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 journalArticlepeer-review

35 Citations (Scopus)
164 Downloads (Pure)


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.

Original languageEnglish
Pages (from-to)1009-1022
Number of pages14
JournalPlant Journal
Issue number4
Early online date5 Sept 2020
Publication statusPublished - Nov 2020


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

ASJC Scopus subject areas

  • Genetics
  • Plant Science
  • Cell Biology


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