1 Institute of Plant Genetics and Developmental Biology, College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China.
2 College of Agriculture, Northeast Agricultural University, Harbin 150030, China; and.
Phytopathology. 2019 Jul;109(7):1171-1183. doi: 10.1094/PHYTO-07-18-0225-R. Epub 2019 May 15.
The global regulator Crp-like protein (Clp) is positively involved in the production of virulence factors in some of the spp. However, the functional importance of Clp in pv. has not been investigated previously. Here, we showed that deletion of led to significant reduction in the virulence of pv. in soybean, which was highly correlated with the drastic reductions in carbohydrates utilization, extracellular polysaccharide (EPS) production, biofilm formation, cell motility, and synthesis of cell wall degrading enzymes (CWDEs). These significantly impaired properties in the mutant were completely rescued by a single-copy integration of the wild-type into the mutant chromosome via homologous recombination. Interestingly, overexpression of in the wild-type strain resulted in significant increases in cell motility and synthesis of the CWDEs. To our surprise, significant reductions in carbohydrates utilization, EPS production, biofilm formation, and the protease activity were observed in the wild-type strain overexpressing , suggesting that Clp also plays a negative role in these properties. Furthermore, quantitative reverse transcription polymerase chain reaction analysis suggested that was positively regulated by the diffusible signal factor-mediated quorum-sensing system and the HrpG/HrpX cascade. Taken together, our results reveal that Clp functions as both activator and repressor in multiple biological processes in pv. that are essential for its full virulence.
全球调节蛋白 Crp 样蛋白(Clp)正向参与一些 spp. 的毒力因子的产生。然而,Clp 在 pv. 中的功能重要性以前尚未被研究过。在这里,我们表明,删除 导致 pv. 在大豆中的毒力显著降低,这与碳水化合物利用、细胞外多糖(EPS)产生、生物膜形成、细胞运动和细胞壁降解酶(CWDE)合成的急剧减少高度相关。这些在 突变体中严重受损的特性通过同源重组将野生型 单个拷贝整合到突变体染色体中完全得到了挽救。有趣的是,在野生型菌株中过表达 导致细胞运动和 CWDE 合成显著增加。令我们惊讶的是,在过表达 的野生型菌株中观察到碳水化合物利用、EPS 产生、生物膜形成和蛋白酶活性的显著降低,表明 Clp 也在这些特性中起负作用。此外,定量逆转录聚合酶链反应分析表明, 受可扩散信号因子介导的群体感应系统和 HrpG/HrpX 级联正向调控。总之,我们的结果表明,Clp 在 pv. 中的多个生物学过程中既作为激活剂又作为抑制剂发挥作用,这些过程对其完全毒力至关重要。