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1.
Biosci Biotechnol Biochem ; 83(2): 318-321, 2019 Feb.
Article in English | MEDLINE | ID: mdl-30345896

ABSTRACT

Dimethyl sulfoxide (DMSO) is a dipolar aprotic solvent widely used in biological assays. Here, we observed that DMSO enhanced the hypo-osmotically induced increases in the concentration of Ca2+ in cytosolic and nucleic compartments in the transgenic cell-lines of tobacco (BY-2) expressing aequorin.


Subject(s)
Calcium/metabolism , Cell Nucleus/metabolism , Cytosol/metabolism , Dimethyl Sulfoxide/administration & dosage , Nicotiana/metabolism , Osmotic Pressure , Aequorin/metabolism , Cell Compartmentation , Luminescence , Plants, Genetically Modified , Nicotiana/cytology
2.
Ann Bot ; 122(5): 849-860, 2018 11 03.
Article in English | MEDLINE | ID: mdl-29579139

ABSTRACT

Background and Aims: Methanol is a volatile organic compound released from plants through the action of pectin methylesterases (PMEs), which demethylesterify cell wall pectins. Plant PMEs play a role in developmental processes but also in responses to herbivory and infection by fungal or bacterial pathogens. However, molecular mechanisms that explain how methanol could affect plant defences remain poorly understood. Methods: Using cultured cells and seedlings from Arabidopsis thaliana and tobacco BY2 expressing the apoaequorin gene, allowing quantification of cytosolic Ca2+, a reactive oxygen species (ROS) probe (CLA, Cypridina luciferin analogue) and electrophysiological techniques, we followed early plant cell responses to exogenously supplied methanol applied as a liquid or as volatile. Key Results: Methanol induces cytosolic Ca2+ variations that involve Ca2+ influx through the plasma membrane and Ca2+ release from internal stores. Our data further suggest that these Ca2+ variations could interact with different ROS and support a signalling pathway leading to well known plant responses to pathogens such as plasma membrane depolarization through anion channel regulation and ethylene synthesis. Conclusions: Methanol is not only a by-product of PME activities, and our data suggest that [Ca2+]cyt variations could participate in signalling processes induced by methanol upstream of plant defence responses.


Subject(s)
Arabidopsis/physiology , Calcium/metabolism , Ethylenes/metabolism , Nicotiana/physiology , Plant Growth Regulators/metabolism , Aequorin/metabolism , Apoproteins/metabolism , Arabidopsis/drug effects , Cell Membrane/physiology , Cells, Cultured , Cytosol/metabolism , Methanol/administration & dosage , Plant Proteins/metabolism , Reactive Oxygen Species/metabolism , Recombinant Proteins/metabolism , Seedlings/drug effects , Seedlings/physiology , Nicotiana/drug effects
3.
Biosci Biotechnol Biochem ; 80(8): 1546-54, 2016 Aug.
Article in English | MEDLINE | ID: mdl-27149194

ABSTRACT

Indole-3-acetic acid (IAA) is the major natural auxin involved in the regulation of a variety of growth and developmental processes such as division, elongation, and polarity determination in growing plant cells. It has been shown that dividing and/or elongating plant cells accompanies the generation of reactive oxygen species (ROS) and a number of reports have suggested that hormonal actions can be mediated by ROS through ROS-mediated opening of ion channels. Here, we surveyed the link between the action of IAA, oxidative burst, and calcium channel activation in a transgenic cells of rice expressing aequorin in the cytosol. Application of IAA to the cells induced a rapid and transient generation of superoxide which was followed by a transient increase in cytosolic Ca(2+) concentration ([Ca(2+)]c). The IAA-induced [Ca(2+)]c elevation was inhibited by Ca(2+) channel blockers and a Ca(2+) chelator. Furthermore, ROS scavengers effectively blocked the action of IAA on [Ca(2+)]c elevation.


Subject(s)
Calcium/metabolism , Indoleacetic Acids/pharmacology , Oryza/drug effects , Plant Cells/drug effects , Plant Growth Regulators/pharmacology , Reactive Oxygen Species/metabolism , Aequorin/genetics , Aequorin/metabolism , Calcium Channel Blockers/pharmacology , Calcium Chelating Agents/pharmacology , Cations, Divalent , Cell Culture Techniques , Egtazic Acid/pharmacology , Gene Expression , Genes, Reporter , Ion Transport , Luminescent Measurements , Oryza/genetics , Oryza/metabolism , Plant Cells/metabolism , Plants, Genetically Modified , Respiratory Burst/drug effects , Signal Transduction , Verapamil/pharmacology
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