Involvement of the Plasma Membrane Ca²⁺-ATPase in the Short-Term Response of Arabidopsis thaliana Cultured Cells to Oligogalacturonides

Abstract

Treatment of Arabidopsis thaliana cells with oligogalacturonides (OG) initiates a transient production of reactive oxygen species (ROS), the concentration of which in the medium peaks after about 20 min of treatment. The analysis of OG effects on Ca²⁺ fluxes shows that OG influence both Ca²⁺ influx and Ca²⁺ efflux (measured as ⁴⁵Ca²⁺ fluxes) in a complex way. During the first 10 - 15 min, OG stimulate Ca²⁺ influx and decrease its efflux, while at successive times of treatment, OG cause an increase of Ca²⁺ efflux and a slight decrease of its influx. Treatment with sub-µM concentrations of eosin yellow (EY), which selectively inhibits the Ca²⁺-ATPase of plasma membrane (PM), completely prevents the OG-induced increase in Ca²⁺ efflux. EY also suppresses the transient feature of OG-induced ROS accumulation, keeping the level of ROS in the medium high. The biochemical analysis of PM purified from OG-treated cells indicates that treatment with OG for 15 to 45 min induces a significant decrease in Ca²⁺-ATPase activation by exogenous calmodulin (CaM), and markedly increases the amount of CaM associated with the PM. During the same time span, OG do not influence the expression of At-ACA8, the main isoform of PM Ca²⁺-ATPase in suspension-cultured A. thaliana cells, and of CaM genes. Overall, the reported results demonstrate that the PM Ca²⁺-ATPase is involved in the response of plant cells to OG and is essential in regulation of the oxidative burst.

Key words

Arabidopsis thaliana cells - Ca²⁺ fluxes - calmodulin - plasma membrane Ca²⁺-ATPase - oligogalacturonides - oxidative burst

References

• 1 Askerlund P., Sommarin M.. Calcium efflux transporters in higher plants. Smallwood, M., Knox, J. P., and Bowles, D. J., eds. Membranes: Specialized Functions in Plants. Oxford, UK; BIOS Scientific Publishers Ltd (1996): 281-299
• 2 Atkinson M. M., Keppler L. D., Orlandi E. W., Baker C. J., Mischke C. F.. Involvement of plasma membrane calcium influx in bacterial induction of the K⁺/H⁺ and hypersensitive responses in tobacco.  Plant Physiol.. (1990);  92 215-221
• 3 Bach M., Schnitzler I. P., Seitz H. U.. Elicitor-induced changes in Ca²⁺ influx, K⁺ efflux, and 4-hydroxybenzoic acid synthesis in protoplasts of Daucus carota L..  Plant Physiol.. (1993);  103 407-412
• 4 Beffagna N., Romani G., Sforza M. C.. H⁺ fluxes at plasmalemma level: in vivo evidences for a significant contribution of the Ca²⁺- ATPase and for the involvement of its activity in the abscisic acid - induced changes in Egeria densa leaves.  Plant Biol.. (2000);  2 168-175
• 5 Blume B., Nurnberger T., Nass N., Scheel D.. Receptor-mediated increase in cytoplasmic free calcium required for activation of pathogen defence in parsley.  Plant Cell. (2000);  12 1425-1440
• 6 Blumwald E., Aharon G. S., Lam B. C. H.. Early signal transduction pathways in plant-pathogen interactions.  Trends Plant Sci.. (1998);  3 342-346
• 7 Bonza C., Carnelli A., De Michelis M. I., Rasi-Caldogno F.. Purification of the plasma membrane Ca²⁺-ATPase from radish seedlings by calmodulin-agarose affinity chromatography.  Plant Physiol.. (1998);  116 845-851
• 8 Bonza M. C., Morandini P., Luoni L., Geisler M., Palmgren M. G., De Michelis M. I.. Cloning of At-ACA8, a plasma membrane localized, calmodulin regulated calcium-ATPase of Arabidopsis thaliana and localization of its calmodulin binding domain at the N-terminus.  Plant Physiol.. (2000);  123 1495-1505
• 9 Braam J., Davis R. W.. Rain-, wind- and touch-induced expression of calmodulin and calmodulin-related genes in Arabidopsis. .  Cell. (1990);  60 357-364
• 10 Brandt P. C., Vanaman T. C.. Calmodulin and ion flux regulation. Van Eldi, K. L. and Watterson, D. M., eds. Calmodulin and Signal Transduction. Harcourt, USA; Brace & Company Publishers Academic Press (1998): 397-471
• 11 Carnelli A., De Michelis M. I., Rasi-Caldogno F.. Plasma membrane Ca²⁺- ATPase of radish seedlings: I. Biochemical characteristics using ITP as substrate.  Plant Physiol.. (1992);  98 1196-1201
• 12 Cervone F., Costorne R., Leckie F. D., De Lorenzo G.. Perception of fungal elicitors and signal transduction. Aducci, P., ed. Signal Transduction in Plants. Basel; Birkhauser (1997): 153-177
• 13 Chandra S., Low P. S.. Measurement of Ca²⁺ fluxes during elecitation of the oxidative burst in aqueorin-transformed tobacco cells.  J. Biol. Chem.. (1997);  272 28274-28280
• 14 Collinge M., Trewavas A. J.. The location of calmodulin in the pea plasma membrane.  J. Biol. Chem.. (1989);  262 8865-8872
• 15 De Lorenzo G., D'Ovidio R., Cervone F.. The role of polygalacturonase-inhibiting proteins (PGIPs) in defense against pathogenic fungi.  Annu. Rev. Phytopathol.. (2001);  39 313-335
• 16 De Michelis M. I., Carnelli A., Rasi-Caldogno F.. The Ca-pump of the plasma membrane of Arabidopsis thaliana: characteristics and sensitivity to fluorescein derivatives.  Bot. Acta. (1993);  106 20-25
• 17 Ebel J., Cosio E. G.. Elicitors of plant defense responses.  Int. Rev. Cytol.. (1994);  148 1-36
• 18 Evans D. E., Askerlund P., Boyce J. M., Briars S. A., Coates J., Cooke D. T., Theodouolu F. L.. Studies on higher plant calmodulin stimulated ATPase. Cooke, D. T. and Clarkson, D. T., eds. Transport and Receptor Proteins of Plant Membranes. New York; Plenum Press (1992): 39-53
• 19 Felle H., Tretyn A., Wagner G.. The role of the plasma-membrane Ca²⁺-ATPase in Ca²⁺ homeostasis in Sinapis alba root hairs.  Planta. (1992);  188 306-313
• 20 Geisler M., Axelsen K., Harper J. F., Palmgren M. G.. Molecular aspects of higher plant P-type Ca²⁺-ATPases.  Biochim. Biophys. Acta. (2000);  1465 52-78
• 21 Gelli A., Higgins V. J., Blumwald E.. Activation of plasma membrane Ca²⁺-permeable channels by race-specific fungal elicitors.  Plant Physiol.. (1997);  113 269-279
• 22 Heo W. D., Lee S. H., Kim M. C., Kim J. C., Chung W. S., Chun H. J., Lee K. J., Park C. Y., Park H. C., Choi J. Y., Cho M. J.. Involvement of specific calmodulin isoforms in salicylic acid-independent activation of plant disease resistance responses.  Proc. Natl. Acad. Sci. USA. (1999);  96 766-771
• 23 Jiang Z-Y., Woolland A. C. S., Wolff S. P.. Hydrogen peroxide production during experimental protein glycation.  FEBS Lett.. (1990);  268 69-71
• 24 Knight H.. Calcium signalling during abiotic stress in plants.  Int. Rev. Cytol.. (2000);  195 269-324
• 25 Laemmli U. K.. Cleavage of structural proteins during the assembly of the head of the bacteriophage T4.  Nature. (1970);  227 680-685
• 26 Lam C.-H. B., Xing T., Higgins V. J., Blumwald E.. Effect of race-specific elicitors of Cladosporium fulvum on the tomato plasma membrane Ca²⁺-ATPase.  Physiol. and Mol. Plant Pathol.. (1998);  52 309-321
• 27 Lamb C., Dixon R. A.. The oxidative burst in plant-disease resistance.  Ann. Rev. Plant Physiol. Plant Biol. Mol.. (1997);  48 251-275
• 28 Lecourieux D., Mazars C., Pauly N., Ranjeva R., Pugin A.. Analysis and effects of cytosolic free calcium increases in response to elicitors in Nicotiana plumbaginifolia cells.  Plant Cell. (2002);  14 2627-2641
• 29 Luoni L., Bonza M. C., De Michelis M. I.. H⁺/Ca²⁺ exchange driven by the plasma membrane Ca²⁺-ATPase of Arabidopsis thaliana reconstituted in proteoliposomes after calmodulin-affinity purification.  FEBS Lett.. (2000);  482 225-230
• 30 Markwell M. A. K., Haas S. M., Bicher L. L., Tolbert N. E.. Modification of the Lowry procedure to simplify protein determination in membrane and lipoprotein samples.  Anal. Biochem.. (1978);  87 206-210
• 31 Mathieu Y., Kurkdjian A., Xia H., Guern J., Koller A., Spiro M. D., O'Neill M., Albersheim P., Darvill A.. Membrane responses induced by oligogalacturonides in suspension cultured tobacco cells.  Plant J.. (1991);  1 333-343
• 32 Messiaen J., Van Cutsem P.. Pectic signal transduction in carrot cells: membrane, cytosolic and nuclear responses induced by oligogalacturonides.  Plant Cell Physiol.. (1994);  35 677-689
• 33 Messiaen J., Read N. D., Van Cutsem P., Trewavas A. J.. Cell wall oligogalacturonides increase cytosolic free Ca²⁺ in carrots protoplasts.  J. Cell Sci.. (1993);  104 365-371
• 34 Murashige T., Skoog F.. A revised medium for rapid growth and bio assay with tobacco tissue cultures.  Physiol. Plant.. (1962);  15 473-497
• 35 Navazio L., Moscatiello R., Bellincampi D., Baldan B., Meggio F., Brini M., Bowler C., Mariani P.. The role of calcium in oligogalacturonide-activated signalling in soybean cells.  Planta. (2002);  215 596-605
• 36 Olbe M., Sommarin M.. The spinach plasma membrane calcium pump is a 120 kDa polypeptide regulated by calmodulin binding to a terminal region.  Physiol. Plant.. (1998);  103 35-44
• 37 Olivari C., Meanti C., De Michelis M. I., Rasi-Caldogno F.. Fusicoccin binding to its plasma membrane receptor and the activation of the plasma membrane H⁺-ATPase. IV. Fusicoccin induces the association between the plasma membrane H⁺-ATPase and the fusicoccin receptor.  Plant Physiol.. (1998);  116 529-537
• 38 Rasi-Caldogno F., Carnelli A., De Michelis M. I.. Controlled proteolysis activates the plasma membrane Ca²⁺-pump of higher plants.  Plant Physiol.. (1993);  103 385-390
• 39 Rasi-Caldogno F., Carnelli A., De Michelis M. I.. Identification of the plasma membrane Ca²⁺-ATPase and of its autoinhibitory domain.  Plant Physiol.. (1995);  108 105-113
• 40 Rasi-Caldogno F., Pugliarello M. C., De Michelis M. I.. The Ca²⁺ transport ATPase of plant plasma membrane catalyzes a nH⁺/Ca²⁺ exchange.  Plant Physiol.. (1987);  83 994-1000
• 41 Robinson C., Larsson C., Buckhout T. J.. Identification of a calmodulin-stimulated (Ca + Mg)-ATPase in a plasma membrane fraction isolated from maize (Zea mays). .  Physiol. Plant.. (1988);  72 177-194
• 42 Sanders D., Pelloux J., Brownlee C., Harper J. F.. Calcium at the crossroads of signaling.  Plant Cell Supplement. (2002);  2002 S401-S417
• 43 Sze H., Liang F., Hwang I., Curran A. C., Harper J. F.. Diversity and regulation of plant Ca²⁺- pumps: Insights from expression in yeast.  Annu. Rev. Plant Physiol. Plant Mol. Biol.. (2000);  51 433-462
• 44 Tavernier E., Wendehenne D., Blein J.-P., Pugin A.. Involvement of free calcium in action of cryptogein, a proteinaceous elicitor of hypersensitive reaction in tobacco cells.  Plant Physiol.. (1995);  48 83-89
• 45 Trewavas A. J.. Le calcium, c'est la vie: calcium makes waves.  Plant Physiol.. (1999);  120 1-6
• 46 Williams L. E., Schueler S. B., Briskin D. P.. Further characterization of the red beet plasma membrane Ca²⁺-ATPase using GTP as an alternative substrate.  Plant Physiol.. (1990);  92 747-754
• 47 Zielinski R. E.. Calmodulin and calmodulin-binding proteins in plants.  Annu. Rev. Plant Physiol. Plant. Mol. Biol.. (1998);  49 697-725
• 48 Zimmermann S., Nurnberger T., Frachisse J. M., Wirtz W., Guern J., Hedrich R., Scheel D.. Receptor-mediated activation of a plant Ca²⁺-permeable ion channel involved in pathogen defence.  Proc. Natl. Acad. Sci. USA. (1997);  94 2751-2755

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