[发明专利]同时检测小麦抗赤霉病基因与抗白粉病基因的多重PCR标记引物组及其应用

说明书

本申请公开了同时检测小麦抗赤霉病基因与抗白粉病基因的多重PCR标记引物组及其应用，该引物组包括；核苷酸序列如SEQ ID NO.1所示的前引物F1、核苷酸序列如SEQ ID NO.2所示的前引物F2、核苷酸序列如SEQ ID NO.3所示的后引物R1、核苷酸序列如SEQ ID NO.4所示的后引物R2；以该引物组为引物对小麦基因组DNA进行PCR扩增，可以同时检测小麦抗赤霉病基因Fhb1和小麦携带抗白粉病基因Pm21；该检测扩增效率较高，可提高分子标记辅助选择的准确性与效率。

技术领域

本申请涉及分子标记领域，特别是一种同时检测小麦抗赤霉病基因Fhb1与抗白粉病基因Pm21的多重PCR标记及应用。

背景技术

小麦是最重要的粮食作物之一，其生产安全对社会经济稳定具有重要意义。小麦生产易受多种病害影响，由禾谷镰刀菌(Fusarium graminearum)引起的赤霉病与禾布氏白粉菌(Blumeria graminis f.sp.tritici)引起的白粉病严重影响小麦的产量及品质(Parry D W,Jenkinson P,Mcleod L.Fusarium Ear Blight(Scab)in Small GrainCereals-a Review.Plant Pathology,1995,44:207-238；Zhao Z,Sun H,Song W,Lu M,Huang J,Wu L,Wang X,Li H.Genetic analysis and detection of the geneMlLX99onchromosome 2BL conferring resistance to powdery mildew in the wheat cultivarLiangxing 99.TheoreticalApplied Genetics,2013,126:3081-3089.)。在我国，小麦赤霉病过去主要发生于长江中下游麦区、华南冬麦区和东北春麦区，近年来扩展至黄淮麦区、北方麦区、西南麦区和西北麦区，而小麦白粉病在我国的大多数冬小麦产区均有发生。

抗赤霉病基因Fhb1是目前公认的抗性稳定且效应最大的位点，在小麦抗赤霉病育种中应用最为广泛，也最为成功(Xie GQ,Zhang MC,Chakraborty S,Liu CJ.The effectof 3BS locus of Sumai 3on Fusarium head blight resistance in Australianwheats.Animal Production Science.2007,47:603-607)，目前其基因克隆工作已经完成(Li G,Zhou J,Jia H,Gao Z,Fan M,Luo Y,Zhao P,Xue S,Li N,Yuan Y,Ma S,Kong Z,JiaL,An X,Jiang G,Liu W,Cao W,Zhang R,Fan J,Xu X,Liu Y,Kong Q,Zheng S,Wang Y,QinB,Cao S,Ding Y,Shi J,Yan H,Wang X,Ran C,Ma Z.Mutation of a Histidine-RichCalcium-Binding-Protein Gene in Wheat Confers Resistance to Fusarium HeadBlight.Nature Genetics,2019,51:1106-1112；Su Z,Bernardo A,Tian B,Chen H,WangS,Ma H,Cai S,Liu D,Zhang D,Li T,Trick H,St.Amand P,Yu J,Zhang Z,Bai G.ADeletion Mutation in Tahrc Confers Fhb1 Resistance to Fusarium Head Blight inWheat.Nature Genetics,2019,51:1099-1105)，诊断标记得到广泛应用(朱展望,徐登安,程顺和,高春保,夏先春,郝元峰,何中虎.中国小麦品种抗赤霉病基因fhb1的鉴定与溯源.作物学报,2018,44:473-482)。南京农业大学细胞遗传研究所通过染色体工程技术创制了高抗白粉病的小麦-簇毛麦T6VS/6AL整臂易位系，该易位系携带来源于簇毛麦的抗白粉病基因被命名为Pm21，它是目前国际上公认的抗性最强、抗谱最广的抗白粉病基因(Cao A,Xing L,Wang X,Yang X,Wang W,Sun Y,Qian C,Ni J,Chen Y,Liu D,Wang X,ChenP.Serine/threonine kinase gene Stpk-V,a key member of powdery mildewresistance gene Pm21,confers powdery mildew resistance in wheat.Proceedingsof the National Academy of Sciences of the United States of America,2011,108:7727-7732；Chen PD,Qi LL,Zhou B,Zhang SZ,Liu DJ.Development and molecularcytogenetic analysis of wheat-Haynaldia villosa6VS/6AL translocation linesspecifying resistance to powdery mildew.TheoreticalApplied Genetics,1995,91:1125-1128)，其克隆工作也已完(He H,Zhu S,Zhao R,Jiang Z,Ji Y,Ji J,Qiu D,Li H,Bie T.Pm21,Encoding a Typical Cc-Nbs-Lrr Protein,Confers Broad-SpectrumResistance to Wheat Powdery Mildew Disease.Molecular Plant,2018,11:879-882；Xing L,Hu P,Liu J,Witek K,Zhou S,Xu J,Zhou W,Gao L,Huang Z,Rui-qi Z,Wang X,Chen P,Wang H,Jones J,Havrankova M,Vrána J,J,Dolezel J,Tian Y,CaoA.Pm21 from Haynaldia Villosa Encodes a Cc-Nbs-Lrr That Confers PowderyMildew Resistance in Wheat.Molecular Plant,2018,11)，Bie等(Bie T,Zhao R,Zhu S,Chen S,Cen B,Zhang B,Gao D,Jiang Z,Chen T,Wang L,Wu R,He H.Development andcharacterization of marker MBH1 simultaneously tagging genes Pm21 and PmVconferring resistance to powdery mildew in wheat.Molecular Breeding,2015,35：189)成功开发出Pm21的诊断标记MBH1。