Figure 2.

Vector construction and genetic transformation. (A) Schematic diagram of the expression vector pAtHKT1 + GFP. Npt II, neomycin phosphotransferase II gene; 35S, cauliflower mosaic virus 35S promoter; AtHKT1, Arabidopsis thaliana high affinity K⁺-transporter gene; GFP, green fluorescent protein gene; Hind III, Bam H I, Sma I, and Xho I are restriction endonuclease recognition sites. (B) Verification of the plasmid pAtHKT1 + GFP by PCR amplification of AtHKT1 gene, Npt II gene and GFP gene. M, DNA Marker V; N, PCR products of ddH₂O. (C) Formation of shoots directly from leaf discs of the tobacco cultivar CV87 after 4 weeks of growing in the selective medium (MS medium containing 0.50 mg L⁻¹ 6-BA + 0.05 mg L⁻¹ NAA) supplemented with 100.00 mg L⁻¹ kanamycin and 250.00 mg L⁻¹ carbenicillin, and (D) formation of roots about 7 days in the selective rooting medium (MS medium containing 100.00 mg L⁻¹ kanamycin and 200.0 mg L⁻¹cephalosporin). 1, non-transgenic tobacco plant in MS medium; 2, non-transgenic control plant in selective rooting medium; 3 to 5, transgenic lines in rooting selective medium. The grouping of gels cropped from same part of the same gel.