Identification of candidate MYB transcription factors that influence CslF6 expression in barley grain

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dc.contributor.authorGarcia-Gimenez, G.
dc.contributor.authorSchreiber, M.
dc.contributor.authorDimitroff, G.
dc.contributor.authorLittle, A.
dc.contributor.authorSingh, R.
dc.contributor.authorFincher, G.B.
dc.contributor.authorBurton, R.A.
dc.contributor.authorWaugh, R.
dc.contributor.authorTucker, M.R.
dc.contributor.authorHouston, K.
dc.date.issued2022
dc.description.abstract(1,3;1,4)-β-Glucan is a non-cellulosic polysaccharide required for correct barley grain fill and plant development, with industrial relevance in the brewing and the functional food sector. Barley grains contain higher levels of (1,3;1,4)-β-glucan compared to other small grain cereals and this influences their end use, having undesirable effects on brewing and distilling and beneficial effects linked to human health. HvCslF6 is the main gene contributing to (1,3;1,4)-β-glucan biosynthesis in the grain. Here, the transcriptional regulation of HvCslF6 was investigated using an in-silico analysis of transcription factor binding sites (TFBS) in its putative promoter, and functional characterization in a barley protoplast transient expression system. Based on TFBS predictions, TF classes AP2/ERF, MYB, and basic helix-loop-helix (bHLH) were over-represented within a 1,000 bp proximal HvCslF6 promoter region. Dual luciferase assays based on multiple HvCslF6 deletion constructs revealed the promoter fragment driving HvCslF6 expression. Highest HvCslF6 promoter activity was narrowed down to a 51 bp region located -331 bp to -382 bp upstream of the start codon. We combined this with TFBS predictions to identify two MYB TFs: HvMYB61 and HvMYB46/83 as putative activators of HvCslF6 expression. Gene network analyses assigned HvMYB61 to the same co-expression module as HvCslF6 and other primary cellulose synthases (HvCesA1, HvCesA2, and HvCesA6), whereas HvMYB46/83 was assigned to a different module. Based on RNA-seq expression during grain development, HvMYB61 was cloned and tested in the protoplast system. The transient over-expression of HvMYB61 in barley protoplasts suggested a positive regulatory effect on HvCslF6 expression.
dc.description.statementofresponsibilityGuillermo Garcia-Gimenez, Miriam Schreiber, George Dimitroff, Alan Little, Rohan Singh, Geoffrey B. Fincher, Rachel A. Burton, Robbie Waugh, Matthew R. Tucker, and Kelly Houston
dc.identifier.citationFrontiers in Plant Science, 2022; 13:1-18
dc.identifier.doi10.3389/fpls.2022.883139
dc.identifier.issn1664-462X
dc.identifier.issn1664-462X
dc.identifier.orcidLittle, A. [0000-0003-0107-7643]
dc.identifier.orcidBurton, R.A. [0000-0002-0638-4709]
dc.identifier.orcidTucker, M.R. [0000-0003-4661-6700]
dc.identifier.urihttps://hdl.handle.net/2440/136676
dc.language.isoen
dc.publisherFrontiers Media
dc.relation.grantARC
dc.rights© 2022 Garcia-Gimenez, Schreiber, Dimitroff, Little, Singh, Fincher, Burton, Waugh, Tucker and Houston. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
dc.source.urihttps://doi.org/10.3389/fpls.2022.883139
dc.subject(1,3;1,4)-β-glucan
dc.subjectMYB
dc.subjectbarley
dc.subjectcell wall
dc.subjecttranscription factors
dc.titleIdentification of candidate MYB transcription factors that influence CslF6 expression in barley grain
dc.typeJournal article
pubs.publication-statusPublished online

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