ACTIVE CALCIUM TRANSPORT IN PLASMA MEMBRANE VESICLES FROM DEVELOPING COTYLEDONS OF COMMON BEAN

ACTIVE CALCIUM TRANSPORT IN PLASMA MEMBRANE VESICLES FROM DEVELOPING COTYLEDONS OF COMMON BEAN 1 corresp first-author Section of Plant Physiology, Zhejiang Agricultural University Section of Plant Physiology, Zhejiang Agricultural University 1 Institute of Nuclear Agricultural Sciences, Zhejiang Agricultural University, Hangzhou 310029 Institute of Nuclear Agricultural Sciences, Zhejiang Agricultural University, Hangzhou 310029 Plasma membrane vesicles were prepared from the developing cotyledons of common bean (Phaseolus vulgaris L cv Diyundou) by aqueous two-phase partitioning and characterized as to their purity by assaying marker enzymes for other membranes. The putative plasma membrane fraction was minimally contaminated by membranes other than plasma membrane and hence was of high purity. It exhibited a Ca 2+ -dependent ATPase activity, which was inhibited by 1 µ mol/L EB and promoted by calcium ionophore A23187. Such an activity was responsible for the observed ATP-dependent 45 Ca 2+ uptake into inside-out plasma membrane vesicles. This process was stimulated by 0.6 µ mol/L CaM and 20 µ mol/L IAA but inhibited by 2 µ mol/L ABA and abolished by A23187. Possible role of cytoplasmic Ca 2+ in mediating phytohormones activity is discussed. 45 Ca 2+ uptake Plasma membrane vesicles Phytohormones Developing cotyledons of common bean Tracer techniques REFERENCES Rasmussen H , Barrett P Q . Physiol Rev , 1984 ; 64 : 938 - 984 Physiol Rev Poovaiah B W , Reddy A S N . CRC Crit Rev Plant Sci , 1987 ; 6 : 47 - 103 CRC Crit Rev Plant Sci Briskin D P . Plant Physiol , 1990 ; 94 : 397 - 400 Plant Physiol Thomas T H . In: Bopp M , ed. Plant growth substances 1985 . Berlin Heidelberg : Springer-Verlag , 1986 : 350 - 359 Plant growth substances 1985 Huang Jianzhong , Chen Ziyuan . Proceedings of Sino-International Colloquium on Soilless Culture'94 . Hangzhou : China Agricultural Sci Tech Press , 1994 : 192 - 193 Hodges T K , Mills D . Methods Enzymol , 1986 ; 118 : 41 - 54 Methods Enzymol Huang Jianzhong . H + /sucrose symport in plasma membrane vesicles from developing cotyledons of Phaseolus vulgaris and its regulation by plant hormones . Ph D Thesis , Zhejiang Agric Univ , 1992 H+/sucrose symport in plasma membrane vesicles from developing cotyledons of Phaseolus vulgaris and its regulation by plant hormones Blum W , Key G , Weiler E W . Physiol Plant , 1988 ; 72 : 279 - 287 Physiol Plant Lanzetta P A , Alvarez L J , Reinach P S . Anal Biochem , 1979 ; 100 : 95 - 97 Anal Biochem Hodges T K , Leonard R T . Methods Enzymol , 1974 ; 32 : 392 - 406 Methods Enzymol Tolbert N E . Methods Enzymol , 1974 ; 31 ( part A ): 734 - 746 Methods Enzymol Sandelius A S , Penel C , Auderset G et al . Plant Physiol , 1986 ; 81 : 177 - 185 Plant Physiol Bush D R , Sze H . Plant Physiol , 1986 ; 80 : 549 - 555 Plant Physiol Allen J D , Parsons S M . Anal Biochem , 1979 ; 92 : 22 - 30 Anal Biochem Bradford M M . Anal Biochem , 1976 ; 72 : 248 - 254 Anal Biochem Williams L E , Nelson S J , Hall J L . Planta , 1990 ; 182 : 532 - 539 Planta Larsson C , Widell S , Kjellbom P . Methods Enzymol , 1987 ; 148 : 558 - 568 Methods Enzymol Mandala S , Taiz L . J Biol Chem , 1986 ; 261 : 12850 - 12855 J Biol Chem Gallagher S R , Leonard R T . Plant Physiol , 1982 ; 70 : 1335 - 1340 Plant Physiol Cocucci M C . Plant Sci , 1986 ; 47 : 21 - 27 Plant Sci Rasi-Caldogno F , Pugliarello M C , de Michelis M I . Plant Physiol , 1987 ; 83 : 994 - 1000 Plant Physiol Blatt M R , Thiel G . Ann Rev Plant Physiol Plant Mol Biol , 1993 ; 44 : 543 - 567 Ann Rev Plant Physiol Plant Mol Biol Wang M , van Duijn B , Schram A W . FEBS Lett , 1991 ; 278 : 69 FEBS Lett *This work was supported in part by a grant from the Foundation of Natural Sciences of Zhejiang Province 1994-08-02 1995-02-01 2021-02-05 1995 Nuclear Science and Techniques 1 6