Overexpression of vacuolar proton pump ATPase(V-H^+-ATPase) subunits B, C and H confers tolerance to salt and saline-alkali stresses in transgenic alfalfa(Medicago sativa L.)  

作　　者：WANG Fa-wei WANG Chao SUN Yao WANG Nan LI Xiao-wei DONG Yuan-yuan YAO Na LIU Xiu-ming CHEN Huan CHEN Xi-feng WANG Zhen-min LI Hai-yan 

机构地区：[1]College of Life Sciences, Jilin Agricultural University/Engineering Research Center of Bioreactor and Pharmaceutical Development, Ministry of Education [2]Jilin Technology Innovation Center for Soybean Region, Jilin Agricultural University

出　　处：《Journal of Integrative Agriculture》2016年第10期2279-2289,共11页农业科学学报（英文版）

基　　金：supported by the National Natural Science Foundation of China (31271746, 31401403, 31501366,31201237)

摘　　要：The vacuolar proton pump ATPase（V-H^＋-ATPase）, which is a multi-subunit membrane protein complex, plays a major role in the activation of ion and nutrient transport and has been suggested to be involved in several physiological processes, such as cell expansion and salt tolerance. In this study, three genes encoding V-H^＋-ATPase subunits B（Sc VHA-B, Gen Bank： JF826506）, C（Sc VHA-C, Gen Bank： JF826507） and H（Sc VHA-H, Gen Bank： JF826508） were isolated from the halophyte Suaeda corniculata. The transcript levels of Sc VHA-B, Sc VHA-C and Sc VHA-H were increased by salt, drought and saline-alkali treatments. V-H^＋-ATPase activity was also examined under salt, drought and saline-alkali stresses. The results showed that V-H^＋-ATPase activity was correlated with salt, drought and saline-alkali stress. Furthermore, V-H^＋-ATPase subunits B, C and H（Sc VHA-B, Sc VHA-C and Sc VHA-H） from S. corniculata were introduced separately into the alfalfa genome. The transgenic alfalfa was verified by Southern and Northern blot analysis. During salt and saline-alkali stresses, transgenic lines carrying the B, C and H subunits had higher germination rates than the wild type（WT）. More free proline, higher superoxide dismutase（SOD） activity and lower malondialdehyde（MDA） levels were detected in the transgenic plants under salt and saline-alkali treatments. Moreover, the Sc VHA-B transgenic lines showed greater tolerance to salt and saline-alkali stresses than the WT. These results suggest that overexpression of Sc VHA-B, Sc VHA-C and Sc VHA-H improves tolerance to salt and saline-alkali stresses in transgenic alfalfa.The vacuolar proton pump ATPase（V-H^＋-ATPase）, which is a multi-subunit membrane protein complex, plays a major role in the activation of ion and nutrient transport and has been suggested to be involved in several physiological processes, such as cell expansion and salt tolerance. In this study, three genes encoding V-H^＋-ATPase subunits B（Sc VHA-B, Gen Bank： JF826506）, C（Sc VHA-C, Gen Bank： JF826507） and H（Sc VHA-H, Gen Bank： JF826508） were isolated from the halophyte Suaeda corniculata. The transcript levels of Sc VHA-B, Sc VHA-C and Sc VHA-H were increased by salt, drought and saline-alkali treatments. V-H^＋-ATPase activity was also examined under salt, drought and saline-alkali stresses. The results showed that V-H^＋-ATPase activity was correlated with salt, drought and saline-alkali stress. Furthermore, V-H^＋-ATPase subunits B, C and H（Sc VHA-B, Sc VHA-C and Sc VHA-H） from S. corniculata were introduced separately into the alfalfa genome. The transgenic alfalfa was verified by Southern and Northern blot analysis. During salt and saline-alkali stresses, transgenic lines carrying the B, C and H subunits had higher germination rates than the wild type（WT）. More free proline, higher superoxide dismutase（SOD） activity and lower malondialdehyde（MDA） levels were detected in the transgenic plants under salt and saline-alkali treatments. Moreover, the Sc VHA-B transgenic lines showed greater tolerance to salt and saline-alkali stresses than the WT. These results suggest that overexpression of Sc VHA-B, Sc VHA-C and Sc VHA-H improves tolerance to salt and saline-alkali stresses in transgenic alfalfa.

关 键 词：vacuolar proton pump salt tolerance saline-alkali tolerance alfalfa 

分 类 号：S54[农业科学—作物学]