花生SSR核心引物筛选及育成品种DNA指纹图谱构建  

Selection of core SSR markers and identification of fingerprint on peanut cultivars

作　　者：任小平[1] 郑艳丽[1] 黄莉[1] 陈玉宁[1] 周小静[1] 陈伟刚[1] 雷永[1] 晏立英[1] 万丽云[1] 廖伯寿[1] 姜慧芳[1] 

机构地区：[1]农业部油料作物生物学与遗传育种重点实验室/中国农业科学院油料作物研究所,湖北武汉430062

出　　处：《中国油料作物学报》2016年第5期563-571,共9页Chinese Journal of Oil Crop Sciences

基　　金：国家科技支撑计划(2013BAD01B03);国家自然科学基金(31571713和31461143022);农作物种质资源保护项目(NB2010-2130135-28B);国家花生产业技术体系建设项目(CARS-14-种质资源评价)

摘　　要：根据SSR引物在遗传连锁图上的位置,首先选择200对均匀分布在染色体上,且通过聚丙烯酰胺凝胶电泳银染技术筛选表现为条带清晰、可重复的单位点引物,再利用毛细管电泳技术筛选等位变异数≥4个、引物的PIC值≥0.4、杂合度≤0.1的引物,并参考其所在染色体位置,获得60对具有丰富的多态性、广泛的代表性、均匀分布的SSR引物,同时普通引物和荧光引物都具有较好的扩增效果,作为花生品种构建指纹图谱的核心引物。60对SSR引物在100份材料中共扩增出352个多态性等位位点,引物扩增的等位位点均值为5.87;每对引物可区分的基因型数目均值是6.35;引物的多态性信息指数（PIC值）均值是0.54。高多态性的SSR引物占66.67%,SSR引物杂合度都在0.06以下。品种间的遗传相似系数变幅为0.530~0.683。构建了100份花生的指纹图谱,每一条指纹都具有唯一性,可标识一个品种,为全国花生品种及资源的DNA指纹数据库的构建奠定基础。According to the location of SSR primers in genetic linkage map,200 pairs of primers were selected with uniform distribution on the chromosome. Then,through polyacrylamide gel electrophoresis and silver stain technology,120 primers were chosen with clear bands and repeating. Finally,using capillary electrophoresis technology to screen,primers with uniform distribution on the chromosome were selected with the alleles number more than 4,and polymorphism information content（PIC） value higher than 0. 4,and heterozygosity being lower than0. 1 primers. 60 of primers were selected to identify fingerprint of peanut cultivars as core primers,which had abundant polymorphism,broad representative and uniform distribution of genome,with good amplification effect in both common primers and fluorescent primers. Among 60 core primers,352 polymorphic alleles were amplified in100 peanut varieties with the average polymorphic alleles being 5. 87,and the average number of genotypes distinguished being 6. 35,and the mean PIC being 0. 54. Primers accounted for 66. 67% with high polymorphism primers,and heterozygosity was below 0. 06. The genetic similarity coefficient among cultivars ranged from 0. 530 to0. 683. Fingerprint establishment of 100 peanut varieties provided a foundation for the construction of DNA fingerprint database of peanut varieties and resources.

关 键 词：花生 SSR荧光标记 毛细管电泳技术 指纹图谱 

分 类 号：S565.203[农业科学—作物学]