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Azorhizobium caulinodans c-di-GMP phosphodiesterase Chp1 involved in motility, EPS production, and nodulation of the host plant

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Abstract

Establishment of the rhizobia-legume symbiosis is usually accompanied by hydrogen peroxide (H2O2) production by the legume host at the site of infection, a process detrimental to rhizobia. In Azorhizobium caulinodans ORS571, deletion of chp1, a gene encoding c-di-GMP phosphodiesterase, led to increased resistance against H2O2 and to elevated nodulation efficiency on its legume host Sesbania rostrata. Three domains were identified in the Chp1: a PAS domain, a degenerate GGDEF domain, and an EAL domain. An in vitro enzymatic activity assay showed that the degenerate GGDEF domain of Chp1 did not have diguanylate cyclase activity. The phosphodiesterase activity of Chp1 was attributed to its EAL domain which could hydrolyse c-di-GMP into pGpG. The PAS domain functioned as a regulatory domain by sensing oxygen. Deletion of Chp1 resulted in increased intracellular c-di-GMP level, decreased motility, increased aggregation, and increased EPS (extracellular polysaccharide) production. H2O2-sensitivity assay showed that increased EPS production could provide ORS571 with resistance against H2O2. Thus, the elevated nodulation efficiency of the ∆chp1 mutant could be correlated with a protective role of EPS in the nodulation process. These data suggest that c-di-GMP may modulate the A. caulinodans-S. rostrata nodulation process by regulating the production of EPS which could protect rhizobia against H2O2.

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Acknowledgements

We thank Professors Haiping Cheng and Claudine Elmerich for the helpful and insightful comments on an earlier version of the manuscript. This work was financed by the Strategic Priority Research Program of the Chinese Academy of Sciences (XDA23050102),the NSFC-Shandong Joint Fund Key Projects (U1806206), National Natural Science Foundation of China (31870020 and 31570063).

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Correspondence to Zhihong Xie.

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Sun, Y., Liu, Y., Liu, X. et al. Azorhizobium caulinodans c-di-GMP phosphodiesterase Chp1 involved in motility, EPS production, and nodulation of the host plant. Appl Microbiol Biotechnol 104, 2715–2729 (2020). https://doi.org/10.1007/s00253-020-10404-6

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  • DOI: https://doi.org/10.1007/s00253-020-10404-6

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