利用抑制差减杂交筛选稻瘟菌继代接种菌株差异表达基因

Screening genes related to adaptive variation in Magnaporthe oryza by suppression subtracted hybridization

稻瘟菌是重要的水稻病原微生物,通过各种变异机制提高自身对水稻的致病力。通过继代接种可使稻瘟菌发生适应性变异,从而增强致病力。为了解相关适应性变异的分子机制,利用稻瘟菌原始菌株与在水稻上继代接种的第四代菌株构建了抑制消减杂交文库。经过PCR验证及测序分析,共计获得168条EST非冗余序列。拼接后,Unigene的数量为110个。利用不同数据库,进行BLASTX分析,获得了与这些序列推测蛋白相似的功能蛋白,分析蛋白功能注释表明这些蛋白主要与合成和信号传导相关。GO分析表明相关蛋白涉及生命活动的多个方面,但主要集中于细胞生长发育、核酸结合及外界刺激的感知及传导。

Rice blast fungus is one of the most important rice pathogens,whose pathogenicity on rice can be enhanced through several ways.Successive inoculation can make their virulence improved,which is called adaptive variation.In order to understand the molecular mechanisms related to adaptive mutation,a suppression subtracted hybridization library was constructed using the original strain and the fourth generation strain after successive inoculation on rice.After verification by PCR and sequence analysis,total of 168 non-redundant EST sequences were obtained.Alignment analysis led to 110 Unigenes.BLASTX analysis against different databases was used to reveal the functions of these proteins deduced from EST sequences,which were obtained from sequence analysis of the suppression subtracted hybridization library.KEGG analysis showed that 20 Unigenes can be classified into metabolic pathways.Annotation of the proteins showed that these proteins were primarily associated with the synthesis and signal transduction.GO analysis revealed that the proteins involved in many aspects of life activities,but mainly in cell growth,nucleic acid binding and the perception of external stimuli and signal transduction.