Role of nitric oxide dependence on nitric oxide synthase-like activity in the water stress signaling of maize seedling

Nitric oxide (NO) has been known as an important signal in plant antioxidative defense but its production and roles in water stress are less known. The present study investigated whether NO dependence on a NO synthase-lika (NOS) activity is involved in the signaling of drought-induced protective responses in maize seedlings. NOS activity, rate of NO release and drought responses were analyzed when NO donor sodium nitroprusside (SNP), NO scavenger c-PTIO (2-(4-carboxyphenyl)-4,4,5,5-tetramathylimidazoline-1-oxyl-3-oxide) and NOS inhibitor L-NAME (NG-nitro-L-arginine methyl ester) were applied to both detached maize leaves and whole plants. Both NOS activity and the rate of NO release increased substantially under dehydration stress. The high NOS activity induced by c-PTIO as NO scavenger and NO accumulation Inhibited by NOS inhibitor L-NAME In dehydration-treated maize seedlings Indicated that most NO production under water deficit stress may be generated from NOS-like activity. After dehydration stress for 3 h, detached maize leaves pretreated with NO donor SNP maintained more water content than that of control leaves pretreated with water. This result was consistent with the decrease in the transpiration rate of SNP-treated leaves subjected to drought treatment for 3 h. Membrane permeability, a cell injury index, was lower in SNP-trested maize leaves under dehydration stress for 4 h when compared with the control leaves. Also, superoxide dismutsse (SOD) activity of SNP combined drought treatment maize leaves was higher than that of drought treatment alone, indicating that exogenous NO treatment alleviated the water loss and oxidative damage of maize leaves under water deficit stress. When c-PTIO as a specific NO scavenger was applied, the effects of applied SNP were overridden. Treatment with L-NAME on leaves also led to higher membrane permeability, higher transpiration rate and lower SOD activities than those of control leaves, indicating that NOS-like activity was involved in the antioxidative defense under water stress. These results suggested that NO dependence on NOS-like activity serves as a signaling component in the induction of protective responses and is associated with drought tolerance in maize seedlings.     

Nitric oxide is involved in dehydration/drought tolerance in Poncirus trifoliata seedlings through regulation of antioxidant systems and stomatal response

Nitric oxide (NO) is a component of the repertoire of signals implicated in plant responses to environmental stimuli. In the present study, we investigated the effects of exogenous application of NO-releasing donor sodium nitroprusside (SNP) and nitric oxide synthase inhibitor N ^G-nitro-l-arginine-methyl ester (l-NAME) on dehydration and drought tolerance of Poncirus trifoliata. The endogenous NO level was enhanced by SNP pretreatment, but decreased by l-NAME, in the hydroponic or potted plants with or without stresses. Under dehydration, leaves from the SNP-treated hydroponic seedlings displayed less water loss, lower electrolyte leakage and reactive oxygen species accumulation, higher antioxidant enzyme activities and smaller stomatal apertures as compared with the control (treated with water). In addition, pretreatment of the potted plants with SNP resulted in lower electrolyte leakage, higher chlorophyll content, smaller stomatal conductance and larger photosynthetic rate relative to the control. By contrast, the inhibitor treatment changed these physiological attributes or phenotypes in an opposite way. These results indicate that NO in the form of SNP enhanced dehydration and drought tolerance, whereas the inhibitor makes the leaves or plants more sensitive to the stresses. The stress tolerance by NO might be ascribed to a combinatory effect of modulation of stomatal response and activation of the antioxidant enzymes. Taken together, NO is involved in dehydration and drought tolerance of P. trifoliata, implying that manipulation of this signal molecule may provide a practical approach to combat the environmental stresses.     

Drought-induced proline accumulation is uninvolved with increased nitric oxide,which alleviates drought stress by decreasing transpiration in rice

Accumulation of proline is trusted to be an adaptive response of plants against drought stress, and exogenous application of nitric oxide (NO) enhances proline accumulation in Cu-treated algae. In order to investigate whether NO works as a necessary signaling molecule in drought-induced proline accumulation in rice leaves, effects of drought stress on endogenous NO content and proline accumulation were studied in rice leaves, using sodium nitroprusside (SNP, a NO donor) and 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO, a NO scavenger). The results showed that drought treatment increased both endogenous NO and proline contents in rice leaves, while foliar spray of various concentrations of SNP failed to induce proline accumulation in the leaves of well-watered rice and foliar spray of cPTIO failed to inhibit proline accumulation in the leaves of drought-stressed rice. These results indicate that increase of endogenous NO is dispensable for proline accumulation in the leaves of rice under drought stress. Further studies indicate that exogenous application of NO alleviates drought-induced water loss and ion leakage by decreasing transpiration rate of rice leaves.     

一氧化氮是脱落酸诱导杨树叶片气孔关闭的信号分子

研究了外源NO和ABA对杨树气孔运动调节作用．结果表明,外源NO和ABA都能诱导杨树离体叶片气孔关闭,且具有剂量效应,NO可加强ABA诱导气孔关闭的作用．NO清除剂(c—PTIO)可大大减弱NO和ABA对气孔关闭的诱导作用．证实了NO参与ABA调控气孔开闭运动过程,不同浓度NO供体SNP和ABA处理杨树离体叶片,SOD活性变化不明显,POD活性受到显著抑制．杨树叶片粗酶液的体外实验表明,不同浓度SNP对POD活性的抑制呈明显的浓度及时间效应;而ABA对POD活性则几乎没有影响．本研究证明,NO调节ABA诱导的树木气孔关闭作用,是ABA诱导树木气孔关闭的一种重要信号分子．     

Hydraulic and Chemical Responses of Citrus Seedlings to Drought and Osmotic Stress

In this work we investigated the function of abscisic acid (ABA) as a long-distance chemical signal communicating water shortage from the root to the shoot in citrus plants. Experiments indicated that stomatal conductance, transpiration rates, and leaf water potential decline progressively with drought. ABA content in roots, leaves, and xylem sap was also increased by the drought stress treatment three- to sevenfold. The addition of norflurazon, an inhibitor of ABA biosynthesis, significantly decreased the intensity of the responses and reduced ABA content in roots and xylem fluid, but not in leaves. Polyethylene glycol (PEG)-induced osmotic stress caused similar effects and, in general, was counteracted only by norflurazon at the lowest concentration (10%). Partial defoliation was able to diminish only leaf ABA content (22.5%) at the highest PEG concentration (30%), probably through a reduction of the active sites of biosynthesis. At least under moderate drought (3–6 days without irrigation), mechanisms other than leaf ABA concentration were required to explain stomatal closure in response to limited soil water supply. Measurements of xylem sap pH revealed a progressive alkalinization through the drought condition (6.4 vs. 7.1), that was not counteracted with the addition of norflurazon. Moreover, in vitro treatment of detached leaves with buffers iso-osmotically adjusted at pH 7.1 significantly decreased stomatal conductance (more than 30%) as much as 70% when supplemented with ABA. Taken together, our results suggest that increased pH generated in drought-stressed roots is transmitted by the xylem sap to the leaves, triggering reductions in shoot water loss. The parallel rise in ABA concentration may act synergistically with pH alkalinization in xylem sap, with an initial response generated from the roots and further promotion by the stressed leaves.     

植物激素ABA在水分胁迫下的功能及信号途径

植物根系感知外界水分胁迫刺激,诱导ABA生物合成。ABA既可诱导气孔关闭或抑制气孔开放,以降低植物的蒸腾失水,又可影响植物根系发育,以抵御水分胁迫。本文就植物激素ABA及其下游信号H2O2、NO以及Ca2＋等在植物生长调节方面的研究进展进行概述,以构建水分胁迫下植物生长自我调控的可能模式。     

H_2O_2作为根源信号介导盐胁迫诱导的蚕豆气孔关闭反应

H2O2作为信号分子可被多种胁迫诱导产生并在细胞内积累,进而参与调节植物的抗逆反应。文章通过远红外热成像观察等实验发现,根部NaCl胁迫可诱导蚕豆气孔关闭,叶片温度上升,叶片内Na＋和H2O2含量增加,蒸腾流汁液中H2O2浓度升高。另外,NaCl可直接诱导离体蚕豆根产生H2O2,却不能影响表皮条内H2O2含量。NaCl胁迫条件下产生的蒸腾流汁液可直接诱导表皮条气孔关闭,该过程可被抗氧化剂抗坏血酸（AsA）所逆转。这些结果表明,H2O2作为盐胁迫的根源信号,可能通过维管系统运输参与调节蚕豆气孔的关闭反应。     

Stomatal response characteristics of Tradescantia virginiana grown at high relative air humidity

Plants produced at high relative air humidity (RH) show poor control of water loss after transferring to low RH, a phenomenon which is thought to be due to their stomatal behaviour. The stomatal anatomy and responses of moderate (55%) and high (90%) RH grown Tradescantia virginiana plants to treatments that normally induce stomatal closure, i.e. desiccation, abscisic acid (ABA) application and exposure to darkness were studied using attached or detached young, fully expanded leaves. Compared with plants grown at moderate RH the transpiration rate, stomatal conductance and aperture of high RH grown plants measured at the same condition (40% RH) were, respectively, 112, 139 and 132% in light and 141, 188 and 370% in darkness. Besides the differences in stomatal size (guard cell length was 56.7 and 73.3 µm for moderate and high RH grown plants, respectively), there was a clear difference in stomatal behaviour. The stomata responded to desiccation, ABA and darkness in both moderate and high RH grown plants, but the high variability of stomatal closure in high RH grown plants was striking. Some stomata developed at high RH closed in response to darkness or to a decrease in relative water content to the same extent as did stomata from moderate RH grown plants, whereas others closed only partly or did not close at all. Evidently, some as yet unidentified physiological or anatomical changes during development disrupt the normal functioning of some stomata in leaves grown at high RH. The failure of some stomata to close fully in response to ABA suggests that ABA deficiency was not responsible for the lack of stomatal closure in response to desiccation.     

Overexpressing <Emphasis Type="Italic">SgNCED1</Emphasis> in Tobacco Increases ABA Level,Antioxidant Enzyme Activities,and Stress Tolerance

Abscisic acid (ABA) regulates plant adaptive responses to various environmental stresses. 9-cis-epoxycarotenoid dioxygenase (NCED) is the key enzyme of ABA biosynthesis in higher plants. A NCED gene, SgNCED1, was overexpressed in transgenic tobacco plants which resulted in 51–77% more accumulation of ABA in leaves. Transgenic tobacco plants decreased stomatal conductance, transpiration rate, and photosynthetic rate but induced activities of superoxide dismutase (SOD), catalase (CAT), and ascorbate-peroxidase (APX). Hydrogen peroxide (H₂O₂) and nitric oxide (NO) in leaves were also induced in the transgenic plants. Compared to the wild-type control, the transgenic plants improved growth under 0.1 M mannitol-induced drought stress and 0.1 M NaCl-induced salinity stress. It is suggested that the ABA-induced H₂O₂ and NO generation upregulates the stomatal closure and antioxidant enzymes, and therefore increases drought and salinity tolerance in the transgenic plants.     

外源ABA、NO和H2O2对葱莲花瓣及叶片表皮气孔开闭的影响

以葱莲(Zephyranthes candida)为材料,研究不同浓度外源脱落酸、硝普钠(sodium nitroprusside,SNP)及过氧化氢对花瓣和叶片表皮气孔开闭的影响,以期为三者在切花保鲜中的应用提供新的依据。实验结果表明,10~1000 μmol/L脱落酸和硝普钠均能不同程度地引起花瓣和叶片表皮气孔关闭,且花瓣气孔较叶片气孔有更高的敏感性。过氧化氢对叶片表皮气孔开闭的影响大于对花瓣气孔的影响,花瓣表皮的气孔孔径仅在1000 μmol/L处理时变化显著。这说明在外源信号物质延缓切花衰老的过程中,花瓣表皮气孔的运动也可能起到了一定的作用。适当外源信号物质处理能诱导花瓣表皮气孔关闭,从而使花瓣的蒸腾作用减小,维持植物体内水势,延缓切花衰老。