[Mutant construction and characterization of hfq in Mesorhizobium huakuii 7653R].

微生物学报 Pub Date : 2017-01-04
Chuncao Ma, Xuejuan Zhou, Fuli Xie, Youguo Li
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Abstract

Objective: We studied the functions and characteristics of hfq gene in Mesorhizobium huakuii 7653R in adverse environment and symbiotic with its host plant.

Methods: The hfq mutant of 7653R was constructed via homologous recombination with small cloned fragments on suicide plasmids pK19mob to insert target gene. We applied 7653RΔhfq to characterize stress tolerance and symbiosis with host plant, in comparison with the complementary strains 7653R △hfq-C and the wild type.

Results: Mutant 7653RΔhfq presented lower growth rate, and higher mortality after heat shock-pretreated than that of the wild type, as well as the decreasing adaptability under the stress of 4.5% ethanol and 50 mmol H2O2. The defection of hfq affected the expression of some sRNAs in 7653R. Moreover, the mutant displayed significant reduced nodulation ability and nitrogenase activity compared with the wild type.

Conclusion: As a crucial post transcriptional regulatory factor, hfq plays an important role in Mesorhizobium Huakuii 7653R on both processes of stress resistance and symbiosis with the host plant Astragalus sinicus L.

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[华奎中干酪根菌7653R hfq突变体的构建与鉴定]。
目的:研究华葵中根菌7653R在逆境环境下及其与寄主植物共生时hfq基因的功能和特性。方法:在自杀质粒pK19mob上同源重组克隆小片段,构建7653R hfq突变体,插入目的基因。利用7653RΔhfq与互补菌株7653R△hfq-C和野生型进行比较,分析菌株的耐受性和与寄主植物的共生关系。结果:突变体7653RΔhfq经热休克预处理后的生长率低于野生型,死亡率高于野生型,在4.5%乙醇和50 mmol H2O2胁迫下的适应性下降。hfq的缺失影响了7653R中部分srna的表达。此外,与野生型相比,突变体的结瘤能力和氮酶活性显著降低。结论:hfq作为一个重要的转录后调控因子,在华葵中根菌7653R的抗逆性和与寄主植物黄芪的共生过程中都起着重要的作用。
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期刊介绍: Acta Microbiologica Sinica(AMS) is a peer-reviewed monthly (one volume per year)international journal,founded in 1953.It covers a wide range of topics in the areas of general and applied microbiology.The journal publishes original papers,reviews in microbiological science,and short communications describing unusual observations. Acta Microbiologica Sinica has been indexed in Index Copernicus (IC),Chemical Abstract (CA),Excerpt Medica Database (EMBASE),AJ of Viniti (Russia),Biological Abstracts (BA),Chinese Science Citation Database (CSCD),China National Knowledge Infrastructure(CNKI),Institute of Scientific and Technical Information of China(ISTIC),Chinese Journal Citation Report(CJCR),Chinese Biological Abstracts,Chinese Pharmaceutical Abstracts,Chinese Medical Abstracts and Chinese Science Abstracts.
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