短密木霉与白术根腐病菌的空间互作及相关蛋白的克隆与功能研究

道地药材白术根腐病严重，目前主要靠农药和轮作，但收效一般。课题组通过前期研究，从白术根部筛选到短密木霉菌Trichoderma brevicompactum，实验发现其有良好的抑菌及促生作用，但对致病菌拮抗机制、互作模式以及分子机理尚不明确。显然，木霉在诱导白术抗病过程中，定会分泌一些疏水蛋白小分子作为MAMPs效应分子与白术互作。据此我们提出，在植物-微生物-土壤的微生态系统中，短密木霉会改变根际微生物区系，在根表大量定殖以防病菌入侵，并通过微生物相关分子模式诱导植株产生抗病性。因此，本项目拟用qPCR和MiSeq研究木霉对根际微生物区系的影响；同时构建木霉和镰刀菌的GFP/RFP穿梭载体，利用激光共聚焦明确其定殖及互作细节；并用基因敲除、瞬时干扰、RNA-seq等克隆其控制定殖的疏水蛋白及植株互作蛋白，研究其功能，揭示诱导植株免疫发生的分子途径，为白术可持续发展及菌肥开发奠定理论基础。

The famous authentic herbs Atractylides macrocephala is disturbed by theroot rot，which is one of the important factors affecting the yield and quality of A. macrocephala. It has been confirmed that Trichoderma could prevent A. macrocephala root rot. In our previous experiments, a highly effective biocontrol fungi, Trichoderma brevicompactum, was screened and found to have good bacteriostatic and growth-promoting effects and to effectively prevent root decay. However, the antagonistic mechanism and molecular mechanism of interaction in the root system of T. brevicompactum and Fusarium oxysporum is not clear. Accordingly, we propose that in the micro-ecosystem of plant-microbe-soil, T. brevicompactum may alter the rhizosphere microbial flora, form a bacterial membrane at the root surface to prevent pathogen invasion and induce plant-derived antimicrobial activity through Microbe-associated molecular pattern (MAMPs). Based on the above hypothesis, some techniques including qPCR and MiSeq will be used to study the effect of T. brevicompactum on rhizosphere microbial flora and fluorescent labeling technology will be used to make interaction of T. brevicompactum and F. oxysporum visualization. The hydrophobic proteins and the plants interacting protein related with colonization of T. brevicompactum will be screened by Knockout, RNAi and RNA-seq. In this way, the molecular pathways of the plant immunity induced by T. brevicompactum was revealed systematically, and a more effective and theoretical support were established for the sustainable development of A. macrocephala and the exploitation of microbial fertilizer.