Shelud'ko Andrei, Volokhina Irina, Mokeev Dmitry, Telesheva Elizaveta, Yevstigneeva Stella, Burov Andrei, Tugarova Anna, Shirokov Alexander, Burigin Gennady, Matora Larisa, Petrova Lilia
Institute of Biochemistry and Physiology of Plants and Microorganisms, Saratov Scientific Centre of the Russian Academy of Sciences (IBPPM RAS), 13 Prospekt Entuziastov, Saratov, 410049, Russia.
World J Microbiol Biotechnol. 2023 Oct 10;39(12):336. doi: 10.1007/s11274-023-03785-z.
Azospirillum alphaproteobacteria, which live in the rhizosphere of many crops, are used widely as biofertilizers. Two-component signal transduction systems (TCSs) mediate the bacterial perception of signals and the corresponding adjustment of behavior facilitating the adaptation of bacteria to their habitats. In this study, we obtained the A. baldaniorum Sp245 mutant for the AZOBR_150176 gene, which encodes the TCS of the hybrid histidine kinase/response sensory regulator (HSHK-RR). Inactivation of this gene affected bacterial morphology and motility. In mutant Sp245-HSHKΔRR-Km, the cells were still able to synthesize a functioning polar flagellum (Fla), were shorter than those of strain Sp245, and were impaired in aerotaxis, elaboration of inducible lateral flagella (Laf), and motility in semiliquid media. The mutant showed decreased transcription of the genes encoding the proteins of the secretion apparatus, which ensures the assembly of Laf, Laf flagellin, and the repressor protein of translation of the Laf flagellin's mRNA. The study examined the effects of polyethylene glycol 6000 (PEG 6000), an agent used to simulate osmotic stress and drought conditions. Under osmotic stress, the mutant was no longer able to use collective motility in semiliquid media but formed more biofilm biomass than did strain Sp245. Introduction into mutant cells of the AZOBR_150176 gene as part of an expression vector led to recovery of the lost traits, including those mediating bacterial motility under mechanical stress induced by increased medium density. The results suggest that the HSHK-RR under study modulates the response of A. baldaniorum Sp245 to mechanical and osmotic/water stress.
固氮螺菌属α-变形菌生活在多种作物的根际,被广泛用作生物肥料。双组分信号转导系统(TCSs)介导细菌对信号的感知以及相应的行为调整,促进细菌适应其生存环境。在本研究中,我们获得了编码杂合组氨酸激酶/应答传感调节器(HSHK-RR)的TCS的AZOBR_150176基因的拜氏固氮螺菌Sp245突变体。该基因的失活影响了细菌的形态和运动性。在突变体Sp245-HSHKΔRR-Km中,细胞仍能合成有功能的极鞭毛(Fla),但比Sp245菌株的细胞短,且在趋氧性、诱导性侧鞭毛(Laf)的形成以及在半液体培养基中的运动性方面存在缺陷。该突变体显示出编码分泌装置蛋白的基因转录减少,而分泌装置负责Laf的组装、Laf鞭毛蛋白以及Laf鞭毛蛋白mRNA翻译的阻遏蛋白。本研究考察了用于模拟渗透胁迫和干旱条件的聚乙二醇6000(PEG 6000)的作用。在渗透胁迫下,该突变体不再能够在半液体培养基中进行群体运动,但形成的生物膜生物量比Sp245菌株更多。将AZOBR_150176基因作为表达载体的一部分导入突变体细胞后,导致丧失的性状得以恢复,包括那些介导细菌在培养基密度增加引起的机械应力下运动的性状。结果表明,所研究的HSHK-RR调节拜氏固氮螺菌Sp245对机械和渗透/水分胁迫的反应。