Properties of the Signal Transduction Pathway in the Blue Light Response of Stomatal Guard Cells of Vicia faba and Commelina benghalensis :

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Blue light-dependent proton extrusion in guard cell protoplasts from Vicia faba and light-dependent stomatal opening in the epidermis of Commelina benghalensis are inhibited by the calmodulin (CaM) antagonist, N-(6-aminohexyl)-5-chloro-1-naphthalenesulfononamide (W-7) and the myosin light chain kinase (MLCK) inhibitor, 1-(5-iodonaphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazepine (ML-7) [Shimazaki, K., Kinoshita, T. and Nishimura, M. (1992) Plant Physiol. 99: 1416] . We now suggest that the inhibition occurs in the blue light signaling pathway without affecting the proton pump. Addition of fusicoccin (FC), an activator of H^+-ATPase, to the protoplasts and the epidermis whose blue light-dependent proton extrusion and light-dependent stomatal opening had been inhibited by W-7 and ML-7, induced both proton extrusion and stomatal opening, respectively. Blue light-dependent proton extrusion was inhibited by K-252a, a wide-range inhibitor of protein kinases, and KT5926, a selective inhibitor of MLCK. FC induced proton extrusion in the presence of K-252a and KT5926. In contrast, phenylmercuric acetate (PMA), carbonyl cyanide-m-chlorophenylhydrazone (CCCP) and N.N'-dicyclohexylcarbodiimide (DCCD) inhibited both the proton extrusion and stomatal opening, but FC did not induce the responses. These results suggest that W-7, ML-7, K-252a and KT5926 inhibit the signal transduction process by which the perception of blue light is transduced into activation of the proton pump in guard cells, and that MLCK or MLCK-like protein is involved in the blue light response of stomata. The possibility that calcium-dependent, calmodulin independent protein kinase [Harper, J.F. et al. (1991) Science 252: 951] functions rather than MLCK in the blue light response of stomata should be noted, however.

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