水稻稻瘟病菌基因组中疏水蛋白的分布与聚类分析  

Distribution and Clustering Analysis of Hydrophobins in the Whole Genome of Rice Blast Fungus Magnaporthe Grisea

作　　者：吴毅歆[1,2] 范成明[3] 何月秋[1] 

机构地区：[1]云南农业大学农业生物多样性与病虫害控制教育部重点实验室,昆明650201 [2]云南省农业科学院经济作物研究所,昆明650205 [3]中国农业科学院作物科学研究所,北京100081

出　　处：《分子植物育种》2009年第5期978-984,共7页Molecular Plant Breeding

基　　金：农业部948项目(2006-G61);国家863计划项目(2002AA245041)共同资助

摘　　要：病原菌中的疏水蛋白在病原菌与寄主植物互作的过程中起着重要的作用。本研究利用BLAST工具从NCBI数据库公布的58个疏水蛋白序列中筛选得到18条稻瘟病病原菌基因组中具有潜在疏水蛋白功能的ORF,选取其中的11条ORF设计特异引物,检测其在17个稻瘟病病原菌中的分布状况。同时,利用序列比对工具ClustalX和Mega3.0软件的邻接法,对筛选的58条已知疏水蛋白序列构建疏水蛋白的系统进化树。研究结果表明:经PCR检测,在11对引物中有4对引物高度专一且所得片断与预期大小基本一致,6对引物扩增不特异,1对引物扩增无特异条带。通过系统进化树分析得知,疏水蛋白间的平均分离度为0.62,筛选的58条疏水蛋白可分为两大类:ClusterⅠ和ClusterⅡ,分别与真菌疏水蛋白Ⅰ型和Ⅱ型相对应;所有的担子菌、稻瘟菌和绿僵菌中的疏水蛋白构成ClusterⅠ,其它真菌的疏水蛋白构成ClusterⅡ。本试验的研究将为进一步明确稻瘟病病原菌中的疏水蛋白分布状况,以及相关基因的功能验证奠定基础。Hydrophobins play an important role in the interaction of pathogenic bacteria and its host plants. In this research, We acquired 18 ORFs with potential hydrophobin function in the whole genome of Magnaporthe grisea which screened from 58 hydrophobin sequence from NCBI database by using BLAST tool, and 11 ORFs of which was chosed to design specific primers, then we detected the distribution of these primers in 17 rice blast fungus. At the same time, we also reconstructed the phylogenic tree of 58 hydrophobin sequences by using N-J of Clustal X and tested bootstrap of Mega3.0. The results showed that there were 11 primers were detected by PCR and 4 of which obtained specific bands which consistent with the anticipation, 6 of which had no specific band, 1 primer without any specific band. The results of phyletic evolution analyzed showed that average disparity index （DI） a- mong hydrophobin was 0.62, and 58 hydrophobins were divied into two ClusterS： Ⅰ and Cluster Ⅱ, which accord with class I and class Ⅱ of fungal hydrophobins, respectively. All the hydrophobins from the fungi of Basidiomycotina, M. grisea and M. anisopliae belongs to Cluster Ⅰ , the other fungal hydrophobins belongs to Cluster Ⅱ. At last our research would lay a foundation for further clarifying the distribution of M. grisea and for validating the function of its related genes.

关 键 词：稻瘟病菌 疏水蛋白 系统进化 特异PCR 聚类分析 

分 类 号：S435.111.41[农业科学—农业昆虫与害虫防治]