Extended Data Fig. 7: Analysis of CRISPR/Cas9-mediated PHBMT1 knockout hybrid aspens.
From: Monolignol acyltransferase for lignin p-hydroxybenzoylation in Populus
a, Genotyping of the transgenic lines with selection marker NPT II gene. The experiment was repeated twice independently with the same results. Marker,Thermo Scientific GeneRuler 1 kb DNA ladder. b, Immunoblots with anti-PHBMT1 antibody on the crude proteins from the WT, non-edited control and PHBMT1 knockout transgenic lines. The immunoblots against anti-actin antibody served as the loading control. The original blots are presented in Supplemental Fig. 5a and c. This experiment was repeated twice independently with similar results. c, Growth phenotype of 2-month-old hybrid aspens. Bar = 4 cm. d, Growth phenotype of 6-month-old plants. Bar = 30 cm. e, The basal stem (30 cm) of the 6-month-old WT, non-edited control, and PHBMT1 knockout lines. f, Total acetyl bromide soluble lignin content in the stem of 2-month-old plantlets. Data are presented as means ± S. D. of three biological replicates. Asterisk indicates significant difference compared to the WT, * P < 0.05 (Student’s t-test, with two-tailed distribution, two-sample unequal variance). P = 3.76×10−4 (g1-8), 3.03×10−4 (g1-9), and 0.03 (g1-10). CWR, cell wall residues. g, Stem height of 3- and 5-month-old plants. h, Basal stem diameter of 3- and 5-month-old plants. i, Woody biomass yield of 6-month-old hybrid aspens. j, The calculated wood density of the basal stems of 6-month-old plants. k, l, Total acetyl bromide lignin content (k) and monomeric composition (l) of 6-month-old basal stems. CWR, cell wall residues. G, Guaiacyl monomer. S, Syringyl monomer. The data in (g–l) are presented as means ± S. D. of three biological replicates for WT, Ctrl, g1-9, g1-11 and five biological replicates for g1-8. No statistical differences were found between the knockout and the WT and/or non-edited control (Ctrl) plants.
