GA3 and BAP phytohormone seed priming enhances germination and PEG induced drought stress tolerance in soybean by triggering the expression of osmolytes, antioxidant enzymes and related genes at the early seedling growth stages

Abstract

Drought stress strongly affects seed germination and seedling development during the early stages of plant growth in soybean (Glycine max L.). Seed priming with phytohormone (PH) is known to regulate various physiological and biochemical processes that govern plant growth and yield under optimal and stress conditions. In the present study, the effects of seed priming with Gibberellic acid (GA₃) (0.28 mM)_, 6-Benzyl aminopurine (BAP) (20 mM) and Methyl Jasmonate (MJ) (2.5 mM) was studied during germination and at early seedling growth stages in JS-335 (drought sensitive) and MAUS-71 (moderately drought tolerant) soybean varieties under PFG induced drought stress conditions. The seed priming effects were studied in terms of germination percentage and rate (Timson Index), accumulation of osmolytes (proline and glycine betaine), antioxidant enzymes activity (superoxide dismutase (SOD), ascorbate peroxidase (APX) and catalase (CAT)), and the relative expression of genes viz., Cu/Zn-superoxide dismutase copper chaperone (SOD-CCS) , iron-superoxide dismutase (SODB2) , betaine aldehyde dehydrogenase (BADH2) and dehydration-responsive element binding protein (DREB2A). The germination percentage of non-primed soybean seeds decreased under drought stress but significantly increased upon GA₃ and BAP priming. Interestingly, seed priming with MJ completely inhibited the germination in both the varieties. GA₃ and BAP seed priming increased proline and glycine betaine accumulation under PEG induced drought stress in both the soybean varieties. The results showed that the activities of SOD, APX and CAT were significantly enhanced in the leaves of seedlings raised from GA₃ and BAP-primed seeds, as compared to the non-primed seeds under the drought stress conditions. Isozyme profiling of SOD and APX revealed that the GA₃ and BAP seed priming treatments enhanced the drought-inducible expression of SOD isozymes rather than inducing novel isoforms. In case of APX isozymes, GA₃ seed priming appears to preserve APX isozyme diversity in both varieties, and BAP seed priming elevated the expression of APX isoforms in both soybean varieties under the drought conditions. In response to the drought stress, relative expression of the genes associated with superoxide dismutase and glycine betaine viz., SOD CCS, SODB2, BADH2 and DREB2A was markedly upregulated in the leaves of seedlings raised from the GA₃ and BAP primed seeds in both the soybean varieties. The global correlation analysis designated that the osmolyte Proline, antioxidant enzyme APX and CAT, and SOD CCS, SODB2, BADH2 and DREB2A genes were potent regulator for drought stress tolerance mechanism in soybean. This study illuminates the intricate interplay of GA₃ and BAP hormonal seed priming during the germination and early seedling stages in soybean plants under the drought stress. This finding emphasizes the critical consideration of genotype specific reactions to the hormonal seed priming under PEG induced drought stress conditions.