Securing reproductive function in mungbean grown under high temperature environment with exogenous application of proline

Escalating temperatures are adversely impacting the production potential of various cool- and warm-season crops, such as Mungbean, therefore effective strategies are required to improve heat tolerance of various crops. Mungbean, a summer season food legume, is seriously affected at temperatures more than 35/25 °C, especially at its reproductive stage, resulting in pollen infertility to induce loss of flowers and potential pods. Proline (Pro), a well-researched stress-related molecule, has been implicated in determining pollen fertility, but its involvement in affecting reproductive function under heat stress is not reported so far. In the present study, it was hypothesised that depletion of endogenous Pro in reproductive components of the flowers of heat-stressed Mungbean plants might impair the reproductive function. To test this hypothesis, Mungbean genotypes (heat tolerant and heat-sensitive), growing in outdoor environment (32.5/17.5 ± 1 °C mean day/night temperature), until on the onset of flowering (30 days after sowing) were subjected to mild heat stress (MS; 40/28 °C) and high heat stress (HS; 45/33 °C), in the absence or presence of 5 mM proline treatment, applied as soil drenching and foliar spray, 2 days before imposition of heat stress. In MS plants, the endogenous Pro showed a significant increase in leaves, anthers, pollen and ovules, while in SS plants, a marked reduction was noticed. In later case, the activity of proline synthesising enzymes (pyrolline-5-carboxylate synthase and pyrroline-5-carboxylate reductase) declined severely, along with a proline catabolism enzyme (proline dehydrogenase) suggesting disruption in proline metabolism in vegetative and reproductive components. This was associated with considerable decrease in pollen germination, stigma receptivity and ovule viability in heat-stressed plants. Simultaneously, leaf tissue showed high damage to cell membranes, leaf water status, stomatal conductance and cellular respiration. Photosynthetic ability (Chlorophyll, Photo system II function), carbon fixation (RuBisCo activity) and assimilation processes (sucrose synthesis and its hydrolysis) were significantly inhibited, in heat-stressed (HS) plants, which impacted the pod number, pod and seed weight per plant. Pro treatment, especially to HS plants resulted in appreciable increase in its endogenous concentration in vegetative and reproductive parts, which significantly improved the pollen fertility as well as stigma and ovule function. At the same time, stress damage to leaves was reduced significantly, leaf water status and chlorophyll were significantly higher, as a result the carbon fixation and assimilation capacity improved notably to increase the pod set, filled pod number, pod weight and seed weight per plants, suggesting a vital role of proline in enhancing the thermo-tolerance. The effects of Pro treatment were more pronounced in heat-sensitive genotype.