小麦条锈菌看家基因SNP引物的开发及多态性位点研究

Development and Polymorphic Loci Research of SNP Primers from House-keeping Genes of Puccinia striiformis f. sp. tritici

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。

The objective of this studywas to develop single nucleotide polymorphism （SNP） primers from sequences of house-keeping genes of Puccinia striiformis f. sp. tritici （Pst） based on NCBI database. The sequence of 15 genes were searched from NCBI database, and 22 pairs of primers were designed by Primer Premier 5.0 software. These primers were used to carry out PCR amplification and sequencing of DNA samples of Pst isolates, the results of which were compared with the standard sequences, so as to screen out SNP primers of these genes. Among the candidate genes, 5 pairs of SNP primers of 4 genes including Citrate synthase （Cs）, Heat sock protein 90 kD （lisp）, Ubiquitin-activating enzyenme E1 （Uba）, and Ubiquitin-conjugating enzyme E2 （Ubc） expressed polymorphism. The number of SNP loci in Pst that could be detected by the 4 genes was 4, 6 to 7, 4, and 7, respectively. All of them were phylogenetically informative loci. Moreover, the optimized PCR reaction system and conditions were explored. Using the obtained primers to study population structure of Pst could reveal the pathogen＇s origin, evolution, migration, and provide information for the disease control.