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。     

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.     

植物细胞质膜H+-ATPase的结构与功能

植物细胞质膜H+-ATPase属于P型质子泵。由该酶产生的跨膜电化学梯度是物质跨膜运输的原初动力。研究表明,质膜H+-ATPase与植物的生长发育密切相关,被称为植物细胞的“主宰酶”。近年,关于该酶的生化特性,基因表达与调控以及结构与功能等方面的研究取得重要进展。对质膜H+-ATPase的生化特性,分子结构,调节机制和生理功能等进行了综述。     

质膜转运蛋白及其与植物耐盐性关系研究进展

植物细胞质膜有两种主要功能：(1)溶质运输(进出细胞),溶质运输主要由转运蛋白完成;(2)信号传导,即接收信号并引发细胞生理生化响应。盐分过多对植物的伤害主要是离子毒害。质膜转运蛋白活性对环境变化能做出迅速响应。本文简要叙述了植物细胞质膜转运蛋白类型、分子特性、生理功能及其活性调节。介绍了植物细胞质膜H+_ATPase、质膜氧化还原系统、质膜离子载体和离子通道对盐胁迫的响应及其这些响应与植物耐盐性之间的关系。     

植物质膜H+-ATPase的研究进展

质膜H -ATPase参与植物细胞的物质跨膜转运、细胞的伸长生长、气孔的开闭以及植物对环境胁迫的响应等生理过程,是植物生命活动的“主宰酶”。其活性调节涉及激素、环境因子等多种因素,可发生在转录、翻译和酶分子等多级水平。因此,在植物生长发育过程中,质膜H -ATPase活性的调节对生理活动起重要作用。本文就植物质膜H -ATPase的结构特征、生理功能、活性变化及其调节机理等的研究进展进行综述,以进一步揭示该酶的生理功能及其调节机理与植物生命活动过程的关系。     

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。     

豌豆根胞质V1-ATPase的鉴定

利用免疫印迹、免疫电镜和ATP水解活性的测定对豌豆(Pisum sativum L.)根细胞胞质中V1-ATPase复合物的存在进行鉴定.用兔抗绿豆V-type H+-ATPase 的A、B亚基的抗体进行的immuno-blotting和胶体金电镜结果都表明,胞质中存在有A、B亚基.活性测定结果进一步表明胞质具有ATP水解活性.这些结果说明豌豆根胞质具有有活性的V1-ATPase复合物.这是首次直接证明植物中有胞质V1-ATPase的存在.     

不同生境下木麻黄脯氨酸含量和质膜ATPase活性的研究

采集生长于恶劣环境和中生环境的普通木麻黄(Casuarina)小枝,超速离心提取粗质膜制剂后,用两相系统法纯化得到质膜微囊,研究不同生境下木麻黄的质膜ATPase活性,并测定木麻黄小枝的游离脯氨酸含量。实验结果表明:同一生境中的木麻黄ATPase活性相对一致,而同一树种木麻黄不同生境下质膜微囊H~+-ATPase、Ca~(2+)-ATPase、K~+-ATPase活性有显著差异,表现出以渗透胁迫为主的恶劣环境下的木麻黄质膜微囊ATPase活性和木麻黄细胞内游离脯氨酸明显高于中生环境下生长的木麻黄。说明普通木麻黄在干旱和盐胁迫下能调整生理生化过程来提高其质膜ATPase活性和增加细胞内脯氨酸含量提高渗透调节能力以保证其在恶劣环境的正常生长。     

NaCl胁迫对宁夏枸杞幼苗根系质膜和液泡膜H+-ATPase活性的影响

以宁夏枸杞为材料,研究NaCl胁迫下枸杞幼苗组织含水量、干物质重量、无机离子分布、质膜透性、丙二醛 (MDA)含量及幼根质膜、液泡膜H -ATPase活性和耐盐性之间的关系。结果表明,在0．3％NaCl胁迫下,干物质重量、质膜透性、MDA含量略有增加,组织含水量略有下降;随着NaCl胁迫强度的增加,Na 、Cl-含量逐渐增加, K 含量则呈现下降的趋势;质膜透性增大与MDA含量升高呈显著正相关;质膜和液泡膜H -ATPase活性逐渐增加,且液泡膜H -ATPase活性显著高于质膜H -ATPase活性,表明枸杞具有较强的抗盐性。     

K~ 营养对NaCl胁迫下盐地碱蓬生长及叶片液泡膜V-H~ -ATPase和V-H~ -PPase活性的影响(英文)

对溶液培养的盐地碱蓬 (SuaedasalsaL .)幼苗进行不同浓度NaCl胁迫并改变培养液中K 浓度 ,以了解K 营养对NaCl胁迫下盐地碱蓬幼苗生长及叶片液泡膜V_H _ATPase、V_H _PPase活性的影响。提高培养液K 浓度可明显增加盐胁迫下碱蓬植株的鲜重、干重 ,促进盐地碱蓬叶片及根部组织K 积累。盐地碱蓬叶片液泡膜V_H _ATPase至少由A、B、C、D、E及c亚基组成 ,其表达量在缺K 处理 (12 μmol/LK )下随盐胁迫浓度的增加而减小 ,而在正常K (6mmol/L)培养下则随盐胁迫浓度的增加而增加 ;盐地碱蓬叶片液泡膜V_H _PPase分子量为 72kD ,在缺K 和正常K 供应情况下 ,V_H _PPase均有较高表达。V_H _ATPase及V_H _PPase活性变化与其亚基表达量变化基本成正相关。结果表明 :K 对盐生植物碱蓬的耐盐性有重要作用 ,盐胁迫下 ,K 可能参与了V_H _ATPase和V_H _PPase活性调控     

甘氨酸甜菜碱增强青菜抗盐性的作用(英文)

通过对青菜 (BrassicachinensisL .)叶面喷施甜菜碱 ,发现其易于为叶片所吸收并运至其他部位。一定浓度范围内的甜菜碱可明显增强青菜对盐胁迫的抗性。甜菜碱可显著降低盐胁迫下叶和根中Na 的累积 ,这种降低主要是根系对Na 、K 的选择性吸收能力增强所致。盐胁迫下甜菜碱导致根系质膜H ATPase活性提高了 45.1 % ,据此推测甜菜碱降低植株中Na 的累积很可能部分由于促进根系质膜的主动排Na 过程。另外 ,甜菜碱对抗盐性的增强还体现在对叶片质膜和叶绿素的稳定作用和对脯氨酸合成的促进。     

豌豆根胞质V1—ATPase的鉴定

利用免疫印迹,免疫电镜和ATP水解活性的测定对豌豆（Pisum sativum L.)根细胞胞质中V1-ATPase复合物的存在进行鉴定。用兔抗绿豆V-typeH^ -ATPase的A,B亚基的抗体进行的immuno-blotting和胶体金电镜结果都表明,胞质中存在有A,B亚基。活性测定结果进一步表明胞质具有ATP水解活性,这些结果说明豌豆根胞质具有活性的V1-ATPase复合物。这是首次直接证明植物中有胞质V1-ATPase的存在。     

NaCl胁迫下大麦根系液泡膜H~ -ATPase活性与膜流动性的关系(英文)

用 5 0～ 2 0 0mmol/LNaCl处理 2d后 ,大麦 (HordeumvulgareL .)品种“滩引 2号”(耐盐性强 )根的液泡膜H _ATPase活性增强 ,6 0 0mmol/LNaCl处理下酶活性下降 ;“科品 7号”(耐盐性弱 )在 5 0～ 10 0mmol/LNaCl处理 2d后根的液泡膜H _ATPase活性增强 ,2 0 0～ 6 0 0mmol/LNaCl处理下酶活性随盐浓度增加而降低。 5 0～ 2 0 0mmol/LNaCl处理下“滩引 2号”根的液泡膜流动性下降 ,6 0 0mmol/LNaCl处理下膜流动性明显增大 ;盐胁迫下液泡膜膜脂脂肪酸不饱和度下降时 ,膜流动性下降 ,反之则膜流动性上升。由此推断高盐胁迫下液泡膜膜脂脂肪酸不饱和度上升而引起膜流动性上升可能是引起H _ATPase活性下降的原因之一。     

Protective Effects of Glycinebetaine on Brassica chinensis Under Salt Stress (in English)

Brassica chinensis L. were foliarly applied with glycinebetaine (GB), as this species is unable to synthesis GB and sensitive to osmotic stress such as salt. The exogenous GB was easily absorbed and transported by the leaf of B. chinensis . Its application (0-20 mmol/L) enhanced the plant tolerance to salt stress. The treatment of 15 mmol/L GB significantly decreased the Na+ accumulation in leaf and root under NaCl stress. This difference in accumulating Na+ and K+ is caused by higher selectivity of root absorption. Furthermore, GB increased H+-ATPase activity of root plasma membrane evidently. This result strongly suggested that in root the decreased Na+ accumulation was caused by the GB accumulation that enhanced the extrusion of Na+ from the cell in some way through plasma membrane transporter, e.g. Na+/H+ antiport driven by H+-ATPase. The GB application was also found to stabilize the plasma membrane, to decrease the loss of chlorophyll, and to stimulate the osmosis induced proline response under salt stress.     

NaCl胁迫下大麦根系液泡膜H+-ATPase活性与膜流动性的关系

用50～200 mmol/L NaCl处理2 d后,大麦(Hordeum vulgare L.)品种"滩引2号"(耐盐性强)根的液泡膜H+-ATPase活性增强,600 mmol/L NaCl处理下酶活性下降;"科品7号"(耐盐性弱)在50～100 mmol/L NaCl处理2 d后根的液泡膜H+-ATPase活性增强,200～600 mmol/L NaCl处理下酶活性随盐浓度增加而降低.50～200 mmol/L NaCl处理下"滩引2号"根的液泡膜流动性下降,600 mmol/L NaCl处理下膜流动性明显增大;盐胁迫下液泡膜膜脂脂肪酸不饱和度下降时,膜流动性下降,反之则膜流动性上升.由此推断高盐胁迫下液泡膜膜脂脂肪酸不饱和度上升而引起膜流动性上升可能是引起H+-ATPase活性下降的原因之一.     

Yeast plasma membrane Ena1p ATPase alters alkali-cation homeostasis and confers increased salt tolerance in tobacco cultured cells

In plants, the plasma membrane Na(+)/H(+) antiporter is the only key enzyme that extrudes cytosolic Na(+) and contributes to salt tolerance. But in fungi, the plasma membrane Na(+)/H(+) antiporter and Na(+)-ATPase are known to be key enzymes for salt tolerance. Saccharomyces cerevisiae Ena1p ATPase encoded by the ENA1/PMR2A gene is primarily responsible for Na(+) and Li(+) efflux across the plasma membrane during salt stress and for K(+) efflux at high pH and high K(+). To test if the yeast ATPase would improve salt tolerance in plants, we expressed a triple hemagglutinin (HA)-tagged Ena1p (Ena1p-3HA) in cultured tobacco (Nicotiana tabacum L.) cv Bright Yellow 2 (BY2) cells. The Ena1p-3HA proteins were correctly localized to the plasma membrane of transgenic BY2 cells and conferred increased NaCl and LiCl tolerance to the cells. Under moderate salt stress conditions, the Ena1p-3HA-expressing BY2 clones accumulated lower levels of Na(+) and Li(+) than nonexpressing BY2 clones. Moreover, the Ena1p-3HA expressing BY2 clones accumulated lower levels of K(+) than nonexpressing cells under no-stress conditions. These results suggest that the yeast Ena1p can also function as an alkali-cation (Na(+), Li(+), and K(+)) ATPase and alter alkali-cation homeostasis in plant cells. We conclude that, even with K(+)-ATPase activity, Na(+)-ATPase activity of the yeast Ena1p confers increased salt tolerance to plant cells during salt stress.     

Effects of Salt Stress on the Activity and the Amount of Tonoplast H+-ATPase from Pea Roots

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.     

Herbivore exposure alters ion fluxes and improves salt tolerance in a desert shrub

Plants have evolved complex mechanisms that allow them to withstand multiple environmental stresses, including biotic and abiotic stresses. Here, we investigated the interaction between herbivore exposure and salt stress of Ammopiptanthus nanus, a desert shrub. We found that jasmonic acid (JA) was involved in plant responses to both herbivore attack and salt stress, leading to an increased NaCl stress tolerance for herbivore-pretreated plants and increase in K⁺/Na⁺ ratio in roots. Further evidence revealed the mechanism by which herbivore improved plant NaCl tolerance. Herbivore pretreatment reduced K⁺ efflux and increased Na⁺ efflux in plants subjected to long-term, short-term, or transient NaCl stress. Moreover, herbivore pretreatment promoted H⁺ efflux by increasing plasma membrane H⁺-adenosine triphosphate (ATP)ase activity. This H⁺ efflux creates a transmembrane proton motive force that drives the Na⁺/H⁺ antiporter to expel excess Na⁺ into the external medium. In addition, high cytosolic Ca²⁺ was observed in the roots of herbivore-treated plants exposed to NaCl, and this effect may be regulated by H⁺-ATPase. Taken together, herbivore exposure enhance s A. nanus tolerance to salt stress by activating the JA-signalling pathway, increasing plasma membrane H ⁺ - ATPase activity, promoting cytosolic Ca²⁺ accumulation, and then restricting K⁺ leakage and reducing Na⁺ accumulation in the cytosol.     

K+营养对NaCl胁迫下盐地碱蓬生长及叶片液泡膜V-H+-ATPase和V-H+-PPase活性的影响

对溶液培养的盐地碱蓬(Suaeda salsa L.)幼苗进行不同浓度NaCl胁迫并改变培养液中K+浓度,以了解K+营养对NaCl胁迫下盐地碱蓬幼苗生长及叶片液泡膜V-H+-ATPase、V-H+-PPase活性的影响.提高培养液K+浓度可明显增加盐胁迫下碱蓬植株的鲜重、干重,促进盐地碱蓬叶片及根部组织K+积累.盐地碱蓬叶片液泡膜V-H+-ATPase至少由A、B、C、D、E及c亚基组成,其表达量在缺K+处理(12 μmol/L K+)下随盐胁迫浓度的增加而减小,而在正常K+(6 mmol/L)培养下则随盐胁迫浓度的增加而增加;盐地碱蓬叶片液泡膜V-H+-PPase分子量为72 kD,在缺K+和正常K+供应情况下,V-H+-PPase均有较高表达.V-H+-ATPase及V-H+-PPase活性变化与其亚基表达量变化基本成正相关.结果表明: K+对盐生植物碱蓬的耐盐性有重要作用,盐胁迫下,K+可能参与了V-H+-ATPase和V-H+-PPase活性调控.