与番茄灰叶斑病抗病基因Sm连锁的CAPS标记开发  

Development of CAPS markers closely linked with Sm gene of tomato resistant to gray leaf spot

作　　者：郝耀港 宋建军[1] 李珂 李振侠[1] HAO Yaogang;SONG Jianjun;LI Ke;LI Zhenxia(College of Food and Biology,Heibei University of Science and Technology,Shijiazhuang 050018,Hebei,China)

机构地区：[1]河北科技大学食品与生物学院,石家庄050018

出　　处：《中国瓜菜》2023年第4期47-55,共9页China Cucurbits And Vegetables

基　　金：河北省省级科技计划(21326309D)。

摘　　要：利用与番茄灰叶斑病Sm基因连锁的RFLP标记,设计开发了一个CAPS标记,用于抗番茄灰叶斑病育种。以已知抗病基因型的番茄种质为材料,参照与Sm基因紧密连锁的RFLP标记设计特异引物,进行PCR扩增,产物克隆测序分析寻找特异性酶切位点,开发了一个与抗病基因Sm紧密连锁的CAPS标记,命名为T280。利用特异引物T280进行PCR扩增,供试材料均扩增出1条655 bp的特异条带。经限制性内切酶Mme I酶切后,抗病纯合材料仍保留1条655 bp的特异条带,感病纯合材料产生206 bp和449 bp的特异条带,抗病杂合材料产生206 bp、449 bp和655 bp的特异条带。利用不同遗传背景的番茄材料对该标记进行验证,结果发现,能够区分抗病杂合、抗病纯合和感病纯合材料。开发的CAPS标记稳定可靠,是一个比较理想的共显性CAPS标记,可用于Sm基因的分子标记辅助育种。Taking advantage of the RFLP marker closely linked to the resistant gene Sm to tomato gray leaf spot,we designed and developed CAPS marker to tomato gray leaf spot disease breeding.Using the known genotype tomato germplasm resources as materials,designed the specific primers according to the RFLP markers closely linked to gene Sm,the experiment conducted PCR amplification,the product was cloned and sequenced to search for specific restriction sites.A CAPS marker named T280 closely linked to the resistant gene Sm was successfully developed.Using T280 specific primer to PCR amplification,all materials amplified a specific band with length of 655 bp.After digested by restriction endonuclease Mme I,homozygous resistant material still contained a specific band with length of 655 bp,homozygous susceptible material produced two specific bands with lengths of 206 bp and 449 bp,and heterozygous resistant materials produced three specific bands with lengths of 206 bp,449 bp and 655 bp.Verified T280 marker through the different genetic background tomato materials,the study showed T280 marker was stable and reliable,could succesfully distinguish heterozygous resistant materials,homozygous resistant materials and homozygous susceptible materials,and a relatively idea co-dominant marker.Therefor,it can be used in gene Sm marker assisting breeding.

关 键 词：番茄 灰叶斑病 Sm基因 分子标记辅助育种 

分 类 号：S641.2[农业科学—蔬菜学]