title: Cell wall composition and digestibility alterations in Brachypodium distachyon achieved through reduced expression of the UDP-arabinopyranose mutase.
abstract: Nucleotide-activated sugars are essential substrates for plant cell-wall carbohydrate-polymer biosynthesis. The most prevalent grass cell wall (CW) sugars are glucose (Glc), xylose (Xyl), and arabinose (Ara). These sugars are biosynthetically related via the UDP-sugar interconversion pathway. We sought to target and generate UDP-sugar interconversion pathway transgenic Brachypodium distachyon lines resulting in CW carbohydrate composition changes with improved digestibility and normal plant stature. Both RNAi-mediated gene-suppression and constitutive gene-expression approaches were performed. CWs from 336 T0 transgenic plants with normal appearance were screened for complete carbohydrate composition. RNAi mutants of BdRGP1, a UDP-arabinopyranose mutase, resulted in large alterations in CW carbohydrate composition with significant decreases in CW Ara content but with minimal change in plant stature. Five independent RNAi-RGP1 T1 plant lines were used for in-depth analysis of plant CWs. Real-time PCR analysis indicated that gene expression levels for BdRGP1, BdRGP2, and BdRGP3 were reduced in RNAi-RGP1 plants to 15-20% of controls. CW Ara content was reduced by 23-51% of control levels. No alterations in CW Xyl and Glc content were observed. Corresponding decreases in CW ferulic acid (FA) and ferulic acid-dimers (FA-dimers) were observed. Additionally, CW p-coumarates (pCA) were decreased. We demonstrate the CW pCA decrease corresponds to Ara-coupled pCA. Xylanase-mediated digestibility of RNAi-RGP1 Brachypodium CWs resulted in a near twofold increase of released total carbohydrate. However, cellulolytic hydrolysis of CW material was inhibited in leaves of RNAi-RGP1 mutants. Our results indicate that targeted manipulation of UDP-sugar biosynthesis can result in biomass with substantially altered compositions and highlights the complex effect CW composition has on digestibility.
authors.affiliation: U.S. Dairy Forage Research Center, United States Department of Agriculture - Agricultural Research Service, Madison WI, USA.
authors.forename: David M
authors.initials: DM
authors.lastname: Rancour
authors.affiliation: U.S. Dairy Forage Research Center, United States Department of Agriculture - Agricultural Research Service, Madison WI, USA.
authors.forename: Ronald D
authors.initials: RD
authors.lastname: Hatfield
authors.affiliation: U.S. Dairy Forage Research Center, United States Department of Agriculture - Agricultural Research Service, Madison WI, USA.
authors.forename: Jane M
authors.initials: JM
authors.lastname: Marita
authors.affiliation: Department of Genetics, University of Georgia, Athens GA, USA.
authors.forename: Nicholas A
authors.initials: NA
authors.lastname: Rohr
authors.affiliation: Department of Genetics, University of Georgia, Athens GA, USA.
authors.forename: Robert J
authors.initials: RJ
authors.lastname: Schmitz
keywords: Brachypodium distachyon; biomass; grass; hydroxycinnamates; nucleotide–sugar; plant cell wall