The effects of NaCl stress on the H +-ATPase, H +-PPase activity and lipid composition of plasma membrane(PM) and tonoplast(TP) vesicles isolated from roots and leaves of two soybean cultivars(Glycine max L.) differ...The effects of NaCl stress on the H +-ATPase, H +-PPase activity and lipid composition of plasma membrane(PM) and tonoplast(TP) vesicles isolated from roots and leaves of two soybean cultivars(Glycine max L.) differing in salt tolerance(Wenfeng7, salt-tolerant; Union, salt-sensitive) were investigated. When Wenfeng7 was treated with 0.3%(W/V) NaCl for 3 d, the H +-ATPase activities in PM and TP from roots and leaves exhibited a reduction and an enhancement, respectively. The H +-PPase activity in TP from roots also increased. Similar effects were not observed in roots of Union. In addition, the increases of phospholipid content and ratios of phospholipid to galactolipid in PM and TP from roots and leaves of Wenfeng7 may also change membrane permeability and hence affect salt tolerance.展开更多
Under NaCl stress for 2 d, H+-ATPase activity increased, and H+-PPase activity decreased in the tonoplast of salt-tolerant barley ( Hordeum vulgare L. cv. 'Tanyin 2') roots. La3+ (1 mmol/L), an inhibitor of Ca...Under NaCl stress for 2 d, H+-ATPase activity increased, and H+-PPase activity decreased in the tonoplast of salt-tolerant barley ( Hordeum vulgare L. cv. 'Tanyin 2') roots. La3+ (1 mmol/L), an inhibitor of Ca2+ channel in plasma membrane, and EGTA (5 mmol/L), a Ca2+ chelator, inhibited this NaCl-induced increase in H+-ATPase activity but stimulated the H+-PPase activity. Treatment of barley roots with CaM antagonist (trifluoperazine, TFP, 20 mumol/L) also diminished the increase of H+-ATPase activity induced by NaCl. La3+, TFP or La3+ + TFP increased Na+ uptake and decreased K+ and Ca2+ uptake in barley roots under NaCl stress. These results suggested that the activation of tonoplast H+-ATPase and the regulation of Na+ and K+ uptake under NaCl stress may be related to Ca2+-CaM system.展开更多
To study the function and adaptive mechanism of tonoplast H + ATPase under salt stress, pea ( Pisum sativum L.) seedlings were treated with different concentrations of salt (100-250 mmol/L NaCl) and with 100 mmol...To study the function and adaptive mechanism of tonoplast H + ATPase under salt stress, pea ( Pisum sativum L.) seedlings were treated with different concentrations of salt (100-250 mmol/L NaCl) and with 100 mmol/L NaCl for different days (1-3 d). The ATP hydrolytic activity and the proton transport activity and the changes of the amount of tonoplast H + ATPase (subunit A) were measured. ATP hydrolytic activity of H + ATPase prepared from plants treated with 250 mmol/L NaCl was reduced by about 25% compared to that of control plants, but that of stressed plants treated with 100 mmol/L and 200 mmol/L NaCl was unchanged. The activity from plants treated with 100 mmol/L NaCl for up to 3 d was lower than that of control plants by 20%. But the proton transport activity was increased under the same salt stresses as above. These results showed that the changes of the hydrolytic activity and the proton transport activity were not in proportion and salt stress may cause the change of the coupling ratio of H + transport activity to ATP hydrolysis. The protein amount kept unchanged and reduced a little only when pea was treated with 100 mmol/L NaCl for 3 d. These results indicated that salinity stimulated the increase of the pump efficiency of the V_ATPase from pea roots, which was due to the change of the coupling ratio, but not due to the increase of ATP hydrolysis and the amount of V_ATPase.展开更多
For understanding the function of tonoplast protein in plant cell signal pathway, we have identified an integral protein kinase activity from the highly purified tonoplast isolated from maize ( Zea mays L.) root by...For understanding the function of tonoplast protein in plant cell signal pathway, we have identified an integral protein kinase activity from the highly purified tonoplast isolated from maize ( Zea mays L.) root by a new nonradioactive method in which a color labeled peptide was used as substrate. The protein kinase was Ca 2+ _dependent and CaM and phosphatidylserine_independent, like the calmodulin_like domain protein kinase (CDPK) in many plants. The optimal pH value and Ca 2+ concentration were 6.5 and 10 μmol/L, respectively. According to the optimal pH value and the effect of detergent, it could be inferred that the active site of this protein kinase is oriented toward the cytoplasm. Zn 2+ had no obvious effect on its activity, indicating that this protein kinase has no zinc_finger domain that exists in some mammalian protein kinases. At the same time, when tonoplast proteins were prephosphorylated in the presence of Ca 2+ and ATP, both the ATP_hydrolysis and the proton_transport activity of vacuolar H +_ATPase were stimulated. This stimulation could be reversed by an alkaline_phosphatase. These results indicate that a Ca 2+ _dependent protein kinase was located in the tonoplast, and a Ca 2+ _dependent phosphorylation, probably caused by this kinase, activated the vacuolar H +_ATPase activity. These results are helpful for further research on the function of CDPK in the course of signal transduction in plants.展开更多
The growth of barley (Hordeum vulgare L.) seedling is inhibited by 300 mmol/L NaCl. When 20 mg/L GSH is present in the hydroponic culture solution with NaCl, root length, root and shoot dry weigh, chlorophyll as well ...The growth of barley (Hordeum vulgare L.) seedling is inhibited by 300 mmol/L NaCl. When 20 mg/L GSH is present in the hydroponic culture solution with NaCl, root length, root and shoot dry weigh, chlorophyll as well as K+ contents are increased, Na+ content is decreased. At the same time the activities of H+-ATPase and H+-PPase associated with tonoplast vesicles isolated from leaves are stimulated, and electrolytic leakage are diminished by exogenous GSH.展开更多
文摘The effects of NaCl stress on the H +-ATPase, H +-PPase activity and lipid composition of plasma membrane(PM) and tonoplast(TP) vesicles isolated from roots and leaves of two soybean cultivars(Glycine max L.) differing in salt tolerance(Wenfeng7, salt-tolerant; Union, salt-sensitive) were investigated. When Wenfeng7 was treated with 0.3%(W/V) NaCl for 3 d, the H +-ATPase activities in PM and TP from roots and leaves exhibited a reduction and an enhancement, respectively. The H +-PPase activity in TP from roots also increased. Similar effects were not observed in roots of Union. In addition, the increases of phospholipid content and ratios of phospholipid to galactolipid in PM and TP from roots and leaves of Wenfeng7 may also change membrane permeability and hence affect salt tolerance.
文摘Under NaCl stress for 2 d, H+-ATPase activity increased, and H+-PPase activity decreased in the tonoplast of salt-tolerant barley ( Hordeum vulgare L. cv. 'Tanyin 2') roots. La3+ (1 mmol/L), an inhibitor of Ca2+ channel in plasma membrane, and EGTA (5 mmol/L), a Ca2+ chelator, inhibited this NaCl-induced increase in H+-ATPase activity but stimulated the H+-PPase activity. Treatment of barley roots with CaM antagonist (trifluoperazine, TFP, 20 mumol/L) also diminished the increase of H+-ATPase activity induced by NaCl. La3+, TFP or La3+ + TFP increased Na+ uptake and decreased K+ and Ca2+ uptake in barley roots under NaCl stress. These results suggested that the activation of tonoplast H+-ATPase and the regulation of Na+ and K+ uptake under NaCl stress may be related to Ca2+-CaM system.
文摘To study the function and adaptive mechanism of tonoplast H + ATPase under salt stress, pea ( Pisum sativum L.) seedlings were treated with different concentrations of salt (100-250 mmol/L NaCl) and with 100 mmol/L NaCl for different days (1-3 d). The ATP hydrolytic activity and the proton transport activity and the changes of the amount of tonoplast H + ATPase (subunit A) were measured. ATP hydrolytic activity of H + ATPase prepared from plants treated with 250 mmol/L NaCl was reduced by about 25% compared to that of control plants, but that of stressed plants treated with 100 mmol/L and 200 mmol/L NaCl was unchanged. The activity from plants treated with 100 mmol/L NaCl for up to 3 d was lower than that of control plants by 20%. But the proton transport activity was increased under the same salt stresses as above. These results showed that the changes of the hydrolytic activity and the proton transport activity were not in proportion and salt stress may cause the change of the coupling ratio of H + transport activity to ATP hydrolysis. The protein amount kept unchanged and reduced a little only when pea was treated with 100 mmol/L NaCl for 3 d. These results indicated that salinity stimulated the increase of the pump efficiency of the V_ATPase from pea roots, which was due to the change of the coupling ratio, but not due to the increase of ATP hydrolysis and the amount of V_ATPase.
文摘For understanding the function of tonoplast protein in plant cell signal pathway, we have identified an integral protein kinase activity from the highly purified tonoplast isolated from maize ( Zea mays L.) root by a new nonradioactive method in which a color labeled peptide was used as substrate. The protein kinase was Ca 2+ _dependent and CaM and phosphatidylserine_independent, like the calmodulin_like domain protein kinase (CDPK) in many plants. The optimal pH value and Ca 2+ concentration were 6.5 and 10 μmol/L, respectively. According to the optimal pH value and the effect of detergent, it could be inferred that the active site of this protein kinase is oriented toward the cytoplasm. Zn 2+ had no obvious effect on its activity, indicating that this protein kinase has no zinc_finger domain that exists in some mammalian protein kinases. At the same time, when tonoplast proteins were prephosphorylated in the presence of Ca 2+ and ATP, both the ATP_hydrolysis and the proton_transport activity of vacuolar H +_ATPase were stimulated. This stimulation could be reversed by an alkaline_phosphatase. These results indicate that a Ca 2+ _dependent protein kinase was located in the tonoplast, and a Ca 2+ _dependent phosphorylation, probably caused by this kinase, activated the vacuolar H +_ATPase activity. These results are helpful for further research on the function of CDPK in the course of signal transduction in plants.
文摘The growth of barley (Hordeum vulgare L.) seedling is inhibited by 300 mmol/L NaCl. When 20 mg/L GSH is present in the hydroponic culture solution with NaCl, root length, root and shoot dry weigh, chlorophyll as well as K+ contents are increased, Na+ content is decreased. At the same time the activities of H+-ATPase and H+-PPase associated with tonoplast vesicles isolated from leaves are stimulated, and electrolytic leakage are diminished by exogenous GSH.
