基于转录组的剑麻SSR标记开发与筛选  

SSR Marker Development and Primer Screening from the RNA-Seq Data of Sisal

作　　者：张燕梅[1] 李俊峰[1] 杨子平[1] 鹿志伟 陆军迎[1] 周文钊[1] ZHANG Yanmei;LI Junfeng;YANG Ziping;LU Zhiwei;LU Junying;ZHOU Wenzhao(South Subtropical Crops Research Institute,Chinese Academy of Tropical Agricultural Sciences/Zhanjiang City Key Laboratory for Tropical Crops Genetic Improvement,Zhanjiang,Guangdong 524091,China)

机构地区：[1]中国热带农业科学院南亚热带作物研究所/湛江市热带作物遗传改良重点实验室,广东湛江524091

出　　处：《热带作物学报》2021年第5期1261-1266,共6页Chinese Journal of Tropical Crops

基　　金：广东省甘蔗剑麻产业技术体系创新团队(No.2019KJ104-03);国家麻类产业技术体系项目(No.CARS-16);中国热带农业科学院基本科研业务费专项资金(No.1630062019021)。

摘　　要：本研究基于剑麻转录组测序获得的70110条Unigene序列,采用MISA 1.0软件查找SSR位点,利用Primer 3.0设计SSR引物,通过聚丙烯酰胺凝胶电泳技术对其中的100对SSR引物有效性进行验证。总计获得了13175个SSR位点,SSR的分布频率为15.61%。70110条Unigene序列总计包括60种重复基元,其中单核苷酸重复为主导重复类型(37.96%),其次是二核苷酸重复和三核苷酸重复,比例分别为32.92%和27.90%,四核苷酸、五核苷酸、六核苷酸重复所占比例总计为1.21%。A/T、AG/CT和AGG/CCT分别为单核苷酸、二核苷酸和三核苷酸重复的优势基元。100对SSR引物中有68对引物可扩增出目标产物,其中18对引物在6份剑麻种质中表现出多态性,该结果为利用SSR分子标记技术开展剑麻种质资源鉴定、遗传多样性分析等奠定基础。In this study,the high-throughput sequencing of the transcriptome of sisal was used to seek SSR loci and develop SSR markers by MISA 1.0 and Primer 3.0 softwares,respectively,and preliminary primer verification and polymorphic primer analysis were performed by vertical acrylamide gels.A total of 13175 of SSR loci were found in the 70110 unigene sequences.The SSR distribution frequency was 15.61%.In total,60 repeat elements were obtained,among them,mono-nucleotides were the domiant repeat motif(37.96%),followed by di-nucleotides(32.92%),and tri-nucleotides repeats(27.90%),the other repeat types occupied 1.21%in all.The most dominant mono-nucleotide,di-nucleotide and tri-nucleotide repeat motif was A/T,AG/CT and AAG/TTC,respectively.Among the 100 pairs of SSR primers randomly selected from the designed SSR primers of Rema No.1,sixty-eight pairs of primers could amplify the expected size bands,and eighteen primers showed polymorphism among six sisal germplasms.The obtained primers would provide effective molecule markers for genetic diversity analysis and germplasm identification of sisal.

关 键 词：剑麻 转录组测序 SSR 

分 类 号：S563.8[农业科学—作物学]