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利用细菌编码RNase的barnase基因及其特异抑制剂编码基因barstar构成的双元组分系统,把本实验室构建的具有稻瘟菌(Magnarorth rrisea)诱导活性的嵌合启动子与 barnase基因融合,再与由CaMV 355启动子驱动的barstar的构件融合,构建植物表达载体pWBNBS和pPBNBS,转化水稻,并对转基因水稻进行了抗稻瘟病和白叶枯病的检测.结果表明,接种稻瘟菌后,转基因水稻植株中barnase基因受诱导表达;在稻瘟菌侵染诱导下,barnase的表达超过barstar的抑制,导致转基因水稻叶片出现类似过敏感应,表现出对稻瘟病的抗性;转基因水稻对白叶枯病表现出一定的抗性.这些结果说明,通过该双元组分系统策略获得的转基因水稻具有明显的抗致病性真菌和一定程度的抗细菌危害的较广谱抗性。
Using the binary system composed of the barnase gene encoding bacterial RNase and its specific inhibitor barstar, the chimeric promoter with Magnarorth rrisea-induced activity was fused with barnase gene And the components of barstar driven by CaMV 355 promoter were fused to construct plant expression vectors pWBNBS and pPBNBS and transformed into rice.The resistance to rice blast and bacterial leaf blight of transgenic rice was tested.The results showed that after inoculation with rice blast fungus, The expression of barnase gene was induced in transgenic rice plants. The expression of barnase was more than that of barstar induced by Infection of Magnaporthe grisea, resulting in allergen-like responses in transgenic rice leaves and showed resistance to blast. The results showed that the transgenic rice obtained by the strategy of the two component system has obvious resistance to pathogenic fungi and a certain degree of broad spectrum resistance to bacterial damage.