Involvement of nitric oxide in the signal transduction of salicylic acid regulating stomatal movement

The effects and the relationship between sali-cylic acid(SA)and nitric oxide(NO) on Vicia faba L.stomatal movement were studied.The results here showed that exogenous SA and NO induced stomatal closure,100μmol/L SA induced a rapid and striking NO increase in the cytosol of guard cells.This phenomenon was largely prevented by 2000μmol/L 2-phenyl-4,4,5,5-tetramethylimidazoline-l-oxyl-3-oxide(PTIO),a specific NO scavenger,and 25μmol/L N^G-nitro-L-Arg-methyl eater (L-NAME),an inhibitor of NO synthase(NOS) in mammalian cells that also inhibits plant NOS.In addition,SA-induced stomatal closure was largely prevented by PTIO and L-NAME.These results provide evidence that guard cells generate NO in response to SA via NOS-like activity,and that such NO production is required for full stomatal closure in response to SA.H-(1,2,4)-oxadiazole-[4,3-α]quinoxalin-l-one(ODQ),an inhibitor of guanylate cyclase,and nicotinamide,an antagonist of cADPR production,inhibited the effects of SA-and NO-induced stomatal closure.It suggests that both cGMP and cADPR might mediate the signal transduction of SA and NO-induced stomatal closure.     

Protein tyrosine phosphatases involved in signaling of the ABA-induced H2O2 generation in guard cells of Vicia faba L.

Although protein tyrosine phosphatases (PTPases) play an important role in signal transduction in animal cells, little is known about the function of PTPases in higher plants. Hydrogen peroxide (H2O2) and mitogen-activated protein kinases (MAPKs) are the critical components of ABA signaling pathway in guard cells. PTPase is an important regulator of MAPK, which is believed to mediate ABA-induced H2O2 generation in guard cells of Vicia faba L. Here, we investigate the possible role of PTPases in stomatal movement process. Phenylarsine oxide (PAO), a specific inhibitor of PTPases, could prevent ABA or H2O2-induced stomatal closure of Vicia faba L; furthermore, it could promote opening of the stomata closed by ABA or H2O2. The activity of PTPases can be effectively inhibited by PAO and H2O2. DTT had no effect on the PAO-induced inhibition of PTPases activity, but it could relieve the inhibition of H2O2 on PTPases activity. PAO could also inhibit the ABA-induced H2O2 generation in guard cells of V‘wia faba L. These results suggested that PTPases is a critical signaling component in ABA-induced stomatal closure, and serve as targets for H2O2 lying on the signaling pathways downstream of ABA induced H2O2 generation.     

外源过氧化氢对蚕豆气孔运动的影响

运用外源过氧化氢(H2O2)处理、光镜观察的方法研究了H2O2对蚕豆气孔运动的影响．结果表明，H2O2诱导气孔关闭的最适浓度和最佳时间分别为100μmol／L和3h．H2O2促进气孔关闭的作用可被抗坏血酸和过氧化氢酶(CAT)逆转．还就内源H2O2在光、暗调控的气孔运动中的作用进行了讨论．     

Role of acetylcholine on plant root-shoot signal transduction

The role of accetylcholine (ACh) on plant root-shoot communication was investigated using the root-split system of Vicia faba L.In the experiments,slight osmotic stress caused the decrease of ACh content in root tips and the xylem sap transported up per time unit from root tip to the shool when the water potential of the shoot was kept unchanged .It also caused the decrease of ACh content in the abaxial epidermis,The decrease was highly correlative to the changes of transpiration rate,suggesting that the decrase of ACh content probably functions as a signal to regulate stomatal behavior,The effect of osmotic stress might be mainly through the inhibition of the ACh synthesis in root tip ;thus further influences the ACh content in root tip ,xylem sap and abaxial epidermis and resulting in the changes of stomata behavior,These results provide new evdence that plants transduce positive and negative signals among roots and shoot to coordinate stomatal behavior and adapt to variable environments.     

Regulation between nitric oxide and MAPK signal transduction in mammals

Nitric oxide (NO) is an important biological messenger in the regulation of tissue homeostasis. It exhibits a wide range of effects during physiological and pathophysiological processes. Typical beneficial properties of NO include the regulation of vascular tone,the protection of cells against apoptosis, the modulation of immune responses, and the killing of microbial pathogens. On the other hand,NO may cause severe vasodilation and myocardial depression during bacterial sepsis or act as a cytotoxic and tissue-damaging molecule in autoimmune diseases. Mitogen-activated protein kinase (MAPK) is a family of serine/threonine protein kinases that are widely distributed in mammalian cells. MAPK cascade plays pivotal roles in gene expression, cell proliferation, differentiation, neuronal survival and programmed cell death under a variety of experimental conditions. MAPKs transduce the signal for the cellular response to extracellular stresses or stimuli. The relation between them, however, has never been reviewed. Based on our researches and other reports in the field, we review their reciprocal regulatory functions.     

Rearrangements of microtubule cytoskeleton in stomatal closure of Arabidopsis induced by nitric oxide

NO (nitric oxide), known as a key signal molecule in plant, plays important roles in regulation of stomatal movement. In this study, microtubule dynamics and its possible mechanism in the NO signal pathway were investigated. The results were as follows: (i) In vivo stomatal aperture assays revealed that both vinblastine (microtubule-disrupting drug) and SNP (exogenous NO donor) prevented stomatal opening in the light, and vinblastine even could enhance the inhibitory effect of SNP, whereas taxol (a microtubule-stabilizing agent) was able to reduce this effect; (ii) microtubules in the opening Arabi- dopsis guard cells expressing GFP:α-tubulin-6 (AtGFP:α-tubulin-6) were organized in parallel, straight and dense bundles, radiating from the ventral side to the dorsal side, and most of them were localized perpendicularly to the ventral wall; (iii) under the same environmental conditions, treated with SNP for 30 min, the radial arrays of microtubules in guard cells began to break down, twisted partially and be- came oblique or exhibited a random pattern; (iv) furthermore, the involvement of cytosolic Ca2 in this event was tested. Stomatal aperture assays revealed that BAPTA-AM (a chelator of Ca2 ) greatly sup- pressed the effect of NO on stomatal closure; however, it did not show the same function on stomatal closure induced by vinblastine. When BAPTA-AM was added to the SNP-pretreated solution, the SNP-induced disordered microtubulue cytoskeleton in guard cells underwent rearrangement in a time-dependent manner. After 30 min of treatment with BAPTA-AM, the cortical microtubules resumed the original radial distribution, almost the same as the control. All this indicates that NO may promote rearrangement of microtubule cytoskeleton via elevation of [Ca2 ]cyt (free Ca2 concentration in the cy- toplasm), finally leading to stomatal closure.     

Involvement of carbon monoxide produced by heme oxygenase in ABA-induced stomatal closure in Vicia faba and its proposed signal transduction pathway

Carbon monoxide (CO) has recently proven to be an important bioactive or signaling molecule in mammalian cells. Its effects are mainly mediated by nitric oxide (NO) and cyclic GMP (cGMP). In Vicia faba leaves, CO production and heme oxygenase (HO) activity, an important CO synthetic enzyme, are first reported to increase in response to ABA treatment, which could result in stomatal closure. Inter- estingly, ABA-induced stomatal closure in V. faba guard cells is partially blocked when the synthetic CO inhibitor ZnPP, or the CO/NO scavenger Hb is added. Furthermore, we show that, exogenously applied CO donor, hematin, and CO aqueous solution not only result in the enhancement of CO release, but also time-dependently induce stomatal closure, and the latter is mimicked by the application of an NO donor SNP. The above-mentioned stomatal closure effects are differentially reversed by the addition of tungstate, a potent inhibitor of NO synthetic enzyme nitrate reductase (NR), the specific NO scavenger cPTIO, ZnPP, or Hb. During treatment for 4 h, SNP, 0.01% CO aqueous solution or hematin significantly triggers NO synthesis, whereas cPTIO, or tungstate approximately fully inhibits NO fluorescence. Ad- ditionally, application of the GC inhibitor ODQ blocks CO-induced stomatal closure. This inhibition could be reversed when 8-Br-cGMP is added. Thus, the above results suggest that CO produced by HO is involved in ABA-induced stomatal closure, and NO and cGMP may function as downstream interme- diates in the CO signaling responsible for stomatal closure.     

Ca2+在H2O2促进蚕豆气孔关闭过程中的作用

利用表皮生物分析法，通过表皮条缓冲液中加CaCl2研究了Ca＾2＋在H2O2促进蚕豆气孔关闭过程中的作用．研究发现在不同浓度的H2O2溶液中加入0．05mol/L和0．005mol/L CaCl2均能加强H2O2对蚕豆气孔关闭的促进作用，H2O2和Ca^2 浓度越高，气孔开度减小越明显．推测在H2O2促进蚕豆气孔关闭过程中，保卫细胞质中Ca^2 浓度升高时的来源可能是质外体中的Ca^2＋．     

一氧化氮的细胞内信号转导

本文从一氧化氮/环鸟苷酸途径;激活环氧化酶;调节蛋白激酶C;影响转录因子等多个方面对一氧化氮(NO)的细胞内信号转导途径进行了阐述.     

水分胁迫对蚕豆（Vicia faba L.）光合作用及产量的影响

以重庆市的蚕豆当家品种香珠豆为材料,通过人工水分胁迫处理,发现随土壤干旱程度的增加,蚕豆叶片的净光合速度下降,光饱和点降低,光补偿点提高,CO2补偿点提高,光呼吸增大;生长减弱,同化产物的积累减少,最终表现为植株的株高、根长、鲜干重、叶面积、根瘤的数量与重量、开花数、结荚数都明显下降,根冠比则上升。这是导致生物量积累减少的主要原因。     

Identification of integrin-like in guard cells ofVicia faba

We addressed the existence and localization of integrin-like in guard cells ofVicia faba by using a probe of polyclonal antibody against the human integrin (α_vβ₃/β₅). Western blot results showed that three integrins-like of about 47.3, 43.7 and 41.1 ku were detected from the preparation of membrane fragments of purified guard cell protoplasts. Further research with immunofluorescent scanning microscopy indicated that those integrins-like were localized on plasma membrane of guard cells, most nearing the dorsal wall, which is consistent with the reception of signals from epidermal cells to guard cells. Thus our results indicate, for the first time, that integrins-like are present at guard cell plasma membrane ofVicia faba.     

低浓度二氧化硫引起蚕豆叶片过氧化氢累积及几种清除剂变化的研究

低浓度ＳＯ２（０．２６０、０．１３１ｍｇ／ｍ３）处理蚕豆植株，引起叶片内Ｈ２Ｏ２含量升高，Ｈ２Ｏ２含量的升高与ＳＯＤ活性升高有关。同时体内清除Ｈ２Ｏ２的过氧化物酶和抗坏血酸过氧化物酶活性上升，而抗坏血酸含量下降。     

Involvement of Ca<Superscript>2+</Superscript>/CaM in the signal transduction of acetylcholine regulating stomatal movement

It has been known that the neurotransmitter acetylcholine (ACh) also exists in plants and is able to regulate the movement of stomata.In another aspect,Ca^2 /CaM as the second messengers have a critical role of signal transduction in stomatal guard-cell,Here we showed that Ca^2 /CaM were also involved in the ACh regulated stomatal movement,In the medium containing Ca^2 ,the Ca^2 channel blockers (NIF and Ver)and CaM inhibitors (TFP and W7 ) could neutralize the ACh induced stomatal opening,however,they are ineffective in the medium containing K^ ,Those results indicated that Ca^2 /CaM were involved in the signal transduction pathway of ACh regulating stomatal movement.     

Na2CO3胁迫对蚕豆幼苗几种生理生化指标的影响

用Na2CO3处理蚕豆幼苗,处理浓度为25,50,75,100mmol／L,胁迫6天后取样,测定植株鲜干重、肉质化程度、细胞膜透性、丙二醛含量、脯氨酸含量叶绿素含量等生理指标．结果表明,一定浓度的Na2CO3促进了蚕豆幼苗鲜干重、肉质化程度、叶绿素含量的增加,100mmol／LNa2CO3处理明显抑制了蚕豆幼苗的生长．Na2CO3浓度超过75mmol／L时,蚕豆幼苗MDA含量急剧上升,质膜透性也明显增加,脯氨酸含量增加明显．     

叶面施硼对蚕豆生物效应的影响

用不同浓度的硼酸溶液在蚕豆不同生长期进行叶面喷施.结果表明,在蚕豆生长的幼苗期、现蕾期和开花期叶面喷施0.1%-0.2%的硼酸溶液能促进蚕豆的生长和产量的增加.可能施硼后导致蚕豆叶片的SOD活性和CAT活性的增强,同时降低了O2-生成速率和MDA含量.     

Ethylene-induced nitric oxide production and stomatal closure in <Emphasis Type="Italic">Arabidopsis thaliana</Emphasis> depending on changes in cytosolic pH

We investigated changes in cytosolic pH and nitric oxide (NO) during ethylene-induced stomatal closure in Arabidopsis thaliana using pharmacological, laser scanning confocal microscopy (LSCM), and spectrophotography techniques. Treatment with ethephon (a direct source of ethylene when applied to plants) and 1-aminocycloaminopropane-1-carboxylic acid (ACC, an ethylene precursor) resulted in a rapid accumulation of NO and cytosolic alkalinization in guard cells. Acetic acid (a weak acid) and sodium orthovanadate (NaVO3; a plasmalemma H+-ATPase inhibitor) reduced stomatal closure induced by ethylene and blocked ethylene-induced activity of nitrate reductase. However, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO), a NO scavenger, had no effect. These results suggest that NO production is downstream of the rise in cytosolic pH in A. thaliana.     

蚕豆(Vicia faba L.)对不同水分胁迫的光合适应性研究

以重庆市的蚕豆当家品种之一的大白豆为材料,通过人工水分胁迫处理,发现随干旱程度的增加,蚕豆叶片的叶绿素a、叶绿素b含量和叶绿素总降低,Hill反应活力下降,净光合速率下降,同化产物的积累减少,最终表现为植株生物量的降低,水分胁迫下植株净光合速率下降,光饱和点降低,光补偿点提高,CO2补偿点提高,光呼吸增大,生长减弱,这时导致生物量积累减少的主要原因。     

干旱条件下蚕豆光合和生理特性变化研究

通过盆栽控制水分试验方法,对土壤适宜含水量(90%fθ)、轻度干旱(70%θf)、中度干旱(50%θf)和重度干旱(30%fθ)4种水分条件下蚕豆叶片光合和生理特性进行了系统研究.结果表明:随着干旱程度加重,蚕豆叶片净光合速率(Pn)、蒸腾速率(Tr)和气孔导度(Gs)受到了明显抑制,细胞间隙CO2浓度(Ci)则先升高后下降再上升;随着胁迫时间的延长,蚕豆叶片细胞膜透性、丙二醛(MDA)含量和渗透调节物质含量明显增加,叶片内的相对含水量和叶绿素含量减少,保护酶系统(SOD,POD,CAT)活性随着胁迫时间的延长,总体表现为先升高后降低.     

盐胁迫对蚕豆(Vicia faba L.)叶绿素a和b含量的影响

用不同的盐浓度对蚕豆（Ｖicia faba L.)处理的实验表明：叶绿素含量随盐浓度的增大而有所降低,而叶绿素a/b则无明显变化。当ＮaCl浓度升至１％时,叶绿素的含量在处理后的第三天呈现“增－降”现象,在１．５％ＮaCl浓度下,“增－降”尤为显著。这种叶绿素含量的变化势必影响到叶片叶绿素含量比来判断植物抗盐性大小。     

蚕豆（Vicia faba L.）叶片蒸腾速率的因子分析

利用LI－1600稳态气孔计测定了蚕叶片蒸腾速率的日变化,以及水分胁迫下蒸腾速率的变化,探讨了环境因子对蚕豆叶片蒸腾速率的影响,结果表明,光量子通量密度是影响蒸腾速率的主要气象因子,土壤含水量影响叶片相对含水量,并通过气孔扩散阻力来影响蒸腾速率。