基于代谢组学技术集成的九州虫草种质资源评价及虫草素生物合成的转录组、蛋白质组学研究

利用代谢组学技术，从九州虫草两性世代发育过程的角度探讨代谢物种类和数量的变化规律，开展种质资源评价。拟优化待测样本的提取及预处理方法，实现液质和气质关键分析技术的集成，针对菌丝体和子实体的宽极性代谢物分离分析，建立以虫草素为代表的潜在标志物色谱-质谱数据库，筛选虫草素高产菌株；利用转录组学技术，开展菌丝体和子实体高通量测序、注释/比对，寻找虫草素生物合成的候选基因簇；利用蛋白质组iTRAQ定量质谱技术，筛选世代交替中的差异蛋白表达，推测虫草素生物合成途径、关键作用酶及调控机制，应用微阵列芯片及Western-blot技术初步验证候选蛋白，完成合成生物学化学基础研究。. 种质资源评价和虫草素合成的化学生物学研究，对虫草核心种质的构建、遗传多样性及稳定性、优异基因的定位、功能描述等工作具有重大的理论和实践意义。

The evaluation of germplasm resource of Cordyceps kyushuensis was carried out with the metabolomics technology, from a standpoint of alternation of generations, to summary the changing rules of metabolite. The proposed integrated technology of were established based on sample extraction and pretreatment methods, research methods of GC-MS and LC-MS/MS, to separate and analysis the wide polar metabolites of the mycelium and fruiting body. The potential key metabolites were identificated to establish the appropriate biomarkers library. RNA-Sequence technology and iTRAQ technology were employed to make a comparative transcriptome and proteomic research of cordyceps in the life history evolution. The possible biosynthesis pathway, regulation mechanism and crucial enzymes of cordycepin was speculated and proved by protein micro-array chip and Western blot, laying a solid foundation for the followed study of synthetic biology.This topic research had a profound theoretical and practical significance on construction of core collection, genetic diversity and stability, gene mapping and function discription.