In order to analyze the population structure of Puccinia striiformis f.sp.tritici(Pst),SNPs primers of Pst were developed from DNA sequences of nine house-keeping genes Chs,Act,Mapk1,Tub,Cdc2,Rd,Sm,Ls and Ef-1α obtai...In order to analyze the population structure of Puccinia striiformis f.sp.tritici(Pst),SNPs primers of Pst were developed from DNA sequences of nine house-keeping genes Chs,Act,Mapk1,Tub,Cdc2,Rd,Sm,Ls and Ef-1α obtained from GenBank.Eight of which were from Pst and one from P.graminis f.sp.tritici(Pgt).Thirteen pairs of primers were designed and screened based on at least 30 isolates of Pst obtained from diverse locations.Three of them were polymorphic namely Mapl351S/Mapl683 A,Cd28S/Cd352 A and Efl37S/Ef531 A.Polymorphic loci analysis based on 149 Pst isolates from 5 provinces indicated that the three primers had good polymorphism.Cd28S/Cd352 A had 8 polymorphic loci,three of them were phylogenetically informative.Efl37S/Ef531 A had 6 polymorphic loci and 4 of them were informative.Mapl351S/Mapl683 A had 8 polymorphic loci and 4 were informative.The 3 primers were used for analyzing Pst population and revealed the ancestral origin,phylogeny relation of haplotypes,genetic differentiation of the population,gene flow,and the evolution and migration relation between Pst populations thereby.The findings indicate that the 3genes SNP primers can be used for Pst population genetic structure analysis.展开更多
利用NCBI数据库进行小麦条锈菌看家基因SNP引物开发,从NCBI搜索到15个基因的序列,利用Primer Premier 5.0软件设计了22对引物,利用其进行条锈菌DNA的PCR扩增、测序,并与标准序列比对,从而筛选出SNP引物。在候选基因中,柠檬酸盐合酶(Cs)...利用NCBI数据库进行小麦条锈菌看家基因SNP引物开发,从NCBI搜索到15个基因的序列,利用Primer Premier 5.0软件设计了22对引物,利用其进行条锈菌DNA的PCR扩增、测序,并与标准序列比对,从而筛选出SNP引物。在候选基因中,柠檬酸盐合酶(Cs)、热休克蛋白hsp90(Hsp)、泛素活化酶E1(Uba)及泛素结合酶E2(Ubc)4个基因的5对SNP引物表现多态性,能检测到条锈菌SNP位点依次为4、6~7、4和7个,均为系统发生信息位点,并摸索出优化的PCR反应体系及条件。利用开发的引物研究条锈菌群体结构,可揭示病原菌起源、进化、传播,为病害治理提供依据。展开更多
基金Financial support by the Ministry of Science and Technology(2011CB100403)the Ministry of Agriculture(200903035)+1 种基金National Natural Science Foundation(31260417)Yunnan Academy of Agricultural Sciences(2007yaas07)
文摘In order to analyze the population structure of Puccinia striiformis f.sp.tritici(Pst),SNPs primers of Pst were developed from DNA sequences of nine house-keeping genes Chs,Act,Mapk1,Tub,Cdc2,Rd,Sm,Ls and Ef-1α obtained from GenBank.Eight of which were from Pst and one from P.graminis f.sp.tritici(Pgt).Thirteen pairs of primers were designed and screened based on at least 30 isolates of Pst obtained from diverse locations.Three of them were polymorphic namely Mapl351S/Mapl683 A,Cd28S/Cd352 A and Efl37S/Ef531 A.Polymorphic loci analysis based on 149 Pst isolates from 5 provinces indicated that the three primers had good polymorphism.Cd28S/Cd352 A had 8 polymorphic loci,three of them were phylogenetically informative.Efl37S/Ef531 A had 6 polymorphic loci and 4 of them were informative.Mapl351S/Mapl683 A had 8 polymorphic loci and 4 were informative.The 3 primers were used for analyzing Pst population and revealed the ancestral origin,phylogeny relation of haplotypes,genetic differentiation of the population,gene flow,and the evolution and migration relation between Pst populations thereby.The findings indicate that the 3genes SNP primers can be used for Pst population genetic structure analysis.
文摘利用NCBI数据库进行小麦条锈菌看家基因SNP引物开发,从NCBI搜索到15个基因的序列,利用Primer Premier 5.0软件设计了22对引物,利用其进行条锈菌DNA的PCR扩增、测序,并与标准序列比对,从而筛选出SNP引物。在候选基因中,柠檬酸盐合酶(Cs)、热休克蛋白hsp90(Hsp)、泛素活化酶E1(Uba)及泛素结合酶E2(Ubc)4个基因的5对SNP引物表现多态性,能检测到条锈菌SNP位点依次为4、6~7、4和7个,均为系统发生信息位点,并摸索出优化的PCR反应体系及条件。利用开发的引物研究条锈菌群体结构,可揭示病原菌起源、进化、传播,为病害治理提供依据。
