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Arabidopsis Ca2+-dependent protein kinase CPK3 mediates relationship of putative inositol triphosphate receptor with slow-type anion channel

  • Original Papers
  • Published:
Biologia Plantarum

Abstract

It has been suggested in Arabidopsis thaliana (L.) Heynh. cv. Columbia that, contrary to 30 μM abscisic acid (ABA), 20 μM ABA induces guard cell Ca2+ mobilization through activating phosphoinositide-specific phospholipase C (PI-PLC)-dependent inositol 1,4,5-triphosphate (IP3) production. Here, it was investigated whether Ca2+-dependent protein kinase, CPK3 or CPK6 would mediate ABA-induced stomatal closure downstream of IP3 production. In the knockout cpk3-1 mutant, the PLC inhibitor (U73122) adjusted 20 μM ABA-induced stomatal closure to the extent observed in the knockout cpk6-1 and cpk3-1cpk6-1 mutants and the wild type, whereas, in the wild type, the inhibitor of IP3-induced Ca2+ mobilization, xestospongin C (XeC), adjusted this closure to the extent observed in the cpk3-1 mutant. The Ca2+ buffer, EGTA and XeC positively interacted with the slow anion channel blocker, anthracene-9-carboxylic acid (9-AC) to inhibit 20 μM ABA-induced stomatal closure, which was suppressed in the dexamethasone-inducible AtPLC1 antisense transgene or the knockout cpk3-1, cpk6-1, cpk3-1cpk6-1 and NADPH oxidase atrbohD/F mutants. Discrete concentrations of 9-AC or another slow anion channel blocker (probenecid) negatively interacted with the Ca2+ buffer, BAPTA or the inhibitor of cyclic ADP-ribose-induced Ca2+ mobilization, ruthenium red, to inhibit 30 μM ABAinduced stomatal closure in the wild type but not in the cpk6-1, cpk3-1cpk6-1 and atrbohD/F mutants. Based on so far revealed features of the tested compounds and plant materials, interpretation of the results confirmed that guard cell ABA concentration discriminates between two Ca2+ mediations and outlined that one of them sequentially implicates CPK6, PLC1, a putative IP3 receptor homologue, CPK3, and the slow anion channel, whereas the other one excludes AtPLC1-dependent IP3 production and CPK3.

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Abbreviations

ABA:

abscisic acid

AGI No.:

Arabidopsis gene identifier number

ARC:

ADP-ribosyl cyclase

cADPR:

cyclic ADP-ribose

CDPK:

Ca2+-dependent protein kinase

CFTR:

cystic fibrosis transmembrane conductance regulator

Dex:

dexamethasone

Gα:

GTP-binding protein α-subunit

G protein:

GTP-binding protein

IP3 :

inositol 1,4,5-triphosphate

MRP:

multidrug resistance-associated protein

MTX:

methotrexate

PI-PLC:

phosphoinositide-specific phospholipase C

Proben:

probenecid

RRed:

ruthenium red

R-type:

rapid-type anion channel

S-type:

slow-type anion channel

U73122:

1,6-[17β-3-methoxyestra-1,3,5(10)-trien-17-yl]aminohexyl-1H-pyrrole-2,5-dione

XeC:

xestospongin C

9-AC:

anthracene-9-carboxylic acid

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Acknowledgements

This work was supported by an IFCPAR grant. The author gratefully thanks Prof. Chua (The Rockefeller University, New York, USA) for his gift of the dexamethasone-inducible AtPLC1 antisense transgene of A. thaliana and Prof. Schroeder (University of California, La Jolla, USA) for his gift of the cpk3-1, cpk6-1, cpk3-1cpk6-1 and atrbohD/F mutants of A. thaliana.

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Cousson, A. Arabidopsis Ca2+-dependent protein kinase CPK3 mediates relationship of putative inositol triphosphate receptor with slow-type anion channel. Biol Plant 55, 507–521 (2011). https://doi.org/10.1007/s10535-011-0117-4

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