When the 7_d old barley ( Hordeum vulgare L.) seedlings were treated with different concentrations of NaCl for 3 d, the levels of the noncovalently conjugated polyamines (PAs) in the plasma membrane and tonoplast v...When the 7_d old barley ( Hordeum vulgare L.) seedlings were treated with different concentrations of NaCl for 3 d, the levels of the noncovalently conjugated polyamines (PAs) in the plasma membrane and tonoplast vesicles and the covalently conjugated PAs in the membrane proteins were promoted by NaCl of low concentrations and suppressed by NaCl of high concentrations. Among the noncovalently conjugated PAs in the vesicles, spermidine (Spd) level was the most abundant, while putrescine (Put) content was predominant among the covalently conjugated PAs, accounted for 40%-70%, 35%-60%, respectively. In addition, the TLC (thin_layer chromatography) profiles of the benzoylated PAs presented an unknown polyamine with Rf =0.92 (X 0.92 ), which conjugated covalently and noncovalently in root tonoplast and its content changed as well as Spd with NaCl treatment. The total PA contents in the roots were higher than that in the leaves, and the types and contents of covalently and noncovalently conjugated PAs in the tonoplast were higher than those in the plasma membrane. The results showed that the above two PAs associated with the membrane might be essential in salt adaption of cells and the maintenance of membrane function.展开更多
We have mapped the expression of the tonoplast intrinsic protein （TIP） gene family members in Arabidopsis seeds by fluorescent protein tagging of their genomic sequences and confocal microscopy. Three isoforms （TIP...We have mapped the expression of the tonoplast intrinsic protein （TIP） gene family members in Arabidopsis seeds by fluorescent protein tagging of their genomic sequences and confocal microscopy. Three isoforms （TIP1;1, TIP2;1, and TIP2;2） have distinct patterns of expression in maternal tissues （outer integument and placento-chalazal region）. Two isoforms, TIP3;1 and the previously uncharacterized TIP3;2, are the only detectable TIPs in embryos during seed maturation and the early stages of seed germination. Throughout these developmental stages, both isoforms co-locate to the tono- plast of the protein storage vacuoles, but also appear to label the plasma membrane. Plasma membrane labeling is specific to TIP3;1 and TIP3;2, is independent of the position of the fluorescent protein tag, and appears to be specific to early seed maturation and early germination stages. We discuss these results in the context of the predicted distribution of aquaporins in Arabidopsis seeds.展开更多
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
Both enhanced were H^+ transport activities of tonoplast vesicles isolated from roots of the salt-tolerant mutant and wild type of wheat with treatment of NaCI, but the activity of the mutant was significantly higher ...Both enhanced were H^+ transport activities of tonoplast vesicles isolated from roots of the salt-tolerant mutant and wild type of wheat with treatment of NaCI, but the activity of the mutant was significantly higher than that of wild type. H^+ transport activity was indicated as the stable value of fluorescence quenching per mg membrane proteins. The H^+ transport activities dependent on ATP of the mutant and wild type were 1099 and 558 respectively and their activities dependent on PPi were 358 and 228 separately.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
Brassinosteroid(BR)is a vital plant hormone that regulates plant growth anddevelopment.BRASSINAZOLE RESISTANT1(BZR1)is a key transcription factor in BR signaling,and its nucleocytoplasmic localization is crucial for B...Brassinosteroid(BR)is a vital plant hormone that regulates plant growth anddevelopment.BRASSINAZOLE RESISTANT1(BZR1)is a key transcription factor in BR signaling,and its nucleocytoplasmic localization is crucial for BR signaling.However,the mechanisms that regulate BzR1 nucleocytoplasmic distribution and thus the homeostasis of BR signaling remain largely unclear.The vacuole is the largest organelle in mature plantcells and plays a key role in maintenance of cell ular pH,storage of intracellular substances,and transport ofions.In this study,weuncovered anovel mechanismof BR signaling homeostasis regulatedbythe vacuolar H+-ATPase(V-ATPase)and BZR1 feedback loop.Our results revealed that the vha-a2 vha-a3 mutant(vha2,lacking V-ATPase activity)exhibits enhanced BR signaling with increased total amount of BZR1,nuclearlocalized BZR1,and the ratio of BZR1/phosphorylated BZR1 in the nucleus.Further biochemical assays revealed that VHA-a2 and VHA-a3 of V-ATPase interact with the BZR1 protein through a domain that is conserved across multiple species.VHA-a2 and VHA-a3 negatively regulate BR signaling by interacting with BzR1 and promoting its retention in the tonoplast.Interestingly,a series of molecular analyses demonstrated that nuclear-localized BZR1 could bind directlyto specific motifs in the promoters of VHA-a2 andVHAa3topromote their expression.Taken together,these results suggest that V-ATPase and BzR1 mayforma feedback regulatory loop to maintain thehomeostasis of BR signaling in Arabidopsis,providing new insights into vacuole-mediated regulation of hormone signaling.展开更多
Effect of nitrate and ammonium on the activity of tonoplast pyrophosphatase (V-PPase) was investigated in the roots of tomato ( Lycopersicon esculentum L.). The results showed that the activity of V-PPase was increase...Effect of nitrate and ammonium on the activity of tonoplast pyrophosphatase (V-PPase) was investigated in the roots of tomato ( Lycopersicon esculentum L.). The results showed that the activity of V-PPase was increased by ammonium nutrition, compared with nitrate nutrition. The H+ transport of tonoplast vesicles by V-PPase was also stimulated by ammonium nutrition. The result of Western blot indicated that the protein amount of V-PPase was increased by ammonium nutrition.展开更多
文摘When the 7_d old barley ( Hordeum vulgare L.) seedlings were treated with different concentrations of NaCl for 3 d, the levels of the noncovalently conjugated polyamines (PAs) in the plasma membrane and tonoplast vesicles and the covalently conjugated PAs in the membrane proteins were promoted by NaCl of low concentrations and suppressed by NaCl of high concentrations. Among the noncovalently conjugated PAs in the vesicles, spermidine (Spd) level was the most abundant, while putrescine (Put) content was predominant among the covalently conjugated PAs, accounted for 40%-70%, 35%-60%, respectively. In addition, the TLC (thin_layer chromatography) profiles of the benzoylated PAs presented an unknown polyamine with Rf =0.92 (X 0.92 ), which conjugated covalently and noncovalently in root tonoplast and its content changed as well as Spd with NaCl treatment. The total PA contents in the roots were higher than that in the leaves, and the types and contents of covalently and noncovalently conjugated PAs in the tonoplast were higher than those in the plasma membrane. The results showed that the above two PAs associated with the membrane might be essential in salt adaption of cells and the maintenance of membrane function.
文摘We have mapped the expression of the tonoplast intrinsic protein （TIP） gene family members in Arabidopsis seeds by fluorescent protein tagging of their genomic sequences and confocal microscopy. Three isoforms （TIP1;1, TIP2;1, and TIP2;2） have distinct patterns of expression in maternal tissues （outer integument and placento-chalazal region）. Two isoforms, TIP3;1 and the previously uncharacterized TIP3;2, are the only detectable TIPs in embryos during seed maturation and the early stages of seed germination. Throughout these developmental stages, both isoforms co-locate to the tono- plast of the protein storage vacuoles, but also appear to label the plasma membrane. Plasma membrane labeling is specific to TIP3;1 and TIP3;2, is independent of the position of the fluorescent protein tag, and appears to be specific to early seed maturation and early germination stages. We discuss these results in the context of the predicted distribution of aquaporins in Arabidopsis seeds.
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Both enhanced were H^+ transport activities of tonoplast vesicles isolated from roots of the salt-tolerant mutant and wild type of wheat with treatment of NaCI, but the activity of the mutant was significantly higher than that of wild type. H^+ transport activity was indicated as the stable value of fluorescence quenching per mg membrane proteins. The H^+ transport activities dependent on ATP of the mutant and wild type were 1099 and 558 respectively and their activities dependent on PPi were 358 and 228 separately.
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.
基金the National Natural Science Foundation of China(32070342 and 32270339)the Agri-X Interdisciplinary Fund of Shanghai Jiao Tong University(Agri-X20200204)+1 种基金the Bio-X Interdisciplinary Fund of Shanghai Jiao Tong University(20CX-04)the Scientific and Technological Innovation Funds of Shanghai Jiao Tong University(19X160020009).
文摘Brassinosteroid(BR)is a vital plant hormone that regulates plant growth anddevelopment.BRASSINAZOLE RESISTANT1(BZR1)is a key transcription factor in BR signaling,and its nucleocytoplasmic localization is crucial for BR signaling.However,the mechanisms that regulate BzR1 nucleocytoplasmic distribution and thus the homeostasis of BR signaling remain largely unclear.The vacuole is the largest organelle in mature plantcells and plays a key role in maintenance of cell ular pH,storage of intracellular substances,and transport ofions.In this study,weuncovered anovel mechanismof BR signaling homeostasis regulatedbythe vacuolar H+-ATPase(V-ATPase)and BZR1 feedback loop.Our results revealed that the vha-a2 vha-a3 mutant(vha2,lacking V-ATPase activity)exhibits enhanced BR signaling with increased total amount of BZR1,nuclearlocalized BZR1,and the ratio of BZR1/phosphorylated BZR1 in the nucleus.Further biochemical assays revealed that VHA-a2 and VHA-a3 of V-ATPase interact with the BZR1 protein through a domain that is conserved across multiple species.VHA-a2 and VHA-a3 negatively regulate BR signaling by interacting with BzR1 and promoting its retention in the tonoplast.Interestingly,a series of molecular analyses demonstrated that nuclear-localized BZR1 could bind directlyto specific motifs in the promoters of VHA-a2 andVHAa3topromote their expression.Taken together,these results suggest that V-ATPase and BzR1 mayforma feedback regulatory loop to maintain thehomeostasis of BR signaling in Arabidopsis,providing new insights into vacuole-mediated regulation of hormone signaling.
文摘Effect of nitrate and ammonium on the activity of tonoplast pyrophosphatase (V-PPase) was investigated in the roots of tomato ( Lycopersicon esculentum L.). The results showed that the activity of V-PPase was increased by ammonium nutrition, compared with nitrate nutrition. The H+ transport of tonoplast vesicles by V-PPase was also stimulated by ammonium nutrition. The result of Western blot indicated that the protein amount of V-PPase was increased by ammonium nutrition.
