Targeted mutation of barley (1,3;1,4)-β-glucan synthases reveals complex relationships between the storage and cell wall polysaccharide content

Guillermo Garcia-Gimenez, Abdellah Barakate, Pauline Smith, Jennifer Stephens, Shi F. Khor, Monika S. Doblin, Pengfei Hao, Antony Bacic, Geoffrey B. Fincher, Rachel A. Burton, Robbie Waugh, Matthew R. Tucker (Lead / Corresponding author), Kelly Houston (Lead / Corresponding author)

Research output: Contribution to journalArticle

Abstract

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 knock-out lines whereas cslf9 knock-out lines had similar (1,3;1,4)-β-glucan content to WT. However, cslf9 mutants showed changes in the abundance of other cell wall-related monosaccharides compared to WT. Thousand grain weight (TGW), grain length, width and surface area were altered in cslf6 knock-outs and to a lesser extent TGW in cslf9 knock-outs. 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.

Original languageEnglish
JournalPlant Journal
DOIs
Publication statusE-pub ahead of print - 5 Sep 2020

Keywords

  • barley
  • cell walls
  • gene editing
  • CRISPR/Cas9
  • (1,3;1,4)‐β‐glucan

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    Garcia-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)-β-glucan synthases reveals complex relationships between the storage and cell wall polysaccharide content. Plant Journal. https://doi.org/10.1111/tpj.14977