Methane alleviates alfalfa cadmium toxicity via decreasing cadmium accumulation and reestablishing glutathione homeostasis
Abstract Although methane (CH 4 ) generation triggered by some environmental stimuli, displays the protective response against oxidative stress in plants, whether and how CH 4 regulates plant tolerance against cadmium stress is largely unknown. Here, we discovered that cadmium (Cd) stimulated the production of CH 4 in alfalfa root tissues. The pretreatment with exogenous CH 4 could alleviate seedling growth inhibition. Less amounts of Cd accumulation was also observed. Consistently, in comparison with Cd stress alone, miR159 transcript was down-regulated by CH 4 , and expression levels of its target gene ABC transporter was increased. By contrast, miR167 transcript was up-regulated, showing a relatively negative correlation with its target gene Nramp6. Meanwhile, Cd-triggered redox imbalance was improved by CH 4 , evidenced by the reduced lipid peroxidation and hydrogen peroxide accumulation, as well as the induction of representative antioxidant genes. Further results showed that Cd-triggered decrease of the ratio of reduced/oxidized (homo)glutathione was rescued by CH 4 . Additionally, CH 4 -triggered alleviation of seedling growth was sensitive to a selective inhibitor of glutathione biosynthesis. Overall, above results revealed that CH 4 -alleviated Cd accumulation at least partially, required the modulation of heavy metal transporters via miR159 and miR167 . Finally, the role of glutathione homeostasis elicited by CH 4 was preliminarily suggested. Highlights Cd induced CH 4 production in alfalfa seedling roots. Cd accumulation in alfalfa seedling roots is decreased by CH 4 . CH 4 -modulated heavy metal transporters via miR159 and miR167. CH 4 -modulated GSH decreased Cd accumulation. Graphical abstract [DISPLAY OMISSION]
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