BWMK1互作基因BWIP4对粳稻的转化

Transformation of the BWMK1-interacting gene BWIP4

通过酵母双杂交体系,获得了7个BWMK1互作基因BWIP1～BWIP7.从中选取BWIP4作进一步的功能分析,构建了BWIP4-RNAi表达载体pANDA-BWIP4和超表达载体NCGR-1300-BWIP4,利用农杆菌介导法转化粳稻日本晴,获得了35个干涉株系和136个超表达转基因株系,对经PCR、GUS检测的阳性植株进一步进行RT-PCR检测,干涉株系中BWIP4的表达量低于日本晴,而超表达株系中BWIP4的表达量高于日本晴.

BWMK1,a first reported rice MAPK gene,was transcriptionally activated by blast fungus infection and wounding.To make a further understanding about the BWMK1-mediated defense signaling,we identified 7 BWMK1 interacting genes by yeast two-hybrid screening,designate BWIP1~BWIP7,and selected BWIP4 for further functional characterization.Through Agrobacterium mediated transformation method,we generated overexpression and RNAi transgenic plant using a japonica cultivar nipponbare as a receptor.The GUS assay and PCR results showed that the target gene had been intergrated into receptor.Further expression analysis was confirmed by RT-PCR.These transgenic lines provided us a good material for in-deepth function analysis of BWIP4.