茎瘤固氮根瘤菌环二鸟苷酸代谢相关基因的功能鉴定

Characterization of diguanylate cyclase metabolism-related gene in Azorhizobium caulinodans ORS571

摘要:

[目的]考察茎瘤固氮根瘤菌ORS571中c-di-GMP合成酶AZC-2412的编码基因缺失的突变表型，初步探究其功能机理。[方法]本实验构建基于cre-loxp重组酶系统的根瘤菌基因敲除系统，以及采用三亲接合技术构建突变株。测定野生型和突变株的生长速率、趋化能力、胞外多糖产量、生物膜形成等表型。[结果]突变株与野生型生长速率几乎相同。与野生型相比突变株由于细胞内c-di-GMP水平降低，胞外多糖、生物膜产量等均有所下降。[结论]实验表明，环二鸟苷酸合成酶AZC-2412缺失，使得c-di-GMP水平降低，对胞外多糖生成、细菌的运动能力、生物膜的形成、细胞絮凝、与植物的互作等均有调控作用。

Abstract:

[Objective] To study the phenotypes of Azorhizobium caulinodans ORS571 mutant strain lacking the diguanylate cyclase AZC-2412, and to explore its functional mechanism. [Methods] We constructed the gene knockout system of rhizobia based on the homologous recombination of cre-loxp, and the mutant strain was constructed by the triparental conjugation. Growth rate, chemotactic ability, exopolysaccharide production, biofilm formation and other phenotypes of the wild type and mutant strains were studied. [Results] The mutant strain had almost the same growth rate as the wild type. Compared with the wild type, the mutant strain impaired in exopolysaccharide production, chemotaxis behavior and biofilm formation due to lower levels of c-di-GMP in cells. [Conclusion] The deletion of the diguanylate cyclase AZC-2412 reduces level of c-di-GMP levels. It had a regulatory effect on the exopolysaccharide production, bacterial motility, biofilm formation, cell flocculation of Azorhizobium caulinodans ORS571 and its interaction with host plant.