Better salinity tolerance in tetraploid vs diploid volkamer lemon seedlings is associated with robust antioxidant and osmotic adjustment mechanisms

Tetraploids are usually more tolerant to environmental stresses than diploids. Citrus plants face numerous abiotic stresses, including salinity, which negatively affect growth and yield. Double diploid citrus rootstocks have been shown to be more tolerant to abiotic stresses than their diploid relatives. In this study, we evaluated the antioxidative and osmotic adjustment mechanisms of diploid (2x) and double diploid (4x) volkamer lemon (Citrus volkameriana Tan. and Pasq.) rootstocks, which act against salt stress (75 and 150 mM). Results indicated that, under salt stress, all physiological variables (photosynthesis, stomatal conductance, transpiration rate, and leaf greenness) decreased, and these decreases were more noticeable in 2x plants than in 4x plants. On the other hand, accumulation of oxidative markers (malondialdehyde and hydrogen peroxide) was greater in the leaves and roots of 2x seedlings than in 4x seedlings. Similarly, the activities of antioxidative enzymes (peroxidase, ascorbate peroxidase, glutathione reductase, and catalase) were higher in the leaves and roots of 4x plants than in 2x plants. However, superoxide dismutase activity was higher in the roots of 2x seedlings than 4x seedlings. Double diploid plants affected by salt stress accumulated more osmolytes (i.e. proline and glycine betaine) in their leaves and roots than that by 2x plants. Total protein content, antioxidant capacity, and total phenolic content were also higher in 4x plants than 2x plants under salinity. At 150 mM, both 2x and 4x plants showed more symptoms of stress than those at 75 mM. Sodium content was the highest in the roots of 2x plants and in the leaves of 4x plants, while chloride content peaked in the leaves of 2x plants and in the roots of 4x plants. Overall, our results demonstrate that the active antioxidative defence mechanisms of 4x plants increase their tolerance to salinity compared to their corresponding 2x relatives. Thus, the use of newly developed tetraploid rootstocks may be a strategy for enhancing crop production in saline conditions.