Oxidative Signals in Tobacco Increase Cytosolic Calcium

Adam Huw Price, A Taylor, S J Ripley, A Griffiths, A J Trewavas, M R Knight

Research output: Contribution to journalArticle

296 Citations (Scopus)

Abstract

Tobacco (Nicotiana plumbaginifolia) seedlings genetically transformed to express apoaequorin were incubated in h-coelenterazine to reconstitute the calcium-sensitive luminescent protein aequorin. Treatment of these seedlings with hydrogen peroxide resulted in a transient burst of calcium-dependent luminescence lasting several minutes. Even though the hydrogen peroxide stimulus was persistent, the change in cytosolic free calcium concentration ([Ca2+](cyt)) was transient, suggesting the presence of a refractory period. When seedlings were pretreated with hydrogen peroxide, there was no increase in [Ca2+](cyt) upon a second application, which confirmed the refractory character of the response. Only when the two treatments were separated by 4 to 8 hr was full responsiveness recovered. However, treatment with hydrogen peroxide did not inhibit mobilization of [Ca2+](cyt) induced by either cold shock or touching, suggesting that these three signals mobilize different pools of intracellular calcium. To examine whether [Ca2+](cyt) is regulated by the redox state of the cytoplasm, we pretreated seedlings with buthionine sulfoximine (to modify cellular glutathione levels) and inhibitors of ascorbate peroxidase. These inhibitors modify the hydrogen peroxide-induced transients in [Ca2+](cyt), which is consistent with their effects on the cellular prooxidant/antioxidant ratio. Treatment with hydrogen peroxide that elicited [Ca2+](cyt) increases also brought about a reduction in superoxide dismutase enzyme activity. This reduction could be reversed by treatment with the calcium channel blocker lanthanum. This indicates that there is a role for calcium in plant responses to oxidative stress.

Original languageEnglish
Pages (from-to)1301-1310
Number of pages10
JournalThe Plant Cell
Volume6
Issue number9
Publication statusPublished - Sep 1994

Keywords

  • PLANT CHEMI-LUMINESCENCE
  • CELL
  • INHIBITION
  • DEFENSE

Cite this

Price, A. H., Taylor, A., Ripley, S. J., Griffiths, A., Trewavas, A. J., & Knight, M. R. (1994). Oxidative Signals in Tobacco Increase Cytosolic Calcium. The Plant Cell, 6(9), 1301-1310.

Oxidative Signals in Tobacco Increase Cytosolic Calcium. / Price, Adam Huw; Taylor, A ; Ripley, S J ; Griffiths, A ; Trewavas, A J ; Knight, M R .

In: The Plant Cell, Vol. 6, No. 9, 09.1994, p. 1301-1310.

Research output: Contribution to journalArticle

Price, AH, Taylor, A, Ripley, SJ, Griffiths, A, Trewavas, AJ & Knight, MR 1994, 'Oxidative Signals in Tobacco Increase Cytosolic Calcium', The Plant Cell, vol. 6, no. 9, pp. 1301-1310.
Price AH, Taylor A, Ripley SJ, Griffiths A, Trewavas AJ, Knight MR. Oxidative Signals in Tobacco Increase Cytosolic Calcium. The Plant Cell. 1994 Sep;6(9):1301-1310.
Price, Adam Huw ; Taylor, A ; Ripley, S J ; Griffiths, A ; Trewavas, A J ; Knight, M R . / Oxidative Signals in Tobacco Increase Cytosolic Calcium. In: The Plant Cell. 1994 ; Vol. 6, No. 9. pp. 1301-1310.
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AB - Tobacco (Nicotiana plumbaginifolia) seedlings genetically transformed to express apoaequorin were incubated in h-coelenterazine to reconstitute the calcium-sensitive luminescent protein aequorin. Treatment of these seedlings with hydrogen peroxide resulted in a transient burst of calcium-dependent luminescence lasting several minutes. Even though the hydrogen peroxide stimulus was persistent, the change in cytosolic free calcium concentration ([Ca2+](cyt)) was transient, suggesting the presence of a refractory period. When seedlings were pretreated with hydrogen peroxide, there was no increase in [Ca2+](cyt) upon a second application, which confirmed the refractory character of the response. Only when the two treatments were separated by 4 to 8 hr was full responsiveness recovered. However, treatment with hydrogen peroxide did not inhibit mobilization of [Ca2+](cyt) induced by either cold shock or touching, suggesting that these three signals mobilize different pools of intracellular calcium. To examine whether [Ca2+](cyt) is regulated by the redox state of the cytoplasm, we pretreated seedlings with buthionine sulfoximine (to modify cellular glutathione levels) and inhibitors of ascorbate peroxidase. These inhibitors modify the hydrogen peroxide-induced transients in [Ca2+](cyt), which is consistent with their effects on the cellular prooxidant/antioxidant ratio. Treatment with hydrogen peroxide that elicited [Ca2+](cyt) increases also brought about a reduction in superoxide dismutase enzyme activity. This reduction could be reversed by treatment with the calcium channel blocker lanthanum. This indicates that there is a role for calcium in plant responses to oxidative stress.

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