的亲缘关系,并进行种质资源鉴定,判断同名异物和同物异名品种。[方法] 以柿近缘种君迁子、油柿、浙江柿、美洲柿、金枣柿及柿变种野柿共计9份材料作为对照,从已发表的柿属植物SSR引物中,筛选出多态性较高的,对来自河南省不同地区的柿主栽品种(18份)、农家品种(55份)及野生资源(20份),共计93份资源进行遗传多样性分析。[结果] 利用17对SSR引物对102份材料进行PCR扩增,共得到159个不同的DNA片段(即条带),平均每对引物9.35个,范围为5~14个,扩增条带最多的引物有2对,分别来自位点ssrDK11/DQ097479和ssrDK14/DQ097482,最少的来自位点8125/DC592401; 所得到的所有条带均具有多态性; 共得到508个谱带类型(即带型),平均每对引物29.88个,范围为8~53个,最多的引物对来自位点ssrDK11/DQ097479,而最少的则来自位点5553/DC585710; 特异带型总数为267个,平均每对引物15.71个,范围为2~37个,最多的依然来自位点ssrDK11/DQ097479,最少的同样是5553/DC585710; 特异带型比率73.68%~25.00%; 种质鉴定率1.96%~36.27%; 多态性信息含量(PIC)平均为0.839 7,范围是0.466 7~0.964 7; Shannon信息指数(I)平均为2.658 6,范围为1.112 1~3.596 7; 平均观察杂合度(Ho)为0.777 4,范围是0.147 1~0.960 8。综合对比,ssrDK11/DQ097479、ssrDK14/DQ097482和mDp17/EF567410这3个位点遗传变异程度最高。主坐标分析与聚类分析结果基本一致:柿及其近缘种彼此之间具有明显的遗传差异; 柿种下所有资源单独聚为一大类,能够与近缘种区别开; 与其他近缘种相比,美洲柿与柿的亲缘关系相对较远。扩增的位点可鉴别所有供试资源,并可判断部分同名异物或同物异名品种。[结论] 河南省柿种质资源遗传变异程度较大,杂合程度高,具有较高水平的遗传多样性; 本研究所用方法可有效应用于柿种质资源鉴定。
Abstract:[Objective] The aim of the present study was to elucidate the genetic diversity and relationship among the germplasm accessions of Diospyros kaki in Henan province, meanwhile to identify the germplasms especially those of synonyms and homonyms. [Method] The genetic diversity among 102 germplasms including 93 accessions of D. kaki (18 major cultivars, 55 local varieties and 20 wild persimmons) originating from different areas in Henan and nine materials used as control which belong to five relative species i.e., D. lotus, D. oleifera, D. glaucifolia, D. virginiana and Jinzaoshi (D. spp.), and D. kaki var. sylvestris were analyzed based on the polymorphic SSR (simple sequence repeat) primers selected from those developed for Diospyros in previous reports. [Result] A total of 159 polymorphic DNA fragments i.e., bands, were obtained from the analysis of genetic variation among all 102 accessions by 17 pairs of SSR primers with an average of 9.35 bands per pair of primers ranging from 5 (8125/DC592401) to 14 (ssrDK11/DQ097479 and ssrDK14/DQ097482). The number of band patterns was 508 in total with an average of 29.88 per pair of primers ranging from 8 (5553/DC585710) to 53 (ssrDK11/DQ097479). The total number of unique band patterns was 267 with an average of 15.71 per pair of primers ranging from 2 (5553/DC585710) to 37 (ssrDK11/DQ097479). The rate of unique band patterns ranged from 73.68% to 25.00% and the rate of germplasm identified ranged from 1.96% to 36.27%. The average value of polymorphism information content (PIC) was 0.839 7 ranging from 0.466 7 to 0.964 7 and the average Shannon‘s information index (I) was 2.658 6 ranging from 1.112 1 to 3.596 7. Observed heterozygosity (Ho) ranged from 0.147 1 to 0.960 8 with an average value of 0.777 4. The highest genetic diversity level was found in three loci ssrDK11/DQ097479, ssrDK14/DQ097482 and mDp17/EF567410 by comprehensive comparison. The result obtained from principal coordinate analysis was roughly the same as that from cluster analysis: There were obvious genetic differences among different species. All germplasm accessions of D. kaki were divided into the same group which was clearly different from the samples of other closely related species. Compared with the other related species, the relationship between D. kaki and D. virginiana was relatively distant. All germplasm accessions tested including some homonymous or synonymous cultivars were identified by the 17 loci. [Conclusion] The results in present study revealed high level of genetic diversity among D. kaki germplasm accessions in Henan province with large variations and high heterozygosity. Moreover, the method was proved to be efficient in germplasm identification of D. kaki.