Fig 1.
Phenotypic Variation in Seed Oil Content in High-Oil and Low-Oil Peanut Cultivars, Expression Pattern of AhLPAT2 in Eight Seed Developmental Stages of Them.
A. Comparison of seed oil content from high-oil cultivar Zhonghua12 and low-oil Te21. Values are average seed oil content ± SE (n = 3). Asterisks indicate a significant difference between the compared pairs at p < 0.01. B. Transcript abundance of AhLPAT2 in different stages of developing peanut seeds. Total RNA was prepared from developing seeds of Zhonghua12 and Te21 at 10 to 80 DAF. qRT-PCR was performed to compare the overall transcript abundance of AhLPAT2 in Zhonghua12 and Te21. Gene expression levels are shown relative to the expression of peanut AhUbiquitin gene in each sample. Values are means ± SE (n = 3).Asterisks indicate significant differences between the wild-type and transgenic lines at p < 0.01 (**) and p < 0.05 (*).
Fig 2.
Subcellular Localization of the AhLPAT2–GFP Fusion Protein in Onion Epidermal Cells.
The top panels show the expression vector containing AhLPAT2 and the GFP reporter gene. AhLPAT2 tagged with GFP in the C-terminus was inserted into the pEGFP vector between the KpnI and XmaI restriction sites. LB, left border; RB, right border; PNOS and TNOS are the promoter and polyadenylation signal of the nopaline synthase gene, respectively; NptII neomycin phosphotransferase II, CaMV 35S promoter. Bottom panels show the expression of GFP and AhLPAT2-GFP. (A) GFP fluorescence of onion epidermal cells expressing GFP. (B) Bright field image of (A). (C) Cell nuclei counterstained with DAPI. (D) Merged image of (A), (B) and (C). (E) GFP fluorescence of onion epidermal cells expressing AhLPAT2-GFP. (F) Bright field image of (E). (G) Cell nuclei counterstained with DAPI. (H) Merged image of (E), (F) and (G). Scale bars = 50 μm.
Fig 3.
AhLPAT2 Gene Expression in Young Siliques of the Selected Transgenic Lines Overexpressing AhLPAT2.
Two transgenic T3 lines, FNH1-21 and FNH2-2, were used. Total RNA was prepared from developing siliques of transgenic lines. Gene expression levels are shown relative to the expression of AtActin7 in each sample. Values are means ± SE (n = 3).
Fig 4.
Effect of AhLPAT2 Overexpression on Seed Storage Reserve Content of Independent Homozygous T3 Lines.
A. Seeds per silique. B. Numbers of siliques per plant. C. 100-seed weight. D. Seed yield per plant. Values are means ± SE (n = 10). Asterisks indicate significant differences between the wild-type and transgenic lines at p < 0.01 (**).
Fig 5.
Effect of AhLPAT2 Overexpression on Seed Oil and Protein Contents of Independent Homozygous T3 Lines.
A. Seed oil content as a percentage. B. Oil yield per plant. C. Seed protein content. Values are means ± SE of measurements of individual plants (n = 10). Asterisks indicate a significant difference between the wild-type and transgenic lines at p < 0.01 (**)and p < 0.05 (*).
Fig 6.
Effect of AhLPAT2 Overexpression on FA Content and Composition in Independent Homozygous Seeds.
A. Total FA content. B. Main FA profiles. C. The ratio of unsaturated to saturated FAs. D. The ratio of 18- to 20-carbon FA. Values are means ± SE (n = 10). Asterisks indicate a significant differences between the wild-type and transgenic lines at p < 0.01 (**) and p < 0.05 (*).
Fig 7.
Altered Expression of Genes Involved in the FAs and TAG Biosynthesis Pathway.
Values are means ± SE (n = 3). Total RNA was prepared from developing siliques of transgenic lines. Gene expression levels are shown relative to the expression of AtActin7 in each sample. The transcription level of each gene in the wildtype was set as 1.0.Asterisks indicate significant differences between the wild-type and transgenic lines at p < 0.01 (**) and p < 0.05 (*).
Fig 8.
Altered Expression of Genes Involved in Sucrose Metabolism and Glycolysis.
Values are means ± SE (n = 3). Total RNA was prepared from developing siliques of transgenic lines. Gene expression levels are shown relative to the expression of AtActin7 in each sample. The transcription level of each gene in the wild type was set as 1.0.Asterisks indicate significant differences between the wild-type and transgenic lines at p < 0.01 (**) and p < 0.05 (*).