Identification and partial characterization of two cysteine proteases from castor bean leaves (Ricinus communis L.) activated by wounding and methyl jasmonate stress

Abstract

Jasmonates are signaling molecules that play key roles in wound response and regulate the biosynthesis of many defensive proteins, including proteases. In this study, we investigate the effects of wounding and methyl jasmonate (MJ) application on the protein expression pattern of Ricinus communis L. leaves and on proteolytic activity. Gelatin zymography demonstrated that both MJ and mechanical wounding induce alterations in the proteolytic pattern of castor bean leaves (R. communis L.). Expression of two cysteine proteases (38 and 29 kDa) was induced by the treatments employed; however, MJ induced a higher protease level than mechanical wounding during the stress period (24, 48, and 72 h). The increase in protease activity mirrors the decline in soluble protein content and rubisco degradation that may indicate initiation of senescence in castor plants. The 29 kDa protease has an acidic optimal pH; whereas the 38 kDa protease has a neutral optimum activity. Both proteases were almost completely inhibited by E-64 and cystatin. The significant induction of these proteins by MJ suggests a possible role of cysteine proteases in leaf senescence as well as their involvement in regulating both the wound response and MJ in castor bean plants.