BrlA抑制食线虫真菌紫色紫孢菌营养菌丝生长机理研究

It is very important to study the life cycle switching to totally understanding the infection mechanisms of the pathogenic fungi to their hosts. Purpureocillium lavendulum (formally Paecilomyces lilacinus) is an important fungus for nematode bio-control. Our previous study revealed that BrlA, the central regulating gene for conidiation, can inhibit vegetative growth in the fungus. In this project, we would like to continue our research based on this phenomenon. Firstly, we want to construct an induced expression system of BrlA. Then we will use comparative transcriptome analysis and random mutation screening to find BrlA downstream genes involved in inhibiting vegetative growth. These genes were then confirmed by gene knockout, complementation, overexpression, or GFP fusing expression. Finally, we will confirm whether the pathways relative to cell starvation or vegetative growth, such as TOR, autophagy, or PkaA, are regulated by BrlA. after complete this project, we will explain how BrlA inhibit vegetative growth in Purpureocillium lavendulum, more comprehensively understand the mechanism of lifestyle switching and infection of hosts.

研究病原真菌的生活史转化调控对于理解真菌侵染宿主的分子机制具有重要意义。紫色紫孢菌（原名淡紫拟青霉）是重要的线虫生防菌。课题组研究发现紫色紫孢菌产孢核心调控转录因子BrlA可以抑制营养菌丝的生长。截止目前，BrlA的表达是经何种途径抑制菌丝的生长没有报道。本项目拟在前期研究的基础上构建BrlA诱导表达菌株，并以该菌株作为出发菌株通过比较转录组的方法发现BrlA调控的抑制营养菌丝生长的基因，同时对该菌株进行农杆菌介导的Ti质粒随机插入突变，筛选不受BrlA抑制的突变株，鉴定突变的基因并通过基因敲除、回补、过表达和荧光定位等验证相关基因的功能。通过分析BrlA诱导表达时TOR，PkaA等通路是否被激活来确定它们是否参与BrlA抑制菌丝生长的调控。通过本项研究能够深入理解BrlA抑制营养菌丝生长的分子机制，为阐明食线虫真菌生活史转化的调控和侵染宿主的分子机制奠定基础。

研究病原真菌的生活史转化调控对于理解真菌侵染宿主的分子机制具有重要意义。紫色紫孢菌（原名淡紫拟青霉）是重要的线虫生防菌。课题组研究发现紫色紫孢菌产孢核心调控转录因子BrlA可以抑制营养菌丝的生长。截止目前，BrlA的表达是经何种途径抑制菌丝的生长没有报道。本项目在前期研究的基础上构建BrlA诱导表达菌株，并以该菌株作为出发菌株通过比较转录组的方法发现BrlA调控的抑制营养菌丝生长的基因，同时对该菌株进行农杆菌介导的Ti质粒随机插入突变，筛选不受BrlA抑制的突变株。研究结果表明brlA抑制紫色紫孢菌营养菌丝生长的机制主要体现在：brlA对氨基酸代谢，核糖体合成、细胞骨架合成，TCA循环等过程，分子伴侣hsp90可能在其中起到重要作用。通过本项研究能够深入理解BrlA抑制营养菌丝生长的分子机制，为阐明食线虫真菌生活史转化的调控和侵染宿主的分子机制奠定基础。