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Functional testing of a PF02458 homologue of putative rice arabinoxylan feruloyl transferase genes in Brachypodium distachyon

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Abstract

Main conclusion

We show that changing the expression of a putative feruloyl transferase gene belonging to the BAHD acyl-transferase family alters the levels of cell wall esterified ferulates and diferulates in Brachypodium distachyon cell walls.

While the potential of grass cell walls for biofuel production has been realized, the technology for lignocellulosic biomass conversion for the production of ethanol is still inefficient because of structural mechanisms that plants have evolved to make the cell wall recalcitrant to enzymatic attack. One of these mechanisms in grasses involves the esterification of arabinoxylans in the cell wall with ferulic acid via an ester linkage to arabinose side chains on xylans. These ferulates undergo oxidative coupling reactions to form ferulate dimers, thus crosslinking polysaccharides. Arabinoxylan feruloylation is an important factor that determines cell wall recalcitrance because it directly cross-links xylans and because ferulates act as nucleating sites for the formation of lignin and for the linkage of lignin to the xylan/cellulose network. Here we report on the effects of changing the expression of Bradi2g43520 (BdAT1), a homologue of the rice feruloyl transferase gene Os01g42880 belonging to the Pfam PF02458 family, in Brachypodium distachyon. Down regulation in several independent RNAi::BdAT1 lines, resulted in up to a 35 % reduction of ferulate levels in both leaves and stems compared to control plants, over 2–3 generations of selfing. In contrast, overexpression of putative BdAT1 resulted in an increase of up to 58 and 47 % of ferulate levels in leaves and stems, respectively, compared to control plants and analyzed over 2–3 generations of selfing. These findings suggest that Bradi2g43520 may be a good candidate for feruloylation of AX in Brachypodium.

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Abbreviations

AFT:

Arabinoxylan feruloyl-transferase

AIR:

Isolated cell walls

Ara:

Arabinose

AX:

Arabinoxylan

cDNA:

Complementary DNA

EST:

Expressed sequence tags

FA:

Ferulic acid

FAEA:

Ferulic acid esterase

GAPDH:

Glyceraldehyde-3-phosphate dehydrogenase

HCAs:

Hydroxycinnamic acids

HPAEC:

High performance anion exchange chromatography

HPLC:

High performance liquid chromatography

PAHBAH:

ρ-hydroxybenzoic acid hydrazide

pCA:

p-coumarate

qRT-PCR:

Quantitative reverse-transcription polymerase chain reaction

RNAi:

RNA interference

SamDC:

S-adenosylmethionine decarboxylase

TFA:

Trifluoroacetic acid

UBC18:

Ubiquitin-conjugating enzyme 18

CaMV 35S:

Cauliflower mosaic virus (CaMV) 35S promoter

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Acknowledgments

The authors acknowledge the USDA-DOE Plant Feedstock Genomics Research Program (ER64701) for funding and Prof. Phillip Morris for useful discussions on the manuscript.

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  1. Department of Biology, The Pennsylvania State University, 208 Mueller Laboratory, University Park, PA, 16802, USA

    Marcia M. de O. Buanafina, Howard W. Fescemyer, Mandeep Sharma & Erica A. Shearer

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  1. Marcia M. de O. Buanafina
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  2. Howard W. Fescemyer
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Correspondence to Marcia M. de O. Buanafina.

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Buanafina, M.M.d., Fescemyer, H.W., Sharma, M. et al. Functional testing of a PF02458 homologue of putative rice arabinoxylan feruloyl transferase genes in Brachypodium distachyon . Planta 243, 659–674 (2016). https://doi.org/10.1007/s00425-015-2430-1

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