基于转录组信息的坛紫菜EST-SSR标记的开发及新品系SW-81的种质鉴定  

Development of EST-SSR marker based on transcriptome and the research of germplasm identification for the new strain(SW-81)of Pyropia haitanensis

作　　者：费勤娇 丁洪昌 张鹏[3] 王铁杆[3] 严兴洪[1,2] FEI Qinjiao;DING Hongchang;ZHANG Peng;WANG Tiegan;YAN Xinghong(Key Laboratory of Exploration and Utilization of Aquatic Genetic Resources,Ministry of Education,Shanghai Ocean University,Shanghai 201306,China;Shanghai Engineering Research Center of Aquaculture,Shanghai Ocean University,Shanghai 201306,China;Zhejiang Mariculture Research Institute,Wenzhou 325005,Zhejiang,China)

机构地区：[1]上海海洋大学水产种质资源发掘与利用教育部重点实验室,上海201306 [2]上海海洋大学上海水产养殖工程技术研究中心,上海201306 [3]浙江省海洋水产养殖研究所,浙江温州325005

出　　处：《上海海洋大学学报》2021年第3期464-474,共11页Journal of Shanghai Ocean University

基　　金：浙江省农业(水产)新品种选育重大科技专项(2016C02055-6);国家重点研发计划“蓝色粮仓科技创新”重点专项(2018YFD0900606);国家自然科学基金(31072208);江苏省科技计划(现代农业)重点项目(BE2018335)。

摘　　要：基于坛紫菜(Pyropia haitanensis)转录组序列信息进行EST-SSR标记的开发,并利用多态性SSR标记构建DNA指纹图谱用于坛紫菜新品系SW-81的种质鉴定。结果显示:转录组数据中共检测出9033个微卫星位点,其中三核苷酸重复类型为绝对优势基元,占比高达95%;经Primer 5.0软件对微卫星位点进行引物设计,并从中随机合成187对微卫星引物进行有效性分析,最终获得92对有效引物,其中42对为多态性EST-SSR标记,多态扩增率约为22.46%;利用开发的多态性EST-SSR标记构建了适用于5个坛紫菜品系的DNA指纹图谱,为新品系SW-81的种质鉴定提供有效的分子检测手段。上述结果表明:坛紫菜基因组中有着丰富的微卫星位点,且同一位点的基因序列在不同品系间存在变异,表现为扩增产物的多态性。基于EST-SSR标记的多态性分析可为坛紫菜种质资源的管理与鉴定提供有效的分子检测手段。EST-SSR markers were developed based on the transcriptome of Pyropia haitanensis and DNA fingerprinting was constructed using polymorphic SSR markers for the germplasm identification of the new strain SW-81 of Pyropia haitanensis.Experimental results:9033 microsatellite loci were detected in the transcriptome data,of which the trinucleotide repeat motif was the absolute dominant type,accounting for 95%;Designing primers for microsatellite loci by Primer 3.0 software,and 187 EST-SSR primers were synthesized randomly to analyze the effectiveness,and the 92 pairs of effective primers were finally obtained,of which 42 pairs were polymorphism EST-SSR markers,with the polymorphism amplification rate 22.46%;DNA fingerprints were constructed using the polymorphism EST-SSR marker,which was suitable for 5 strains of P.haitanensis,providing an effective molecular detection method for the germplasm identification of the new strain SW-81.The results above showed that there were abundant microsatellite loci in the genome of P.haitanensis,and the gene sequence of the same loci varied among different strains,showing the polymorphism of amplification products.The polymorphism analysis based on EST-SSR marker can provide an effective means of molecular detection for the management and identification of germplasm resources.

关 键 词：坛紫菜 EST-SSR 指纹图谱 种质鉴定 

分 类 号：S917[农业科学—水产科学]