草业学报2018，Vol.27Issue(2)：124-134,11.DOI:10.11686/cyxb2017144
磁珠富集法开发蕨麻SSR标记引物
Development of microsatellite primers in Potentilla anserina by magnetic beads enrichment
摘要
Abstract
Simple sequence repeat (SSR) markers have been used extensively in studies on the heredity of flora and fauna because they are widely distributed throughout the genome,codominant,stable,and polymorphic.The genomic DNA of Potentilla anserina was digested with the restriction enzymes EcoR Ⅰ and Mse Ⅰ and the digested fragments were hybridized with biotinylated (AG)15 and (GT)15 probes.The fragments containing SSR genomic loci were adsorbed onto magnetic beads coated with streptavidin.Then,by elution,PCR amplification,cloning,and sequencing,a collection of SSRs was obtained.A total of 236 effective clones was acquired.Eighty sequences were randomly selected for sequencing,and 68 of them (85％) were found to be sin gle sequences.Forty sequences including SSR loci were selected and 40 primers were developed using primer design soft (Primer 5).Finally,20 pairs of primers with rich polymorphism and stable amplification were obtained through preliminary screening.The success rate of amplification was 50％.A preliminary analysis using the 20 pairs of primers amplified 338 polymorphic sites,with an average of 17 sites per primer pair.The mean values of effective allele number (Ne),polymorphism information content (PIC),Nei's gene diversity (H),and Shannon--Weaver index (I) were 1.2440,0.8996,0.1768,and 0.3078,respectively.These 20 pairs of SSR primers provide novel molecular markers for research on the genetic diversity of P.anserina.关键词
蕨麻/微卫星/磁珠富集/分子标记Key words
Potentilla anserina/simple sequence repeat/magnetic beads enrichment/molecular marker引用本文复制引用
富贵,李军乔,包锦渊,白世俊,韦梅琴..磁珠富集法开发蕨麻SSR标记引物[J].草业学报,2018,27(2):124-134,11.基金项目
“青海省高原作物种质资源创新与利用”国家重点实验室培育基地开放课题(2014-10),科技部农业科技成果转化基金项目(2010GB2G00514),国家自然科学基金项目(31660425,30607026,30660019)和青海省自然科学基金项目(2012-Z-907)资助. （2014-10）
