Abstract
The study aimed to decipher the impact of multiple drought stress on wheat. To that effect, Geumgangmil, PL 337 (1AL.1RS), PL 371 (1BL.1RS), and PL 257 (1DL.1RS) seedlings were subjected to four treatments: G1 (control), G2 (stressed thrice with rewatering), G3 (stressed twice with rewatering), and G4 (single stressful event). The findings provided a comprehensive framework of drought-hardening effect at physiological, biochemical, and gene expression levels of drought-stressed wheat genotypes. The treatments resulted in differentially higher levels of malondialdehyde (MDA), hydrogen peroxide (H(2)O(2)), soluble sugar, and proline accumulation, and reduced relative water content (RWC) in wheat plants. Photosynthetic pigment (chlorophyll and carotenoid) levels, the membrane stability index (MSI), and shoot biomass decreased dramatically and differently across genotypes, particularly in G3 and G4 compared to G2. The activity of antioxidant enzymes [ascorbate peroxidase (APX), superoxide dismutase (SOD), and catalase (CAT)] increased with the duration and severity of drought treatment. Furthermore, the relative expression of DREB, LEA, HSP, P5CS, SOD1, CAT1, APX1, RBCL, and CCD1 genes was higher in G2 than in other treatments. Drought hardening increased drought tolerance and adaptability in plants under G2 by enhancing growth and activating defensive mechanisms at the physio-biochemical and molecular levels. The findings of the study indicated that early drought stress exposure-induced acclimation (hardening), which enhanced tolerance to subsequent drought stress in wheat seedlings. The findings of this study will be useful in initiating a breeding program to develop wheat cultivars with improved drought tolerance. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-021-02991-6.