安徽农业科学Issue(2)：188-191,4.
细菌性条斑病菌JH01诱导水稻抗病均一化差减文库的构建
Construction of a Normalized Subtractive cDNA Library of Rice Inoculated with Xanthomonas oryzae pv.oryzacola JH01
摘要
Abstract
[Objective] A normalized subtractive cDNA library of rice inoculated with Xanthomonas oryzae pv.oryzacola JH01 was constructed,the expression analysis of biological information and disease resistance related genes was conducted on obtained differential expression of ESTs.[Method] In this study,using seedling leaves of rice ‘IR26’ (Tester) and ‘ Liangyoupeijiu’ (Driver) as experimental materials,a subtracted cDNA library of rice inoculated by Xanthomonas oryzae JH01was constructed by the method based on subtractive hybridization of the DSN (duplex-specific nuclease) mediated subtractive hybridization.The quality of the subtractive library was detected by PCR amplification using primers pLB-F and pLB-R.[Result] The results demonstrated that the sizes of inserted fragments in the vector were between 0.5-2.0 kb,that the average size was about 1 kb,and that 95％ of the library clones were recombinants.504 clones were randomly selected,and 98 genes were obtained after clearing pollution,redundant,low quality sequences,and 59 genes were found to have homologous genes in analysis by BLAST.Gene ontology showed that 59 genes were related with resistance signals transduction,elementary metabolic,hypersensitive response,defense response et al.The other 39 sequences had no similar genes,which need further study.By real-time fluorescent quantitative PCR,OsNDPK4 isolated from the library was found to be involved in defense responses in rice induced by Xanthomonas oryzae JH01.[Conclusion] The above suggests that the constructed cDNA library of rice is of good quality and of use for isolation of interacting proteins.关键词
细菌性条斑病/水稻/双链特异性核酸酶(DSN)/均一化差减杂交/荧光定量PCRKey words
Bacterial leaf streak/Rice/Duplex-specific nuclease (DSN)/Normalized subtractive hybridization/Real-tine fluorescence quantitative PCR分类
农业科技引用本文复制引用
凌丹燕,路梅..细菌性条斑病菌JH01诱导水稻抗病均一化差减文库的构建[J].安徽农业科学,2016,(2):188-191,4.基金项目
浙江省自然科学基金项目(Y3110194). （Y3110194）
