Pharmacological, laser scanning confocal microscopic （LSCM）, and spectrophotographic approaches were used to study the roles of hydrogen sulfide （H2S） and nitric oxide （NO） in signaling transduction of stomatal ...Pharmacological, laser scanning confocal microscopic （LSCM）, and spectrophotographic approaches were used to study the roles of hydrogen sulfide （H2S） and nitric oxide （NO） in signaling transduction of stomatal movement in response to ethylene in Viciafaba L. Ethylene treatment resulted in the dose-dependent stomatal closure under light, and this effect was blocked by the inhibitors of H2S biosynthesis in V. faba L. Additionally, ethylene induces H2S generation and increases L-/D-cysteine desulfhydrase （pyridoxalphosphate-dependent enzyme） activity in leaves of V. faba L. Inhibitors of H2S biosynthesis have no effect on the ethylene-induced stomatal closure, NO accumulation, and nitrate reductase （NR） activity in guard cells or leaves of II. faba L. Moreover, the ethylene-induced increase of H2S levels and L-/D- cysteine desulfhydrase activity declined when NO generation was inhibited. Therefore, we conclude that H2S and NO probably are involved in the signal transduction pathway of ethylene-induced stomatal closure. H2S may represent a novel component downstream of NO in the ethylene-induced stomatal movement in V. faba L.展开更多
Nitric oxide (NO) and Jasmonic acid (JA) are two key signaling molecules involved in many and diverse biological pathways in plants. Growing evidence suggested that NO signaling interacts with JA signaling. In this wo...Nitric oxide (NO) and Jasmonic acid (JA) are two key signaling molecules involved in many and diverse biological pathways in plants. Growing evidence suggested that NO signaling interacts with JA signaling. In this work, Our experiment showed that NO exists in guard cell of Vicia faba L., and NO is involved in signal transduction of JA- induced stomata closuring: (ⅰ) JA enhances NO synthesis in guard cell; (ⅱ) both JA and NO induced stomatal closure, and had dose response to their effects; (ⅲ) there are synergetic correlation between JA and lower NO concentration in regu- lation of stomatal movement; (ⅳ) JA-induced stomatal closure was largely prevented by 2-phenyl-4,4,5,5-tetrame- thylimidazoline-1-oxyl-3-oxide (PTIO), a specific NO scav- enger. An inhibitor of NO synthase (NOS) in mammalian cells, NG-nitro-L-Arg-methyl eater (L-NAME) also inhibits plant NOS, repressing JA-induced NO generation and JA-induced stomatal closure. We presumed that NO mainly comes from NOS after JA treatment.展开更多
The regulation of stomatal movement is one of the most important signaling networks in plants. The H+-ATPase at the plasma membrane of guard cells plays a critical role in the stomata opening, while there are some con...The regulation of stomatal movement is one of the most important signaling networks in plants. The H+-ATPase at the plasma membrane of guard cells plays a critical role in the stomata opening, while there are some conflicting results regarding the effectiveness of the plasma membrane H+-ATPase inhibitor, vanadate, in inhibiting stomata opening. We observed that 2 mmol/L vanadate hardly inhibited light-stimulated stomata opening in epidermal peels of Vicia faba L., but significantly inhibited dark- and ABA-induced stomatal closure. These results cannot be explained with the previous findings that H+-ATPase was inhibited by vanadate. In view of the fact that vanadate is an inhibitor of protein tyrosine phosphatases (PTPases), we investigated whether the stomatal movement regulated by vanadate is through the regulation of PTPase. As expected, phenylarsine oxide (PAO), a specific inhibitor of PTPase, has very similar effects and even more effective than vanadate. Typical PTPase activity was found in guard cells of V. faba; moreover, the phosphatase activity could be inhibited by both vanadate and PAO. These results not only provide a novel explanation for conflicting results about vanadate modulating stomatal movement, but also provide further evidence for the involvement of PTPases in modulating signal transduction of stomatal movement.展开更多
The effect of salicylic acid (SA) on stomatal aperture of 3～4 weeks’ Vicia faba L. under different conditions was investigated. It was shown that stomatal closure responded to SA after 3 hours treatment. The concent...The effect of salicylic acid (SA) on stomatal aperture of 3～4 weeks’ Vicia faba L. under different conditions was investigated. It was shown that stomatal closure responded to SA after 3 hours treatment. The concentration and pH of SA solution at different media affeeted SA to different extents. The degree of effects was concentration-dependent. With increasing SA concentration, the promotion effect on stomatal closure was enhanced. At 10 -3 mol L -1 SA exerted the most effect. The effect of SA was dependent on the pH of the solution. In the range of 10 -6 ～10 -3 mol L -1 as pH decreased, the effect of SA on stomatal closure was enhanced. SA 10 -4 mol L -1 in distilled water at pH 4 the percentage of inhibition on the stomatal aperture was 67%, while at pH 7 the percentage of inhibition was only 20%. The degrees of SA effect on stomatal aperture in distilled water、MES buffer solution and citric acid buffer solution were different. In MES buffer the effect of SA was lower than that in distilled water, or in citric acid buffer.展开更多
In animal cells, actions of acetylcholine depend on its binding to specific receptors. Activation of nicotinic acetylcholine receptor can affect the membrane permeability to ions directly. As concerned about acetylcho...In animal cells, actions of acetylcholine depend on its binding to specific receptors. Activation of nicotinic acetylcholine receptor can affect the membrane permeability to ions directly. As concerned about acetylcholine regulating stomatal movement, nicotinic acetylcholine receptor may also be involved. Experiments with nicotinic acetylcholine receptor antagonist (D\|tubocurarine) and agonist (nicotine) showed that nicotinic acetylcholine receptor was involved in acetylcholine inducing stomatal opening. In addition, these effects of D\|tubocurarine and nicotine on stomatal movement can only be found in medium containing K\++ while no effects can be found in medium containing Ca 2+ . The result by Western blotting showed that at least α\| and β\|subunits of nicotinic receptor existed in microsomes of guard cell protoplasts. It was found that the distribution of nicotinic acetylcholine receptor was mainly on the surface of protoplasts as shown by FITC labelled α\|bungarotoxin. The results above show that nicotinic acetylcholine receptor exists in guard cells and mediates acetylchlone to induce stomatal opening.展开更多
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, 100 mmol/...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, 100 mmol/L SA induced a rapid and striking NO increase in the cytosol of guard cells. This phenomenon was largely pre- vented by 200 mmol/L 2-phenyl-4,4,5,5-tetramethylimida- zoline-1-oxyl-3-oxide (PTIO), a specific NO scavenger, and 25 mmol/L NG-nitro-L-Arg-methyl eater (L-NAME), an in- hibitor 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-a]quinoxalin-1-one (ODQ), an inhibitor of guanylate cyclase, and nicotinamide, an antago- nist 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.展开更多
基金supported by the National Natural Science Foundation of China (30970288 and 31170237)the National Science Foundation of Shandong Province, China(ZR2010CM024)the Foundation of State Key Laboratory of Plant Physiology and Biochemistry, China(SKLPPBKF11001)
文摘Pharmacological, laser scanning confocal microscopic （LSCM）, and spectrophotographic approaches were used to study the roles of hydrogen sulfide （H2S） and nitric oxide （NO） in signaling transduction of stomatal movement in response to ethylene in Viciafaba L. Ethylene treatment resulted in the dose-dependent stomatal closure under light, and this effect was blocked by the inhibitors of H2S biosynthesis in V. faba L. Additionally, ethylene induces H2S generation and increases L-/D-cysteine desulfhydrase （pyridoxalphosphate-dependent enzyme） activity in leaves of V. faba L. Inhibitors of H2S biosynthesis have no effect on the ethylene-induced stomatal closure, NO accumulation, and nitrate reductase （NR） activity in guard cells or leaves of II. faba L. Moreover, the ethylene-induced increase of H2S levels and L-/D- cysteine desulfhydrase activity declined when NO generation was inhibited. Therefore, we conclude that H2S and NO probably are involved in the signal transduction pathway of ethylene-induced stomatal closure. H2S may represent a novel component downstream of NO in the ethylene-induced stomatal movement in V. faba L.
基金This work was supported by the National Natural Science Foundation of China(Grant No.30370141)Doctor's Fund of College&University Education(Grant No.2002001918)Fund of Education Department of Shandong Province(Grant No.J04C13).
文摘Nitric oxide (NO) and Jasmonic acid (JA) are two key signaling molecules involved in many and diverse biological pathways in plants. Growing evidence suggested that NO signaling interacts with JA signaling. In this work, Our experiment showed that NO exists in guard cell of Vicia faba L., and NO is involved in signal transduction of JA- induced stomata closuring: (ⅰ) JA enhances NO synthesis in guard cell; (ⅱ) both JA and NO induced stomatal closure, and had dose response to their effects; (ⅲ) there are synergetic correlation between JA and lower NO concentration in regu- lation of stomatal movement; (ⅳ) JA-induced stomatal closure was largely prevented by 2-phenyl-4,4,5,5-tetrame- thylimidazoline-1-oxyl-3-oxide (PTIO), a specific NO scav- enger. An inhibitor of NO synthase (NOS) in mammalian cells, NG-nitro-L-Arg-methyl eater (L-NAME) also inhibits plant NOS, repressing JA-induced NO generation and JA-induced stomatal closure. We presumed that NO mainly comes from NOS after JA treatment.
基金国家自然科学基金，Doctor's Fund of College and University Education
文摘The regulation of stomatal movement is one of the most important signaling networks in plants. The H+-ATPase at the plasma membrane of guard cells plays a critical role in the stomata opening, while there are some conflicting results regarding the effectiveness of the plasma membrane H+-ATPase inhibitor, vanadate, in inhibiting stomata opening. We observed that 2 mmol/L vanadate hardly inhibited light-stimulated stomata opening in epidermal peels of Vicia faba L., but significantly inhibited dark- and ABA-induced stomatal closure. These results cannot be explained with the previous findings that H+-ATPase was inhibited by vanadate. In view of the fact that vanadate is an inhibitor of protein tyrosine phosphatases (PTPases), we investigated whether the stomatal movement regulated by vanadate is through the regulation of PTPase. As expected, phenylarsine oxide (PAO), a specific inhibitor of PTPase, has very similar effects and even more effective than vanadate. Typical PTPase activity was found in guard cells of V. faba; moreover, the phosphatase activity could be inhibited by both vanadate and PAO. These results not only provide a novel explanation for conflicting results about vanadate modulating stomatal movement, but also provide further evidence for the involvement of PTPases in modulating signal transduction of stomatal movement.
文摘The effect of salicylic acid (SA) on stomatal aperture of 3～4 weeks’ Vicia faba L. under different conditions was investigated. It was shown that stomatal closure responded to SA after 3 hours treatment. The concentration and pH of SA solution at different media affeeted SA to different extents. The degree of effects was concentration-dependent. With increasing SA concentration, the promotion effect on stomatal closure was enhanced. At 10 -3 mol L -1 SA exerted the most effect. The effect of SA was dependent on the pH of the solution. In the range of 10 -6 ～10 -3 mol L -1 as pH decreased, the effect of SA on stomatal closure was enhanced. SA 10 -4 mol L -1 in distilled water at pH 4 the percentage of inhibition on the stomatal aperture was 67%, while at pH 7 the percentage of inhibition was only 20%. The degrees of SA effect on stomatal aperture in distilled water、MES buffer solution and citric acid buffer solution were different. In MES buffer the effect of SA was lower than that in distilled water, or in citric acid buffer.
文摘In animal cells, actions of acetylcholine depend on its binding to specific receptors. Activation of nicotinic acetylcholine receptor can affect the membrane permeability to ions directly. As concerned about acetylcholine regulating stomatal movement, nicotinic acetylcholine receptor may also be involved. Experiments with nicotinic acetylcholine receptor antagonist (D\|tubocurarine) and agonist (nicotine) showed that nicotinic acetylcholine receptor was involved in acetylcholine inducing stomatal opening. In addition, these effects of D\|tubocurarine and nicotine on stomatal movement can only be found in medium containing K\++ while no effects can be found in medium containing Ca 2+ . The result by Western blotting showed that at least α\| and β\|subunits of nicotinic receptor existed in microsomes of guard cell protoplasts. It was found that the distribution of nicotinic acetylcholine receptor was mainly on the surface of protoplasts as shown by FITC labelled α\|bungarotoxin. The results above show that nicotinic acetylcholine receptor exists in guard cells and mediates acetylchlone to induce stomatal opening.
基金the National KeyBasic Research Special Funds of China (Grant No. G1999011700)
文摘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, 100 mmol/L SA induced a rapid and striking NO increase in the cytosol of guard cells. This phenomenon was largely pre- vented by 200 mmol/L 2-phenyl-4,4,5,5-tetramethylimida- zoline-1-oxyl-3-oxide (PTIO), a specific NO scavenger, and 25 mmol/L NG-nitro-L-Arg-methyl eater (L-NAME), an in- hibitor 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-a]quinoxalin-1-one (ODQ), an inhibitor of guanylate cyclase, and nicotinamide, an antago- nist 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.
