The soybean GmPUB21-interacting protein GmDi19-5 responds to drought and salinity stresses via an ABA-dependent pathway

Drought-induced protein 19(Di19) is a Cys2/His2 zinc-finger protein that functions in plant growth and development and in tolerance to abiotic stresses.Gm PUB21,an E3 ubiquitin ligase,negatively regulates drought and salinity response in soybean.We identified potential interaction target proteins of Gm PUB21by yeast two-hybrid c DNA library screening,Gm Di19-5 as a candidate.Bimolecular fluorescence complementation and glutathionine-S-transferase pull-down assays confirmed the interaction between Gm Di19-5 and Gm PUB21.Gm Di19-5 was induced by Na Cl,drought,and abscisic acid(ABA) treatments.Gm Di19-5 was expressed in the cytoplasm and nucleus.Gm Di19-5 overexpression conferred hypersensitivity to drought and high salinity,whereas Gm Di19-5 silencing increased drought and salinity tolerance.Transcripts of ABA-and stress response-associated genes including Gm RAB18 and Gm DREB2A were downregulated in Gm Di19-5-overexpressing plants under drought and salinity stresses.ABA decreased the protein level of Gm Di19-5 in vivo,whereas Gm PUB21 increased the decrease of Gm Di19-5 after exogenous ABA application.The accumulation of Gm PUB21 was also inhibited by Gm Di19-5.We conclude that Gm PUB21 and Gm Di19-5 collaborate to regulate drought and salinity tolerance via an ABA-dependent pathway.