Vos K, Braeken K, Fauvart M, Ndayizeye M, Verhaert J, Zachurzok S, Lambrichts I, Michiels J
Centre of Microbial and Plant Genetics, Katholieke Universiteit Leuven, 3001 Heverlee, Belgium.
Environ Microbiol. 2007 Jul;9(7):1665-74. doi: 10.1111/j.1462-2920.2007.01284.x.
Rhizobium etli is a Gram-negative root-colonizing soil bacterium capable of fixing nitrogen while living in symbiosis with its leguminous host Phaseolus vulgaris. A genome-wide screening for R. etli symbiotic mutants revealed a R. etli operon encoding an oligopeptide ABC-transporter (Opt), two redA homologous genes and one redB gene. Expression analysis showed this opt operon to be transcribed both under free-living and symbiotic conditions and expression levels were demonstrated to be growth-phase-dependent. Plants nodulated by R. etli opt mutants showed a reduced symbiotic nitrogen fixation activity (approximately 50% reduction). Growth experiments with opt mutants in the presence of oligopeptides as the sole nitrogen source confirmed the involvement of the opt genes in oligopeptide uptake. Further phenotypic analysis of the opt mutants revealed them to display an enhanced resistance to the oligopeptide antibiotic bacitracin, an increased susceptibility to the beta-lactam antibiotic ampicillin and a decreased osmotolerance. In conclusion, our results demonstrate that the opt operon plays a crucial role during symbiosis and stress resistance.
费氏中华根瘤菌是一种革兰氏阴性的、定殖于根部的土壤细菌,能够在与其豆科宿主菜豆共生时固氮。对费氏中华根瘤菌共生突变体进行全基因组筛选,发现了一个费氏中华根瘤菌操纵子,其编码一个寡肽ABC转运蛋白(Opt)、两个redA同源基因和一个redB基因。表达分析表明,该opt操纵子在自由生活和共生条件下均有转录,且表达水平呈生长阶段依赖性。由费氏中华根瘤菌opt突变体结瘤的植物表现出共生固氮活性降低(约降低50%)。以寡肽作为唯一氮源对opt突变体进行生长实验,证实了opt基因参与寡肽摄取。对opt突变体的进一步表型分析表明,它们对寡肽抗生素杆菌肽的抗性增强,对β-内酰胺抗生素氨苄青霉素的敏感性增加,且渗透压耐受性降低。总之,我们的结果表明,opt操纵子在共生和抗逆过程中起关键作用。