Cell wall chemical analyses and enzymatic saccharification assay of OsCHS1-TDNA, OsCHI-CRISPR-a, and OsCHIL-CRISPR-a and their isogenic wild-types
| Cell wall chemotypes . | WT1 (cv. Zhonghua 11) . | OsCHS1-TDNA (cv. Zhonghua 11) . | WT2 (cv. Nipponbare) . | OsCHI-CRISPR-a (cv. Nipponbare) . | OsCHIL-CRISPR-a (cv. Nipponbare) . |
|---|---|---|---|---|---|
| Lignin content | |||||
| Klason lignin (mg/g CWR) | 152.0 ± 5.5 | 137.7 ± 9.3 | 120.4 ± 6.3 | 80.0 ± 10.9** | 135.6 ± 5.5* |
| Lignin composition by thioacidolysis | |||||
| Total monomer yield (µmol/g CWR) | 157.9 ± 9.7 | 161.2 ± 14.3 | 67.2 ± 0.2 | 56.4 ± 3.8* | 81.0 ± 3.5* |
| S/(S + G + H) (mol%) | 35.7 ± 2.7 | 34.7 ± 2.4 | 43.0 ± 1.0 | 42.8 ± 0.4 | 35.2 ± 0.6** |
| G/(S + G + H) (mol%) | 59.9 ± 3.0 | 62.7 ± 2.2 | 52.6 ± 0.7 | 54.3 ± 0.4* | 60.5 ± 0.6** |
| H/(S + G + H) (mol%) | 4.4 ± 0.4 | 2.7 ± 0.3** | 4.4 ± 0.3 | 2.9 ± 0.1** | 4.3 ± 0.0* |
| S/G | 0.6 ± 0.1 | 0.6 ± 0.1 | 0.8 ± 0.0 | 0.8 ± 0.0 | 0.6 ± 0.0** |
| Cell wall-bound hydroxycinnamates | |||||
| pCA (mg/g CWR) | 25.8 ± 0.6 | 25.6 ± 1.6 | 21.5 ± 3.5 | 15.0 ± 2.9 | 24.3 ± 1.2 |
| FA (mg/g CWR) | 3.6 ± 0.2 | 3.3 ± 0.3 | 2.6 ± 0.2 | 2.1 ± 0.1 | 2.7 ± 0.1 |
| Enzymatic saccharification efficiency | |||||
| Glc released after 6 h (mg/g CWR) | 198.0 ± 14.8 | 208.9 ± 3.4 | 226.3 ± 18.7 | 202.1 ± 21.2 | 266.8 ± 47.1 |
| Glc released after 24 h (mg/g CWR) | 211.9 ± 11.5 | 219.0 ± 3.8 | 239.3 ± 16.4 | 210.4 ± 19.3 | 275.8 ± 39.7 |
| Glc released after 6 h (g/g glucan) | 464.8 ± 33.4 | 475.6 ± 9.3 | 552.8 ± 42.0 | 531.5 ± 43.3 | 623.8 ± 116.0 |
| Glc released after 24 h (g/g glucan) | 496.3 ± 24.6 | 498.6 ± 13.1 | 584.7 ± 39.8 | 553.5 ± 36.4 | 644.7 ± 98.7 |
| Cell wall chemotypes . | WT1 (cv. Zhonghua 11) . | OsCHS1-TDNA (cv. Zhonghua 11) . | WT2 (cv. Nipponbare) . | OsCHI-CRISPR-a (cv. Nipponbare) . | OsCHIL-CRISPR-a (cv. Nipponbare) . |
|---|---|---|---|---|---|
| Lignin content | |||||
| Klason lignin (mg/g CWR) | 152.0 ± 5.5 | 137.7 ± 9.3 | 120.4 ± 6.3 | 80.0 ± 10.9** | 135.6 ± 5.5* |
| Lignin composition by thioacidolysis | |||||
| Total monomer yield (µmol/g CWR) | 157.9 ± 9.7 | 161.2 ± 14.3 | 67.2 ± 0.2 | 56.4 ± 3.8* | 81.0 ± 3.5* |
| S/(S + G + H) (mol%) | 35.7 ± 2.7 | 34.7 ± 2.4 | 43.0 ± 1.0 | 42.8 ± 0.4 | 35.2 ± 0.6** |
| G/(S + G + H) (mol%) | 59.9 ± 3.0 | 62.7 ± 2.2 | 52.6 ± 0.7 | 54.3 ± 0.4* | 60.5 ± 0.6** |
| H/(S + G + H) (mol%) | 4.4 ± 0.4 | 2.7 ± 0.3** | 4.4 ± 0.3 | 2.9 ± 0.1** | 4.3 ± 0.0* |
| S/G | 0.6 ± 0.1 | 0.6 ± 0.1 | 0.8 ± 0.0 | 0.8 ± 0.0 | 0.6 ± 0.0** |
| Cell wall-bound hydroxycinnamates | |||||
| pCA (mg/g CWR) | 25.8 ± 0.6 | 25.6 ± 1.6 | 21.5 ± 3.5 | 15.0 ± 2.9 | 24.3 ± 1.2 |
| FA (mg/g CWR) | 3.6 ± 0.2 | 3.3 ± 0.3 | 2.6 ± 0.2 | 2.1 ± 0.1 | 2.7 ± 0.1 |
| Enzymatic saccharification efficiency | |||||
| Glc released after 6 h (mg/g CWR) | 198.0 ± 14.8 | 208.9 ± 3.4 | 226.3 ± 18.7 | 202.1 ± 21.2 | 266.8 ± 47.1 |
| Glc released after 24 h (mg/g CWR) | 211.9 ± 11.5 | 219.0 ± 3.8 | 239.3 ± 16.4 | 210.4 ± 19.3 | 275.8 ± 39.7 |
| Glc released after 6 h (g/g glucan) | 464.8 ± 33.4 | 475.6 ± 9.3 | 552.8 ± 42.0 | 531.5 ± 43.3 | 623.8 ± 116.0 |
| Glc released after 24 h (g/g glucan) | 496.3 ± 24.6 | 498.6 ± 13.1 | 584.7 ± 39.8 | 553.5 ± 36.4 | 644.7 ± 98.7 |
Values refer to means ± standard deviations from biologically independent plants. Asterisks indicate significant differences between mutant line and its isogenic wild type (Student’s t test, n = 3, *P < 0.05,
P < 0.01).
Cell wall chemical analyses and enzymatic saccharification assay of OsCHS1-TDNA, OsCHI-CRISPR-a, and OsCHIL-CRISPR-a and their isogenic wild-types
| Cell wall chemotypes . | WT1 (cv. Zhonghua 11) . | OsCHS1-TDNA (cv. Zhonghua 11) . | WT2 (cv. Nipponbare) . | OsCHI-CRISPR-a (cv. Nipponbare) . | OsCHIL-CRISPR-a (cv. Nipponbare) . |
|---|---|---|---|---|---|
| Lignin content | |||||
| Klason lignin (mg/g CWR) | 152.0 ± 5.5 | 137.7 ± 9.3 | 120.4 ± 6.3 | 80.0 ± 10.9** | 135.6 ± 5.5* |
| Lignin composition by thioacidolysis | |||||
| Total monomer yield (µmol/g CWR) | 157.9 ± 9.7 | 161.2 ± 14.3 | 67.2 ± 0.2 | 56.4 ± 3.8* | 81.0 ± 3.5* |
| S/(S + G + H) (mol%) | 35.7 ± 2.7 | 34.7 ± 2.4 | 43.0 ± 1.0 | 42.8 ± 0.4 | 35.2 ± 0.6** |
| G/(S + G + H) (mol%) | 59.9 ± 3.0 | 62.7 ± 2.2 | 52.6 ± 0.7 | 54.3 ± 0.4* | 60.5 ± 0.6** |
| H/(S + G + H) (mol%) | 4.4 ± 0.4 | 2.7 ± 0.3** | 4.4 ± 0.3 | 2.9 ± 0.1** | 4.3 ± 0.0* |
| S/G | 0.6 ± 0.1 | 0.6 ± 0.1 | 0.8 ± 0.0 | 0.8 ± 0.0 | 0.6 ± 0.0** |
| Cell wall-bound hydroxycinnamates | |||||
| pCA (mg/g CWR) | 25.8 ± 0.6 | 25.6 ± 1.6 | 21.5 ± 3.5 | 15.0 ± 2.9 | 24.3 ± 1.2 |
| FA (mg/g CWR) | 3.6 ± 0.2 | 3.3 ± 0.3 | 2.6 ± 0.2 | 2.1 ± 0.1 | 2.7 ± 0.1 |
| Enzymatic saccharification efficiency | |||||
| Glc released after 6 h (mg/g CWR) | 198.0 ± 14.8 | 208.9 ± 3.4 | 226.3 ± 18.7 | 202.1 ± 21.2 | 266.8 ± 47.1 |
| Glc released after 24 h (mg/g CWR) | 211.9 ± 11.5 | 219.0 ± 3.8 | 239.3 ± 16.4 | 210.4 ± 19.3 | 275.8 ± 39.7 |
| Glc released after 6 h (g/g glucan) | 464.8 ± 33.4 | 475.6 ± 9.3 | 552.8 ± 42.0 | 531.5 ± 43.3 | 623.8 ± 116.0 |
| Glc released after 24 h (g/g glucan) | 496.3 ± 24.6 | 498.6 ± 13.1 | 584.7 ± 39.8 | 553.5 ± 36.4 | 644.7 ± 98.7 |
| Cell wall chemotypes . | WT1 (cv. Zhonghua 11) . | OsCHS1-TDNA (cv. Zhonghua 11) . | WT2 (cv. Nipponbare) . | OsCHI-CRISPR-a (cv. Nipponbare) . | OsCHIL-CRISPR-a (cv. Nipponbare) . |
|---|---|---|---|---|---|
| Lignin content | |||||
| Klason lignin (mg/g CWR) | 152.0 ± 5.5 | 137.7 ± 9.3 | 120.4 ± 6.3 | 80.0 ± 10.9** | 135.6 ± 5.5* |
| Lignin composition by thioacidolysis | |||||
| Total monomer yield (µmol/g CWR) | 157.9 ± 9.7 | 161.2 ± 14.3 | 67.2 ± 0.2 | 56.4 ± 3.8* | 81.0 ± 3.5* |
| S/(S + G + H) (mol%) | 35.7 ± 2.7 | 34.7 ± 2.4 | 43.0 ± 1.0 | 42.8 ± 0.4 | 35.2 ± 0.6** |
| G/(S + G + H) (mol%) | 59.9 ± 3.0 | 62.7 ± 2.2 | 52.6 ± 0.7 | 54.3 ± 0.4* | 60.5 ± 0.6** |
| H/(S + G + H) (mol%) | 4.4 ± 0.4 | 2.7 ± 0.3** | 4.4 ± 0.3 | 2.9 ± 0.1** | 4.3 ± 0.0* |
| S/G | 0.6 ± 0.1 | 0.6 ± 0.1 | 0.8 ± 0.0 | 0.8 ± 0.0 | 0.6 ± 0.0** |
| Cell wall-bound hydroxycinnamates | |||||
| pCA (mg/g CWR) | 25.8 ± 0.6 | 25.6 ± 1.6 | 21.5 ± 3.5 | 15.0 ± 2.9 | 24.3 ± 1.2 |
| FA (mg/g CWR) | 3.6 ± 0.2 | 3.3 ± 0.3 | 2.6 ± 0.2 | 2.1 ± 0.1 | 2.7 ± 0.1 |
| Enzymatic saccharification efficiency | |||||
| Glc released after 6 h (mg/g CWR) | 198.0 ± 14.8 | 208.9 ± 3.4 | 226.3 ± 18.7 | 202.1 ± 21.2 | 266.8 ± 47.1 |
| Glc released after 24 h (mg/g CWR) | 211.9 ± 11.5 | 219.0 ± 3.8 | 239.3 ± 16.4 | 210.4 ± 19.3 | 275.8 ± 39.7 |
| Glc released after 6 h (g/g glucan) | 464.8 ± 33.4 | 475.6 ± 9.3 | 552.8 ± 42.0 | 531.5 ± 43.3 | 623.8 ± 116.0 |
| Glc released after 24 h (g/g glucan) | 496.3 ± 24.6 | 498.6 ± 13.1 | 584.7 ± 39.8 | 553.5 ± 36.4 | 644.7 ± 98.7 |
Values refer to means ± standard deviations from biologically independent plants. Asterisks indicate significant differences between mutant line and its isogenic wild type (Student’s t test, n = 3, *P < 0.05,
P < 0.01).
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