Arabidopsis abi1-1 and abi2-1 phosphatase mutations reduce abscisic acid-induced cytoplasmic calcium rises in guard cells.
Elevations in cytoplasmic calcium ([Ca(2)+](cyt)) are an important component of early abscisic acid (ABA) signal transduction. To determine whether defined mutations in ABA signal transduction affect [Ca(2)+](cyt) signaling, the Ca(2)+-sensitive fluorescent dye fura 2 was loaded into the cytoplasm of Arabidopsis guard cells. Oscillations in [Ca(2)+](cyt) could be induced when the external calcium concentration was increased, showing viable Ca(2)+ homeostasis in these dye-loaded cells. ABA-induced [Ca(2)+](cyt) elevations in wild-type stomata were either transient or sustained, with a mean increase of approximately 300 nM. Interestingly, ABA-induced [Ca(2)+](cyt) increases were significantly reduced but not abolished in guard cells of the ABA-insensitive protein phosphatase mutants abi1 and abi2. Plasma membrane slow anion currents were activated in wild-type, abi1, and abi2 guard cell protoplasts by increasing [Ca(2)+](cyt), demonstrating that the impairment in ABA activation of anion currents in the abi1 and abi2 mutants was bypassed by increasing [Ca(2)+](cyt). Furthermore, increases in external calcium alone (which elevate [Ca(2)+](cyt)) resulted in stomatal closing to the same extent in the abi1 and abi2 mutants as in the wild type. Conversely, stomatal opening assays indicated different interactions of abi1 and abi2, with Ca(2)+-dependent signal transduction pathways controlling stomatal closing versus stomatal opening. Together, [Ca(2)+](cyt) recordings, anion current activation, and stomatal closing assays demonstrate that the abi1 and abi2 mutations impair early ABA signaling events in guard cells upstream or close to ABA-induced [Ca(2)+](cyt) elevations. These results further demonstrate that the mutations can be bypassed during anion channel activation and stomatal closing by experimental elevation of [Ca(2)+](cyt).
Allen
G J
GJ
Department of Biology, Center for Molecular Genetics, University of California-San Diego, La Jolla, California 92093-0116, USA. gallen2biomail.ucsd.edu
Kuchitsu
K
K
Chu
S P
SP
Murata
Y
Y
Schroeder
J I
JI
D000040:Abscisic Acid / Q000494:pharmacology; D017360:Arabidopsis / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D029681:Arabidopsis Proteins; D020013:Calcium Signaling / Q000187:drug effects / Q000235:genetics; D005456:Fluorescent Dyes; D016257:Fura-2; D017343:Genes, Plant; D008954:Models, Biological; D009154:Mutation; D010749:Phosphoprotein Phosphatases / Q000235:genetics* / Q000378:metabolism*; D010937:Plant Growth Regulators / Q000494:pharmacology
D029681:Arabidopsis Proteins; D005456:Fluorescent Dyes; D010937:Plant Growth Regulators; D000040:Abscisic Acid; C087434:ABI1 protein, Arabidopsis; C106443:ABI2 protein, Arabidopsis; D010749:Phosphoprotein Phosphatases; D016257:Fura-2