Identification of wheat genotypes with high tolerance to combined salt and waterlogging stresses using biochemical and morpho-physiological insights at the seedling stage

Developing crop varieties with combined salinity and waterlogging tolerance is essential for sustainable agriculture and food security in regions affected by these stresses. This process requires an efficient method to rapidly and accurately assess the tolerance of multiple genotypes to these stresses. Our study examined the use of a pot trial in combination with the assessment of multiple traits to assess the tolerance of 100 wheat (Triticum aestivum L.) genotypes sourced from around the world to these combined stresses. The stresses were imposed on the plants using 100 mM NaCl and by submerging the root systems of the plants in their bathing solutions. The data gathered were subjected to principal component analysis (PCA), and an integrated score (IS) for each genotype was calculated based on multiple morpho-physiological traits; the score was used to rank the genotypes with respect to tolerance or susceptibility. There were significant differences among the 100 wheat genotypes in terms of the relative reductions in their growth parameters and chlorophyll contents, suggesting a rich, genetic diversity. To assess the accuracy of this methodology and to gain insight into the causes of tolerance or susceptibility, the five most tolerant (Misr4 (W85), Corack (W41), Kzyl-Sark (W94), Hofed (W57), BAW-1157 (W14)), and two least tolerant (Livingstong (W60) and Sunvale (W73)) genotypes were selected based on their IS and PCA analysis. These genotypes were then grown hydroponically with and without salinity stress. The data from this second trial were again subjected to PCA, and their IS were calculated; there was reasonable agreement in the ranking of the genotypes between the two trials. The most tolerant genotype (W85; Misr4 from Egypt) and most susceptible genotype (W73; Sunvale from Australia) were then examined in further detail in a third trial. Plants of Misr4 (W85) had lower Na⁺/K⁺ ratios, higher superoxide dismutase, peroxidase, catalase, and ascorbate peroxidase activities, and higher glutathione concentrations. As a result, plants of Misr4 (W85) had lower concentrations of reactive oxygen species (H₂O₂ and O₂^•−) and malondialdehyde than those of Sunvale (W73). This study offers an efficient methodology for the assessment of multiple sources of germplasm for stress tolerance. It has also identified germplasm that can be used for future breeding work and for further research on the mechanisms of tolerance and susceptibility to combined salinity and waterlogging stresses.