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<skos:definition xml:lang="en">Lignin determination in dry matter after hydrolysis by acetyl bromide. The protocol for lignin determination by acetyl bromide (Lignin) was adapted from Fukushima and Hatfield (2001). Internode samples were lyophilized after sampling and crushed (<100 µm) with mixer mill (Retsch MM301). Cell wall residues (CWR, i.e. structural carbohydrates) were prepared from 100 mg of the powder weighted precisely. Samples were washed twice in 5 ml of distilled water at 80°C. After centrifugation (10 minutes, 10000 rpm), the pellet was rinsed twice in 5 ml of absolute ethanol for 15 min at 80°C, then rinsed twice in 5 ml of acetone at room temperature for 10 min and left to dry under a fume hood overnight at room temperature. CWR was weighed to calculate the percentage of CWR in dry matter. Lignin from the prepared cell wall residue (5 mg +/- 1 mg) was solubilized in 1 ml (V1) of acetyl bromide solution (acetyl bromide/acetic acid (1/4, v/v)) in a glass vial at 55°C for 2.5 hours under shaking. Samples were then let to cool down at room temperature and 1.2 ml of NaOH 2M/acetic acid (9/50, v/v) was added in the vial. 0.1 mL (V2) of this sample was transferred in 300 µl of 0.5M hydroxylamine chlorhydrate and mixed with 1.4 ml of acetic acid. The absorbance (A280) of the samples was measured at 280 nm.</skos:definition>
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<skos:definition xml:lang="en">Lignin determination in dry matter after hydrolysis by acetyl bromide. The protocol for lignin determination by acetyl bromide (Lignin) was adapted from Fukushima and Hatfield (2001). Internode samples were lyophilized after sampling and crushed (<100 µm) with mixer mill (Retsch MM301). Cell wall residues (CWR, i.e. structural carbohydrates) were prepared from 100 mg of the powder weighted precisely. Samples were washed twice in 5 ml of distilled water at 80°C. After centrifugation (10 minutes, 10000 rpm), the pellet was rinsed twice in 5 ml of absolute ethanol for 15 min at 80°C, then rinsed twice in 5 ml of acetone at room temperature for 10 min and left to dry under a fume hood overnight at room temperature. CWR was weighed to calculate the percentage of CWR in dry matter. Lignin from the prepared cell wall residue (5 mg +/- 1 mg) was solubilized in 1 ml (V1) of acetyl bromide solution (acetyl bromide/acetic acid (1/4, v/v)) in a glass vial at 55°C for 2.5 hours under shaking. Samples were then let to cool down at room temperature and 1.2 ml of NaOH 2M/acetic acid (9/50, v/v) was added in the vial. 0.1 mL (V2) of this sample was transferred in 300 µl of 0.5M hydroxylamine chlorhydrate and mixed with 1.4 ml of acetic acid. The absorbance (A280) of the samples was measured at 280 nm.</skos:definition>
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