Increase in cellulose accumulation and improvement of saccharification by overexpression of arabinofuranosidase in rice

PLoS One. 2013 Nov 4;8(11):e78269. doi: 10.1371/journal.pone.0078269. eCollection 2013.

Abstract

Cellulosic biomass is available for the production of biofuel, with saccharification of the cell wall being a key process. We investigated whether alteration of arabinoxylan, a major hemicellulose in monocots, causes an increase in saccharification efficiency. Arabinoxylans have β-1,4-D-xylopyranosyl backbones and 1,3- or 1,4-α-l-arabinofuranosyl residues linked to O-2 and/or O-3 of xylopyranosyl residues as side chains. Arabinose side chains interrupt the hydrogen bond between arabinoxylan and cellulose and carry an ester-linked feruloyl substituent. Arabinose side chains are the base point for diferuloyl cross-links and lignification. We analyzed rice plants overexpressing arabinofuranosidase (ARAF) to study the role of arabinose residues in the cell wall and their effects on saccharification. Arabinose content in the cell wall of transgenic rice plants overexpressing individual ARAF full-length cDNA (OsARAF1-FOX and OsARAF3-FOX) decreased 25% and 20% compared to the control and the amount of glucose increased by 28.2% and 34.2%, respectively. We studied modifications of cell wall polysaccharides at the cellular level by comparing histochemical cellulose staining patterns and immunolocalization patterns using antibodies raised against α-(1,5)-linked l-Ara (LM6) and β-(1,4)-linked d-Xyl (LM10 and LM11) residues. However, they showed no visible phenotype. Our results suggest that the balance between arabinoxylan and cellulose might maintain the cell wall network. Moreover, ARAF overexpression in rice effectively leads to an increase in cellulose accumulation and saccharification efficiency, which can be used to produce bioethanol.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabinose / metabolism
  • Biofuels
  • Cell Wall / chemistry
  • Cell Wall / metabolism*
  • Cellulose / chemistry
  • Cellulose / metabolism*
  • Gene Expression Regulation, Plant*
  • Glucose / metabolism
  • Glycoside Hydrolases / classification
  • Glycoside Hydrolases / genetics*
  • Glycoside Hydrolases / metabolism
  • Immunohistochemistry
  • Oryza / genetics*
  • Oryza / metabolism
  • Phylogeny
  • Plant Proteins / classification
  • Plant Proteins / genetics*
  • Plant Proteins / metabolism
  • Plants, Genetically Modified
  • Substrate Specificity
  • Xylans / chemistry
  • Xylans / metabolism*

Substances

  • Biofuels
  • Plant Proteins
  • Xylans
  • Cellulose
  • arabinoxylan
  • Arabinose
  • Glycoside Hydrolases
  • alpha-N-arabinofuranosidase
  • Glucose

Grants and funding

This work was supported by a Grant-in-Aid for Scientific Research on Priority Areas (18075004 to H. Iwai), a Grant-in-Aid for Scientific Research on Innovative Areas (24114006 to S. Satoh and H. Iwai) and grants from the Ministry of Agriculture, Forestry and Fisheries of Japan (Genomics for Agricultural Innovation, GMA0007 to S. Satoh) and AMR0001 to H. Ichikawa, and Green Technology Project, EF1004 to H. Ichikawa. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.