高杆野生稻PR2同源序列的分离与序列分析  

Molecular Cloning and Sequence Analysis of PR2-Like Sequences in Oryza alta

作　　者：王德彬[1] 韦靖鸾[1] 张丹[1] 刘新琼[1] 刘学群[1] 王春台[1] 张向明[1] 

机构地区：[1]中南民族大学生命科学学院生物技术国家民委重点实验室,武汉430074

出　　处：《武汉植物学研究》2009年第6期674-678,共5页Journal of Wuhan Botanical Research

基　　金：国家自然科学青年基金项目(30600400);武汉市青年科技晨光计划资助项目(200750731302)

摘　　要：根据已知植物病程相关蛋白基因β-1，3-葡聚糖酶基因（PR2）的保守结构域设计2对简并引物，从高杆野生稻基因组DNA中分离出3条防卫基因类似物（defense—genes analogues，DGAs），其中2条具有通读的ORF，另一条提前出现终止密码子。对这3条序列在NCBI上进行同源性搜索发现，在核苷酸水平这3条序列均与水稻的β-1，3-葡聚糖酶基因具有90％～93％的同源性，与已知大麦、小麦、高梁、黑麦、燕麦、玉米等其它植物的β-1，3-葡聚糖酶基因具有69％-81％的同源性。在氨基酸水平与水稻、大麦、小麦、黑麦的β-1，3-葡聚糖酶具有60％～93％的同源性。对具有通读ORF的2条序列RD1-GG6和RD1-GG12进行表达分析，发现经水杨酸（SA）诱导后表达量明显提高。Two sets of degenerate primers, which were designed based on the conservative domain of known plant pathogenesis-related protein gene β-1,3-glucanase,were used to isolated defense-genes analogues （DGAs） from genomic DNA of the Oryza alta. The desired bands were purified from the gel, and then cloned by T/A cloning. After sequencing and analyzing by alignment, three DGAs with uninterrupted open reading frames （ORFs） were obtained, except for one with immature stop codon. At the nucleic acid level,sequence identity of three DGAs with the β-1,3-glucanase genes of Oryza sativa ranged from 90% to 93% , and with the β-1,3-glucanase genes from Hordeum vulgate, Triticum aestivum, Sorghum bicolor, Secale cereale,Avena sativa and Zea mays ranged from 69% to 81%. At the amino acid level,the results showed that both two DGAs RD1-GG6 and RD1-GG12 were highly homologous with β-1,3-glucanase amino acid sequences of Oryza sativa, Hordeum vulgare, Triticum aestivum, Secale cereale with 60% - 93%. More importantly,expression level of RD1-GG6 and RD1-GG12 were upregulated at 3,12,24 and 48 h after salicylic acid （SA） treatment.

关 键 词：高杆野生稻 防卫基因类似物 Β-1 3-葡聚糖酶 

分 类 号：Q943.2[生物学—植物学] S511.9[农业科学—作物学]