College of Life Sciences, State Key Laboratory of Crop Stress Biology in Arid Areas, Northwest Agriculture and Forestry University, Yangling, Shaanxi, 712100, China.
Appl Microbiol Biotechnol. 2017 Apr;101(7):2967-2978. doi: 10.1007/s00253-017-8114-y. Epub 2017 Jan 17.
Mesorhizobium alhagi, a legume-symbiont soil bacterium that forms nodules with the desert plant Alhagi sparsifolia, can produce large amounts of exopolysaccharide (EPS) using mannitol as carbon source. However, the role of EPS in M. alhagi CCNWXJ12-2, an EPS-producing rhizobium with high salt resistance, remains uncharacterized. Here, we studied the role of EPS in M. alhagi CCNWXJ12-2 using EPS-deficient mutants constructed by transposon mutagenesis. The insertion sites of six EPS-deficient mutants were analyzed using single primer PCR, and two putative gene clusters were found to be involved in EPS synthesis. EPS was extracted and quantified, and EPS production in the EPS-deficient mutants was decreased by approximately 25 times compared with the wild-type strain. Phenotypic analysis revealed reduced salt resistance, antioxidant capacity, and cell motility of the mutants compared with the wild-type strain. In conclusion, our results indicate that EPS can influence cellular Na content and antioxidant enzyme activity, as well as play an important role in the stress adaption and cell motility of M. alhagi CCNWXJ12-2.
根瘤菌 Mesorhizobium alhagi 是一种与沙漠植物骆驼刺共生的土壤细菌,能以甘露醇为碳源大量合成胞外多糖(EPS)。然而,高耐盐 EPS 产生菌 M. alhagi CCNWXJ12-2 中 EPS 的作用尚不清楚。本研究通过转座子诱变构建 EPS 缺陷突变株,研究了 EPS 在 M. alhagi CCNWXJ12-2 中的作用。采用单引物 PCR 分析了 6 株 EPS 缺陷突变株的插入位点,发现有两个假定的基因簇参与 EPS 合成。提取并定量 EPS,结果显示 EPS 缺陷突变株的 EPS 产量比野生型菌株降低了约 25 倍。表型分析表明,突变株的耐盐性、抗氧化能力和细胞迁移能力均低于野生型菌株。综上所述,本研究结果表明 EPS 可影响细胞内 Na 含量和抗氧化酶活性,并在 M. alhagi CCNWXJ12-2 的应激适应和细胞迁移中发挥重要作用。