Characterization of transgenic alfalfa lines overexpressing MsSN1 . (A) Schematic representation of the T-DNA region of binary vector pART-35S::MsSN1 containing the MsSN1 gene under the CaMV 35S promoter. Relevant restriction enzymes used in plasmid construction and Southern Blot analysis are shown. RB: right border; CaMV 35S: promoter; Topo: region derived from pCR2.1-TOPO vector; UTRs: untranslated regions derived from the native MsSN1 gene; os-t: octopine synthase terminator; pnos-nptII-nos-t: kanamycin cassette (where, pnos and nos-t are nopaline synthase promoter and terminator, respectively); LB: left border. (B) Real-Time RT-PCR assays of alfalfa transgenic lines (S1-S3) and control untransformed plants (wt). All values are log means ± SEM (n = 3). Asterisks indicate a statistically significant difference (Tukey: ***p < 0.001).