转录因子AP1调控灵芝酸生物合成中ROS信号和钙信号的交互作用机制

Ganoderma lucidum is a famous medicinal fungi in China. Ganoderic acid is its main medicinal ingredient. Applicants earlier studies have found that transcription factor AP1 regulates the biosynthesis of Ganoderma lucidum. At the same time, ROS signal and calcium signal are involved in the regulation process. But its regulatory mechanism is not clear. In this study, we will investigate the crosstalk between ROS signaling and calcium signaling on the biosynthesis of Ganoderma lucidum. Further we will screen the ROS signal and the calcium-related genes in the transcription factor AP1 silence strain. We also will screen the new ROS signal related genes and calcium-related genes as candidate genes by transcriptome difference analysis. Based on the transcription factor AP1 silencing strain, the overexpression strain or gene silencing strain of the candidate gene was constructed. The ROS content, calcium ion content and the biosynthesis of Ganoderma lucidum were studied, and the Role of the crosstalk between ROS signal and calcium signal was analyzed in transcription factor AP1 silence strain in the regulation of Ganoderma lucidum biosynthesis. These researches will help to further elucidate the mechanism of GA biosynthesis network regulation, and provide theoretical basis for improving content of GA in G. lucidum using breeding or cultivating methods. At the same time, it will provide reference for studying on other terpenoid compounds biosynthesis in other Chinese medicine.

灵芝是我国著名药用菌。灵芝酸是其主要药用成分。申请者前期研究发现，转录因子AP1调控灵芝酸的生物合成。同时，ROS信号和钙信号均参与到该调控过程，但是其调控机制尚不清楚。本研究拟通过考察ROS信号与钙信号的交互作用对灵芝酸生物合成的影响，进一步筛选转录因子AP1调控下的ROS信号和钙信号相关的已知基因以及通过转录组差异分析筛选新的ROS信号和钙信号相关的基因作为候选基因。在转录因子AP1沉默菌株的基础上，构建候选基因的过表达菌株或基因沉默菌株，并考察该菌株的ROS含量，钙离子含量以及灵芝酸生物合成的变化，进而解析ROS信号和钙信号的交互作用在转录因子AP1调控灵芝酸生物合成中到的分子机制。研究结果将进一步阐明转录因子AP1调控灵芝酸生物合成的分子机制和调控网络，为进一步通过育种或栽培方法提高灵芝酸含量提供理论基础，加深人们对药用菌次生代谢调控的了解，为其它中药中萜烯类化合物研究提供借鉴。

灵芝是我国著名药用菌，灵芝酸是其主要药用成分。申请者在前期研究转录因子AP1调控灵芝酸生物合成基础上，对转录因子AP1影响的ROS信号和钙信号交互作用调控灵芝酸生物合成的机制开展了研究。全面考察转录因子AP1基因沉默菌株中ROS信号和钙信号的变化，进一步考察转录因子AP1基因沉默引起的ROS变化、钙信号变化与灵芝酸生物合成的关系。分析转录因子AP1基因沉默菌株中ROS信号和钙信号在AP1调控灵芝酸过程中的相互作用关系。分析转录组数据，筛选与ROS信号途径和钙信号途径相关的差异表达基因，筛选到一些候选基因，克隆并研究了基因功能、表达特性等，并构建了相关双突变体菌株，并考察了其对灵芝酸以及ROS含量的影响。此外，筛选了转录因子AP1下游调控ROS信号和钙信号相关途径的靶基因。构建了AP1下游调控的靶基因PLC、Gpx的沉默菌株，并考察菌株灵芝酸含量。该研究结果为通过育种或栽培方法提高灵芝酸含量提供新的靶基因，同时拓展了转录因子AP1调控灵芝酸生物合成的分子机制和调控网络，加深人们对药用菌次生代谢调控的理解。