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Proteomic Techniques for Plant–Fungal Interactions

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Plant Fungal Pathogens

Part of the book series: Methods in Molecular Biology ((MIMB,volume 835))

Abstract

Proteomics is a key technique that is helping elucidate many complex biological processes. The analysis of plant–pathogen interactions using proteomics is complicated by the presence of the proteomes of two species, but is benefiting from the developing maturity and power of these techniques. More and more pathogen genomes are being sequenced, so fungal proteomics is reaching its full potential and remains the chosen technology to unravel the molecular pathways of pathogenicity and resistance. In this chapter, we suggest proteomic strategies that have proved successful on various plant-interacting fungal species. Several protein extraction methods are described. For adequate quantitative analyses of protein abundances, we recommend either separation using two-dimensional gel electrophoresis or labelling with isobaric tags followed by two-dimensional HPLC separation. Proteins of interest are then identified using mass spectrometry. Identified proteins can assist in refining genome annotations, otherwise known as proteogenomics.

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Abbreviations

2-DE:

Two-dimensional electrophoresis

2-ME:

2-Mercaptoethanol

APS:

Ammonium persulfate

CAs:

Carrier Ampholites (IPG buffers from Bio-Rad for instance)

CHAPS:

3-[3-(Cholamidopropyl)dimethylammonio]-1-propanesulfonate

DMSO:

Dimethyl sulfoxide

DTT:

Dithiothreitol

EDTA Na2 :

Ethylenediaminetetraacetic acid disodium salt

IEF:

Isoelectric focusing

IPG:

Immobilised pH gradient

iTRAQ:

Isobaric tag for relative and absolute quantitation

LC:

Liquid chromatography

MALDI-ToF/ToF:

Matrix assisted laser desorption ionisation- time of flight/time of flight

MeOH:

Methanol

MMTS:

Methyl methanethiosulfonate

PMSF:

Phenylmethanesulfonide fluoride

PVP:

Polyvinylpyrrolidone

SCX:

Strong cation exchange

SDS:

Sodium dodecyl sulphate

SPE:

Solid phase extraction

TCA:

Trichloroacetic acid

TCEP:

Tris-(2-carboxyethyl)-phosphine-HCl

TCEP:

Tris(2-carboxyethyl)phosphine

TEAB:

Triethylammonium bicarbonate

TEMED:

Tetramethylethylenediamine

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Author information

Authors and Affiliations

  1. Research School of Biology, The Australian National University, Canberra, ACT, Australia

    Delphine Vincent, Liam Cassidy & Peter S. Solomon

  2. Australian Centre for Necrotrophic Fungal Pathogens, Environment and Agriculture, Curtin University, Perth, WA, Australia

    Kar-Chun Tan

  3. Department of Environment and Agriculture, Australian Centre for Necrotrophic Fungal Pathogens, Curtin University, Perth, ACT, Australia

    Richard P. Oliver

Authors
  1. Delphine Vincent
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  2. Kar-Chun Tan
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  3. Liam Cassidy
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  4. Peter S. Solomon
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  5. Richard P. Oliver
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Corresponding author

Correspondence to Richard P. Oliver .

Editor information

Editors and Affiliations

  1. Northern Crops Science Laboratory, USDA-ARS, North 18th Street 1307, Fargo, 58105-5677, North Dakota, USA

    Melvin D. Bolton

  2. Lab. Phytopathology, Wageningen University, Droevendaalsesteeg 1, Wageningen, 6708 PB, Netherlands

    Bart P.H.J. Thomma

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Vincent, D., Tan, KC., Cassidy, L., Solomon, P.S., Oliver, R.P. (2012). Proteomic Techniques for Plant–Fungal Interactions. In: Bolton, M., Thomma, B. (eds) Plant Fungal Pathogens. Methods in Molecular Biology, vol 835. Humana Press. https://doi.org/10.1007/978-1-61779-501-5_5

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  • DOI: https://doi.org/10.1007/978-1-61779-501-5_5

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-500-8

  • Online ISBN: 978-1-61779-501-5

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