Figure 1.

Plant Growth and Development of Brachypodium Pro35S:BdTGD1 RNAi Lines.

(A) and (B) Relative BdTGD1 mRNA levels in different tissues of wild-type Bd21-3 (A) and 4-week-old Pro35S:BdTGD1RNAi lines (B). Different Brachypodium tissues were harvested as described in Supplemental Figures 1A to 1F. Total RNA was isolated and relative mRNA levels were determined by quantitative RT-PCR using BdUBC18 as an internal control. Different tissues from one plant were pooled together for one sample. The error bars represent sd; n = three independent plants. Significant differences are indicated (Student’s t test): *P < 0.05 and **P ≤ 0.01. B, H, and Q represent three independent Pro35S:BdTGD1 RNAi lines.

(C) Developmental phenotypes of Pro35S:BdTGD1 RNAi lines. B, H, and Q represent three independent Pro35S:BdTGD1 RNAi lines.

(D) FA composition of MGDG of 4-week-old Brachypodium leaf blades. The molar percentages of individual FAs relative to total FAs in MGDG were quantified using GC-FID of FAMEs. The error bars represent sd; n = three independent plants. Significant differences are indicated (Student’s t test): *P < 0.05 and **P ≤ 0.01. B, H, and Q represent three independent Pro35S:BdTGD1 RNAi lines.

(E) FA composition of DGDG of 4-week-old Brachypodium leaf blades. Molar percentages of individual FAs relative to total FAs in DGDG were quantified by GC-FID of FAMEs. The error bars represent sd; n = three independent plants. Significant differences are indicated (Student’s t test): *P < 0.05 and **P ≤ 0.01. B, H, and Q represent three independent Pro35S:BdTGD1 RNAi lines.

(F) Analysis of glycolipids. A thin-layer chromatogram of polar lipids is shown. Total lipids were extracted form 4-week-old Brachypodium leaf blades. Glycolipids were visualized by α-naphthol staining. The arrow indicates TGDG; + indicates lipid preparation 24 h after 1M MgCl₂ infiltration; − indicates lipid samples from leaf blades without Mg²⁺ treatment. B, H, and Q represent three independent Pro35S:BdTGD1 RNAi lines.