北细辛内生真菌A.h-Fs-1降解马兜铃酸Ⅰ的分子机制

含马兜铃酸药材的合理减毒是目前急需解决的关键问题，传统的减毒方法难以达到完全去毒的效果，而内生菌与宿主互惠共生，能利用自身优势分解转化有毒成分。申请者前期以马兜铃酸Ⅰ为唯一碳源，从北细辛根状茎中首次筛选分离出具有降解马兜铃酸Ⅰ的内生真菌A.h-Fs-1，但其降解机制及活性还未阐明。本项目拟以发现的降解菌A.h-Fs-1为研究对象，通过多组学分析得到其降解马兜铃酸Ⅰ的途径和候选功能基因，克隆基因全长并通过异源表达系统验证候选功能基因的催化功能。再利用标记后的A.h-Fs-1回染北细辛，分析A.h-Fs-1对北细辛药材中马兜铃酸含量的影响，进一步验证其活性。本项目是申请者博士后研究工作的拓展和创新，通过栽培过程的活体减毒研究为有毒中药减毒提供新思路，对细辛的安全使用具有重要意义。

The rational detoxification of Chinese medicine containing aristolochic acidⅠis a key issue that needs to be solved urgently. Traditional attenuation methods are difficult to achieve the effect of complete detoxification. However, endophytic fungus and the host symbiosis mutually and can use their own advantages to decompose and transform toxic components. In the early stage, the applicant used aristolochic acid I as the sole carbon source, and for the first time screened and isolated the endophytic fungus A.h-Fs-1 that can degrade aristolochic acid I from the rhizomes of Asarum heterotropoides. But its degradation mechanism and activity are still not clarified. This project intends to use the discovered degrading bacterium A.h-Fs-1 as the research object. Through multi-omics analysis, to obtain the pathway and candidate functional genes to degrade aristolochic acid I. Clone the full length of the candidate gene and verify the catalytic function through a heterologous expression system. The labeled A.h-Fs-1 was re-stained with Asarum heterotropoides, and the effect on the content of aristolochic acid I was analyzed to further verify its activity. This project is an extension and innovation of the applicant's post-doctoral research work. Through the in vivo detoxification research of the cultivation process, it provides new ideas for the attenuation of toxic traditional Chinese medicines, which is of great significance to the safe use of Asarum heterotropoides.