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摘要：稻瘟病菌(Magnaporthe oryzae)引起的稻瘟病是水稻上的毁灭性病害之一，造成水稻产量大幅度降低，严重威胁全球的粮食安全。蛋白磷酸酶在真核细胞中的基因调控及信号传导等过程中具有重要作用[1]。在酿酒酵母中有研究报道，Ptps作为一类蛋白磷酸酶参与MAPK信号途径的反馈调节作用[2]，但是其在丝状真菌中的功能还鲜有报道,为了解析蛋白磷酸酶MoPtp1、MoPtp2稻瘟病菌生长发育及致病中的作用机制，通过同源重组的方法对蛋白磷酸酶MoPtp1、MoPtp2 进行了基因敲除并且进行了生物表型分析。结果显示，与野生型Guy11相比，突变体∆MoPtp2产孢量和致病能力均显著下降，而∆MoPtp1产孢量和致病能力并无显著性变化。利用PEG介导的原生质体转化方法，获得互补菌株∆MoPtp1/MoPTP11、∆MoPtp2/MoPTP2，结果显示互补菌株的生长、产孢、致病力与野生型Guy11基本一致。上述结果表明， MoPtp1、MoPtp2在调控稻瘟病菌的生长、分化及致病力过程中具有重要作用。研究结果对深入了解稻瘟病菌的致病分子机制及开发新型农药提供潜在靶标具有重要的理论指导意义。25650
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毕业论文关键词：稻瘟病菌； 蛋白磷酸酶；致病性；MoPtp2
Functional Analysis of Phosphorase MoPtp1 and MoPtp2 in Magnaporthe oryzae
Abstract：Rice blast caused by Magnaporthe oryzae is one of the important diseases of rice. Rice blast can lead to a significant reduction in rice yield and a serious threat to food security in China. Thestudies of Saccharomyces cerevisiae have reported that Ptps is a class of protein phosphatase involved in MAPK feedback regulation, but its function in the filamentous fungi is still rarely reported, to analyze the protein phosphatase MoPtp1, MoPtp2 in rice blast fungus growth and development Pathogenicity of the protein phosphatase MoPtp1, MoPtp2 was genetically knocked out by homologous recombination, and the phenotypic analysis was carried out. The results showed that the growth of the mutant ΔMoPtp2 showed slowed down, and the sporulation and pathogenicity were significantly decreased compared with the wild type Guy11，nevertheless, ΔMoPtp1 had no significant changes in these ways . The resultsshwed that the growth, sporulation and pathogenicity of the complementary strains were consistent with Guy11 ,which is obtained by using the PEG-mediated method to These results show that MoPtp1 and MoPtp2 control the growth and pathogenesis of Magnaporthe grisea by regulating the synthesis of protein phosphatase. Theses results of the study have important theoretical significance for deeply understanding the pathogenic molecular mechanism of rice blast fungus and the development of potential targets for the development of new pesticides.
Key words: Magnaporthe oryzae；Protein phosphatase；pathogenicity；,MoPtp2
目 录
摘要1
关键词1
Abstract1
Key words1
引言1
1 材料与方法2
1.1材料...2
1.1.1项目采用的菌株和培养基2
1.1.2项目采用的酵母及其培育条件3
1.2 方法3
1.2.1构建目的基因敲除载体3
1.2.2稻瘟病菌的原生质体的转化3
1.2.3目的基因序列与pYF11载体的酵母转化3
1.2.3.1制备酵母感受态细胞.4
1.2.3.2 酵母转化4
1.2.4酵母质粒转化大肠杆菌4
1.2.5 互补菌株的获得4
1.2.6供试菌株营养生长的测定....4
1.2.7供试菌株产孢量的测定4
1.2.8供试菌株致病力的测定4
2 结果与分析.4
2.1敲除载体与互补载体的构建.... ..5
2.2 MoPtp1与MoPtp2不参与调控营养生长...5
2.3 MoPtp2调控稻瘟病菌的产孢过程.....6
2.4 MoPtp1与MoPtp2参与调控稻瘟病菌的致病力...6
3 讨论7
4致谢.8
参考文献.8
稻瘟病菌蛋白磷酸酶MoPtp1, MoPtp2的功能分析