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Published on January 11, 2006; 10.1104/pp.105.074971

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Received December 1, 2005
Returned for revision December 19, 2005
Accepted December 19, 2005

Abscisic Acid Stimulates a Calcium-Dependent Protein Kinase in Grape Berry

Xiang-Chun Yu , Mei-Jun Li , Gui-Feng Gao , Hai-Zhong Feng , Xue-Qing Geng , Chang-Cao Peng , Sai-Yong Zhu , Xiao-Jing Wang , Yuan-Yue Shen , and Da-Peng Zhang *

China State Key Laboratory of Plant Physiology and Biochemistry, China Agricultural University, 100094 Beijing, China

* Corresponding author; email: zhangdp{at}sohu.net.

It has been demonstrated that calcium play a central role in mediating ABA signal, but many of the Ca++-binding sensory proteins as the components of ABA signaling pathway remain to be elucidated. Here, we identified, characterized and purified a 58-kD ABA-stimulated calcium-dependent protein kinase from the mesocarp of grape berries, designated ACPK1 (for Abscisic acid-stimulated calcium-dependent protein kinase1). ABA stimulates ACPK1 in a dose-dependent manner, and the ACPK1 expression and enzyme activities alter accordantly with the endogenous ABA concentrations during fruit development. The ABA-induced ACPK1 stimulation appears to be transient with a rapid effect in 15 min but also with a slow and steady state of induction after 60 min. ABA acts on ACPK1 indirectly and dependently on in vivo state of the tissues. Two inactive ABA isomers (-)-ABA and trans-ABA are ineffective for inducing ACPK1 stimulation, revealing that the ABA-induced effect is stereo-specific to physiological active (+)-ABA. The other phytohormones such as auxin indoleacetic acid, gibberellin GA3, synthetic cytokinin N-benzyl-6-aminopurine and brassinolide are also ineffective in this ACPK1 stimulation. Based on sequencing of the two-dimensional-electrophoresis-purified ACPK1, we cloned ACPK1 gene. The ACPK1 is expressed specifically in grape berry covering a fleshy portion and seeds, and in a developmental-stage-dependent manner. We further showed that ACPK1 is localized in both plasma membranes and chloroplasts/plastids, and positively regulates plasma membrane H+-ATPase in vitro, suggesting that ACPK1 may be involved in ABA signaling pathway.




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