Bennett H P J, Clarke D J
Department of Biology and Biochemistry, Molecular Microbiology Laboratory, University of Bath, Bath BA2 7AY, United Kingdom.
J Bacteriol. 2005 Jan;187(1):77-84. doi: 10.1128/JB.187.1.77-84.2005.
Photorhabdus is a genus of gram-negative Enterobacteriaceae that is pathogenic to insect larvae while also maintaining a mutualistic relationship with nematodes from the family Heterorhabditis, where the bacteria occupy the gut of the infective juvenile (IJ) stage of the nematode. In this study we describe the identification and characterization of a mutation in the pbgE1 gene of Photorhabdus luminescens TT01, predicted to be the fifth gene in the pbgPE operon. We show that this mutant, BMM305, is strongly attenuated in virulence against larvae of the greater wax moth, Galleria mellonella, and we report that BMM305 is more sensitive to the cationic antimicrobial peptide, polymyxin B, and growth in mildly acidic pH than the parental strain of P. luminescens. Moreover, we also show that the lipopolysaccharide (LPS) present on the surface of BMM305 does not appear to contain any O antigen. Complementation studies reveal that the increased sensitivity to polymyxin B and growth in mildly acidic pH can be rescued by the in trans expression of pbgE1, while the defects in O-antigen assembly and pathogenicity require the in trans expression of pbgE1 and the downstream genes pbgE2 and pbgE3. Finally, we show that BMM305 is defective in symbiosis as this mutant is unable to colonize the gut of the IJ stage of the nematode. Therefore, we conclude that the pbgPE operon is required for both pathogenicity and symbiosis in P. luminescens.
光杆状菌属是革兰氏阴性肠杆菌科的一个属,对昆虫幼虫具有致病性,同时还与异小杆线虫科的线虫保持着互利共生关系,细菌占据线虫感染性幼虫(IJ)阶段的肠道。在本研究中,我们描述了发光光杆状菌TT01的pbgE1基因中一个突变的鉴定和特征,该基因预计是pbgPE操纵子中的第五个基因。我们表明,这个突变体BMM305对大蜡螟幼虫的毒力大大减弱,并且我们报告BMM305比发光光杆状菌的亲本菌株对阳离子抗菌肽多粘菌素B更敏感以及在轻度酸性pH条件下生长更受影响。此外,我们还表明BMM表面存在的脂多糖(LPS)似乎不包含任何O抗原。互补研究表明,通过pbgE1的反式表达可以挽救对多粘菌素B的敏感性增加和在轻度酸性pH条件下的生长,而O抗原组装和致病性的缺陷需要pbgE1以及下游基因pbgE2和pbgE3的反式表达。最后,我们表明BMM305在共生方面存在缺陷,因为这个突变体无法定殖在线虫IJ阶段的肠道中。因此,我们得出结论,pbgPE操纵子是发光光杆状菌致病性和共生所必需的。