Xiao Jingfan, Wang Qiyao, Liu Qin, Xu Lili, Wang Xin, Wu Haizhen, Zhang Yuanxing
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai 200237, People's Republic of China.
Appl Microbiol Biotechnol. 2009 May;83(1):151-60. doi: 10.1007/s00253-009-1924-9. Epub 2009 Mar 13.
In this study, rpoS gene was identified from Edwardsiella tarda EIB202 and its functional role was analyzed by using an in-frame deletion mutant rpoS and the complemental strain rpoS (+). Compared with the wild type and rpoS (+), rpoS was impaired in terms of the ability to survive under oxidative stress and nutrient starvation, as well as the resistance to 50% serum of Scophthalmus maximus in 3 h, demonstrating essential roles of RpoS in stress adaptation. The rpoS mutant also displayed markedly increased chondroitinase activity and biofilm formation. Real-time polymerase chain reaction revealed that the expression level of quorum sensing autoinducer synthetase genes luxS and edwI was increased by 3.7- and 2.5-fold in the rpoS mutant strain. Those results suggested that rpoS might be involved in the negative or positive regulation of chondroitinase and biofilm formation, or quorum sensing networks in E. tarda, respectively. Although there were no obvious differences between the wild-type and the rpoS mutant in adherence of epithelioma papulosum cyprini (EPC) cell and in the lethality on fish model, rpoS deletion leads to the drastically reduced capacity for E. tarda to internalize in EPC cells, indicating that RpoS was, while not the main, the factor required for the virulence network of E. tarda.
在本研究中,从迟缓爱德华氏菌EIB202中鉴定出rpoS基因,并通过使用框内缺失突变体rpoS和互补菌株rpoS(+)对其功能作用进行了分析。与野生型和rpoS(+)相比,rpoS在氧化应激和营养饥饿条件下的存活能力以及对3小时内大菱鲆50%血清的抗性方面受损,这表明RpoS在应激适应中起重要作用。rpoS突变体还表现出明显增加的软骨素酶活性和生物膜形成。实时聚合酶链反应显示,群体感应自诱导物合成酶基因luxS和edwI在rpoS突变体菌株中的表达水平分别提高了3.7倍和2.5倍。这些结果表明,rpoS可能分别参与迟缓爱德华氏菌中软骨素酶和生物膜形成或群体感应网络的负调控或正调控。虽然野生型和rpoS突变体在鲤上皮瘤(EPC)细胞黏附以及鱼模型致死性方面没有明显差异,但rpoS缺失导致迟缓爱德华氏菌内化到EPC细胞中的能力大幅降低,这表明RpoS是迟缓爱德华氏菌毒力网络所需的因素,虽不是主要因素。