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PLANT PHYSIOLOGY , Vol 108, Issue 2 713-719, Copyright © 1995 by American Society of Plant Biologists
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DEVELOPMENT AND GROWTH REGULATION |
Expression of Phospholipase D during Castor Bean Leaf Senescence
S. B. Ryu and X. Wang
Department of Biochemistry, Kansas State University, Manhattan, Kansas 66506
Membrane deterioration in plant senescence is commonly associated with
progressive decreases in membrane phospholipid content. This study
investigated the expression and regulation of phospholipase D (PLD; EC
3.1.4.4) during senescence in castor bean (Ricinus communis L. cv Hale)
leaf discs. The rate of leaf senescence was accelerated by 50 [mu]M
abscisic acid and was attenuated by 50 [mu]M cytokinin during incubation at
23[deg]C for up to 5 d. Leaf senescence was indicated by decreases in the
content of total proteins, chlorophyll, and phospholipids. PLD activity in
both membrane-associated and cytosolic fractions showed a gradual increase
in the absence of phytohormones. Abscisic acid stimulated an increase in
membrane-associated PLD and had little effect on the soluble form. On the
other hand, cytokinin retarded the increase in membrane-associated PLD.
Immunoblotting analysis using PLD-specific antibodies revealed that the
changes in PLD activity were correlated with those of PLD protein. Analysis
of PLD by nondenaturing PAGE showed the appearance of a PLD structural
variant, PLD 3, in abscisic acid-treated leaf discs. Northern blotting
analysis using a PLD cDNA probe revealed an increase in PLD mRNA in
senescing leaf discs. These data indicate complex mechanisms for the
regulation of PLD during senescence, which include increases in
membrane-associated PLD, differential expression of PLD isoforms, and
changes in amounts of PLD protein and mRNA. Such controlled expression
points to a role for PLD in membrane deterioration and plant senescence.
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