A Trihelix Family Transcription Factor Is Associated with Key Genes in Mixed-Linkage Glucan Accumulation
[1];
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- Michigan State Univ., East Lansing, MI (United States). Dept. of Pant Biology; Michigan State Univ., East Lansing, MI (United States). Great Lakes Bioenergy Research Center; Michigan State University
- Univ. of Edinburgh, Scotland (United Kingdom). Edinburgh Cell Wall Group, Inst. of Molecular Plant Sciences
- Michigan State Univ., East Lansing, MI (United States). Dept. of Pant Biology; Michigan State Univ., East Lansing, MI (United States). Great Lakes Bioenergy Research Center; Øster Søgade, Copenhagen (Denmark)
- Michigan State Univ., East Lansing, MI (United States). Great Lakes Bioenergy Research Center, and MSU-DOE Plant Research Lab.
- Michigan State Univ., East Lansing, MI (United States). Dept. of Pant Biology; Michigan State Univ., East Lansing, MI (United States). Great Lakes Bioenergy Research Center; Michigan State Univ., East Lansing, MI (United States). Dept. of Biochemistry and Molecular Biology
Mixed-linkage glucan (MLG) is a polysaccharide that is highly abundant in grass endosperm cell walls and present at lower amounts in other tissues. Cellulose synthase-like F (CSLF) and cellulose synthase-like H genes synthesize MLG, but it is unknown if other genes participate in the production and restructuring of MLG. Using Brachypodium distachyon transcriptional profiling data, we identified a B. distachyon trihelix family transcription factor (BdTHX1) that is highly coexpressed with the B. distachyon CSLF6 gene (BdCSLF6), which suggests that BdTHX1 is involved in the regulation of MLG biosynthesis. To determine the genes regulated by this transcription factor, we conducted chromatin immunoprecipitation sequencing (ChIP-seq) experiments using immature B. distachyon seeds and an anti-BdTHX1 polyclonal antibody. The ChIP-seq experiment identified the second intron of BdCSLF6 as one of the most enriched sequences. The binding of BdTHX1 to the BdCSLF6 intron sequence was confirmed using electrophoretic mobility shift assays (EMSA). ChIP-seq also showed that a gene encoding a grass-specific glycoside hydrolase family 16 endotransglucosylase/hydrolase (BdXTH8) is bound by BdTHX1, and the binding was confirmed by EMSA. Radiochemical transglucanase assays showed that BdXTH8 exhibits predominantly MLG:xyloglucan endotransglucosylase activity, a hetero-transglycosylation reaction, and can thus produce MLG-xyloglucan covalent bonds; it also has a lower xyloglucan:xyloglucan endotransglucosylase activity. B. distachyon shoots regenerated from transformed calli overexpressing BdTHX1 showed an abnormal arrangement of vascular tissue and seedling-lethal phenotypes. Finally, these results indicate that the transcription factor BdTHX1 likely plays an important role in MLG biosynthesis and restructuring by regulating the expression of BdCSLF6 and BdXTH8.
- Research Organization:
- Michigan State Univ., East Lansing, MI (United States)
- Sponsoring Organization:
- USDOE Office of Science (SC), Biological and Environmental Research (BER) (SC-23)
- Grant/Contract Number:
- FC02-07ER64494; SC0018409
- OSTI ID:
- 1487019
- Journal Information:
- Plant Physiology (Bethesda), Journal Name: Plant Physiology (Bethesda) Journal Issue: 3 Vol. 178; ISSN 0032-0889
- Publisher:
- American Society of Plant BiologistsCopyright Statement
- Country of Publication:
- United States
- Language:
- English
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