松材线虫虫体特异表达cDNA文库的构建与分析

cDNA Library Construction and Analysis of Differentially Expressed in Mixed Stage Nematode of Bursaphelenchus xylophilus

松材线虫取食和致病相关基因的鉴定是揭示松材线虫病致病机制的重要研究内容。以松材线虫混合虫体为测试方,线虫卵为驱动方,构建二者差减cDNA文库。文库测序结果经序列拼接、功能注释,共获得松材线虫虫体特异表达的有效EST序列354个,拼接后获得34个重叠群,其中15个重叠群获得明确的功能注释,其中MixC12、MixC25、MixC33分别与过氧化物酶、β-1,4内切葡聚糖酶、锌指家族蛋白有较高的同源性,这些基因被认为与植物线虫的取食和致病性密切相关。RT-PCR分析验证了所获得的差减cDNA的代表性。

Identification of nematode feeding and pathogenicity-related genes of Bursaphelenchus xylophilus was critical to reveal the molecular mechanism of the Pine Wood Disease（PWD）.In this study,a suppression subtractive hybridization（SSH） cDNA library was constructed from the mixed stage nematode cDNA of B.xylophilus isolate AN19#F1,which was driven by the egg cDNA of AN19#F1.A total of 34 contigs generated from 354 ESTs after cDNA sequencing were obtained.Among them 15 contigs were functionally annotated.Contigs,MixC12,MixC25 and MixC33 had high homology with Peroxidase,Beta-1,4-endoglucanase,zinc finger family protein in the database,respectively.The genes were considered closely related to the feeding and pathogenicity processes.RT-PCR analysis confirmed the respective of the SSH cDNAs.