关键词: 条锈菌, 病程相关蛋白, 表达模式, 基因克隆, 电子克隆

Abstract:

【Objective】 A stripe rust pathogen induced PR10 gene from wheat was isolated to better understand adult wheat resistance response. 【Method】In silico cloning and RT-PCR techniques were performed to isolate a PR10 gene in wheat induced by stripe rust pathogen. Its DNA sequence structure and conserved domain of encoding protein with basic properties were determined using bioinformatics analysis. The expression patterns of the gene during adult and seedling stage of wheat infected by stripe rust pathogen were investigated using real time quantitative RT-PCR. 【Result】 The PR10 gene designated as TaPR10 was obtained from wheat inoculated with stripe rust pathogen. Open reading frame (ORF) of TaPR10 was 483bp in length, encoding 160 amino acids containing one typical conserved domain of pathogenesis related protein Bet_v_I family, and other four kinds of conserved motifs. TaPR10, without trans-membrane domain or signal peptide sequences, was predicted to locate in cytosol. Multiple alignment analysis based on the amino acids encoded by different PR10 genes from Maize (Zea mays), rice (Oryza sativa), broomcorn (Sorghum bicolor) and wheat (Triticum aestivum) indicated that PR10 was conserved among the four species of plants with about 80% of sequence similarity. DNA sequence of TaPR10 showed that it contained one 84 bp intron with the splicing sites of GT-AT bi-nucleotide sequence between 188 bp and 271 bp. Expression pattern results revealed that TaPR10 was up regulated in both of adult and seedling stages. However, expression in adult stage was higher than that of seedling stage. 【Conclusion】 Gene TaPR10 was firstly isolated and characterized from wheat infected by stripe rust pathogen, which may participate in defense response of adult wheat resistance to stripe rust pathogen.

Key words: stripe rust, pathogenesis-related protein, expression profile, gene cloning, in silico cloning