Targeted mutation of barley (1,3;1,4)-β-glucan synthases reveals complex relationships between the storage and cell wall polysaccharide content.
(1,3;1,4)-β-glucan
CRISPR/Cas9
barley
cell walls
gene editing
Journal
The Plant journal : for cell and molecular biology
ISSN: 1365-313X
Titre abrégé: Plant J
Pays: England
ID NLM: 9207397
Informations de publication
Date de publication:
11 2020
11 2020
Historique:
received:
05
06
2020
revised:
28
07
2020
accepted:
04
08
2020
pubmed:
6
9
2020
medline:
11
5
2021
entrez:
5
9
2020
Statut:
ppublish
Résumé
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.
Substances chimiques
Plant Proteins
0
Polysaccharides
0
beta-Glucans
0
beta-glucan, (1-3)(1-4)-
55965-23-6
Glucosyltransferases
EC 2.4.1.-
cellulose synthase
EC 2.4.1.-
glucan synthase
EC 2.4.1.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1009-1022Subventions
Organisme : European Research Council
ID : 669182
Pays : International
Organisme : Biotechnology and Biological Sciences Research Council
ID : BB/J014869/1
Pays : United Kingdom
Informations de copyright
© 2020 The Authors. The Plant Journal published by Society for Experimental Biology and John Wiley & Sons Ltd.
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