Fig 1.
Construct maps: a. SBP- GUS b. 35S- GUS c. SBP- DGAT1; P. NOS: Nopaline synthase promoter; NPTII: Neomycin phosphotransferase gene; T. NOS: Nopaline synthase terminator; SBP.P: SBP promoter, gusA: β-glucuronidase gene; CaMV 35S.P: CaMV 35S promoter; DGAT1: Diacylglycerol acyl-CoA acyltransferase gene; TerE9: E9 terminator.
Table 1.
Identified cis-acting regulatory elements in SBP using PLANTCARE and PLACE databases.
Fig 2.
Detection of the transgenic tobacco by PCR analysis: a. SBP-DGAT1 transgenic plants; 1: DGAT1 (1135 bp), 2: nptII (1400 bp), and 3: vir (900 bp) are the PCR products that were synthesized by DGAT1, nptII, and vir forward and reverses primers. 1–15: putative SBP-DGAT1 transgenic plants, WT: non-transgenic plant, (+): positive control, (-): negative control. b. SBP-GUS transgenic plants; 1: SBP (574 bp), 2: nptII (700 bp), and 3: vir (700 bp) are the PCR products that were synthesized by SBP, nptII, and vir forward and reverses primers. A- D putative SBP-GUS transgenic plants, (+): positive control. (-): negative control.
Fig 3.
Histochemical GUS analysis of transgenic tobacco plants at T0 generation: a. transgenic plant b. 35S-GUS c. non-transgenic control.
Fig 4.
Histochemical GUS analysis of T1 generation of SBP-GUS transgenic tobacco plants.
a. vegetative leaf b. reproductive leaf c. sepal d. stamen e. petal f. seeds: a.1, b.1, d.1, e.2, and f.2 represented GUS analysis in 35S-GUS samples. a.2, b.2, c.2, d.2, e.1, f.1 represented GUS analysis in non-transgenic plants. a.3, b.3, c.1, d.3, e.3, and f.3 represented GUS analysis in SBP-GUS samples.
Fig 5.
Histochemical GUS analysis in four stages of seed germination: a. endosperm appearance b. radical emerges c. radical growth and d. cotyledon appearance in SBP-GUS, 35S- GUS, and non-transgenic control plants.
Fig 6.
Histochemical GUS analysis in different stages of plant growth (4, 30, and 90 old days) in: a: SBP-GUS, b. 35s-GUS, and c. non-transgenic control plants.
Fig 7.
Functional impact of SBP promoter on mRNA levels of DGAT1 in seed and leaf of the transgenic plant (SBP-DGAT1).
Relative expression levels are expressed as fold changes in blue bars: seeds and in green bars: leaves. Mean values with different letters are significantly different by one-way ANOVA (P < 0.01), n = 6.
Fig 8.
Functional impact of SBP promoter on the seed and leaf oil content (%) of transgenic (SBP-DGAT1) and non-transgenic tobacco lines.
Mean values with different letters are significantly different by one-way ANOVA (P < 0.01), n = 6.
Fig 9.
Functional impact of SBP promoter on fatty acid composition in SBP-DGAT1 transgenic plant seeds.
Mean values with different letters are significantly different by one-way ANOVA (P < 0.01), n = 6.
Fig 10.
Functional impact of SBP promoter in size and weight of transgenic seeds: a. seed size (mm) and b. seed weight (mg). Seed’s size in c. non-transgenic and d. SBP-DGAT1 plants. Bars = 1mm. Mean values with different letters are significantly different by one-way ANOVA (P < 0.01), n = 6.