Investigating the Effect of Foliar Applied Proline on the Photosynthetic Pigments Content and Osmolytes of Pistachio Rootstock UCB1 under Water Stress Conditions

Introduction  UCB-1 hybrid rootstock is propagated from the seed of a controlled cross between a P. atlantica female and a P. integerrima male. Clonal propagation of this rootstock is also necessary since it produces identical genotypes. UCB1 has recently been introduced in Iran. However, little research has been done on this subject. Water stress is one of the most important environmental stresses and occurs for several reasons, including low rainfall, high and low temperatures, salinity, and high intensity of light, among others. Drought stress is a multidimensional stress and causes changes in the physiological, morphological, biochemical, and molecular traits in plants. Proline is also found widely in plants and accumulates in large quantities in response to environmental stresses such as drought. It is the essential amino acids that accumulate in different tissues of the plant, especially in the leaves through the effect of water stress, and that the accumulation of it has a function in the regulation of osmosis in the cell as the proline is increased in the cytoplasm to counterbalance effort osmosis cell sap. Proline is an indicator of water stress tolerance and its increase in the leaf proof that the plant suffered drought stress, also is the way the plant tolerance to drought stress.   Materials and Methods A pot experiment was conducted, to investigate the effectiveness of foliar applied proline in mitigating the concurrent effects of drought stresses on UCB1 rootstock, at greenhouse of Faculty of Agriculture, University of Shahrekord. Experiment was carried out in a completely randomized design with split arrangement having three replications. Chl. and carotenoid contents were determined according to Lichtenthaler (1987). Leaf discs were obtained from expanded leaves of each pot in the morning. The leaf discs were weighed immediately to obtain the fresh weight (FW), and submerged in distilled water for 4 h at 4◦C in dark condition and then weighed to prepare turgor weight (TW). The leaves were dried in a forced-air oven at 70◦C for 24 h, and the dry weight (DW) was recorded. The RWC of samples was calculated using the following equation (Bastam et al., 2012): RWC = [(FW–DW)/(TW–DW)] × 100 Levels of glycine betaine were quantified as described previously by Arakawa et al. (1990). To determine the free-proline concentration, leaves were homogenized in 5 ml of ethanol at 95%. Proline concentration was calculated with a standard curve and expressed as µg g−1fresh mass (Paquin and Lechasseur, 1979).   Results and Discussion The UCB1 proximate analysis in the present study depicted that imposition of drought stress increased the leaf and root proline content and electrolyte leakage. Exogenous application of proline as foliar spray significantly increased the moisture content of leaf and root, RWC, Chl a and total chlorophyll. Exogenous proline application upregulated leaf and root proline contents and decreased the lipid peroxidation (decrease electrolyte leakage), resulting into improvement in chlorophyll contents. 150 mg. l-1 proline application gave maximum alleviation against stress. Numerous reports depict that the exogenous application of proline as a foliar spray can play an important role in enhancing plant tolerance against drought stress, and our results are consistent with them. This ameliorating effect of exogenously applied proline can take the form of osmoprotection, cryoprotection, or protection against reactive oxygen species.For example, in various plant species growing under stress conditions, exogenously supplied proline provided osmoprotection and facilitated plant growth. Normally, proline accumulation in plants, is in response to drought or salinity stress occurs in the cytosol where it contributes substantially to the cytoplasmic osmotic adjustment. It actively takes part in plant osmotic adjustment under stressful environmental conditions. In addition to its role as an osmolyte for osmotic adjustment, it actively takes part to stabilize subcellular structures, biological membranes, proteins, and scavenge free radicals. It also plays a vital role in buffering cellular redox potential under stressful environmental conditions.   Conclusion  In summary, our results showed that, drought induces a decrease in moisture content, RWC, T Chl and carotenoids and an increase in some osmoregulators (proline, glycine betaine, TSC). The most favorable treatment was 150 mg.l-1 proline foliar spray. 150 mg.l-1 proline application gave maximum alleviation against stress. Foliar application increased the moisture content of leaf and root, as well as increased the Chl a, total, RWC and proline content of leaf and root. It can be suggested that the foliar application of proline (150 mg L-1) used as a plants defense factor against drought stress conditions.