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Salinity is a major limiting factor in crop productivity worldwide. Medicago sativa L. is an important fodder crop, broadly cultivated in different environments, and it is moderately tolerant of salinity. Medicago arborea L. is considered a stress-tolerant species and could be an important genetic resource for the improvement of M. sativa’s salt tolerance. The aim of the study was to evaluate the seedling response of M. sativa, M. arborea, and their hybrid (Alborea) to salt shock and salt stress treatments. Salt treatments were applied as follows: salt stress treatment at low dose (50 mM NaCl), gradual acclimatization at 50–100 and 50–100–150 mM NaCl, and two salt shock treatments at 100 and 150 mM NaCl. Growth rates were evaluated in addition to transcriptional profiles of representative genes that control salt uptake and transport (NHX1 and RCI2A), have an osmotic function (P5CS1), and participate in signaling pathways and control cell growth and leaf function (SIMKK, ZFN, and AP2/EREB). Results showed that the studied population of M. sativa and M. arborea performed equally well under salt stress, whereas that of M. sativa performed better under salt shock. The productivity of the studied population of Alborea exceeded that of its parents under normal conditions. Nevertheless, Alborea was extremely sensitive to all initial salt treatments except the low dose (50 mM NaCl). In addition, significantly higher expression levels of all the studied genes were observed in the population of M. arborea under both salt shock and salt stress. On the other hand, in the population of M. sativa, NHX1, P5CS1, and AP2/EREB were highly upregulated under salt shock but to a lesser extent under salt stress. Thus, the populations of M. sativa and M. arborea appear to regulate different components of salt tolerance mechanisms. Knowledge of the different parental mechanisms of salt tolerance could be important when incorporating both mechanisms in Alborea populations.

Seedling Growth and Transcriptional Responses to Salt Shock and Stress in Medicago sativa L., Medicago arborea L., and Their Hybrid (Alborea)