Targeted mutation of barley (1,3;1,4)-beta-glucan synthases reveals complex relationships between the storage and cell wall polysaccharide content
AuthorGarcia-Gimenez, G; Barakate, A; Smith, P; Stephens, J; Khor, SF; Doblin, MS; Hao, P; Bacic, A; Fincher, GB; Burton, RA; ...
Source TitleThe Plant Journal
University of Melbourne Author/sDoblin, Monika
AffiliationSchool of BioSciences
Document TypeJournal Article
CitationsGarcia-Gimenez, G., Barakate, A., Smith, P., Stephens, J., Khor, S. F., Doblin, M. S., Hao, P., Bacic, A., Fincher, G. B., Burton, R. A., Waugh, R., Tucker, M. R. & Houston, K. (2020). Targeted mutation of barley (1,3;1,4)-beta-glucan synthases reveals complex relationships between the storage and cell wall polysaccharide content. PLANT JOURNAL, 104 (4), pp.1009-1022. https://doi.org/10.1111/tpj.14977.
Access StatusOpen Access
Barley (Hordeum vulgare L) grain is comparatively rich in (1,3;1,4)-β-glucan, a source of fermentable dietary fibre that protects against various human health conditions. However, low grain (1,3;1,4)-β-glucan content is preferred for brewing and distilling. We took a reverse genetics approach, using CRISPR/Cas9 to generate mutations in members of the Cellulose synthase-like (Csl) gene superfamily that encode known (HvCslF6 and HvCslH1) and putative (HvCslF3 and HvCslF9) (1,3;1,4)-β-glucan synthases. Resultant mutations ranged from single amino acid (aa) substitutions to frameshift mutations causing premature stop codons, and led to specific differences in grain morphology, composition and (1,3;1,4)-β-glucan content. (1,3;1,4)-β-Glucan was absent in the grain of cslf6 knockout lines, whereas cslf9 knockout lines had similar (1,3;1,4)-β-glucan content to wild-type (WT). However, cslf9 mutants showed changes in the abundance of other cell-wall-related monosaccharides compared with WT. Thousand grain weight (TGW), grain length, width and surface area were altered in cslf6 knockouts, and to a lesser extent TGW in cslf9 knockouts. cslf3 and cslh1 mutants had no effect on grain (1,3;1,4)-β-glucan content. Our data indicate that multiple members of the CslF/H family fulfil important functions during grain development but, with the exception of HvCslF6, do not impact the abundance of (1,3;1,4)-β-glucan in mature grain.
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