Salt stress triggers augmented levels of Na+, K+ and ROS alters salt-related gene expression in leaves and roots of tall wheatgrass (Agropyron elongatum)

Abstract

In turfgrass breeding, competent grass ecotypes are preferably identified for their resistance to salinity condition. This research was designed to explore genes that induce salt resistance (NHX1, NHX2, HKT1;4, SnRK2.4 and NAC9) and their role in physiological modifications of six tall wheatgrass ecotypes (Agropyron elongatum L.). The sites of sample collection were characterized by different levels of salinity, i.e. low (EC: 4 dS m^-1 and pH: 6.5), moderate (EC: 7 dS m^-1 and pH: 6.5) and high (EC: 12 dS m^-1 and pH: 7.5). This study was designed as a split-plot in a randomized complete block where salinity treatments served as the whole-plot factor and ecotypes served as the subplot factor. The ecotypes were screened for their resistance to salinity, based on visual symptoms, salt injury index, physiological features and biochemical parameters. The results revealed that ecotype 'AE5' was most resistant to salinity than other ecotypes, whereas 'AE3' was the most susceptible. To understand why these differences occurred, measurements were aimed at revealing mRNA levels that resulted from genes responsible for salt resistance. Our results demonstrated that salinity-resistant ecotypes showed high expression levels of several genes, i.e. NHX1, NHX2, HKT1;4, SnRK2.4 and NAC9 in the leaves and roots. These results were corroborated by a decrease (by 1.5-2.5 times) in stress markers, namely, superoxide anion (O₂^-), hydrogen peroxide (H₂O₂) and malondialdehyde (MDA), as well as an increase (by 0.5-7 times) in enzymatic and non-enzymatic antioxidant activity in salinity-resistant ecotypes when the plants were exposed to salinity. We observed higher values of initial root length and lateral root density (21% and 18%, respectively) in salinity-resistant ecotypes under salinity condition, compared to other ecotypes. There were lower expression levels of NHX1 and NHX2 in the roots, which were 3.2 and 2.1 times less, respectively, compared to the leaves. This implied that NHX1 and NHX2 expressions can lead to the sequestration of Na⁺ in the leaves during salinity condition. The current research revealed that HKT1;4 was more able to restrict Na ⁺ accumulation, compared to the actions of NHX1 and NHX2 genes. The over-expression of HKT1;4 in 'AE5' allowed a better maintenance of root growth during salinity condition. The expression of NAC9 had an increase of 2.1-fold which correlated with an increase in the amount of antioxidant enzymes. In general, the location of sample collection explained the differences in gene expression, especially regarding the extent to which plants respond to salinity condition. Ultimately, these differences can define physiological features in salinity-resistant and salinity-susceptible ecotypes of tall wheatgrass.

Keywords: Environmental stress; Gene expression; Salinity condition; Turfgrass.