构建黍稷分子遗传图谱SSR引物的筛选  

The SSR Marker Selection of Broomcorn Millet(Panicum miliaceum L.) for Construction of Genetic Linkage Map

作　　者：王银月[1,2] 刘敏轩[2] 陆平[2] 乔治军[3] 杨天育[4] 李海[5] 崔喜艳[1] 

机构地区：[1]吉林农业大学生命科学学院,吉林长春130118 [2]中国农业科学院作物科学研究所,北京100081 [3]山西省农业科学院农作物品种资源研究所,山西太原030031 [4]甘肃省农业科学院作物研究所,甘肃兰州730070 [5]山西省农业科学院高寒区作物研究所,山西太原030031

出　　处：《作物杂志》2014年第4期32-38,共7页Crops

基　　金：农业部谷子糜子产业技术体系(CARS-07-12.5-A1);国家"十二五"科技支撑-地方特色作物种质资源发掘与创新利用(2013BAD01B05-2-3);吉林省科技厅重点科技攻关项目(20130206012NY)

摘　　要：采用SSR-PCR扩增技术,将实验室利用高通量测序与磁珠富集法结合开发得到的1 210对黍稷SSR引物进行筛选,筛选出在会宁大黄糜、陇糜8号、太原55号和会宁野糜子4个黍稷品种之间表现出条带清晰、多态性明显、重复性较好的引物;并通过提取DNA以及PCR过程中退火温度的简单优化来建立比较适宜的黍稷SSR分子标记反应体系。结果表明,在DNA提取过程中将ddH2O的量减少到100μL,增加65℃的水浴时间并增加4%β-巯基乙醇和DNA浓度等可以获得较高质量的DNA。利用优化好的反应体系从1 210对SSR引物中筛选出116对多态性较好的引物,占总数的9.59%,其中,群体一的2个亲本所特有的引物有36对,群体二的2个亲本中特有的引物有97对,2个群体共有的多态性引物有19对;对引物碱基结构分析发现,选用的1 210对引物中有1 173个双碱基重复SSR和36个三碱基重复SSR;筛选到的多态性引物中双碱基重复SSR有102个。三碱基重复SSR有14个。筛选出的多态性SSR标记能用于黍稷的遗传多样性和群体遗传结构研究。Using SSR-PCR, 1 210 primers were screened （which selected from SSR markers developed by highthroughput sequencing combined with bead enrichment） in two research groups of broomcorn millet to select the SSR markers with high polymorphism, repeatability and stability;and establish a more appropriate reaction system for broomcorn millet SSR markers through simple optimization of DNA extraction and the annealing temperature of PC R process. Experiment was analyzed that in the process of extracted DNA, reduce the ddH2 0 to 100μL,increase the time of 65℃ bath and add the 4% β-mercaptoethanol and DNA material to get high quality DNA. Using the reaction system, 116 polymorphie primers were screened from 1 210 primers existing in two research groups, accounted for 9.59% of the total, and 36 primers had specific polymorphism in group one as well as 97 primers had specific polymorphism in group two,19 primers had specific polymorphism in both group one and two. Analysis of the primer base structure showed that there had 1 173 dimers and 36 trimers respectively in 1 210 primers,as well as 102 dimers and 12 trimers in polymorphic markers. The polymorphic SSR markers screened in this study could be used to construct the genetic linkage map and study the genetic diversity for broomcorn millet.

关 键 词：黍稷 SSR标记 引物筛选 遗传图谱 

分 类 号：S516[农业科学—作物学]