Logo of plntphysLink to Publisher's site
Plant Physiol. 1996 Aug; 111(4): 1031–1042.
PMCID: PMC160976

Changes in Stomatal Behavior and Guard Cell Cytosolic Free Calcium in Response to Oxidative Stress.


We have investigated the cellular basis for the effects of oxidative stress on stomatal behavior using stomatal bioassay and ratio photometric techniques. Two oxidative treatments were employed in this study: (a) methyl viologen, which generates superoxide radicals, and (b) H2O2. Both methyl viologen and H2O2 inhibited stomatal opening and promoted stomatal closure. At concentrations [less than or equal to]10-5 M, the effects of methyl viologen and H2O2 on stomatal behavior were reversible and were abolished by 2 mM EGTA or 10 [mu]M verapamil. In addition, at 10-5 M, i.e. the maximum concentration at which the effects of the treatments were prevented by EGTA or verapamil, methyl viologen and H2O2 caused an increase in guard cell cytosolic free Ca2+ ([Ca2+]i), which was abolished in the presence of EGTA. Therefore, at low concentrations of methyl viologen and H2O2, removal of extracellular Ca2+ prevented both the oxidative stress-induced changes in stomatal aperture and the associated increases in [Ca2+]i. This suggests that in this concentration range the effects of the treatments are Ca2+-dependent and are mediated by changes in [Ca2+]i. In contrast, at concentrations of methyl viologan and H2O2 > 10-5 M, EGTA and verapamil had no effect. However, in this concentration range the effects of the treatments were irreversible and correlated with a marked reduction in membrane integrity and guard cell viability. This suggests that at high concentrations the effects of methyl viologen and H2O2 may be due to changes in membrane integrity. The implications of oxidative stress-induced increases in [Ca2+]i and the possible disruption of guard-cell Ca2+ homeostasis are discussed in relation to the processes of Ca2+-based signal transduction in stomatal guard cells and the control of stomatal aperture.

Full Text

The Full Text of this article is available as a PDF (2.1M).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Allan AC, Fricker MD, Ward JL, Beale MH, Trewavas AJ. Two Transduction Pathways Mediate Rapid Effects of Abscisic Acid in Commelina Guard Cells. Plant Cell. 1994 Sep;6(9):1319–1328. [PMC free article] [PubMed]
  • Assmann SM. Signal transduction in guard cells. Annu Rev Cell Biol. 1993;9:345–375. [PubMed]
  • Babbs CF, Pham JA, Coolbaugh RC. Lethal hydroxyl radical production in paraquat-treated plants. Plant Physiol. 1989 Aug;90(4):1267–1270. [PMC free article] [PubMed]
  • Bradley DJ, Kjellbom P, Lamb CJ. Elicitor- and wound-induced oxidative cross-linking of a proline-rich plant cell wall protein: a novel, rapid defense response. Cell. 1992 Jul 10;70(1):21–30. [PubMed]
  • Cadenas E. Biochemistry of oxygen toxicity. Annu Rev Biochem. 1989;58:79–110. [PubMed]
  • Castillo FJ, Heath RL. Ca transport in membrane vesicles from pinto bean leaves and its alteration after ozone exposure. Plant Physiol. 1990 Oct;94(2):788–795. [PMC free article] [PubMed]
  • Chen Z, Malamy J, Henning J, Conrath U, Sánchez-Casas P, Silva H, Ricigliano J, Klessig DK. Induction, modification, and transduction of the salicylic acid signal in plant defense responses. Proc Natl Acad Sci U S A. 1995 May 9;92(10):4134–4137. [PMC free article] [PubMed]
  • Chen Z, Silva H, Klessig DF. Active oxygen species in the induction of plant systemic acquired resistance by salicylic acid. Science. 1993 Dec 17;262(5141):1883–1886. [PubMed]
  • Chien KR, Peau RG, Farber JL. Ischemic myocardial cell injury. Prevention by chlorpromazine of an accelerated phospholipid degradation and associated membrane dysfunction. Am J Pathol. 1979 Dec;97(3):505–529. [PMC free article] [PubMed]
  • Cyr RJ. Microtubules in plant morphogenesis: role of the cortical array. Annu Rev Cell Biol. 1994;10:153–180. [PubMed]
  • Evans DE. PM-type calcium pumps are associated with higher plant cell intracellular membranes. Cell Calcium. 1994 Mar;15(3):241–246. [PubMed]
  • Gilroy S, Fricker MD, Read ND, Trewavas AJ. Role of Calcium in Signal Transduction of Commelina Guard Cells. Plant Cell. 1991 Apr;3(4):333–344. [PMC free article] [PubMed]
  • Gilroy S, Read ND, Trewavas AJ. Elevation of cytoplasmic calcium by caged calcium or caged inositol triphosphate initiates stomatal closure. Nature. 1990 Aug 23;346(6286):769–771. [PubMed]
  • Irving HR, Gehring CA, Parish RW. Changes in cytosolic pH and calcium of guard cells precede stomatal movements. Proc Natl Acad Sci U S A. 1992 Mar 1;89(5):1790–1794. [PMC free article] [PubMed]
  • Johnson CH, Knight MR, Kondo T, Masson P, Sedbrook J, Haley A, Trewavas A. Circadian oscillations of cytosolic and chloroplastic free calcium in plants. Science. 1995 Sep 29;269(5232):1863–1865. [PubMed]
  • Kim M, Hepler PK, Eun SO, Ha KS, Lee Y. Actin Filaments in Mature Guard Cells Are Radially Distributed and Involved in Stomatal Movement. Plant Physiol. 1995 Nov;109(3):1077–1084. [PMC free article] [PubMed]
  • Levine A, Tenhaken R, Dixon R, Lamb C. H2O2 from the oxidative burst orchestrates the plant hypersensitive disease resistance response. Cell. 1994 Nov 18;79(4):583–593. [PubMed]
  • McAinsh MR, Brownlee C, Hetherington AM. Visualizing Changes in Cytosolic-Free Ca2+ during the Response of Stomatal Guard Cells to Abscisic Acid. Plant Cell. 1992 Sep;4(9):1113–1122. [PMC free article] [PubMed]
  • McAinsh MR, Webb AAR, Taylor JE, Hetherington AM. Stimulus-Induced Oscillations in Guard Cell Cytosolic Free Calcium. Plant Cell. 1995 Aug;7(8):1207–1219. [PMC free article] [PubMed]
  • Mirabelli F, Salis A, Vairetti M, Bellomo G, Thor H, Orrenius S. Cytoskeletal alterations in human platelets exposed to oxidative stress are mediated by oxidative and Ca2+-dependent mechanisms. Arch Biochem Biophys. 1989 May 1;270(2):478–488. [PubMed]
  • Munns PL, Leach KL. Two novel antioxidants, U74006F and U78517F, inhibit oxidant-stimulated calcium influx. Free Radic Biol Med. 1995 Mar;18(3):467–478. [PubMed]
  • Murata M, Monden M, Umeshita K, Nakano H, Kanai T, Gotoh M, Mori T. Role of intracellular calcium in superoxide-induced hepatocyte injury. Hepatology. 1994 May;19(5):1223–1228. [PubMed]
  • Orrenius S, Burkitt MJ, Kass GE, Dypbukt JM, Nicotera P. Calcium ions and oxidative cell injury. Ann Neurol. 1992;32 (Suppl):S33–S42. [PubMed]
  • Poovaiah BW, Reddy AS. Calcium and signal transduction in plants. CRC Crit Rev Plant Sci. 1993;12(3):185–211. [PubMed]
  • Price AH, Taylor A, Ripley SJ, Griffiths A, Trewavas AJ, Knight MR. Oxidative Signals in Tobacco Increase Cytosolic Calcium. Plant Cell. 1994 Sep;6(9):1301–1310. [PMC free article] [PubMed]
  • Schroeder JI, Hagiwara S. Repetitive increases in cytosolic Ca2+ of guard cells by abscisic acid activation of nonselective Ca2+ permeable channels. Proc Natl Acad Sci U S A. 1990 Dec;87(23):9305–9309. [PMC free article] [PubMed]
  • Shimazaki K, Zeiger E. Cyclic and Noncyclic Photophosphorylation in Isolated Guard Cell Chloroplasts from Vicia faba L. Plant Physiol. 1985 Jun;78(2):211–214. [PMC free article] [PubMed]
  • Toraason M, Heinroth-Hoffmann I, Richards D, Woolery M, Hoffmann P. H2O2-induced oxidative injury in rat cardiac myocytes is not potentiated by 1,1,1-trichloroethane, carbon tetrachloride, or halothane. J Toxicol Environ Health. 1994 Apr;41(4):489–507. [PubMed]

Articles from Plant Physiology are provided here courtesy of American Society of Plant Biologists


Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...


  • MedGen
    Related information in MedGen
  • PubMed
    PubMed citations for these articles

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...