Assay for NaCl, PEG and ABA stress on SiLEA14 overexpression foxtail millet germination. (A) Diagram of the T-DNA region of the binary vector pCOU-SiLEA14-flag. LB, left border; RB, right border; 35S-Pro, cauliflower mosaic virus 35S promoter; Hpt, hygromycin B phosphotransferase gene; Ubi-pro, maize Ubiquitin promoter; SiLEA14-flag, SiLEA14 gene fused with a flag tag; NOS, nopaline synthase terminator. (B) QRT-PCR analysis of SiLEA14 in transgenic foxtail millet lines (L68, L76 and L78). Actin7 was used as an internal control. (C-D) The phenotype of transgenic and WT foxtail millet under various abiotic stress treatment during the germination stage. The T2 seeds soaked in water (as control) or in water containing 150 mM NaCl, 250 mM NaCl, 10% PEG, 20% PEG and 10 μM ABA for 1 day at 30°C and then placed on the filter paper in a Petri dish wet with the same solutions mentioned above for 4 and 9 days, respectively. This experiment had three replicates, and each experiment comprised 30–40 plants. Bar = 1 cm. (E-F) The shoot length of transgenic and WT foxtail millet germinated under above conditions for 4 and 9 days, respectively. (G-H) The root length of transgenic and WT foxtail millet germinated under above conditions for 4 and 9 days, respectively. Data in B, E-H represent means and standard errors for three biological replicates. Statistical significance was determined by Student’s t-test. *P < 0.05; **P < 0.01.