中文摘要
本项目瞄准植物寄生线虫生防中侵染机理的关键问题，完成了不同类群食线虫真菌侵染性蛋白酶的基因克隆，以及同种中不同侵染力菌株（高毒力、弱毒力和无毒力）的侵染性蛋白酶基因克隆及蛋白质表达、基因敲除、蛋白质荧光定位等功能研究，阐明了侵染过程中关键蛋白酶以及其它不同种类的侵染性酶的作用和功能。完成了丝氨酸蛋白酶结构功能与微生物侵染线虫机理研究，构建了侵染性丝氨酸蛋白酶的三维结构模型，设计相关算法，分析比较这些模型功能关键残基在模型中物理化学特征、空间拓扑形态、分子电位以及各种作用信息，阐明了侵染相关氨基酸关键位点、蛋白酶结构与侵染能力的关系。通过上述研究，揭示食线虫真菌侵染线虫的分子机理，为线虫生防奠定理论基础。 本项目已取得一批原创性成果，在PNAS, FASEB J，Environ Microbiol, PLoS one和BMC Evol Biol等发表SCI论文25篇，两次荣获云南省科学技术奖自然科学类一等奖。申请10项专利,6项获授权。主办1次和参加17次国内外学术研讨会，培养博士生8名，硕士研究生20名。超额完成了项目预期的目标。
英文摘要
This project focused on the most important problems in areas of biocontrol against plant-parasitic nematodes. We have studied the infection process of different types of nematophageous fungi, and then cloned the genes encoding virulence proteases involved in the nematode infection. In our research, gene cloning of different virulence proteases with different abilities (including strong, weak or no activities) to infect nematodes in the same species was especially emphasized. Additionally, a series of functional experiments, such as homologous/heterologous protein expression, gene knock out and fluorescence protein localization, was also performed to understand the roles played by serine protease or the other kinds of virulence enzymes in infection. On the other hand, we have carried out the studies on the serine proteases structure-function in molecular mechanism of microbe infecting nematodes. Batched three-dimensional structural models for various virulence proteases were constructed. Meanwhile, the phsicochemical and stuctural topology properties, molecular surface electronic potential and various atomic interaction for these models were also analyzed and compared. The active sites and substrate association regions in these proteases were identified and further to clarify the relationship between protein conf
结题摘要
本项目瞄准植物寄生线虫生防中侵染机理的关键问题，完成了不同类群食线虫真菌侵染性蛋白酶的基因克隆，以及同种中不同侵染力菌株（高毒力、弱毒力和无毒力）的侵染性蛋白酶基因克隆及蛋白质表达、基因敲除、蛋白质荧光定位等功能研究，阐明了侵染过程中关键蛋白酶以及其它不同种类的侵染性酶的作用和功能。完成了丝氨酸蛋白酶结构功能与微生物侵染线虫机理研究，构建了侵染性丝氨酸蛋白酶的三维结构模型，设计相关算法，分析比较这些模型功能关键残基在模型中物理化学特征、空间拓扑形态、分子电位以及各种作用信息，阐明了侵染相关氨基酸关键位点、蛋白酶结构与侵染能力的关系。通过上述研究，揭示食线虫真菌侵染线虫的分子机理，为线虫生防奠定理论基础。 本项目已取得一批原创性成果，在PNAS, FASEB J，Environ Microbiol, PLoS one和BMC Evol Biol等发表SCI论文25篇，两次荣获云南省科学技术奖自然科学类一等奖。申请10项专利,6项获授权。主办1次和参加17次国内外学术研讨会，培养博士生8名，硕士研究生20名。超额完成了项目预期的目标。