基于等位基因特异性PCR技术检测Sw-5b基因  

Development of a Molecular Marker for Sw-5b Based on Allele-Specific PCR in Tomatoes

作　　者：刘文明[1] 华明艳[1] 宋兰芳[1] 崔少杰[1] 孙海波[1] 金凤媚[1] LIU Wenming;HUA Mingyan;SONG Lanfang;CUI Shaojie;SUN Haibo;JIN Fengmei(Tianjin Academy of Agricultural Sciences,Tianjin 300384,China)

机构地区：[1]天津市农业科学院,天津300384

出　　处：《天津农业科学》2023年第4期1-7,共7页Tianjin Agricultural Sciences

基　　金：天津市科技计划项目(22TDTPJC00600);宝坻区科技计划项目(202102)。

摘　　要：番茄斑萎病毒(Tomato spotted wilt virus,TSWV)是世界范围内危害最大的病毒病之一。为了加快番茄抗TSWV育种进程,利用等位基因特异性PCR(Allele-Specific PCR,AS-PCR)技术建立了与TSWV抗性基因Sw-5b连锁的标记。结果表明:在63℃退火温度下,引入A-C错配的特异性引物rg3-F可以检测出抗性基因型。没有引入错配的特异性引物sg1-F,在60℃退火温度下可以检测出感病基因型。在验证试验中,利用引物Sw5b-f1/r1进行PCR扩增并测序,结果表明利用rg3-F和sg1-F进行AS-PCR,能够有效区分番茄的不同基因型。本研究建立的AS-PCR方法为育种工作者在番茄抗TSWV育种中筛选Sw-5b抗性基因提供了有力的工具。Tomato spotted wilt virus(TSWV)is one of the most devastating viral diseases of tomato worldwide.In order to accelerate the tomato breeding process,a marker linked to the TSWV resistance gene Sw-5b was established by allele-specific PCR.The results showed that the specific rg3-F that introduced an A-C mismatch could distinguish the resistant genotypes from the susceptible genotypes at an annealing temperature of 63℃.Conversely,the specific sg1-F introduced no mismatch and could distinguish the susceptible genotypes from the resistant genotypes at an annealing temperature of 60℃.In the validation experiment,primers Sw-5b-f1/r1 were used for PCR amplification and sequencing,and the results showed that allele specific PCR using rg3-F and sg1-F could effectively distinguish different genotypes of tomato.This method of allele-specific PCR will provide breeders with a powerful tool in the selection of Sw-5b resistance gene in tomato breeding programs.

关 键 词：等位基因特异性PCR SNP 番茄 TSWV 

分 类 号：S641.2[农业科学—蔬菜学]