Functional characterization of barley betaglucanless mutants demonstrates a unique role for CslF6 in (1,3;1,4)-β-D-glucan biosynthesis

Shin Taketa, Takahisa Yuo, Takuji Tonooka, Yoichi Tsumuraya, Yoshiaki Inagaki, Naoto Haruyama, Oscar Larroque, Stephen A. Jobling

Research output: Contribution to journalArticle

44 Citations (Scopus)

Abstract

(1,3;1,4)-β-D-glucans (mixed-linkage glucans) are found in tissues of members of the Poaceae (grasses), and are particularly high in barley (Hordeum vulgare) grains. The present study describes the isolation of three independent (1,3;1,4)-β-D-glucanless (betaglucanless; bgl) mutants of barley which completely lack (1,3;1,4)-β-D-glucan in all the tissues tested. The bgl phenotype cosegregates with the cellulose synthase like HvCslF6 gene on chromosome arm 7HL. Each of the bgl mutants has a single nucleotide substitution in the coding region of the HvCslF6 gene resulting in a change of a highly conserved amino acid residue of the HvCslF6 protein. Microsomal membranes isolated from developing endosperm of the bgl mutants lack detectable (1,3;1,4)-β-D-glucan synthase activity indicating that the HvCslF6 protein is inactive. This was confirmed by transient expression of the HvCslF6 cDNAs in Nicotiana benthamiana leaves. The wild-type HvCslF6 gene directed the synthesis of high levels of (1,3;1,4)-β-D-glucans, whereas the mutant HvCslF6 proteins completely lack the ability to synthesize (1,3;1,4)-β-D-glucans. The fine structure of the (1,3;1,4)-β-D-glucan produced in the tobacco leaf was also very different from that found in cereals having an extremely low DP3/DP4 ratio. These results demonstrate that, among the seven CslF and one CslH genes present in the barley genome, HvCslF6 has a unique role and is the key determinant controlling the biosynthesis of (1,3;1,4)-β-D-glucans. Natural allelic variation in the HvCslF6 gene was found predominantly within introns among 29 barley accessions studied. Genetic manipulation of the HvCslF6 gene could enable control of (1,3;1,4)-β-D-glucans in accordance with the purposes of use.

Original languageEnglish
Pages (from-to)381-392
Number of pages12
JournalJournal of Experimental Botany
Volume63
Issue number1
DOIs
Publication statusPublished - Jan 2012

Fingerprint

glucans
Hordeum
barley
biosynthesis
mutants
Genes
genes
Poaceae
Tobacco
Endosperm
cellulose synthase
Glucans
polyglucosan
Mutant Proteins
proteins
Nicotiana benthamiana
Introns
genetic engineering
Hordeum vulgare
Proteins

Keywords

  • Cell wall
  • grasses
  • Hordeum vulgare
  • mixed-linkage glucans
  • polysaccharide

ASJC Scopus subject areas

  • Plant Science
  • Physiology

Cite this

Functional characterization of barley betaglucanless mutants demonstrates a unique role for CslF6 in (1,3;1,4)-β-D-glucan biosynthesis. / Taketa, Shin; Yuo, Takahisa; Tonooka, Takuji; Tsumuraya, Yoichi; Inagaki, Yoshiaki; Haruyama, Naoto; Larroque, Oscar; Jobling, Stephen A.

In: Journal of Experimental Botany, Vol. 63, No. 1, 01.2012, p. 381-392.

Research output: Contribution to journalArticle

Taketa, Shin ; Yuo, Takahisa ; Tonooka, Takuji ; Tsumuraya, Yoichi ; Inagaki, Yoshiaki ; Haruyama, Naoto ; Larroque, Oscar ; Jobling, Stephen A. / Functional characterization of barley betaglucanless mutants demonstrates a unique role for CslF6 in (1,3;1,4)-β-D-glucan biosynthesis. In: Journal of Experimental Botany. 2012 ; Vol. 63, No. 1. pp. 381-392.
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