MOESM1 of Brassinosteroid overproduction improves lignocellulose quantity and quality to maximize bioethanol yield under green-like biomass process in transgenic poplar
Creators: name: Fan, Chunfen
name: Yu, Hua
name: Qin, Shifei
name: Li, Yongli
name: Alam, Aftab
name: Xu, Changzhen
name: Fan, Di
name: Zhang, Qingwei
name: Wang, Yanting
name: Zhu, Wanbin
name: Peng, Liangcai
name: Luo, Keming
Publisher: name: figshare
Year: 2020
Subjects: subject: Biochemistry
subject: Medicine
subject: Microbiology
subject: FOS: Biological sciences
subject: Genetics
subject: Biotechnology
subject: Chemical Sciences not elsewhere classified
subject: Ecology
subject: Biological Sciences not elsewhere classified
subject: Infectious Diseases
subject: FOS: Health sciences
subject: Plant Biology

description: Additional file 1: Table S1. Information of DET2 genes. Pe: Populus euphratica, Ptr: P. trichocarpa, Me: Manihot esculenta, Jc: Jatropha curcas, Ai: Arachis ipaensis, Vv: Vitis vinifera, Ha: Helianthus annuus, Pb: Pyrus x bretschneideri, At: A. thaliana, Cq: Chenopodium quinoa, Mn: Morus notabilis, Eg: Eucalyptus grandis, Gr: Gossypium raimondii, Gb: Gossypium barbadense, Gh: Gossypium hirsutum, Sl: Solanum lycopersicum, Sc: Solanum chacoense, Os: Oryza sativa, Zm: Zea mays, Sb: Sorghum bicolor. Table S2. Gene primers used for PCR amplification. Table S3. Total pectin and lignin contents (% biomass) in stems of transgenic lines and WT. All data are given as mean ± SD (n = 3, technical replicates). Statistical analyses were performed using Student’s t test as **P &lt; 0.01 and *P &lt; 0.05. Table S4. Wall polymer features in stems of transgenic lines and WT. Rha, Rhamnose; Fuc, Fucose; Ara, Arabinose; Xyl, Xylose; Man, Mannose; Gal, Galactose. Table S5. Characteristic peaks of the FTIR spectra in biomass residues as referred from previous studies. Table S6. Comparison of bioethanol yields obtained in the transgenic poplar plant and other woody plants. Fig. S1 Multiple sequence alignment and phylogenetic analysis of DET2. (a) The phylogenetic relationship of PtoDET2 with other DET2 proteins. (b) Sequence alignments of PtoDET2. The GenBank accession numbers of DET2s were listed in Table S1. Fig. S2 Generation of transgenic poplars. (a) Diagram of the PtoDET2-OE vector. (b) The Hyg levels in the PtoDET2-OE lines. (c) The expression levels of PtoDET2 in the PtoDET2-OE lines. The poplar ubiquitin gene was used as an internal control. (d) Diagram of three CRISPR/Cas9 target sites of PtoDET2. T1, T2 and T3 indicate the positions of sgRNA-targeted sites. (e) Determination of the mutations in the coding region of PtoDET2 generated by the CRISPR/Cas9 system. The text on the right summarizes mutation details in two independent CRISPR/Cas9-generated lines (L11 and L17). Primers are listed in Table S2. Fig. S3 Expression of cell differentiation, expansion and wall biosynthetic genes in PtoDET2 transgenic plants. (a) Cell differentiation genes; (b) cell expansion genes; (c) cellulose biosynthetic genes; (d) hemicellulose biosynthetic genes. Primers are listed in Table S2. The poplar ubiquitin gene was used as an internal control. All data are given as mean ± SD from three biological repeats. Statistical analyses were performed using Student’s t test as **P &lt; 0.01. Fig. S4 Hexoses released from enzymatic hydrolysis after various chemical pretreatments. (a) Hexose yields released from enzymatic (mixed-cellulase) hydrolysis after pretreatments with H2SO4, (b) NaOH, (c) CaO or (d) Na2S + Na2CO3. All data are given as mean ± SD of three technical repeats. Fig. S5 Expression of PtoGH9 genes in transgenic PtoDET2 plants. Primers are listed in Table S2 available as Supplementary Data. The poplar ubiquitin gene was used as an internal control. All data are given as mean ± SD from three technical repeats. Statistical analyses were performed using Student’s t test as **P &lt; 0.01. Fig. S6 Mass balance analysis for bioethanol production during biomass process with green liquor in transgenic poplar lines and WT. Fig. S7 Fourier transform infrared spectra (FTIR) profiling in transgenic poplar lines and WT. Black line as raw material (R) and red line as biomass residue from Na2S + Na2CO3 pretreatment (P). Characteristic peaks of the FTIR spectra were referred in Table S5. Fig. S8 A hypothetical model to demonstrate an integrated approach effective for maximum bioethanol production in lignocellulose-improved transgenic poplar plants overproducing BRs.