The GlnE protein of Azorhizobium caulinodans ORS571 plays a crucial role in the nodulation process of the legume host Sesbania rostrata

Abstract

The GlnE enzyme, functioning as an adenylyltransferase/adenylyl-removing enzyme, plays a crucial role in reversible adenylylation of glutamine synthetase (GS), which in turn regulates bacterial nitrogen assimilation. Genomic analysis of Azorhizobium caulinodans ORS571 revealed an open reading frame encoding a GlnE protein, whose function in the free-living and symbiotic states remains to be elucidated. A glnE deletion mutant retained high GS activity even under nitrogen-rich conditions. However, a reduction in growth was observed for the mutant strain at lower NH₄⁺ concentrations than for the wild-type strain. Furthermore, the ΔglnE mutant strain showed reduced motility on ammonium-containing media. Inactivation of GlnE led to an increase in root adhesion, biofilm formation, and nodulation on Sesbania rostrata. Nevertheless, the nodules induced by the glnE mutant strain were ineffective. In addition, A. caulinodans GlnE played a significant role in enhancing resistance against environmental stresses, such as heat, heavy metals, and cumene hydroperoxide. This study demonstrates that GlnE plays multiple regulatory roles in A. caulinodans beyond nitrogen metabolism and is essential for establishing symbiotic relationships with host plants.