青枯菌中Prt1蛋白对青枯素生物合成的转录调控

青枯素（ralsolamycin）是一种由青枯菌产生的跨界通讯信号，在青枯菌-土壤真菌互作过程中起重要的作用。我们发现prt1基因突变后，青枯素编码基因的表达量显著下调，青枯素的合成量显著减少。那么，Prt1调控青枯素生物合成的机制是什么？进一步EMSA实验显示：Prt1蛋白并不与青枯素编码基因的启动子区域直接结合。由此，我们提出Prt1调控青枯素生物合成的通路上还涉及其它中间调控因子。本项目拟进一步采用转录组测序、基因敲除和互补、qRT-PCR验证、EMSA、DNaseI足迹实验、LC-MS检测和青枯菌-真菌互作分析等方法，旨在鉴定受Prt1直接调控且参与调控青枯素生物合成的中间调控因子，从而揭示Prt1对青枯素生物合成的调控作用和机制。在逐步摸清青枯菌信号调控通路的基础上，也可以为制定更加有效的青枯病防治策略提供理论指导。

Ralsolamycin is one kind of inter-kingdom communication signal, which is produced by Ralstonia solanacearum and plays a key role in inducing interactions between R. solanacearum and fungi. We have found that the encoding gene expression level and production of ralsolamycin were decreased significantly when the prt1 gene was mutated, but the molecular mechanism of Prt1 protein modulating ralsolamycin biosynthesis is still vague. Result of EMSA experiment further demonstrated that Prt1 protein was not bind to the promoter sequence directly. Thus, we infer that other regulatory gene may involve in the ralsolamycin biosynthesis pathway. In this project, to further identify the key genes that modulated by Prt1 and involved in the ralsolamycin biosynthesis, the RNA-seq, gene deletion and functional complementation, qRT-PCR verification, EMSA, DNaseI footprinting assay, LC-MS detection and R. solanacearum-fungi interaction will be conducted. Finally, regulation mechanism of ralsolamycin biosynthesis by Prt1 will be clarified. On the basis of expounding the signal regulation pathway, our results can also provide important guidance for the management of the bacterial wilt.