AUTHOR=Hu Aiqi , Chen Xiaohong , Luo Sha , Zou Qian , Xie Jing , He Donglan , Li Xiaohua , Cheng Guojun 

TITLE=Rhizobium leguminosarum Glutathione Peroxidase Is Essential for Oxidative Stress Resistance and Efficient Nodulation

JOURNAL=Frontiers in Microbiology

VOLUME=12

YEAR=2021

URL=https://www.frontiersin.org/journals/microbiology/articles/10.3389/fmicb.2021.627562

DOI=10.3389/fmicb.2021.627562

ISSN=1664-302X

ABSTRACT=<p>Glutathione (GSH) plays a key role in regulating the cellular Redox Homeostasis, and appears to be essential for initiation and development of root nodules. Glutathione peroxidase (Gpx) catalyzes the reduction of H<sub>2</sub>O<sub>2</sub> and organic hydroperoxides by oxidation of GSH to oxidized GSH (GSSG), which in turn is reduced by glutathione reductase (GR). However, it has not been determined whether the <italic>Rhizobium leguminosarum</italic> Gpx or GR is required during symbiotic interactions with pea. To characterize the role of glutathione-dependent enzymes in the symbiotic process, single and double mutants were made in <italic>gpxA</italic> (encoding glutathione peroxidase) and <italic>gshR</italic> (encoding glutathione reductase) genes. All the mutations did not affect the rhizobial growth, but they increased the sensitivity of <italic>R. leguminosarum</italic> strains to H<sub>2</sub>O<sub>2</sub>. Mutant in GpxA had no effect on intracellular GSH levels, but can increase the expression of the catalase genes. The <italic>gshR</italic> mutant can induce the formation of normal nodules, while the <italic>gpxA</italic> single and double mutants exhibited a nodulation phenotype coupled to more than 50% reduction in the nitrogen fixation capacity, these defects in nodulation were characterized by the formation of ineffective nodules. In addition, the <italic>gpxA</italic> and <italic>gshR</italic> double mutant was severely impaired in rhizosphere colonization and competition. Quantitative proteomics using the TMT labeling method was applied to study the differential expression of proteins in bacteroids isolated from pea root nodules. A total of 27 differentially expressed proteins were identified in these root bacteroids including twenty down-regulated and seven up-regulated proteins. By sorting the down-regulated proteins, eight are transporter proteins, seven are dehydrogenase, deoxygenase, oxidase, and hydrolase. Moreover, three down-regulating proteins are directly involved in nodule process.</p>