Identification and spatio-temporal expression analysis of barley genes that encode putative modular xylanolytic enzymes

Plant Sci. 2021 Jul:308:110792. doi: 10.1016/j.plantsci.2020.110792. Epub 2020 Dec 9.

Abstract

Arabinoxylans are cell wall polysaccharides whose re-modelling and degradation during plant development are mediated by several classes of xylanolytic enzymes. Here, we present the identification and new annotation of twelve putative (1,4)-β-xylanase and six β-xylosidase genes, and their spatio-temporal expression patterns during vegetative and reproductive growth of barley (Hordeum vulgare cv. Navigator). The encoded xylanase proteins are all predicted to contain a conserved carbohydrate-binding module (CBM) and a catalytic glycoside hydrolase (GH) 10 domain. Additional domains in some xylanases define three discrete phylogenetic clades: one clade contains proteins with an additional N-terminal signal sequence, while another clade contains proteins with multiple CBMs. Homology modelling revealed that all fifteen xylanases likely contain a third domain, a β-sandwich folded from two non-contiguous sequence segments that bracket the catalytic GH domain, which may explain why the full length protein is required for correct folding of the active enzyme. Similarly, predicted xylosidase proteins share a highly conserved domain structure, each with an N-terminal signal peptide, a split GH 3 domain, and a C-terminal fibronectin-like domain. Several genes appear to be ubiquitously expressed during barley growth and development, while four newly annotated xylanase and xylosidase genes are expressed at extremely high levels, which may be of broader interest for industrial applications where cell wall degradation is necessary.

Keywords: Arabinoxylan; Barley; Cell wall; Xylanase; Xylosidase.

MeSH terms

  • Amino Acid Sequence
  • Endo-1,4-beta Xylanases / chemistry
  • Endo-1,4-beta Xylanases / genetics*
  • Endo-1,4-beta Xylanases / metabolism
  • Gene Expression Profiling
  • Genes, Plant*
  • Hordeum / enzymology
  • Hordeum / genetics*
  • Phylogeny
  • Plant Proteins / chemistry
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism
  • Sequence Alignment
  • Spatio-Temporal Analysis
  • Xylosidases / chemistry
  • Xylosidases / genetics*
  • Xylosidases / metabolism

Substances

  • Plant Proteins
  • Xylosidases
  • exo-1,4-beta-D-xylosidase
  • Endo-1,4-beta Xylanases