水稻抗稻瘟菌HIGS表达载体的构建及遗传转化

Construction of rice blast resistant HIGS expression vectors for rice genetic transformation

水稻是我国主要粮食作物之一,而稻瘟病是影响水稻安全生产的最主要病害之一。为克服抗病基因的抗病性很快消失的弊端,本研究尝试了宿主诱导的基因沉默(HIGS)技术在创制抗稻瘟病水稻新材料上的可行性。HIGS是新发展起来的以RNAi为基础的抗病技术,即在寄主植物中表达可沉默病原物特定基因的HIGS载体,达到控制病原菌扩展的目的。本研究选取了两个稻瘟菌致病关键基因Nox1和NAC为研究靶标,分别克隆其UTR区和CDS区中的特异区段,利用Gateway技术构建这4个片段HIGS表达载体,再利用农杆菌介导法将各载体分别转化水稻,通过鉴定得到转基因阳性植株,为后续开展该技术在水稻抗稻瘟病方面的深入研究奠定了基础。

Rice blast disease seriously influences the quality and production of rice, the main food crop in China. To overcome the easy breakdown of conventional disease resistant transgenic plants, we used HIGS （host induced gene silencing） to create new kind of transgenic rice plants. By transforming HIGS constructs targeting the viru- lence-related genes of the rice blast fungus Magnaporthe grisea into host plants, HIGS could suppress the fungal de- velopment and virulence. We chose two important genes in M. oryzae and cloned the specific fragments of the genes for HIGS vector construction. Then we transformed the HIGS constructs into rice. This research lays the foundation for molecular breeding of rice blast resistant varieties.