Plant and Cell Physiology Advance Access originally published online on April 23, 2009
Plant and Cell Physiology 2009 50(6):1075-1089; doi:10.1093/pcp/pcp060
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Down-Regulation of PoGT47C Expression in Poplar Results in a Reduced Glucuronoxylan Content and an Increased Wood Digestibility by Cellulase
1Department of Plant Biology, University of Georgia, Athens, GA 30602, USA
2National Exposure Research Laboratory, US Environmental Protection Agency, Athens, GA 30605, USA
*Corresponding author: E-mail, zhye{at}plantbio.uga.edu; Fax, +1-706-542-1805.
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Abstract |
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Xylan is the second most abundant polysaccharide in dicot wood. Unraveling the biosynthetic pathway of xylan is important not only for our understanding of the process of wood formation but also for our rational engineering of wood for biofuel production. Although several glycosyltransferases are implicated in glucuronoxylan (GX) biosynthesis in Arabidopsis, whether their close orthologs in woody tree species are essential for GX biosynthesis during wood formation has not been investigated. In fact, no studies have been reported to evaluate the effects of alterations in secondary wall-associated glycosyltransferases on wood formation in tree species. In this report, we demonstrate that PoGT47C, a poplar glycosyltransferase belonging to family GT47, is essential for the normal biosynthesis of GX and the normal secondary wall thickening in the wood of the hybrid poplar Populus alba x tremula. RNA interference (RNAi) inhibition of PoGT47C resulted in a drastic reduction in the thickness of secondary walls, a deformation of vessels and a decreased amount of GX in poplar wood. Structural analysis of GX using nuclear magnetic resonance (NMR) spectroscopy demonstrated that the reducing end of GX from poplar wood contains the tetrasaccharide sequence, β-D-Xylp-(1
3)-
-L-Rhap-(12)--D-GalpA-(14)-D-Xylp, and that its abundance was significantly decreased in the GX from the wood of the GT47C-RNAi lines. The transgenic wood was found to yield more glucose by cellulase digestion than the wild-type wood, indicating that the GX reduction in wood reduces the recalcitrance of wood to cellulase digestion. Together, these results provide direct evidence demonstrating that the PoGT47C glycosyltransferase is essential for normal GX biosynthesis in poplar wood and that GX modification could improve the digestibility of wood cellulose by cellulase.
Keywords: Biomass recalcitrance - Glucuronoxylan biosynthesis - Glycosyltransferase - Poplar - Secondary wall biosynthesis - Wood formation
Abbreviations: CaMV, cauliflower mosaic virus; fra8, fragile fiber8; GalA, galacturonic acid; GlcA, glucuronic acid; GT, glycosyltransferase; GUS, β-glucuronidase; GX, glucuron-oxylan; irx, irregular xylem; MALDI-TOF MS, matrix-assisted laser-desorption ionization time-of-flight mass spectrometry; MeGlcA, 4-O-methyl-glucuronic acid; NMR, nuclear magnetic resonance; PBS, phosphate-buffered saline; Rha, rhamnose; RNAi, RNA interference; Xyl, xylose.
(Received March 12, 2009; Accepted April 21, 2009)
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