Proline induces heat tolerance in chickpea (Cicer arietinum L.) plants by protecting vital enzymes of carbon and antioxidative metabolism

Abstract

Chickpea is a heat sensitive crop hence its potential yield is considerably reduced under high temperatures exceeding 35°C. In the present study, we evaluated the efficacy of proline in countering the damage caused by heat stress to growth and to enzymes of carbon and antioxidative metabolism in chickpea. The chickpea seeds were raised without (control) and with proline (10μM) at temperatures of 30/25°C, 35/30°C, 40/35°C and 45/40°C as day/ night (12h/12h) in a growth chamber. The shoot and root length at 40/35°C decreased by 46 and 37%, respectively over control while at 45/40°C, a decrease of 63 and 47%, respectively over control was observed. In the plants growing in the presence of 10μM proline at 40/35°C and 45/40°C, the shoot length showed improvement of 32 and 53%, respectively over untreated plants, while the root growth was improved by 22 and 26%, respectively. The stress injury (as membrane damage) increased with elevation of temperatures while cellular respiration, chlorophyll content and relative leaf water content reduced as the temperature increased to 45/40°C. The endogenous proline was elevated to 46μmolg(-1) dw at 40/35°C but declined to 19μmolg(-1) dw in plants growing at 45/40°C that was associated with considerable inhibition of growth at this temperature. The oxidative damage measured as malondialdehyde and hydrogen peroxide content increased manifolds in heat stressed plants coupled with inhibition in the activities of enzymatic (superoxide dismutase, catalase, ascorbate peroxidase, glutathione reductase) and levels of non-enzymatic (ascorbic acid, glutathione, proline) antioxidants. The enzymes associated with carbon fixation (RUBISCO), sucrose synthesis (sucrose phosphate synthase) and sucrose hydrolysis (invertase) were strongly inhibited at 45/40°C. The plants growing in the presence of proline accumulated proline up to 63μmolg(-1) dw and showed less injury to membranes, had improved content of chlorophyll and water, especially at 45/40°C. Additionally, the oxidative injury was significantly reduced coupled with elevated levels of enzymatic and non-enzymatic antioxidants. A significant improvement was also noticed in the activities of enzymes of carbon metabolism in proline-treated plants. We report here that proline imparts partial heat tolerance to chickpea's growth by reducing the cellular injury and protection of some vital enzymes related to carbon and oxidative metabolism and exogenous application of proline appears to have a countering effect against elevated high temperatures on chickpea.