中间锦鸡儿干旱转录组SSR标记的开发及遗传多样性研究  

SSR Marker Development and the Genetic Diversity of Caragana intermedia in Based on Transcriptome under Drought Stress

作　　者：郑如玉[1] 刘坤[1] 杨杞[1] 杨飞芸[1] 杨天瑞[1] 宝力德[1] 王瑞刚[1] 

机构地区：[1]内蒙古农业大学生命科学学院,呼和浩特010018

出　　处：《分子植物育种》2017年第6期2236-2243,共8页Molecular Plant Breeding

基　　金：国家自然科学基金项目(31360169);内蒙古自治区科技创新团队(201503004);内蒙古农业大学科技创新团队(NDTD2013-1)共同资助

摘　　要：利用中间锦鸡儿干旱转录组数据自主开发、筛选具有多态性的SSR标记;在此基础上,选取了内蒙古地区10个居群的中间锦鸡儿种质资源为试验材料,对其遗传多样性展开了系统研究。研究如下:中间锦鸡儿干旱转录组共搜索到404个SSR标记,分布于349条Unigenes序列上,出现频率为14.78%。其中二核苷酸重复是主要的类型,占总SSRs的64.90%;其次是三核苷酸重复,占总SSRs的33.70%;二核苷酸重复基元中以AG/CT和GA/TC为优势重复基元,占总SSRs的29.45%,三核苷酸重复基元以AAG/CTT为主,占总SSRs的5.69%。成功设计了76对引物并筛选出条带清晰、多态性好、重复性好的7对引物进行遗传多样性分析。共检测到29个等位基因(Na),有效等位基因数为1.101~2.007,Nei’s多样性指数为0.254;Shannon多样性指数(I)为0.193~0.709,平均每位点I值为0.482;种群间遗传距离(GD)的范围是0.002 7~0.068 8,遗传一致度(GI)的范围是0.933 5~0.997 3,采用Nei's遗传关系用UPGMA进行聚类,10个居群分为2个亚群,包头市百灵庙镇和乌兰察布市凉城县聚为一个亚群,其余种群聚为另一个亚群。中间锦鸡儿种群间平均近交系数(Fis)大于0且基因流为3.726,遗传距离与地理距离相关系数R^2=0.023。分析表明供试材料的遗传多样性较低,可能是中间锦鸡儿居群受到自然和人为干预较多,遗传交流大所导致的。The transcriptome data of Caragana intermedia under drought was used to develop and screen the polymorphism markers. Ten different populations of C. intermedia as the experimental material in Mongolia were selected, then their genetic diversity and genetic relationship study were carried out. The studies showed that there are 404 SSR markers were found and distributed on the 349 Unigenes sequence with the occurrence frequency of 14.78%. The main frequency type was dinucleotide which accounted for 64.90% of the total SSRs; Then came the trinucleotide frequency with the 33.70% of the total SSRs; the dominant primitive types of dinucleotide were AG/CT and GA/TC accounting for 29.45% of the total SSRs, as the trinucleotide were AAG/CTT with 5.69% of the total SSRs. We designed 76 pairs of synthetic primers and selected 7 pairs of clear primers with perfect polymorphism and repeatability for genetic diversity analysis. We found 29 allele（Na） in which effective alleles was 1.101~2.007,and Nei＇s diversity index was 0.254; Shannon diversity index（I） was 0.193~0.709 with the average I point 0.482;The range of the Genetic distance between populations（GD） was 0.002 7 to 0.068 8, and the genetic identity（GI）was 0.933 5 to 0.997 3. With Nei＇s genetic relationship, we applied the cluster analysis through UPGMA, which10 populations were divided into two sub-groups in Bailing county of Baotou, Liangcheng of Wulanchabu and the others. Average inbreeding coefficient of C. intermedia populations（Fis）0, 3.726 gene flow, genetic distance and geographic distance correlation coefficient R^2=0.023. The analysis results showed that the lower genetic diversity in C.intermedia populations may be caused by the intervention of natural and human and the genetic exchange frequency.

关 键 词：中间锦鸡儿 SSR 遗传多样性 

分 类 号：Q943.2[生物学—植物学] S793.3[农业科学—林木遗传育种]