红鱼干作为我国南海特色水产珍品，加工贮藏过程中游离氨基酸（FAA）变化与镰孢菌T-2毒素污染密切相关，因此，探明FAA调控T-2合成的机制有助于红鱼干质量安全的精准控制。本项目提取红鱼干加工贮藏期样本的∑FAA，添加到镰孢菌Fo17 的培养基中，采用LC-MS/MS分析T-2产量、Real time PCR分析TRI5表达量、形态学表征镰孢菌，找出T-2合成的关键FAA（FAAk）并以其标准品进行产毒验证。同时，利用同源重组技术分别构建TORC1信号系统上游的FAAk响应元件（GTR1和GTR2）和下游的毒素调控开关（SCH9和TAP42）缺失株，分析各元件在T-2合成和TRI5表达过程中的作用，明确与FAAk相关的上游和下游关键元件，并以基因回补技术验证其功能。最后以免疫亲和捕获和沉淀技术解析TORC1蛋白分子上下游目标元件复合体形成中的功能，进而阐明FAAk对T-2合成的级联调控机制。

Dried Lutianus erythropterusis an aquatic specialty in the South China Sea. During its production and storage period, the development of free amino acid (FAA) and T-2 toxin (T-2) contamination are closely related. It is therefore helpful to find out FAA’s regulation mechanism of T-2 synthesis in order to precisely control the quality and safety of dried Lutianus erythropterus. In this project, ∑FAA will be extracted from the dried Lutianus erythropterus samples taken in the production and storage period and will then be added into the substrates of Fusarium Fo17. LC-MS/MS method will be used to analyze the yield of T-2; Real time PCR will be used to analyze TRI5 expression quantity; and phenotypic characteristics will be observed according to morphology. As such, the key FAA (FAAk) relation to T-2 synthesis will be found and its standards can be used to perform T-2 producing verification. In the meantime, homologous recombination method will be used to construe null mutant strains of the upstream response elements (Gtr1 and Gtr2) and the downstream T-2 regulatoryelements (SCH9 and TAP42) in TORC1 signal system respectively, and to analyze the function of each element in the process of T-2 synthesis and TRI5 expression in order to ascertain the key upstream and downstream genes that are related to FAAk. The function of the key genes will be verified through gene recovering technology. Finally, immune affinity capture and precipitation method will be adopted to discover the effect of TORC1 protein molecule on the formationof upstream and downstream target element complex, so as to illustrate the cascade regulatory mechanism of T-2 synthesis induced by FAAk.