天麻作为药材要求天麻素和4-羟基苯甲醇两者总含量大于0.25%，但人工栽培天麻活性成分低而不稳定影响药材质量。本项目前期研究从昭通乌天麻转录组测序数据中筛选出天麻素和4-羟基苯甲醇活性成分生物合成关键酶候选基因单加氧酶和糖基转移酶基因。本研究将通过RACE-PCR克隆这些基因全长cDNA序列, 用原核系统表达并纯化这些基因编码的重组酶进行生化特性分析；再用组成型和诱导型启动子构建这些基因的真核表达载体，通过农杆菌介导法转化蜜环菌获工程蜜环菌，在蜜环菌中过表达单加氧酶和葡糖基转移酶构建一个代谢途径合成天麻活性成分，用前体物质和诱导剂处理工程蜜环菌，分析工程蜜环菌中天麻活性成分含量变化，阐明蜜环菌中天麻活性物质生物合成调控的分子机理。再通过工程蜜环菌接种乌天麻的栽培试验，验证工程菌提高天麻活性成分含量的作用，发展优质天麻栽培新理论和技术，促进天麻产业可持续发展。

Gastrodia elata (G.elata) as a medicinal material requires that the contents of gastrodin and 4- hydroxy benzyl alcohol are more than 0.25%, but the low and unstable active ingredients of the artificial cultivated G. elata may affect the quality of the medicinal materials. The preliminary study of this project selected candidate key enzyme genes in the biosynthesis of active ingredients monooxygenase and glycosyltransferases as well as Armillaria mellea (A. mellea) laccase Unigene from the Zhaotong Gastrodia elata f.glauca transcriptome sequencing analysis. This project would clone the full-length cDNA sequences of these genes through RACE-PCR, express and purify the recombinant enzymes encoded by these genes by using prokaryotic system, and analyze their biochemical characteristics. The eukaryotic expression vectors of these genes would be constructed with the composition and inducible promoter. The engineering A. mellea would be obtained through transformation of these vectors by Agrobacterium mediated method. Over expression of laccase in A. mellea would make efficient use of the wood grown with A.millea. Simultaneous over expression of monooxygenase and glucosyltransferase would construct a metabolic pathway to synthesize the active constituents of G. elata. After the engeering A. mellea treated with precursor and inducer separately, the changes of G. elata active component content would be analyzed to clarify the regulatory molecular mechanism of biosynthesis of G. elata active substances. The effect of the engineering A. mellea control on the active components of G. elata would be made clear through the cultivation test of G. elata inoculated with engineering A. mellea. These would develop a new cultivation theory and technology for high quality G. elata to promote the sustainable development of G. elata industry.