Tracing the location of powdery mildew resistance-related gene Stpk-V by FISH with a TAC clone in Triticum aestivum–Haynaldia villosa alien chromosome lines  

作　　者：YANG XueMing CAO AiZhong SUN YuLei CHEN PeiDu 

机构地区：[1]State Key Laboratory of Crop Genetics and Germplasm Enhancement,Cytogenetics Institute,Nanjing Agricultural University [2]Provincial Key Laboratory of Agrobiology,Institute of Biotechnology,Jiangsu Academy of Agricultural Sciences

出　　处：《Chinese Science Bulletin》2013年第33期4084-4091,共8页

基　　金：supported by the National Basic Research Program of China(2009CB118304);the National Natural Science Foundation of China(31171540,30871519);the Program for New Century Excellent Talents in University(NCET-10-0496);the Independent Innovation of Agricultural Sciences(CX(11)1025);the Sci&Tech Project in Jiangsu Province(BE2011306)

摘　　要：Bacterial artificial chromosomes(BACs)or yeast artificial chromosomes(YACs)containing large inserts as probes for fluorescence in situ hybridization(FISH)have been used in the physical mapping of specific DNA sequences,especially for single-or low-copy sequences.Our earlier study identified Stpk-V,a powdery mildew resistance-related gene located on the 6VS chromosome arm of the wild grass Haynaldia villosa(tribe Triticeae),and obtained several Triticum aestivum–H.villosa alien chromosome lines carrying the Stpk-V gene.However,the precise physical location of the Stpk-V gene on chromosome 6VS is not known.In this study,we used TAC-FISH with TAC15 as the probe coupled with sequential genomic in situ hybridization(GISH)to determine the physical location of the Stpk-V gene in different T.aestivum–H.villosa 6V alien chromosome lines,including addition,substitution and translocation lines.The result indicated that the fraction length of the Stpk-V locus is 0.575±0.035 on the 6V chromosome short arm and this was confirmed by FISH using TAC15 as the probe for tracing the Stpk-V gene in other genetic stocks.The cytological mapping strategies used in this study will be of benefit for tracing the alien gene location in the course of introducing desirable traits from wild species.Bacterial artificial chromosomes （BACs） or yeast artificial chromosomes （YACs） containing large inserts as probes for fluorescence in situ hybridization （FISH） have been used in the physical mapping of specific DNA sequences, especially for single- or low-copy sequences. Our earlier study identified Stpk-V, a powdery mildew resistance-related gene located on the 6VS chromosome arm of the wild grass Haynaldia villosa （tribe Triticeae）, and obtained several Triticum aestivum-H, villosa alien chromosome lines carrying the Stpk-V gene. However, the precise physical location of the Stpk-V gene on chromosome 6VS is not known. In this study, we used TAC-FISH with TAC 15 as the probe coupled with sequential genomic in situ hybridization （GISH） to determine the physical location of the Stpk-V gene in different T. aestivum-H, villosa 6V alien chromosome lines, including addition, substitution and translocation lines. The result indicated that the fraction length of the Stpk-V locus is 0.575±0.035 on the 6V chromosome short arm and this was confirmed by FISH using TACI5 as the probe for tracing the Stpk-V gene in other genetic stocks. The cytological mapping strategies used in this study will be of benefit for tracing the alien gene location in the course of introducing desirable traits from wild species.

关 键 词：异源染色体 相关基因 物理位置 抗白粉病 簇毛麦 小麦族 TAC 跟踪 

分 类 号：S512.1[农业科学—作物学]