Expression Profile of Selected Genes Involved in Na+ Homeostasis and In Silico miRNA Identification in Medicago sativa and Medicago arborea under Salinity Stress

The accumulation of ions due to increased salinity in the soil is one of the major abiotic stressors of cultivated plants that negatively affect their productivity. The model plant, Medicago truncatula, is the only Medicago species that has been extensively studied, whereas research into increased salinity adaptation of two important forage legumes, M. sativa and M. arborea, has been limited. In the present study, the expression of six genes, namely SOS1, SOS3, NHX2, AKT, AVP and HKT1 was monitored to investigate the manner in which sodium ions are blocked and transferred to the various plant parts. In addition, in silico miRNA analysis was performed to identify miRNAs that possibly control the expression of the genes studied. The following treatments were applied: (1) salt stress, with initial treatment of 50 mM NaCl and gradual acclimatization every 10 days, (2) salt shock, with continuous application of 100 mM NaCl concentration and (3) no application of NaCl. Results showed that M. arborea appeared to overexpress and activate all available mechanisms of resistance in conditions of increased salinity, while M. sativa acted in a more targeted way, overexpressing the HKT1 and AKT genes that contribute to the accumulation of sodium ions, particularly in the root. Regarding miRNA in silico analysis, five miRNAs with significant complementarity to putative target genes, AKT1, AVP and SOS3 were identified and served as a first step in investigating miRNA regulatory networks. Further miRNA expression studies will validate these results. Our findings contribute to the understanding of the molecular mechanisms underlying salt-responsiveness in Medicago and could be used in the future for generating salt-tolerant genotypes in crop improvement programs.