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Functional co-expression of a fungal ferulic acid esterase and a β-1,4 endoxylanase in Festuca arundinacea (tall fescue) modifies post-harvest cell wall deconstruction

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Abstract

Main conclusion

Improved post-harvest cell wall deconstruction of tall fescue leaves has been demonstrated by in-planta co-expression of a constitutively expressed ferulic acid esterase together with a senescence-induced β-1,4 endoxylanase.

Tall fescue plants (Festuca arundinacea) constitutively expressing vacuole- or apoplast-targeted ferulic acid esterase from Aspergillus niger were retransformed with a senescence-induced and apoplast-targeted β-1,4 endo-xylanase from Trichoderma reesei. Enzyme activities in co-expressing plants stabilized after repeated vegetative propagation, with xylanase activity in senescent leaves increasing and ferulic acid esterase activity decreasing after tillering. Plants co-expressing both enzymes in the apoplast, with the lowest levels of ferulate monomers and dimers and the lowest levels of cell wall arabinoxylans, released ten times more cell wall hydroxycinnamic acids and five times more arabinoxylan from the cell wall on autodigestion compared to expression of ferulic acid esterase or xylanase alone. These plants also showed a 31 % increase in cellulase-mediated release of reducing sugars, a 5 % point increase in in vitro dry matter digestibility and a 23 % increase in acetyl bromide-soluble lignin. However, plant growth was adversely affected by expressing FAE in the apoplast, giving plants with narrower shorted leaves, and a 71 % decrease in biomass.

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Abbreviations

AIR:

Alcohol insoluble residues,

AX:

Arabinoxylan

XYN2:

β-1,4 endo-xylanase

FA:

Ferulic acid

FAEA:

Ferulic acid esterase

HCAs:

Hydroxycinnamic acids

HPAEC:

Anion exchange chromatography

HPLC:

High performance liquid chromatography

IVDMD:

In vitro dry matter digestibility

LmSee1:

Lolium multiflorum senescence promoter

PAHBAH:

p-Hydroxybenzoic acid hydrazide

pCA:

p-Coumaric acid

TFA:

Trifluoroacetic acid

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Acknowledgments

Funding for this work was provided by the Biotechnology and Biological Sciences Research Council (BBSRC) grant number D10116 and Genencor Inc. and by USDA-DOE Plant Feedstock Genomics Research Program (ER64701). We thank Dr. Paul Robson (IBERS Aberystwyth) for the use of pBKS and Fred Potter (ex IGER Aberystwyth), for advice on aspects of statistical analysis.

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  1. Marcia M. de O. Buanafina

    Present address: Department of Biology, The Pennsylvania State University, 208 Mueller Laboratory, University Park, PA, 16802, USA

Authors and Affiliations

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

    Erica A. Shearer

  2. Institute of Biological, Environmental and Rural Sciences, Aberystwyth University, Plas Gogerddan, Aberystwyth, SY23 3EB, Wales, UK

    Sue Dalton, Tim Langdon & E. Timms-Taravella

  3. Institute of Grassland and Environmental Research, Plas Gogerddan, Aberystwyth, SY23 3EB, Wales, UK

    Marcia M. de O. Buanafina, Sue Dalton, Tim Langdon, E. Timms-Taravella & Phillip Morris

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  1. Marcia M. de O. Buanafina
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Correspondence to Marcia M. de O. Buanafina.

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Buanafina, M.M.d.O., Dalton, S., Langdon, T. et al. Functional co-expression of a fungal ferulic acid esterase and a β-1,4 endoxylanase in Festuca arundinacea (tall fescue) modifies post-harvest cell wall deconstruction. Planta 242, 97–111 (2015). https://doi.org/10.1007/s00425-015-2288-2

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