脱落酸诱导气孔关闭的信号转导研究

气孔保卫细胞是单个细胞水平研究ABA信号转导机制的一 个模式系统。脱落酸（ABA）通过对保卫细胞生理生化状态、胞质Ca2+浓度及其离子通道调节诱导气孔关闭过程。这个过程涉及的因素有：ROS、IP3、cADPR、蛋白质的可逆磷酸化等。     

保卫细胞的ABA信号转导

植物激素脱落酸(ABA)调节植物体多种生理过程,尤其在一些逆境条件下,植物体中ABA大量合成,诱导气孔关闭,从而有效地调控植物体内的水分平衡.尽管人们对ABA诱导气孔关闭作用已得到共识,但有关信号转导的细节还很不清楚.该文简要介绍了研究气孔保卫细胞信号转导途径的相关技术以及与ABA信号转导直接相关的ABA受体、第二信使、蛋白质磷酸化和离子通道调节等方面的最新妍究进展.并在前人研究工作的基础上,勾画出气孔保卫细胞ABA、H2O2的信号转导模式图.     

保卫细胞的ABA信号转导

植物激素脱落酸（ABA）调节植物体多种生理过程,尤其在一些逆境条件下,植物体中ABA大量合成,诱导气孔关闭,从而有效地调控植物体内的水分平衡,尽管人们对ABA诱导气孔关闭作用已得到共识,但有关信号转导的细节还很不清楚,该文简要介绍了研究气孔保卫细胞信号转导途径的相关技术以及与ABA信号转导直接相关的ABA受体,第二信使,蛋白质磷酸化和离子通道调节等方面的最新研究进展,并在前人研究工作的基础上,勾画出气孔保卫细胞ABA,H2O2的信号转导模式图。     

干旱胁迫与ABA 的信号转导

植物经历干旱胁迫时,ABA被普遍认为是一种干旱信号而传递干旱信息。在干旱信号ABA的转导过程中,从ABA的被感知到保卫细胞发生变化引起气孔关闭以及ABA诱导的基因表达都经历了复杂的变化。本文对ABA的信号转导过程进行了综述。     

蚕豆保卫细胞原生质体质膜ABA结合蛋白与气孔对ABA敏感性的关系

在测定蚕豆(Vicia faba L.)保卫细胞原生质体质膜ABA结合蛋白的解离常数(Kd)和最大结合容量的基础上,进一步研究了几种因子(pH、光)对Kd和最大结合容量的影响。结果显示:pH并不改变结合蛋白的Kd值,而仅影响每个原生质体结合的分子数;光、暗处理的结果表明,光可使结合蛋白的Kd值增大,每个原生质体结合的ABA分子数减少,而暗处理可使结合蛋白的Kd值减小及每个原生质体结合的分子数增多,说明黑暗可提高气孔保卫细胞对ABA的敏感性,而光可以降低气孔保卫细胞对ABA的敏感性。因此,光、暗可以通过调节ABA结合蛋白的Kd值来调节气孔对ABA的敏感性。     

干旱胁迫与ABA的信号转导

植物经历干旱胁迫时,ABA被普遍认为是一种干旱信号而传递干旱信息.在干旱信号ABA的转导过程中,从ABA的被感知到保卫细胞发生变化引起气孔关闭以及ABA诱导的基因表达都经历了复杂的变化.本文对ABA的信号转导过程进行了综述.     

保卫细胞中ABA信号调控机制研究进展

脱落酸(ABA)具有调节植物快速响应逆境的重要功能。植物细胞中ABA核心信号通路由ABA受体PYR1/PYLs/ RCARs、A类碱性蛋白磷酸酶PP2Cs和Snf1相关蛋白激酶SnRK2s组成。活性氧(ROS)和Ca²⁺是保卫细胞中的重要第二信使, 调控ABA诱导的气孔关闭。该文对保卫细胞中核心ABA信号蛋白的调控以及ROS和Ca²⁺介导的ABA信号转导等最新研究成果进行综述, 旨在阐明保卫细胞中ABA信号调控机制。     

茉莉酸甲酯诱导保卫细胞气孔关闭的信号转导机制

气孔是由植物器官表面成对的保卫细胞围成的小孔,气孔运动控制气体交换,与植物逆境应答和生长发育等生物学过程密切相关,受多种因子调控,茉莉酸甲酯(MeJA)是其中之一。与ABA类似,MeJA也可诱导气孔关闭,但是其机理尚不清楚。该文综述了近年来MeJA调控气孔运动的信号转导机制进展,包括Ca2+、胞质pH、活性氧和NO等第二信使对气孔开闭的影响以及COI1、JAR1、RCN1和TGG1/2等信号组分之间的调控关系,并讨论了保卫细胞中MeJA与ABA信号途径的相互作用。     

环境因素和ABA对葡萄试管苗气孔开闭的影响

黑暗、低温、低湿、高渗等环境因素和ABA处理,虽能降低葡萄试管苗叶气孔开度,但因长时处于饱和湿度和弱光下的气孔保卫细胞发育不良,造成气孔口过度开放,而保卫细胞胀缩变化的幅度,不足以使这种过度开放的气孔口关闭。通过分步炼苗降低试管苗气孔口的开度后,保卫细胞膨压的变化就能使气孔关闭了。试管苗叶气孔在暗中的关闭率,可作为炼苗适合程度的生理指标。     

植物干旱胁迫下气孔关闭的信号转导

气孔关闭被认为是干旱胁迫的应答事件之一,根源ABA作为其原初信号之一,它向保卫细胞的运输受木质部汁液pH影响。而钙离子和阴离子通道则是ABA诱导气孔关闭的重要第二信使。     

Pyrabactin, an ABA agonist, induced stomatal closure and changes in signalling components of guard cells in abaxial epidermis of Pisum sativum

Pyrabactin, a synthetic agonist of abscisic acid (ABA), inhibits seed germination and hypocotyl growth and stimulates gene expression in a very similar way to ABA, implying the possible modulation of stomatal function by pyrabactin as well. The effect of pyrabactin on stomatal closure and secondary messengers was therefore studied in guard cells of Pisum sativum abaxial epidermis. Pyrabactin caused marked stomatal closure in a pattern similar to ABA. In addition, pyrabactin elevated the levels of reactive oxygen species (ROS), nitric oxide (NO), and cytoplasmic pH levels in guard cells, as indicated by the respective fluorophores. However, apyrabactin, an inactive analogue of ABA, did not affect either stomatal closure or the signalling components of guard cells. The effects of pyrabactin-induced changes were reversed by pharmalogical compounds that modulate ROS, NO or cytoplasmic pH levels, quite similar to ABA effects. Fusicoccin, a fungal toxin, could reverse the stomatal closure caused by pyrabactin, as well as that caused by ABA. Experiments on stomatal closure by varying concentrations of ABA, in the presence of fixed concentration of pyrabactin, and vice versa, revealed that the actions of ABA and pyrabactin were additive. Further kinetic analysis of data revealed that the apparent K(D) of ABA was increased almost 4-fold in the presence of ABA, suggesting that pyrabactin and ABA were competing with each other either at the same site or close to the active site. It is proposed that pyrabactin could be used to examine the ABA-related signal-transduction components in stomatal guard cells as well as in other plant tissues. It is also suggested that pyrabactin can be used as an antitranspirant or as a priming agent for improving the drought tolerance of crop plants.     

Abscisic acid-induced actin reorganization in guard cells of dayflower is mediated by cytosolic calcium levels and by protein kinase and protein phosphatase activities

In guard cells of open stomata under daylight, long actin filaments are arranged at the cortex, radiating out from the stomatal pore. Abscisic acid (ABA), a signal for stomatal closure, induces rapid depolymerization of cortical actin filaments and the slower formation of a new type of actin that is randomly oriented throughout the cell. This change in actin organization has been suggested to be important in signaling pathways involved in stomatal closing movement, since actin antagonists interfere with normal stomatal closing responses to ABA. Here we present evidence that the actin changes induced by ABA in guard cells of dayflower (Commelina communis) are mediated by cytosolic calcium levels and by protein phosphatase and protein kinase activities. Treatment of guard cells with CaCl2 induced changes in actin organization similar to those induced by ABA. Removal of extracellular calcium with EGTA inhibited ABA-induced actin changes. These results suggest that Ca2+ acts as a signal mediator in actin reorganization during guard cell response to ABA. A protein kinase inhibitor, staurosporine, inhibited actin reorganization in guard cells treated with ABA or CaCl2, and also increased the population of cells with long radial cortical actin filaments in untreated control cells. A protein phosphatase inhibitor, calyculin A, induced fragmentation of actin filaments in ABA- or CaCl2-treated cells and in control cells, and inhibited the formation of randomly oriented long actin filaments induced by ABA or CaCl2. These results suggest that protein kinase(s) and phosphatase(s) participate in actin remodeling in guard cells during ABA-induced stomatal closure.     

NO可能作为H2O2的下游信号介导ABA诱导的蚕豆气孔关闭

ABA、H2O2和硝普钠(SNP)均能诱导蚕豆气孔关闭.NO的清除剂c-PTIO可以减轻由ABA或H2O2所诱导的蚕豆气孔关闭的程度,而过氧化氢酶(CAT)则不能减轻NO诱导的气孔关闭程度.激光共聚焦显微检测结果显示,10μmo1/L的ABA处理后,胞内H2O2的产生速率明显高于NO的产生速率;CAT几乎可完全抑制ABA所诱导的DAF的荧光增加;外源H2O2能显著诱导胞内DAF的荧光增加;c-PTIO对ABA诱导的DCF荧光略有促进作用,但外源SNP不能诱导胞内DCF荧光增加.这些结果表明,在ABA诱导气孔关闭过程中,H2O2可能在NO的上游起作用并受NO的负反馈调节.     

Functional Epidermal Cells are Necessary for Abscisic Acid Effects on Guard Cells

Experimental results are presented which show that abscisicacid (ABA) causes stomatal closure only if the stomatal complexis adjacent to live epidermal cells. It is further shown thatABA acts by affecting solute fluxes into and out of epidermaland guard cells. Live epidermal cells function as recipientsfor solutes and thereby assist their movement out of the guardcells. ABA-mediated solute leakage from guard cells alone doesnot suffice to cause stomatal closure within one hour.     

ABA, hydrogen peroxide and nitric oxide signalling in stomatal guard cells

Increased synthesis and redistribution of the phytohormone abscisic acid (ABA) in response to water deficit stress initiates an intricate network of signalling pathways in guard cells leading to stomatal closure. Despite the large number of ABA signalling intermediates that are known in guard cells, new discoveries are still being made. Recently, the reactive oxygen species hydrogen peroxide (H2O2) and the reactive nitrogen species nitric oxide (NO) have been identified as key molecules regulating ABA-induced stomatal closure in various species. As with many other physiological responses in which H2O2 and NO are involved, stomatal closure in response to ABA also appears to require the tandem synthesis and action of both these signalling molecules. Recent pharmacological and genetic data have identified NADPH oxidase as a source of H2O2, whilst nitrate reductase has been identified as a source of NO in Arabidopsis guard cells. Some signalling components positioned downstream of H2O2 and NO are calcium, protein kinases and cyclic GMP. However, the exact interaction between the various signalling components in response to H2O2 and NO in guard cells remains to be established.     

Nitric oxide production occurs after cytosolic alkalinization during stomatal closure induced by abscisic acid

Abscisic acid (ABA) raised the cytosolic pH and nitric oxide (NO) levels in guard cells while inducing stomatal closure in epidermis of Pisum sativum. Butyrate (a weak acid) reduced the cytosolic pH/NO production and prevented stomatal closure by ABA. Methylamine (a weak base) enhanced the cytosolic alkalinization and aggravated stomatal closure by ABA. The rise in guard cell pH because of ABA became noticeable after 6 min and peaked at 12 min, while NO production started at 9 min and peaked at 18 min. These results suggested that NO production was downstream of the rise in cytosolic pH. The ABA-induced increase in NO of guard cells and stomatal closure was prevented by 2-phenyl-4,4,5,5-tetramethyl imidazoline-1-oxyl 3-oxide (cPTIO, a NO scavenger) and partially by N-nitro-L-Arg-methyl ester (L-NAME, an inhibitor of NO synthase). In contrast, cPTIO or L-NAME had only a marginal effect on the pH rise induced by ABA. Ethylene glycol tetraacetic acid (EGTA, a calcium chelator) prevented ABA-induced stomatal closure while restricting cytosolic pH rise and NO production. We suggest that during ABA-induced stomatal closure, a rise in cytosolic pH is necessary for NO production. Calcium may act upstream of cytosolic alkalinization and NO production, besides its known function as a downstream component.     

Anion-Channel Blockers Inhibit S-Type Anion Channels and Abscisic Acid Responses in Guard Cells

The effects of anion-channel blockers on light-mediated stomatal opening, on the potassium dependence of stomatal opening, on stomatal responses to abscisic acid (ABA), and on current through slow anion channels in the plasma membrane of guard cells were investigated. The anion-channel blockers anthracene-9-carboxylic acid (9-AC) and niflumic acid blocked current through slow anion channels of Vicia faba L. guard cells. Both 9-AC and niflumic acid reversed ABA inhibition of stomatal opening in V. faba L. and Commelina communis L. The anion-channel blocker probenecid also abolished ABA inhibition of stomatal opening in both species. Additional tests of 9-AC effects on stomatal aperture in Commelina revealed that application of this anion-channel blocker allowed wide stomatal opening under low (1 mM) KCI conditions and increased the rate of stomatal opening under both low and high (100 mM) KCI conditions. These results indicate that anion channels can function as a negative regulator of stomatal opening, presumably by allowing anion efflux and depolarization, which prohibits ion up-take in guard cells. Furthermore, 9-AC prevented ABA induction of stomatal closure. A model in which ABA activation of anion channels contributes a rate-limiting mechanism during ABA-induced stomatal closure and inhibition of stomatal opening is discussed.     

Cyclic adenosine 5′‐diphosphoribose (cADPR) cyclic guanosine 3′,5′‐monophosphate positively function in Ca2+ elevation in methyl jasmonate‐induced stomatal closure,cADPR is required for methyl jasmonate‐induced ROS accumulation NO production in guard cells

Methyl jasmonate (MeJA) signalling shares several signal components with abscisic acid (ABA) signalling in guard cells. Cyclic adenosine 5′‐diphosphoribose (cADPR) and cyclic guanosine 3′,5′‐monophosphate (cGMP) are second messengers in ABA‐induced stomatal closure. In order to clarify involvement of cADPR and cGMP in MeJA‐induced stomatal closure in Arabidopsis thaliana (Col‐0), we investigated effects of an inhibitor of cADPR synthesis, nicotinamide (NA), and an inhibitor of cGMP synthesis, LY83583 (LY, 6‐anilino‐5,8‐quinolinedione), on MeJA‐induced stomatal closure. Treatment with NA and LY inhibited MeJA‐induced stomatal closure. NA inhibited MeJA‐induced reactive oxygen species (ROS) accumulation and nitric oxide (NO) production in guard cells. NA and LY suppressed transient elevations elicited by MeJA in cytosolic free Ca²⁺ concentration ([Ca²⁺]_cyt) in guard cells. These results suggest that cADPR and cGMP positively function in [Ca²⁺]_cyt elevation in MeJA‐induced stomatal closure, are signalling components shared with ABA‐induced stomatal closure in Arabidopsis, and that cADPR is required for MeJA‐induced ROS accumulation and NO production in Arabidopsis guard cells.