Ree technical replicates of 92 samples grouped in three biological replicates. Means of wild-type controls have been averages of 96 plants unless otherwise stated. Asterisks () indicate statistically important benefits at P 0.05, or as indicated in legends.supplementary InformationThe on line version contains supplementary material readily available at https://doi. org/10.1186/s13068021019051. Further file 1: Figure S1. HCT reactions in crude protein extracts and expression of recombinant HCTs in E. coli. Figure S2. Phylogenetic and structural evaluation from the BAHD loved ones of plant acyltransferases. Figure S3. Lignin deposition and organspecific expression of HCT in wildtype B. distachyon. Figure S4. Building of RNAi vectors for downregulation of Brachypodium HCT genes. Figure S5. HCT1 and HCT2 transcripts in T0 transgenic plants in which HCT1 had been targeted by RNA interference. Figure S6. Lignin content material and composition in T2 generation B. Caspase 10 Inhibitor Compound distachyon lines downregulated in HCT1 or HCT1 and HCT2. Figure S7. Determina tion of lignin molecular weight by gelpermeation chromatography. Table S1. Lignin content material and composition of internodes five and eight of B. distachyon stems harvested at 45 days following germination. Table S2. Indi vidual S:G and H:total lignin monomer ratios of each single and double B. distachyon HCTRNAi lines from T0 and T1 generations. Table S3. Lignin composition and linkage varieties as determined by NMR analysis. Table S4. Primers applied inside the present work. Acknowledgements We acknowledge funding in the University of North Texas to RAD and by the Bioenergy Sciences Center plus the Center for Bioenergy Innovation (Oak Ridge National Laboratory), US Department of Energy (DOE) Bioenergy Analysis Centers supported by the Dopamine Receptor Agonist Storage & Stability Office of Biological and Environmental Investigation within the DOE Office of Science, to RAD and AR. Authors’ contributions JCSY, JBR and RAD contributed towards the idea and design and style; JCSY, JB, LET, LGG, YP and AR produced reagents and/or acquired information. JCSY, JB, LET, LGG, YP, AR and RAD interpreted information; JCSY, JBR and RAD drafted the manuscript. All authors study and approved the final manuscript. Funding This operate was supported by the University of North Texas and by the Bioen ergy Sciences Center plus the Center for Bioenergy Innovation (Oak Ridge National Laboratory), US Division of Energy (DOE) Bioenergy Research Centers supported by the Workplace of Biological and Environmental Research within the DOE Office of Science. Availability of data and supplies All information generated or analyzed during this study are integrated in this published write-up and its supplementary information and facts files. Ethics approval and consent to participate Not applicable. Consent for publication Not applicable.NMR spectra have been acquired on a Bruker Avance III HD 500-MHz spectrometer equipped having a double resonance Prodigy cryoprobe with gradience in Z-direction (Bruker BBO-H F BBO-HD-05 Z). The lignin sample was dissolved in DMSO-d6 as well as a standard Bruker heteronuclear single quantum coherence (HSQC) pulse sequence was employed with all the following acquisition parameters: spectra width 12 ppm in F2 (1H) dimension with 2048 time of domain, 220 ppm in F1 (13C) dimension with 256 time of domain, a 1.5-s delay, a 1JC of 145 Hz, and 64 scans. The central DMSO solvent peak (13C/1H at 39.5/2.49) was applied for chemical shift calibration. Assignments of lignin compositional subunits and interunit linkage had been based on reported contours in HSQC spectra. The relative abundance of signal.