Targeted mutation of barley (1,3;1,4)-beta-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, Kelly Houston
The Plant Journal | WILEY | Published : 2020
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..View full abstract
Awarded by ERC
Awarded by BBSRC
Awarded by Australian Research Council Centre of Excellence in Plant Cell Walls
The authors thank Diane Davidson from the Functional Genomics (FUNGEN) facility, The James Hutton Institute, UK for barley transformation. The authors gratefully acknowledge Dr Alison Roberts and Dr Kathryn Wright, The James Hutton Institute (UK), for technical assistance during immunolabelling experiments and confocal imaging. The authors also thank Edwin R. Lampugnani for discussions. RW and Abdellah Barakate acknowledge support from ERC project 669182 'SHUFFLE' to RW. KH, PS, JS and RW acknowledge support from BBSRC project BB/J014869/1 the Rural & Environment Science & Analytical Services Division of the Scottish Government. RAB, GBF, MRT, MSD and Antony Bacic acknowledge the support of the Australian Research Council Centre of Excellence in Plant Cell Walls Grant CE1101007, and MSD, PH and Antony Bacic acknowledge the support of a startup grant from La Trobe University.