ATPase 与植物抗盐性

 本文综述了高等植物细胞ATPase在盐胁迫下的活性变化及其调控机制。V型H+_ATPase与细胞离子区隔化和植物抗盐性密切相关。盐胁迫提高抗盐植物液泡膜H+_ATPase活性, 主要是通过增加V型H+_ATPase主要功能亚基的基因表达以及蛋白质合成。盐胁迫通常降低质膜H+-ATPase活性, 很可能是由于酶蛋白质合成受阻, 质膜H+-ATPase活性的变化与盐胁迫的强度和时间长短有关。此外, 本 文还对ABA和Ca2+-CaM等胁迫信号物质对ATPase活性的调控及其与植物抗盐性的关系进行了总结。研究ATPase对盐胁迫的响应和调控机制, 有助于阐明植物的盐生境适应机制, 也有利于植物的抗盐育种工作。     

等渗盐胁迫对番茄抗氧化酶和ATP酶及焦磷酸酶活性的影响

用Ca(NO3)2 80 mmol/L和NaCl 120 mmol/L等渗溶液处理番茄幼苗后,细胞质和叶绿体中超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、抗坏血酸过氧化物酶(APX)的活性升高,并且NaCl胁迫的作用明显高于Ca(NO3)2胁迫.Ca(NO3)2处理提高了线粒体中SOD、CAT、APX的活性,而NaCl处理降低了它们的活性.根系质膜H -ATPase、液泡膜H -ATPase、焦磷酸酶(H -PPase)的活性和叶片丙二醛(MDA)及脯氨酸含量在两种盐胁迫后明显增加.NaCl处理对植株生长的抑制程度明显高于Ca(NO3)2处理.     

NaCl胁迫对盐芥质膜和液泡膜ATPase活性的影响

以盐生植物盐芥和中生植物拟南芥幼苗为材料,研究了盐胁迫对它们叶片和根质膜、液泡膜H+-ATPase、Ca2+-ATPases和K+-ATPase活性以及H+-ATPase、Na+/H+ 逆向转运蛋白表达的影响.结果显示:在NaCl胁迫下,盐芥叶片和根质膜的H+-ATPase活性分别比对照显著升高41%～212%和35%～53%,液泡膜的H+-ATPase分别显著升高281%～373%和4%～38%,而拟南芥却比相应对照都显著降低;相同盐浓度胁迫下,盐芥叶片的H+-ATPase活性比根部高4～8倍,盐芥根也远高于拟南芥.在NaCl胁迫下,盐芥叶片和根的液泡膜H+-ATPase蛋白质β亚基含量变化与其酶活性变化趋势一致,质膜Na+/H+ 逆向转运蛋白的表达量与Na+含量变化趋势一致.盐胁迫下盐芥根中Ca2+-ATPases和K+-ATPase活性的增加与根中Ca2+和K+含量呈显著正相关.研究发现,在盐胁迫条件下,盐芥能有效增强H+-ATPase蛋白和Na+/H+逆向转运蛋白表达,显著提高其根系与叶片质膜和液泡膜的H+-ATPase、Ca2+-ATPase和K+-ATPase活性,维持细胞质中较高的Ca2+和K+水平,从而缓解盐胁迫的伤害,增强耐盐性.     

盐胁迫下无花果细胞质膜和液泡膜H+-ATPase活性对脯氨酸积累的影响

盐胁迫降低无花果振荡培养细胞培养液pH ,添加质膜H ATPase活性抑制剂Na3VO4 则抑制盐诱导的培养液pH下降 ,表明盐诱导培养液pH下降主要是细胞质膜H ATPase活性增加的结果。NaCl处理提高活体细胞质膜H ATPase活性 ,而降低膜微囊H ATPase活性。培养液中添加Na3VO4 5 0 μmol/L完全抑制盐胁迫下无花果细胞游离脯氨酸积累 ,但添加更高浓度Na3VO4 ,则提高细胞液泡膜H ATPase活性 ,同时Na3VO4 抑制脯氨酸积累的效应下降 ,暗示盐胁迫下无花果细胞质膜和液泡膜H ATPase共同参与细胞质pH调节 ,影响游离脯氨酸积累。     

盐生车前根液泡膜ATPase对盐胁迫的响应

用蔗糖密度梯度离心法从对照和盐处理的盐生车前根中分离出液泡膜,并对其ＡＴＰａｓｅ某些特性进行了比较研究,蔗糖连续密度梯度离心后ＮＯ＾－３敏感的ＡＴＰａｓｅ活性主要分布在２２％－３５％蔗糖浓度之间,用不连续密度梯度离心法从２２％－３５％界面收集到的膜主要源于液泡膜。因为,这种膜ＡＴＰａｓｅ活性受到Ｃｌ＾－的促进和ＮＯ＾－３的强抑制,而叠氮化钠和正钡酸钠只抑制酶活性的１０％左右,最适ｐＨ８．０。     

液泡膜H~+-PPase与植物耐盐性

文章介绍植物液泡膜H+-PPase的结构、功能及其分子生物学的研究进展,并着重阐述液泡膜H+-PPase在植物耐盐性中的作用。     

液泡膜H+-ATPase在植物的非生物胁迫响应和信号转导中的作用

液泡膜H^＋-ATPase是一种多亚基复合体,在植物受到非生物胁迫后,其对逆境信号的感知转导即做出相应的变化。在Ca^2＋通道、ABA信号通路及盐过敏感途径等信号传递的过程中,都有V-ATPase的参与。文章将对这一领域的研究进展进行介绍。     

盐胁迫对大麦幼苗质膜,液泡膜上共价和非共价结合多胺含量的影响

用不同浓度ＮａＣｌ处理７ｄ龄大麦（Ｈｏｒｄｅｕｍ ｖｕｌｇａｒｅＬ．）幼苗３ｄ。以非共价键和共价键形式分别与质膜和液泡膜微囊及膜蛋白结合的多胺含量受低 度盐的促进而被高浓度盐所抑制。以非共价键形式与膜微囊结合的各种多胺中亚精胺（Ｓｐｄ）含量最高,占膜上多胺总量的４０％－７０％,与膜蛋白共价结合的各种多胺中腐胺（Ｐｕｔ）含量占主导地位,占膜蛋白上多胺总量的３５％－６０％。在根系液泡膜上发现一种含量丰     

外源IAA增强丹波黑大豆抗铝性的生理机制

以铝耐受型丹波黑大豆(RB)幼苗为供试材料,考察了不同浓度铝胁迫下添加外源IAA对RB根尖内源IAA、H2O2、MDA、铝含量、柠檬酸分泌量以及质膜H+-ATPase活性的影响,探讨外源IAA增强RB抗铝能力的生理机制。结果显示:(1)经25、50和200μmol·L-1 AlCl3胁迫处理24h后,RB根尖IAA含量、柠檬酸分泌量和质膜H+-ATPase活性均表现为先上升后下降的趋势,而H2O2、MDA和根尖铝含量却随着铝处理浓度的增加呈显著上升趋势。(2)与单独25、50和200μmol·L-1 AlCl3胁迫处理相比,添加50μmol·L-1外源IAA后使得RB根尖铝、MDA和H2O2含量均显著降低,同时使RB根尖柠檬酸分泌量分别相应增加到单独处理的2.39、1.73和6.85倍,且其相应的质膜H+-ATPase活性也增加了1.09、1.74和1.45倍。研究表明,外源IAA能通过增强丹波黑大豆根尖柠檬酸的分泌量和质膜H+-ATPase活性来提高其对铝胁迫的抗性,明显缓解铝毒害作用。     

植物质膜H+-ATPase响应盐胁迫的分子机制

植物细胞质膜质子泵（PM H^＋-ATPase）有看家酶之称,是由多基因编码的,其主要功能是向细胞营养物质的吸收和离子跨膜运输提供驱动力。文章介绍PM H^＋-ATPase在植物抗盐中的作用及研究进展。     

盐胁迫对小麦根质膜ATPase活性的影响

以小麦为实验材料,研究了盐胁迫对根质膜H^ —ATPase、Ca^2 —ATPase活性及H^ —ATPase蛋白表达的影响。结果显示：50、100、150mmol/L的NaCl处理72h后,小麦根质膜H^ —ATPase、Ca^2 —ATPase活性均降低。100mmol／L NaCl对质膜ATPase活性的抑制程度随处理时间的延长而增强,在处理24h后,H^ —ATPase和Ca^2 —ATPase的活性分别降为对照的72％和75％,而处理72h后,酶活性分别减小到对照的50％和48％。50、100、150mmol／L的NaCl直接作用于提取的质膜微囊,H^ —ATPase的活性分别降低约5％、8％和16％。Western blotting分析结果显示100mmol／L NaCl处理72h后,质膜H^ —ATPase的含量与对照比有所减少。本研究表明：盐胁迫抑制小麦根质膜H^ —ATPase、Ca^2 —ATPase的活性,酶含量的减少可能是盐胁迫导致质膜H^ —ATPase活性降低的原因。     

Salt stress in Mesembryanthemum crystallinum L. cell suspensions activates adaptive mechanisms similar to those observed in the whole plant

A salt-tolerant stable cell-suspension culture from the halophyte Mesembryanthemum crystallinum L. has been established from calli generated from leaves of 6-week-old well-watered plants. Optimal cell growth was observed in the presence of 200 mM NaCl, and within 7 d cells were able to concentrate Na⁺ to levels exceeding those in the growth medium. Accumulation of Na⁺ was paralled by increases in the compatible solute pinitol and myo-inositol methyl transferase (IMT), a key enzyme in pinitol biosynthesis. Increasing concentrations of NaCl stimulated the activities of tonoplast and plasma-membrane H⁺-ATPases. Immunodetection of the ATPases showed that the increased activity was not due to changes in protein amount that could be attributed to treatment conditions. A specific role for these mechanisms in salt-adaptation is supported by the inability of mannitol-induced water stress to elicit the same responses, and the absence of enzyme activity and protein expression associated with Crassulacean acid metabolism in the cells. Results demonstrate that these  M. crystallinum cell suspensions show a halophytic growth response, comparable to that of the whole plant, and thus provide a valuable tool for studying signaling and biochemical pathways involved in salt recognition and response. Received: 18 June 1998 / Accepted: 22 August 1998     

植物感受盐胁迫及相关钙信号的研究进展

盐胁迫对植物的生长和发育造成严重影响,其危害包括渗透胁迫、离子毒害等,严重损害了农业生产和粮食安全。在盐胁迫下,植物相关感受器接受刺激,使得Ca²⁺通过细胞膜以及细胞内钙库膜上打开的Ca²⁺通道进入细胞质基质,导致细胞质内Ca²⁺浓度升高,产生钙信号。钙离子作为重要的第二信使,在植物细胞内和细胞间传递信号,信号往下游传递,在不同生长和发育阶段引起植物一系列的生理响应来应对盐胁迫影响。钙信号主要通过钙调蛋白（CaM）、钙调素样蛋白（CML）、钙依赖性蛋白激酶（CDPK）、钙调磷酸酶B样蛋白（CBL）和CBL互作蛋白激酶（CIPK）感知并将特异的钙信号信息传递到下游;从而激活植物盐胁迫生理响应。本文主要综述植物如何感知盐胁迫刺激,以及钙信号产生与传导机制,并对该研究领域需解决的问题进行了展望。     

Alterations of Intracellular pH in Response to Low Temperature Stresses

+ -ATPase is one of the primary cellular events directly resulting from cold exposure. We demonstrate here that cold-induced inactivation of the proton translocating enzyme is closely linked to the rapid acidification of the cytoplasm and the concomitant alkalization of the vacuoles, suggesting an important role of the enzyme in maintaining homeostasis of the cellular pH in a cold environment. The stability of the vacuolar H⁺-ATPase to cold both in vivo and in vitro is distinctly different between species sensitive and insensitive to cold. These findings provide further insight into the way in which the vacuolar H⁺-ATPase is involved in cold adaptation of plants. In addition, the temperature reduction and the concentration of the cytoplasm as a consequence of freeze-induced dehydration may also result in changes in the cellular pH. In fact, we demonstrate here that the cytoplasm is markedly acidified upon freezing; in particular, in cells of less hardy plants. Freeze-induced acidification is presumably due to changes in the physico-chemical properties of the cytoplasm and the changes in the permeability of the vacuolar membrane both of which result from severe dehydration. The physiological significance of freeze-induced acidification of the cytoplasm is discussed. Received 26 March 1999/ Accepted in revised form 30 March 1999     

Localization of pyrophosphatase in membranes of cauliflower inflorescence cells

Using a polyclonal antiserum specific for the tonoplastic H⁺-pyrophosphatase (tPPase), significant amounts of antigenic polypeptides of the correct molecular mass were detected in Western blots of plasma membrane isolated from cauliflower (Brassica oleracea L.) inflorescence by phase-partitioning and subsequent sucrose density centrifugation. Potassium iodide-stripped plasma membranes continued to give a strong positive signal, indicating that the PPase antigen detected was not a result of contamination through soluble PPase released during homogenisation. The same preparation contained negligible vacuolar (v)H⁺-ATPase activity and the A subunit of the vATPase could not be detected by immunoblotting. Plasma membrane fractions exhibited a proton-pumping activity with ATP as substrate, but such an activity was not measurable with pyrophosphate, although the hydrolysis of this substrate was recorded. By contrast, pyrophosphate supported proton pumping in tonoplast-containing fractions. Immunogold electron microscopy confirmed the presence of PPase at the plasma membrane as well as at the tonoplast, trans Golgi network, and multivesicular bodies. The density of immunogold label was higher at the plasma membrane than at the tonoplast, except for membrane fragments occurring in the lumen of the vacuoles which stained very conspicuously. Received: 29 June 1998 / Accepted: 9 November 1998     

Mobilizing store Ca(2+) in the presence of La(3+) evokes exocytosis in bovine chromaffin cells

The effect on exocytosis of La(3+), a known inhibitor of plasma membrane Ca(2+)-ATPases and Na(+)/Ca(2+) exchangers, was studied using cultured bovine adrenal chromaffin cells. At high concentrations (0.3-3 mM), La(3+) substantially increased histamine-induced catecholamine secretion. This action was mimicked by other lanthanide ions (Nd(3+), Eu(3+), Gd(3+), and Tb(3+)), but not several divalent cations. In the presence of La(3+), the secretory response to histamine became independent of extracellular Ca(2+). La(3+) enhanced secretion evoked by other agents that mobilize intracellular Ca(2+) stores (angiotensin II, bradykinin, caffeine, and thapsigargin), but not that due to passive depolarization with 20 mM K(+). La(3+) still enhanced histamine-induced secretion in the presence of the nonselective inhibitors of Ca(2+)-permeant channels SKF96365 and Cd(2+), but the enhancement was abolished by prior depletion of intracellular Ca(2+) stores with thapsigargin. La(3+) inhibited (45)Ca(2+) efflux from preloaded chromaffin cells in the presence or absence of Na(+). It also enhanced and prolonged the rise in cytosolic [Ca(2+)] measured with fura-2 during mobilization of intracellular Ca(2+) stores with histamine in Ca(2+)-free buffer. The results suggest that the efficacy of intracellular Ca(2+) stores in evoking exocytosis is enhanced dramatically by inhibiting Ca(2+) efflux from the cell.     

Changes of membrane-associated Mg2+-activated ATPase of cucumber roots during calcium starvation

The properties of membrane-associated ATPase of cucumber (Cucumis sativus cv. Seiriki No. 2) roots cultured in a complete medium (complete enzyme) and in a medium lacking Ca²⁺ (Ca²⁺-deficient enzyme) were investigated. The basal activity of membrane-associated ATPase increased during Ca²⁺ starvation, while Mg²⁺-activation of the enzyme decreased and even resulted in inhibition by high Mg²⁺ concentration at the late stage of the Ca²⁺ starvation. The complete enzyme had low basal activity and showed a Mg²⁺-activated hyperbolic reaction curve in relation to ATP concentration. Ca²⁺-deficient enzyme with high basal activity showed a biphasic reaction curve and Mg²⁺-activation was seen only at high ATP concentrations. Activation of membrane-associated ATPase by various cations was decreased or lost during Ca²⁺ starvation. The basal ATPase activity of Ca²⁺-deficient enzyme increased for various substrates including pyrophosphate, p-nitrophenyl phosphate, glucose-6 phosphate, β-glycerophosphate, AMP, ADP and ATP. Mg²⁺-activation was found only for ADP and ATP in both the complete and Ca²⁺-deficient enzymes, but the activation for ATP was greatly reduced by Ca²⁺ starvation. The heat inactivation curves for basal and Mg²⁺-activated ATPase did not differ much between the complete and Ca²⁺-deficient enzyme. The delipidation of membrane-associated enzyme by acetone affected the protein content and the basal activity slightly, but inhibited the Mg²⁺-activated ATPase activity clearly with somewhat different behaviour between the complete and Ca²⁺-deficient enzyme.     

耐盐性植物转基因工程的研究进展

随着分子生物学的迅速发展,已经发现了一系列与植物盐胁迫相关的基因。根据这些基因产物的作用,可以分为两大类:效应分子基因和调控分子基因。根据近年来采用基因工程方法提高植物耐盐性的策略和研究进展进行了概述,同时探讨了目前还存在的一些问题。