Generation of transgenic wheat (Triticum aestivum L.) accumulating heterologous endo-xylanase or ferulic acid esterase in the endosperm

Plant Biotechnol J. 2010 Apr;8(3):351-62. doi: 10.1111/j.1467-7652.2009.00490.x. Epub 2010 Jan 19.

Abstract

Endo-xylanase (from Bacillus subtilis) or ferulic acid esterase (from Aspergillus niger) were expressed in wheat under the control of the endosperm-specific 1DX5 glutenin promoter. Constructs both with and without the endoplasmic reticulum retention signal (Lys-Asp-Glu-Leu) KDEL were used. Transgenic plants were recovered in all four cases but no qualitative differences could be observed whether KDEL was added or not. Endo-xylanase activity in transgenic grains was increased between two and threefold relative to wild type. The grains were shrivelled and had a 25%-33% decrease in mass. Extensive analysis of the cell walls showed a 10%-15% increase in arabinose to xylose ratio, a 50% increase in the proportion of water-extractable arabinoxylan, and a shift in the MW of the water-extractable arabinoxylan from being mainly larger than 85 kD to being between 2 and 85 kD. Ferulic acid esterase-expressing grains were also shrivelled, and the seed weight was decreased by 20%-50%. No ferulic acid esterase activity could be detected in wild-type grains whereas ferulic acid esterase activity was detected in transgenic lines. The grain cell walls had 15%-40% increase in water-unextractable arabinoxylan and a decrease in monomeric ferulic acid between 13% and 34%. In all the plants, the observed changes are consistent with a plant response that serves to minimize the effect of the heterologously expressed enzymes by increasing arabinoxylan biosynthesis and cross-linking.

Publication types

  • Research Support, Non-U.S. Gov't
  • Research Support, U.S. Gov't, Non-P.H.S.

MeSH terms

  • Arabinose / analysis
  • Aspergillus niger / enzymology
  • Bacillus subtilis / enzymology
  • Carboxylic Ester Hydrolases / metabolism*
  • Cell Wall / chemistry
  • Coumaric Acids / analysis
  • Endo-1,4-beta Xylanases / metabolism*
  • Endosperm / enzymology*
  • Plants, Genetically Modified / enzymology
  • Plants, Genetically Modified / genetics
  • Transformation, Genetic
  • Triticum / enzymology*
  • Triticum / genetics
  • Xylans / analysis
  • Xylose / analysis

Substances

  • Coumaric Acids
  • Xylans
  • arabinoxylan
  • Xylose
  • ferulic acid
  • Arabinose
  • Carboxylic Ester Hydrolases
  • feruloyl esterase
  • Endo-1,4-beta Xylanases