Figure 4.

Construction and verification of GmAKT2-overexpressing transgenic soybean plants. (A) T-DNA region of the GmAKT2 overexpression vector. LB, left border; RB, right border; bar, phosphinothricin acetyl transferase gene; P35S, CaMV double 35S promoter; T35S, CaMV 35S terminator; RFP, red fluorescence protein gene. (B) Southern blot analysis of GmAKT2-overexpressing transgenic lines. Oe1, Oe2, Oe3, and Oe4 represent four independent GmAKT2 overexpressing lines. Genomic DNA of 2-week-old T₁ transgenic seedlings and the non-transformed recipient soybean genotype Williams 82 was extracted and digested with HindIII. bar gene was digoxigenin labeled and used as the probe for analysis. (C) Relative GmAKT2 expression in the leaves of the transgenic lines. T₂ generations of Oe1, Oe2, Oe3, and Oe4 and WT were cultured in a hydroponic system. RNA was extracted from the leaves of 2-week-old seedlings. GmAKT2 transcript levels were determined by qRT-PCR. Data represent means of three biological replicates with error bars indicating SD. GmACTIN expression was used as the internal control.