Nitric Oxide Is Involved In Hydrogen Gas-induced Adventitious Rooting In Cucumber By Regulating The Expression And Interaction Of Plasma Membrane H~+-ATPase And 14-3-3 Protein

Hydrogen gas?H₂?and Nitric oxide?NO?as signal molecules,are key factors regulating plant growth and development.Previous results have shown that H₂ and NO promoted adventitious root development in cucumber.But,the mechanism of H₂and NO promoting adventitious root development in cucumber is mainly regulating antioxidant enzymes and increasing the content of NO.In plants,plasma membrane H⁺-ATPase and 14-3-3 protein are ubiquitous,and they are also involved in regulating plant growth and development.Studies have also shown that the interaction between plasma membrane H⁺-ATPase and 14-3-3 protein can regulate various metabolic processes in plants.However,it is not clear whether that the effects of H₂ and NO on plasma membrane H⁺-ATPase and 14-3-3 protein and their interaction in the regulation of adventitious root formation in cucumber.Therefore,in this study,cucumber?xinchun no.4?was used as the experimental material to study the effect of plasma membrane H⁺-ATPase and 14-3-3 protein and their interaction on the formation of adventitious root in cucumber under the condition of NO and H₂,so as to provide a theoretical basis for the signal interaction between NO and H₂.The main results and conclusions are as follows:1.The root number,root length and fresh weight in cucumber treated with HRW or SNP were significantly higher than treated with distilled water?CK?,and 50%hydrogen-rich water?HRW?and 50?M SNP were chosen as being the most effective treatments.2.The root number,root length and fresh weight in cucumber treated with HRW+SNP were significantly higher than treated with HRW or SNP.The root number,root length and fresh weight in cucumber treated with HRW+cPTIO or HRW+tungstate were significantly lower than treated with HRW.The results suggested that NO is involved in H₂-induced adventitious root development in cucumber.3.The protein expression and gene expression of plasma membrane H⁺-ATPase and 14-3-3 protein treated with H₂ or NO were significantly higher than treated with CK,the protein expression and gene expression of plasma membrane H⁺-ATPase and14-3-3 protein treated with H₂+NO were significantly higher than treated with H₂ or NO,cPTIO can reverse the effect of NO on the protein expression and gene expression of plasma membrane H⁺-ATPase and 14-3-3 protein.Meantime,the activities of plasma membrane H⁺-ATPase and H⁺showed the same change.NO is involved in H₂-induced adventitious root development in cucumber by regulating the protein expression and gene expression of plasma membrane H⁺-ATPase and 14-3-3protein.4.Compared with the CK,H₂ or NO treatments significantly promote the interaction between plasma membrane H⁺-ATPase and 14-3-3 protein.H₂+NO treatment significantly promote the interaction between plasma membrane H⁺-ATPase and 14-3-3 protein compared with H₂ or NO treatment.When cPTIO was added to HRW,the effect of NO was reversed on the interaction between plasma membrane H⁺-ATPase and 14-3-3 protein,indicating that NO mag be involved in H₂-induced adventitious root development in cucumber by regulating interaction between plasma membrane H⁺-ATPase and 14-3-3 protein.5.FC is an activator of the protein interaction between plasma membrane H⁺-ATPase and 14-3-3 protein,and AMP is an inhibitor of the protein interaction between plasma membrane H⁺-ATPase and 14-3-3 protein.FC treatment significantly promote the root number,root length and fresh weight in cucumber compared to CK,with 1?M FC being the most effective treatment.Compared to CK,AMP treatment significantly inhibited the root number,root length and fresh weight in cucumber.FC treatment significantly up-regulating the activities of plasma membrane H⁺-ATPase and H⁺,AMP treatment significantly down-regulating the activities of plasma membrane H⁺-ATPase and H⁺,and the protein expression and gene expression of plasma membrane H⁺-ATPase and 14-3-3 protein showed the same change,testifying that NO is involved in H₂-induced adventitious root development in cucumber by regulating interaction between plasma membrane H⁺-ATPase and 14-3-3 protein.