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利用30对多态性良好的SRAP引物对90份山药种质资源进行PCR扩增,构建扩增图谱,共扩增到722个位点,多态性位点581个,多态性比例为80.47%,每对引物组合检测多态性位点3~30个,每对引物能鉴别6~51份山药种质资源;采用DNA数据分析软件对扩增出的多态性位点进行分析,构建山药种质资源方框指纹图谱,该图谱清晰地反映出每对引物能扩增的多态位点数、鉴别资源份数及资源具体编号,资源在该引物组合下所能检测得到的多态位点数及所处的具体位置等信息;从10对SRAP引物组合中挑选出的21个多态性位点,根据谱带的有无转化成的1/0字符串编码,形成山药种质资源DNA数字指纹图谱,该图谱可鉴别区分90份山药种质资源中的82份资源。同时这些指纹图谱可为下一步的山药品种鉴定,种质资源评价、利用,分子标记辅助育种及品种权保护提供技术支撑。
Totally 30 polymorphic SRAP primers were used to amplify 90 accessions of yam germplasm. The amplified sequence was amplified to 722 sites with 581 polymorphic loci and the polymorphic ratio was 80.47 %, 3 to 30 polymorphic loci were detected for each primer pair, and 6 to 51 copies of yam germplasm resources were identified for each pair of primers. The amplified polymorphic sites were analyzed by using DNA data analysis software to construct Yam Germplasm Resources Box fingerprinting, which clearly reflects the number of polymorphic loci that each pair of primers can amplify, the number of identified resources and the specific number of resources, and the number of polymorphisms that the resource can detect under this combination of primers Points and location of the specific information; from 10 pairs of SRAP primer combinations selected 21 polymorphic sites, according to the presence or absence of bands into 1/0 string coding to form yam germplasm resources DNA Digital fingerprinting, which distinguishes 82 of the 90 yams germplasm resources. At the same time, these fingerprints can provide technical support for the next identification of yam varieties, germplasm evaluation, utilization, molecular marker-assisted breeding and breed protection.