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Cytoplasmic acidification as an early phosphorylation-dependent response of tobacco cells to elicitors

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Abstract

Elicitor-induced cytoplasmic pH changes of tobacco (Nicotiana tabacum L. cv. Xanthi) cells grown in suspension cultures were explored under a variety of conditions by using a flexible technique based on the distribution of [14C] benzoic acid between the intracellular and extracellular compartments. Comparison of data obtained by this technique and by 31P-nuclear magnetic resonance spectrometry qualifies the benzoic acid distribution method as a convenient and reliable way to probe cytoplasmic pH variations. Various elicitors shown to induce several defense-related responses in tobacco cells, namely oligogalacturonides of degree of polymerization 7–20, pectolyase from Aspergillus japonicus, Phytophthora megasperma crude elicitors and purified cryptogein, triggered cytoplasmic acidifications differing in intensity and kinetics according to the signal molecule. In contrast, no changes in cytoplasmic protons and external pH were observed in cells treated with short galacturonide oligomers, or with soybean-specific hepta β-glucoside from P. megasperma, which are devoid of elicitor activity in tobacco cells. The oligogalacturonide-induced cytoplasmic acidification was inhibited by two structurally unrelated protein kinase inhibitors, staurosporine and 6-dimethylaminopurine, which both reduced the external alkalinization response to the elicitor. The protein phosphatase inhibitor calyculin A alone behaved as an elicitormimicking molecule in triggering cytoplasmic acidification, again associated with extracellular alkalinization. These results indicate that the increase in the cytoplasmic concentration of protons may be considered as a common early intracellular response of tobacco cells to elicitors, associated with the extracellular alkalinization response and controlled by protein phosphorylation.

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Abbreviations

BA(H):

benzoic acid (protonated form)

6-DMAP:

6-dimethylaminopurine

DP:

degree of polymerization

Mes:

2-(N-morpholino)ethanesulfonic acid

OG:

oligogalacturonide

pHc:

cytoplasmic pH

31P-NMR:

nuclear magnetic resonance spectroscopy of 31P atoms

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Authors and Affiliations

  1. Institut des Sciences Végétales, CNRS, Avenue de la Terrasse, F91198, Gif-sur-Yvette Cedex, France

    Yves Mathieu, Danielle Lapous, Sébastien Thomine, Christiane Laurière & Jean Guern

Authors
  1. Yves Mathieu
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  2. Danielle Lapous
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  3. Sébastien Thomine
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  4. Christiane Laurière
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  5. Jean Guern
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Additional information

The authors thank P. Albersheim (CCRC, Athens, Georgia, USA) for providing the purified oligogalacturonides and the hepta β-glucoside and P. Ricci (INRA, Antibes, France) for providing the purified cryptogein.

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Mathieu, Y., Lapous, D., Thomine, S. et al. Cytoplasmic acidification as an early phosphorylation-dependent response of tobacco cells to elicitors. Planta 199, 416–424 (1996). https://doi.org/10.1007/BF00195734

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