observed in between three h and 27 h in the intolerant cultivar `Vernal'. We speculate

observed in between three h and 27 h in the intolerant cultivar `Vernal’. We speculate that such earlier activation of salt responsive genes and maintenance of a big number of DEGs may possibly be a crucial characteristic for salt tolerance in alfalfa, suggesting alfalfa tolerance is linked with upregulation of important genes from brief term salt pressure. About 60 of DEGs have been assigned to GO categories, while KEGG pathways for less than 30 DEGs have been identified in this study. The DEGs have been primarily involved in metabolic pathways as revealed by KEGG pathway analysis. While specific pathways involved in salt tolerance might be conserved in plant species which include in halophytes, there was nevertheless variation among plant species, cultivars, and tissues [5]. This study demonstrated that transcriptional variation in adaptation to salt stress existsnot only amongst the alfalfa cultivars but also among the diverse tissues. `Ion binding’ (GO:0043167) was significantly enriched in both leaf and root tissues of `Halo’, but not in `Vernal’ below salt stress. This suggested that the genes responsible for `ion binding’ needs to be exceptional for salt tolerance of `Halo’ alfalfa. Consequently, the tissueand genotype-specific salt responsive genes could be useful in identification of salt tolerant genotypes inside the future. Among 13 candidate genes expressed in leaf and root tissues of `Halo’ beneath salt tension (Table 3) in this study, two genes (MS.gene013222 and MS.IL-8 Antagonist Compound gene52595) are responsible for transmembrane protein function. These transmembrane proteins control gateways and selective transport of salt ions to facilitate salt tolerance in plants. Likewise, MS.gene013211, a homologous gene to ribonuclease TUDOR1, is involved in anxiety adaptation and hugely expressed in leaf and root tissues of `Halo’ in our study [30]. MS.gene93979, a homologous gene to NF-X1-type zinc finger protein, is part of mechanisms that regulate growth below salt anxiety and was hugely expressed in leaf and root tissues of `Halo’ in our study [31]. Also, MS.gene029202 (E3 ubiquitin-protein ligase CIP8), MS.gene029203 (F-box/ LRR-repeat protein 4), MS.gene36780 and MS.gene36960 (elongation aspect 1-alpha) were highlyBhattarai et al. BMC Plant Biology(2021) 21:Page ten ofTable four List of 15 salt responsive candidate genes very expressed in leaf tissue of salt tolerant alfalfa cultivar `Halo’Gene ID MS.gene024018 MS.gene029055 MS.gene029201 MS.gene029206 MS.gene037960 MS.gene038586 MS.gene065734 MS.gene07287 MS.gene24098 MS.gene24746 MS.gene36621 MS.gene39381 MS.gene63155 MS.gene81767 MS.geneaNr IDa KHN29288.1 AFK45194.1 AET03044.2 XP_024628388.1 XP_003589866.2 RHN67456.1 XP_013467963.1 XP_003591401.1 PNY14915.1 RHN68722.1 XP_003627058.1 RHN38725.1 RHN41150.1 XP_013467963.1 AIP98334.log2FCb (Leaf) 0h NA 5.2 NA NA NA NA 6.four eight.8 5.1 six.five NA NA NA NA 4.6 3h 8.9 7.three 7.5 four.7 two.7 six.5 9.eight 11.2 5.7 four.9 4.7 6.6 7.3 four.eight 4.eight 27h four.9 5.eight 9.0 5.0 two.7 6.2 6.9 10.five 6.2 five.3 four.five six.five 4.2 three.six four.putative function Monothiol glutaredoxin-S14, chloroplastic [Glycine soja] CDP-diacylglycerol–glycerol-3-phosphate 3-phosphatidyltransferase 2 [Medicago truncatula (barrel medic)] replication protein A 70 kDa DNA-binding subunit C [Medicago t runcatula (barrel medic)] FAD synthetase 1, chloroplastic [Medicago truncatula (barrel medic)] nuclear pore complicated protein NUP1 [Medicago truncatula (barrel medic)] putative minus-end-directed kinesin CYP1 Inhibitor MedChemExpress ATPase [Medicago truncatula] uncharacterized LOC25483798 [Medicago truncatula (barrel medic)] ca