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作 者 ：王德彬;韦靖鸾;张丹;刘新琼;刘学群;王春台;张向明

期 刊 ：武汉植物学研究 2009年 27卷 6期 页码：674-678

关键词：高杆野生稻；防卫基因类似物；beta；-1；3-葡聚糖酶；

Keywords:Oryza alta, Defense-gene analogue(DGA), &beta, -1, 3-glucanase,

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NCB I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-β1,3-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。

Abstract: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 vulgare,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.

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