Table 1.
Recombinant bovine lysozyme yield of transgenic sugarcane culm.
| Bovine lysozyme (BvLzm) expressing line and percentage of plants | BvLzm yield as determined by ELISA | |
|---|---|---|
| BvLzm (mg/kg culm mass) | TSP (%) | |
| Promoter stacking | ||
| 1. Single promoter:BvLzm lines | ||
| pU:BvLzm:single terminator (35ST) lines (43 lines; 114 plants) | 0.08–0.4 (range) | 0.01–0.06 (range) |
| 6.70% | 0.08–0.1 | 0.01–0.015 |
| 40.00% | 0.12–0.18 | 0.02–0.027 |
| 53.30% | 0.2–0.4 | 0.03–0.06 |
| 2. Double promoter:BvLzm lines | ||
| pUD:BvLzm:single terminator (35ST) lines (10 lines; 52 plants) | 0.5–0.7 (range) | 0.07–0.1 (range) |
| 71.00% | 0.5–0.58 | 0.07–0.077 |
| 29.00% | 0.6–0.7 | 0.08–0.1 |
| 3. Triple promoter:BvLzm lines | ||
| pUPE:BvLzm:3′UTR–single terminator (35ST) (10 lines; 32 plants) | 1.0–4.7 (range) | 0.1–0.7 (range) |
| 66.70% | 1.0–2.0 | 0.1–0.3 |
| 20.00% | 2.2–3.2 | 0.33–0.45 |
| 13.30% | 3.5–4.7 | 0.5–0.7 |
| pUDE:BvLzm:3′UTR–single terminator (35ST) (14 lines; 286 plants) | 1.5–6.0 (range) | 0.2–0.8 (range) |
| 27.00% | 1.5–2.0 | 0.2–0.3 |
| 62.00% | 2.2–3.2 | 0.33–0.45 |
| 4.50% | 3.5–4.7 | 0.5–0.7 |
| 6.50% | 5.0–6.0 | 0.7–0.8 |
| 4. Quadruple promoter:BvLzm lines | ||
| pUPBE:BvLzm:3′UTR-single terminator (35ST) (12 lines; 36 plants) | 2.0–6.3 (range) | 0.3–0.9 (range) |
| 33.30% | 2.0–3.5 | 0.3–0.5 |
| 22.20% | 4.0–5.5 | 0.6–0.77 |
| 44.50% | 6.0–6.3 | 0.8–0.9 |
| pUPBE:BvLzm:double terminator (35STNOST) (11 lines; 40 plants) | 2.4–10.0 (range) | 0.3–1.4 (range) |
| 24.20% | 2.4–3.5 | 0.3–0.5 |
| 51.70% | 4.0–5.5 | 0.6–0.77 |
| 24.10% | 6.0–10.0 | 0.8–1.4 |
| Event stacking | ||
| Five promoter:BvLzm lines | ||
| pUDE:BvLzm line + pP:BvLzm + pB:BvLzm (44 lines; 110 plants) | 11.0–82.5 (range) | 1.5–11.5 (range) |
| 12.10% | 11.0–12.4 | 1.5–1.7 |
| 24.20% | 15.9–21.1 | 2.2–2.9 |
| 33.30% | 26.2–32.3 | 3.6–4.5 |
| 18.30% | 59.9–82.5 | 8.3–11.5 |
The percentage (%) of lines with the corresponding BvLzm yield are indicated for each single or stacked promoter: BvLzm construct.
BvLzm, maize codon-optimized bovine lysozyme gene; U, maize ubiquitin 1 promoter; D, sugarcane dirigent16 promoter; P, sugarcane proline rich protein promoter; E, sugarcane elongation factor 1α promoter; B, Sugarcane bacilliform virus promoter; 3′UTR, 3′ untranslated region of Sorghum mosaic virus; 35ST, Cauliflower mosaic virus 35S terminator; NOST, Agrobacterium tumefaciens nopaline synthase terminator; ELISA, enzyme-linked immunosorbent assay; TSP, total soluble protein.