与番茄颈腐根腐病紧密连锁的SCAR标记开发  

Development of SCAR Markers Closely Linked with Fusarium Crown and Root Rot in Tomato

作　　者：程琳 张晓艳 魏美甜 伦尧尧 赵静 王晓武[5] 李传友[6] CHENG Lin;ZHANG Xiaoyan;WEI Meitian;LUN Yaoyao;ZHAO Jing;WANG Xiaowu;LI Chuanyou(Key Laboratory of Protected Vegetable Germplasm Innovation,Ministry of Agriculture and Rural Affairs,Key Laboratory of Protected Vegetable Molecular Breeding,Shandong Province,Vegetable Engineering Technology Research Center of Shandong Province,Shandong Vegetable Seed Industry Group Co.,Ltd.,Shandong Shouguang Vegetable Industry Group Co.,Ltd.of Shouguang Province,Shouguang 262700,Shandong,China;Shouguang Agricultural Technology Center,Shouguang 262700,Shandong,China;Shandong Shouguang Ouyate Vegetable Co.,Ltd.,Shouguang 262700,Shandong,China;Key Laboratory of Biochemistry and Molecular Biology,Colleges and Universities of Shandong Province,College of Biological and Agricultural Engineering,Weifang University,Weifang 261061,Shandong,China;Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China;Institute of Genetics and Developmental Biology,Chinese Academy of Sciences,Beijing 100101,China)

机构地区：[1]农业农村部设施蔬菜种质创新重点实验室,山东省设施蔬菜分子育种省级重点实验室,山东省蔬菜工程技术研究中心,山东寿光蔬菜种业集团有限公司,山东省寿光蔬菜产业集团有限公司,山东寿光262700 [2]寿光市农业技术中心,山东寿光262700 [3]山东寿光欧亚特菜有限公司,山东寿光262700 [4]山东省生物化学与分子生物学高校重点实验室,潍坊学院生物与农业工程学院,山东潍坊261061 [5]中国农业科学院蔬菜花卉研究所,北京100081 [6]中国科学院遗传与发育生物学研究所,北京100101

出　　处：《中国蔬菜》2021年第3期36-42,共7页China Vegetables

基　　金：山东省泰山学者特聘专家项目,寿光市“双百计划”项目。

摘　　要：利用与番茄颈腐根腐病抗性基因Frl紧密连锁的CAPS标记C2-25,开发设计了新的SCAR标记,用于快速检测Frl抗性材料。使用C2-25引物,在25份已知表型的番茄材料中进行PCR,产物单克隆测序后发现,19份纯合抗病材料存在1条完全相同的抗病序列;4份纯合感病材料存在1条完全相同的感病序列;2份杂合抗病材料均存在前述的抗病序列和感病序列。根据抗病、感病序列SNP差异设计引物,最终筛选得到可扩增出370 bp抗病特异片段和520 bp感病特异片段的番茄颈腐根腐病连锁标记R-6F/2R、S-3F/4R。在500株F2材料中进行验证,检测到122株纯合抗病单株、257株杂合抗病单株和121株纯合感病单株。F2材料人工接种鉴定结果显示,473株单株抗性与分子鉴定结果一致,分子标记辅助鉴定的准确率达到94.6%。开发的SCAR标记准确度高、成本低、操作简便,可用于Frl基因的分子标记辅助育种。Taking advantage of the CAPS marker C2-25 closely linked to the resistant gene Frl to tomato Fusarium crown and root rot,this paper developed and designed new SCAR markers to quickly detect the dominant resistant gene Frl.Using C2-25 as the primer,this experiment conducted PCR among the 25 known phenotypic tomato material through monoclonal sequencing,of which 19 homozygous resistant material all containing one same resistant sequence,4 homozygous susceptible material all containing one same susceptible sequence,both 2 heterozyous resistant material containing the above mentioned resistant and susceptible sequences.According to SNP differences between the resistant and susceptible sequences,we designed primers and finally found out primer combination R-6F/2R,S-3F/4R linked to Frl gene and produced 370 bp resistant fragment and 520 bp susceptible fragment.Testing the primer combination among 500 F2 plants,the genotype of 473 plants were consistent with their phenotype,meaning the accuracy rate was 94.6%.This SCAR marker was characterized by high accuracy,low cost and simple operation.Therefor,it can be used in gene Frl marker assisted breeding.

关 键 词：番茄 颈腐根腐病 SCAR标记 分子辅助育种 

分 类 号：S436.412.1[农业科学—农业昆虫与害虫防治]