Molecular characterization and intracellular localization of phospholipase D in castor bean (Ricinus communis L.)

The first eukaryotic phospholipase D cDNA and plant PLD gene were cloned and characterized from castor bean. The cloned PLD cDNA encodes a 92 kDa protein of 808 amino acids. Comparison of the deduced protein sequence of this cloned PLD with the N-terminal amino acid sequence of the purified PLD indicated that a 30 amino acid leader peptide precedes the mature PLD. Sequence comparison with PLD cDNA revealed that the cloned PLD gene consists of four exons and three introns, one of which interrupts the 5;Subcellular distribution of PLD in castor bean leaves and seedlings was examined with immunocytochemistry, immunoblotting and activity assays of PLD in the fractions separated with a two-phase partitioning system. These results showed that PLD was mainly located in vacuoles, and some was associated with plasma membrane and cytoplasmic matrices.;The study of the PLD gene promoter in transgenic tobacco plants also indicated that the 5...