Cloning of an H+-PPase gene from Thellungiella halophila and its heterologous expression to improve tobacco salt tolerance

被引:120
作者
Gao, Feng [1 ]
Gao, Qiang [1 ]
Duan, XiaoGuang [1 ]
Yue, GuiDong [1 ]
Yang, AiFang [1 ]
Zhang, Juren [1 ]
机构
[1] Shandong Univ, Sch Life Sci, Jinan 250100, Peoples R China
关键词
Arabidopsis thaliana; H+-PPase; salt tolerance; Thellungiella halophila;
D O I
10.1093/jxb/erl090
中图分类号
Q94 [植物学];
学科分类号
071001 ;
摘要
An H+-pyrophosphatase (PPase) gene named TsVP involved in basic biochemical and physiological mechanisms was cloned from Thellungiella halophila. The deduced translation product has similar characteristics to H+-PPases from other species, such as Arabidopsis and rice, in terms of bioinformation. The heterologous expression of TsVP in the yeast mutant ena1 suppressed Na+ hypersensitivity and demonstrated the function of TsVP as an H+-PPase. Transgenic tobacco overexpressing TsVP had 60% greater dry weight than wild-type tobacco at 300 mM NaCl and higher viability of mesophyll protoplasts under salt shock stress conditions. TsVP and AVP1, another H+-PPase from Arabidopsis, were heterologously expressed separately in both the yeast mutant ena1 and tobacco. The salt tolerance of TsVP or AVP1 yeast transformants and transgenic tobacco were improved to almost the same level. The TsVP transgenic tobacco lines TL3 and TL5 with the highest H+-PPase hydrolytic activity were studied further. These transgenic tobacco plants accumulated 25% more solutes than wild-type plants without NaCl stress and 20-32% more Na+ under salt stress conditions. Although transgenic tobacco lines TL3 and TL5 accumulated more Na+ in leaf tissues, the malondialdehyde content and cell membrane damage were less than those of the wild type under salt stress conditions. Presumably, compartmentalization of Na+ in vacuoles reduces its toxic effects on plant cells. This result supports the hypothesis that overexpression of H+-PPase causes the accumulation of Na+ in vacuoles instead of in the cytoplasm and avoids the toxicity of excessive Na+ in plant cells.
引用
收藏
页码:3259 / 3270
页数:12
相关论文
共 40 条
[1]   Generic placement of species excluded from Arabidopsis (Brassicaceae) [J].
Al-Shehbaz, IA ;
O'Kane, SL ;
Price, RA .
NOVON, 1999, 9 (03) :296-307
[2]   Abiotic stress and plant genome evolution. Search for new models [J].
Amtmann, A ;
Bohnert, HJ ;
Bressan, RA .
PLANT PHYSIOLOGY, 2005, 138 (01) :127-130
[3]   Salt tolerance conferred by overexpression of a vacuolar Na+/H+ antiport in Arabidopsis [J].
Apse, MP ;
Aharon, GS ;
Snedden, WA ;
Blumwald, E .
SCIENCE, 1999, 285 (5431) :1256-1258
[4]   Cloning and characterization of a wheat vacuolar cation/proton antiporter and pyrophosphatase proton pump [J].
Brini, F ;
Gaxiola, RA ;
Berkowitz, GA ;
Masmoudi, K .
PLANT PHYSIOLOGY AND BIOCHEMISTRY, 2005, 43 (04) :347-354
[5]   Salt, nutrient uptake and transport, and ABA of Populus euphratica;: a hybrid in response to increasing soil NaCl [J].
Chen, SL ;
Li, JK ;
Wang, SS ;
Hüttermann, A ;
Altman, A .
TREES-STRUCTURE AND FUNCTION, 2001, 15 (03) :186-194
[6]   Enhancement of tolerance of abiotic stress by metabolic engineering of betaines and other compatible solutes [J].
Chen, THH ;
Murata, N .
CURRENT OPINION IN PLANT BIOLOGY, 2002, 5 (03) :250-257
[7]   ANION-SENSITIVE, H+-PUMPING ATPASE IN MEMBRANE-VESICLES FROM OAT ROOTS [J].
CHURCHILL, KA ;
SZE, H .
PLANT PHYSIOLOGY, 1983, 71 (03) :610-617
[8]   Antioxidant responses of rice seedlings to salinity stress [J].
Dionisio-Sese, ML ;
Tobita, S .
PLANT SCIENCE, 1998, 135 (01) :1-9
[9]   Vacuolar H+ pyrophosphatases:: from the evolutionary backwaters into the mainstream [J].
Drozdowicz, YM ;
Rea, PA .
TRENDS IN PLANT SCIENCE, 2001, 6 (05) :206-211
[10]   EFFECTS OF SODIUM-CHLORIDE ON TOBACCO PLANTS [J].
FLOWERS, TJ ;
FLOWERS, SA ;
GREENWAY, H .
PLANT CELL AND ENVIRONMENT, 1986, 9 (08) :645-651