Activation of phospholipase PLA2 actvity in Ricinus communis leaves in response to mechanical wounding

Abstract

In order to investigate the defense response in castor bean (Ricinus communis) against predators, we analyzed the effect of mechanical wounding upon the phospholipase A2 (PLA2) activity of leaf extracts. Time course experiments revealed that the highest levels of increased PLA2 activity (ca. two fold) occurred 15 min and 60 min after injury. The induced activities demonstrated high sensitivity towards aristolochic acid (10 mM), a PLA2 inhibitor. Based on SDS-PAGE analysis, the PLA2 activity induced 15 min after wounding migrated with a molecular mass of 40 kDa and was denoted RcPLA2 I. The protein activity induced 60 min after wounding, RcPLA2 II, migrated with a molecular weight of 14 kDa. Furthermore its N-terminal sequence shared homology with PLA2 from elm and rice. The PLA2 enzymes were purified to near homogeneity by a combination of gel filtration and electro-elution of protein bands after native PAGE.