桃(Prunus persica)砧木抗南方根结线虫分子标记开发与利用  

Development and utilization of molecular markers for resistance of peach(Prunus persica)rootstocks to southern root-knot nematodes

作　　者：王慧敏 李勇[2,3] 吴金龙 李文清 曹珂 王新卫[2,3] 王力荣[2,3] WANG Huimin;LI Yong;WU Jinlong;LIWenqing;CAO Ke;WANG Xinwei;WANG Lirong(School of Horticulture and Forestry,Tarim University,Alar 843300,Xinjiang,China;Zhengzhou Fruit Research Institute,Chinese Academy of Agricultural Sciences,Zhengzhou 450009,Henan,China;Institute of Western Agricultural,Chinese Academy of Agricultural Sciences,Changji 831100,Xinjiang,China)

机构地区：[1]塔里木大学园艺与林学学院,新疆阿拉尔843300 [2]中国农业科学院郑州果树研究所,郑州450009 [3]中国农业科学院西部农业研究中心,新疆昌吉831100

出　　处：《果树学报》2024年第7期1429-1437,共9页Journal of Fruit Science

基　　金：国际合作重点研发项目(2023YFE0105400);中国农科院院青创项目(Y2022QC23);河南省优秀青年基金(232300421042);中国农业科学院科技创新工程(西部中心林果团队)。

摘　　要：【目的】南方根结线虫是威胁桃产业绿色发展的地下主要害虫,开发抗性分子标记,对抗性砧木分子育种具有重要意义。【方法】根据前人对桃砧木抗根结线虫的定位结果,在GDR网站peach genome V2.0查询定位区间的候选基因。在5个抗病、5个感病种质中扩增候选基因,并通过DNAMAN、IGV等软件对候选基因进行序列差异分析,开发抗南方根结线虫KASP分子标记,在抗性种质筑波3号与感性种质哈露红的杂交F2群体中对该分子标记进行验证,并与前人开发的SCAR和35 bp indel抗南方根结线虫分子标记的准确性进行比较。【结果】KASP标记结果将基因型划分为3种,分别为抗性纯合型(A),抗性杂合型(B),感性纯合型(C),A∶B∶C=42∶94∶64,与表型符合率为88.5%;SCAR标记检测结果划分为2种,分别为抗病型(A1)和感病型(C1),A1∶C1=135∶65,与表型符合率为87.0%;35 bp indel分子标记分为3种类型,分别为抗性纯合型(A2),抗性杂合型(B2),感性纯合型(C2),A2∶B2∶C2=1∶154∶45,与表型符合率为52.0%。【结论】本研究中开发的KASP标记提高了分子标记选择准确率,对抗南方根结线虫分子育种具有重要意义。【Objective】Meloidogyne incognita is an underground pest threatening the development of peach industry.It is of great significance to develop molecular markers for the resistance to the pest for breeding new rootstocks.【Methods】According to the mapping results of peach rootstocks published by the predecessors,the candidate genes in the mapping interval were queried in GDR website Peach Genome V2.0.The candidate genes were amplified by PCR in five resistant germplasm Nemaguard,Okinawa,Tsukuba 2,Tsukuba 3 and Shouxingtao 1,and five susceptible germplasm Bailey,Kashi 1,Kashi 2,Harrow Blood and Siberian C.The target fragments of PCR products were purified,ligated and sequenced by agarose gel electrophoresis.The hybrid F_(2) population of disease-resistant germplasm Tsukuba 3 and susceptible germplasm Harrow Blood was inoculated with M.incognita,and the phenotypes of the population were investigated three months later to verify the accuracy of the KASP marker,and compared with the accuracy of molecular markers developed by the predecessors for the resistance to Meloidogyne incognita in SCAR and 35 bp indel.【Results】Six candidate genes were found through previous studies,namely Prupe.2G055500,Prupe.2G055600,Prupe.2G055700,Prupe.2G055800,Prupe.2G055900 and Prupe.2G056000.Through sequence comparison,it was found that there were regular variations in the resistant and susceptible varieties of the gene Prupe.2G055500,and there was a 2 bp indel variation(Pp02:6601310 bp,G→GAT)in its intron,and at insertion existed in the resistant varieties,but not in the susceptible varieties.In addition,using IGV software,with v2.0.a1 version as the reference genome,the same results were found in 10 resequencing data of peach germplasm materials.A molecular marker for genotyping was developed by using the above mutation sites.Five resistant germplasm and five susceptible germplasm were detected by this marker.The results showed that FAM and HEX fluorescence signals were simultaneously detected in the resistant germplasm Nemag

关 键 词：桃 KASP标记 南方根结线虫 抗性 

分 类 号：S662.1[农业科学—果树学]