Crosstalk of Nitric Oxide with Ca2+ in Stomatal Movement in Vicia faba Guard Cells

Ma, Y., She, X. and Yang, S. 2013. Cytosolic alkalization-mediated H 2 O 2 and NO production are involved in darkness-induced stomatal closure in Vicia faba. Can. J. Plant Sci. 93: 119–130. Darkness raised cytosolic pH, hydrogen peroxide (H2O2) and nitric oxide (NO) levels in guard cells while inducing Vicia faba stomatal closure. These darkness effects were prevented by weak acid butyric acid, H2O2 modulators ascorbic acid (ASA), catalase (CAT), diphenyleneiodonium (DPI) and NO modulators 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (c-PTIO), NG-nitro-L-arg-methyl ester (L-NAME) respectively. The data suggest that cytosolic alkalization, H2O2 and NO all participate in darkness-induced stomatal closure. During darkness treatment, pH rise became noticeable at 10 min and peaked at 25 min, while H2O2 and NO production increased significantly at 20 min and reached their maximums at 40 min. The H2O2 and NO levels were increased by methylamine in light and decreased by butyric acid in darkness. The results show that cytosolic alkalization induces H2O2 and NO production. ASA, CAT and DPI suppressed NO production by methylamine, c-PTIO and L-NAME prevented H2O2 generation by methylamine. Calcium chelator 1,2-bis (2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid acetoxymethyl ester (BAPTA-AM) and 1,2-bis(2-aminophenoxy)ethane-N,N,N′,N′-tetraacetic acid (BAPTA) restricted darkness-induced alkalization, H2O2 and NO production and stomatal closure. We suggest that cytosolic alkalization is necessary for H2O2 and NO production during darkness-induced stomatal closure. H2O2 mediates NO synthesis by alkalization, and vice versa. Calcium may act upstream of cytosolic alkalization, H2O2 and NO production, besides its known action downstream of H2O2 and NO.

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Ethylene inhibits darkness-induced stomatal closure by scavenging nitric oxide in guard cells of Vicia faba

Functional Plant Biology ◽

10.1071/fp11055 ◽

2011 ◽

Vol 38 (10) ◽

pp. 767 ◽

Cited By ~ 10

Author(s):

Xi-Gui Song ◽

Xiao-Ping She ◽

Juan Wang ◽

Yi-Chao Sun

Keyword(s):

Nitric Oxide ◽

Vicia Faba ◽

Plant Hormone ◽

No Reduction ◽

Enzyme Commission ◽

Stomatal Closure ◽

Guard Cells ◽

Stomatal Opening ◽

Physiological Data ◽

Oxide Synthase

The plant hormone ethylene regulates many aspects of plant growth and development. Despite the well-known relationship between ethylene and stress signalling, the involvement of ethylene in regulating stomatal movement is not completely explored. Here, the role and association between nitric oxide (NO) reduction and the inhibition of darkness-induced stomatal closure by ethylene was studied. Physiological data are provided that both ethylene-releasing compound 2-chloroethylene phosphonic acid (ethephon, ETH) and 1-aminocyclopropane-1-carboxylic acid (ACC), the immediate precursor of ethylene, reduced the levels of NO in Vicia faba L. guard cells, and then induced stomatal opening in darkness. In addition, ACC and ETH not only reduced NO levels in guard cells caused by exogenous NO (derived from sodium nitroprusside, SNP) in light, but also abolished NO that had been generated during a dark period and promoted stomatal opening. Interestingly, 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide (cPTIO) and hemoglobin (Hb), NO scavenger and the potent scavenger of NO/carbon monoxide (CO), respectively, also reduced NO levels by SNP and darkness. However, the above-mentioned effects of ACC and ETH were dissimilar to that of nitric oxide synthase (enzyme commission 1.14.13.39) inhibitor NG-nitro-L-Arg-methyl ester (L-NAME), which could neither reduce NO levels by SNP nor abolish NO that had been generated in the dark. Thus, it is concluded that ethylene reduces the levels of NO in V. faba guard cells via a pattern of NO scavenging, then induces stomatal opening in the dark.

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Role and interrelationship of PTPases and H2O2 in light/dark-regulated stomatal movement in Vicia faba

Australian Journal of Botany ◽

10.1071/bt09017 ◽

2009 ◽

Vol 57 (6) ◽

pp. 486 ◽

Cited By ~ 1

Author(s):

Yuanhua Zhang ◽

Xiaoping She ◽

Guangbin Zhang

Keyword(s):

Vicia Faba ◽

Laser Scanning ◽

Stomatal Closure ◽

Specific Inhibitor ◽

Guard Cells ◽

Stomatal Aperture ◽

Stomatal Movement ◽

Obvious Effect ◽

Exogenous H2o2 ◽

Ptpase Activity

Role and interrelationship of protein tyrosine phosphatases (PTPases) and H2O2 in light/dark-regulated stomatal movement in Vicia faba were investigated by epidermal strip bioassay, laser-scanning confocal microscopy and assays of PTPase activity. Our results indicate that phenylarsine oxide (PAO), a specific inhibitor of PTPases, ascorbic acid (ASA), an important reducing substrate for H2O2 removal, and catalase (CAT), one of the H2O2 scavenging enzymes, did not cause any change of stomatal aperture in light, but remarkably prevented dark-induced stomatal closure. Exogenous H2O2 had no obvious effect on stomatal aperture in the dark, but significantly induced stomatal closure in light. Both PTPase activity in epidermal strips and endogenous H2O2 level in guard cells in the dark were higher than those in light. The results showed that both PTPases and H2O2 mediate light/dark-regulated stomatal movement, that dark-induced stomatal closure requires the activation of PTPases and the enhancement of H2O2 levels in guard cells, and stomatal opening caused by light is associated with the inactivation of PTPases and the reduction of H2O2 levels in guard cells. Additionally, like ASA and CAT, PAO abolished dark-, exogenous H2O2-induced stomatal closure and dichlorofluorescein fluorescence in guard cells, indicating that activation of PTPases can enhance H2O2 levels probably via suppressing the decrease of H2O2 levels in guard cells. On the other hand, similar to PAO, ASA and CAT evidently prevented dark-, exogenous H2O2-induced stomatal closure and obviously inactivated PTPases in the dark. However, exogenous H2O2 significantly activated PTPases in light. The results show that H2O2 can induce activation of PTPases. Taken together, the present results provide evidence that both H2O2 and PTPases are involved in light/dark-regulated stomatal movement, and the interaction between H2O2 and PTPases plays a pivotal role in light/dark signal transduction process in guard cells.

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