Silby Mark W, Nicoll Julie S, Levy Stuart B
Center for Adaptation Genetics and Drug Resistance, Department of Molecular Biology and Microbiology, Tufts University School of Medicine, Boston, Massachusetts 02111, USA.
Appl Environ Microbiol. 2009 Jun;75(12):3872-81. doi: 10.1128/AEM.00017-09. Epub 2009 Apr 24.
Knowledge of the genetic basis for bacterial survival and persistence in soil is a critical component in the development of successful biological control strategies and for understanding the ecological success of bacteria. We found a locus specifying polyphosphate kinase (ppk) and a nonpredicted antisense RNA (iiv8) in Pseudomonas fluorescens Pf0-1 to be necessary for optimal competitive fitness in LB broth culture and sterile loam soil. Pf0-1 lacking ppk and iiv8 was more than 10-fold less competitive against wild-type Pf0-1 in sterile loam soil low in inorganic phosphate. Studies indicated that ppk, and not iiv8, was required for competitive fitness. No role for iiv8 was identified. While a ppk and iiv8 mutant of Pf0-1 did not have increased sensitivity to osmotic, oxidative, and acid stress, it was more sensitive to elevated temperatures in laboratory medium and during growth in sterile soil. ppk was shown to be part of the Pho regulon in P. fluorescens, being upregulated in response to a low external P(i) concentration. Of importance, overproduction of polyphosphate in the soil environment appears to be more deleterious than production of none at all. Our findings reveal a new role for polyphosphate (and the need for proper regulation of its production) in competitive fitness of P. fluorescens in laboratory and soil environments.
了解细菌在土壤中生存和持续存在的遗传基础,是制定成功的生物防治策略以及理解细菌生态成功的关键组成部分。我们发现荧光假单胞菌Pf0-1中一个指定多聚磷酸激酶(ppk)的基因座和一个未预测的反义RNA(iiv8),对于在LB肉汤培养和无菌壤土中实现最佳竞争适应性是必需的。在无机磷酸盐含量低的无菌壤土中,缺乏ppk和iiv8的Pf0-1与野生型Pf0-1相比,竞争力降低了10倍以上。研究表明,竞争适应性需要ppk,而不是iiv8。未发现iiv8有作用。虽然Pf0-1的ppk和iiv8突变体对渗透、氧化和酸胁迫的敏感性没有增加,但在实验室培养基中以及在无菌土壤中生长时,它对高温更敏感。已证明ppk是荧光假单胞菌Pho调节子的一部分,在外部低磷(i)浓度下会上调。重要的是,在土壤环境中多聚磷酸盐的过量产生似乎比根本不产生更有害。我们的研究结果揭示了多聚磷酸盐(以及对其产生进行适当调控的必要性)在荧光假单胞菌在实验室和土壤环境中的竞争适应性方面的新作用。