Drought induces many forms of cysteine proteases not observed during natural senescence

Drought-induced senescence and natural senescence was characterised in the cowpea leaf, with a focus on cysteine proteases. Soluble protein content and ribulose 1,5-bisphosphate carboxylase (Rubisco) content declined as senescence progressed. Endopeptidase activity with Rubisco as a physiological substrate exhibited significant increase at acidic (pH 4.8) than at neutral (pH 7.0) during drought induced senescence and declined during recovery. Natural senescence was associated with a several-fold increase in the endopeptidase activity at both the pHs. Cysteine proteases were analyzed using western blot with polyclonal antibodies raised against papain. Several polypeptides of molecular weights 57, 52, and 43 kDA were recognized by the antibodies, the levels of which showed an increase under water deficit conditions, followed by a decrease during recovery. Three polypeptides of molecular weights 69, 60, and 48 kDa appeared only during the water stress conditions, whereas, during natural senescence, only a single 48 kDa polypeptide with maximum intensity at 9 days after flowering was observed. The results suggests the possibility of distinguishing drought-induced and natural senescence. (C) 1999 Academic Press.