利用SSR荧光标记构建41份山东省苹果资源分子身份证  

Using the Fluorescent Labeled SSR Markers to Establish the Molecular Identity of 41 Malus Germplasms in Shandong Province

作　　者：李慧峰[1] 王涛[1] 冉昆[1] LI Hui-feng;WANG Tao;RAN Kun(Shandong Institute of Pomology,Taian Shandong 271000,China)

机构地区：[1]山东省果树研究所,山东泰安271000

出　　处：《沈阳农业大学学报》2020年第1期70-77,共8页Journal of Shenyang Agricultural University

基　　金：山东省自然科学基金项目(ZR2019MC071);山东省农业良种工程项目(2016LZGC034);山东省农科院农业科技创新工程(CXGC2016A03);国家自然科学基金青年基金项目(31501742)。

摘　　要：山东自古为中国苹果的栽培中心,苹果属植物资源较为丰富,种间杂交类型较多,资源分类难度大,相关研究较为滞后。本研究选取山东省果树研究所苹果资源圃保存的41份资源为试材,利用SSR荧光标记构建分子身份证,旨在探索建立应用分子技术进行资源分类鉴定的技术体系。从SSR富集文库中开发并设计59对引物,从中筛选出10对多态性良好的引物,利用SSR荧光标记毛细管电泳检测扩增条带分子量的方法对41份山东省原产的苹果属资源进行分析,获得相应的等位基因。采用个位数字和小写英文字母对不同等位基因进行编码,按照10对引物从小到大串联排序的方式构建供试样品的分子身份证。试验结果表明:10对SSR引物共检测到139个SSR等位位点,244种基因型,平均每对引物对品种扩增等位位点13.9个,基因型24.4种。利用PopGen32软件计算引物的多态性信息含量,10对引物的平均多态性信息含量为0.85。基于10对SSR引物的扩增结果,采用个位数字和小写英文字母对不同等位基因进行编码,成功构建了41份山东苹果种质资源的分子身份证,为山东省苹果资源的鉴定工作提供了重要依据。As the origin and cultivation center of oriental apples, Shandong is one of the provinces with most of Malus germplasms. Because many Malus germplasms in Shandong province are generated from interspecific hybrid, so the classification is challenging and the research in this field is still insufficient. In our research, 41 Malus germplasms collected from different regions of Shandong province were used to establish molecular identity card(ID) by simple sequence repeat(SSR) markers. The aim of our research is to establish a molecular approach to identify and classify Malus germplasms. Fifty-nine pairs of primers were developed and designed from the SSR enrichment library, and among them 10 pairs of primers with significant polymorphism were screened out. Molecular identity of 41 Malus germplasms from Shandong province were constructed using SSR fluorescent markers. The molecular weight of the amplified bands was analyzed by SSR fluorescence labeled capillary electrophoresis method, and the corresponding amplified banding genotypes were obtained. Single digit and lowercase letters were used to mark these different band genotypes. The molecular identity of the sample was constructed by sequencing the number of10 pairs of primers from smallest to largest in series. Our results showed a total of 139 alleles of SSR and 244 genotypes could be detected by using these 10 pairs SSR primers. About 13.9 alleles and 24.4 genotypes were amplified by each pair primer on average. The polymorphism information was calculated by using PopGen32 software, and we found the average polymorphism of these ten primers was 0.85. The molecular ID codes of 41 tested germplasms were different, which could distinguish all these germplasms. Our research showed the SSR marker-based technology had the merits of reliable, efficient and high-throughput and is convenient to establish the molecular ID for Malus germplasms. Therefore, the molecular identity of 41 Malus germplasms from Shandong province were constructed successfully based on SSR amplifi

关 键 词：种质资源 苹果属 分子身份证 山东省 SSR荧光标记 

分 类 号：S661.1[农业科学—果树学]