藤茶转录组SSR分子标记开发与初步验证  

Development and Identification of SSR Molecular Markers Based on Transcriptome Sequences of Ampelopsis grossedentata

作　　者：许明 曾莉 黄昕颖 汪梅 唐进兰 杨志坚 廖素凤 Xu Ming;Zeng Li;Huang Xinying;Wang Mei;Tang Jinlan;Yang Zhijian;Liao Sufeng(Key Laboratory of Crop Biotechnology,Fujian Province University,College of Agriculture,Fujian Agriculture and Forestry University,Fuzhou,350002)

机构地区：[1]福建农林大学农学院,作物生物技术福建省高校重点实验室,福州350002

出　　处：《分子植物育种》2020年第19期6441-6448,共8页Molecular Plant Breeding

基　　金：国家科技支撑计划(2013BAD01B05);福建农林大学科技创新专项基金(KFA17424A)共同资助。

摘　　要：利用MISA软件对藤茶转录组测序获得的92472条Unigene进行SSR位点信息分析,共检测到单核苷酸至六核苷酸重复类型的SSR位点34420个,分布在27096条Unigene中,出现频率为37.22%,平均分布距离为31.69 kb。SSR重复类型主要以单核苷酸重复为主,占66.11%,其次是二、三核苷酸,各占19.95%和12.52%。34420个SSR位点中共发现77种重复基序,重复次数在5~24次之间,其中出现频率高的基序有A/T、AG/CT、AT/AT、AC/GT、AAG/CTT和AAT/ATT。利用Primer 3.0共设计得到34440对SSR引物,从中随机选取40对引物进行PCR扩增检测,有30对引物扩增出预期大小的条带,其中9对引物在8份藤茶种质材料中表现出多态性。以上结果表明,基于藤茶转录组序列的SSR标记开发是有效可行的,开发的SSR标记为后续藤茶种质资源鉴定及遗传多样性分析等研究提供了科学依据。In this study,a total of 92472 Unigenes from transcriptome of Ampelopsis grossedentata were used for screening of SSR loci using MISA software.34420 SSRs,including the types of 1~6 nucleotide repeats distributed in 27096 Unigenes were identified,with occurring frequency of 37.22%and mean distance of 31.69 kb.Mono-nucleotide repeat was the main type,accounted for as much as 66.11%of all SSRs,followed by tri-nucleotide(19.95%)and di-nucleotide(12.52%)repeat motif.77 repeat motifs with iteration numbers from 5 to 24 were discovered from the 34420 SSR loci,and the most abundant motif were A/T,AG/CT,AT/AT,AC/GT,AAG/CTT and AAT/ATT.A total of 34440 SSR primers were designed using primer 3.0.Randomly 40 pairs of primers were selected for PCR amplification,of which 30 primers could yield the expected PCR products and 9 primers showed polymorphism in8 different A.grossedentata germplasm.The above results indicated that the development of SSR markers based on the transcriptome sequence of A.grossedentata is feasible and effective,and the developed primers will benefit for following germplasms identification and genetic diversity analysis for A.grossedentata.

关 键 词：藤茶(Ampelopsis grossedentata) 转录组 SSR分子标记 开发 

分 类 号：S571.1[农业科学—茶叶生产加工]