Abstract
Proteomics and transcriptomics are established functional genomics tools commonly used to study filamentous fungi. Metabolomics has recently emerged as another option to complement existing techniques and provide detailed information on metabolic regulation and secondary metabolism. Here, we describe broad generic protocols that can be used to undertake metabolomics studies in filamentous fungi.
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References
Fiehn, O. (2002) Metabolomics - The link between genotypes and phenotypes. Plant Mol. Biol. 48, 155–171.
Goodacre, R., Vaidyanathan, S., Dunn, W.B., Harrigan, G.G. & Kell, D.B. (2004) Metabolomics by numbers: Acquiring and understanding global metabolite data. Trends Biotechnol. 22, 245–252.
Hall, R.D. (2006) Plant metabolomics: From holistic hope, to hype, to hot topic. New Phytol. 169, 453–468.
Weckwerth, W. & Morgenthal, K. (2005) Metabolomics: From pattern recognition to biological interpretation. Drug Discov. Today 10, 1551–1558.
Kell, D.B., Brown, M., Davey, H.M., Dunn, W.B., Spasic, I. & Oliver, S.G. (2005) Metabolic footprinting and systems biology: The medium is the message. Nat. Rev. Microbiol. 3, 557–565.
Oliver, S.G., Winson, M.K., Kell, D.B. & Baganz, F. (1998) Systematic functional analysis of the yeast genome. Trends Biotechnol. 16, 373–378.
Hollywood, K., Brison, D.R. & Goodacre, R. (2006) Metabolomics: Current technologies and future trends. Proteomics 6, 4716–4723.
Throck Watson, J. & David Sparkman, O. (2007) Introduction to Mass Spectrometry: Instrumentation, applications and strategies for data interpretation. Wiley, Chichester.
Dunn, W.B. (2008) Current trends and future requirements for the mass spectrometric investigation of microbial, mammalian and plant metabolomes. Phys. Biol. 5, 1–24.
Lowe, R.G., Lord, M., Rybak, K., Trengove, R.D., Oliver, R.P. & Solomon, P.S. (2008) A metabolomic approach to dissecting osmotic stress in the wheat pathogen Stagonospora nodorum. Fungal Genet. Biol. 45, 1479–1486.
Solomon, P.S., Wilson, T.J.G., Rybak, K., Parker, K., Lowe, R.G.T. & Oliver, R.P. (2006) Structural characterisation of the interaction between Triticum aestivum and the dothideomycete pathogen Stagonospora nodorum. Eur. J. Plant Pathol. 114, 275–282.
Solomon, P.S., Rybak, K., Trengove, R.D. & Oliver, R.P. (2006) Investigating the role of calcium/calmodulin-dependent protein kinases in Stagonospora nodorum. Mol. Microbiol. 62, 367–381.
Tan, K.C., Trengove, R.D., Maker, G.L., Oliver, R.P. & Solomon, P.S. (2009) Metabolite profiling identifies the mycotoxin alternariol in the pathogen Stagonospora nodorum. Metabolomics 5, 330–335.
Nielsen, K.F. & Smedsgaard, J. (2003) Fungal metabolite screening: Database of 474 mycotoxins and fungal metabolites for dereplication by standardised liquid chromatography-UV-mass spectrometry methodology. J. Chromatogr. A 1002, 111–136.
Dettmer, K., Aronov, P.A. & Hammock, B.D. (2007) Mass spectrometry-based metabolomics. Mass Spectrom. Rev. 26, 51–78.
Nicholson, J.K. & Wilson, I.D. (2003) Understanding ‘global’ systems biology: Metabonomics and the continuum of metabolism. Nat. Rev. Drug Discov. 2, 668–676.
Sumner, L.W., Mendes, P. & Dixon, R.A. (2003) Plant metabolomics: Large-scale phytochemistry in the functional genomics era. Phytochemistry 62, 817–836.
IpCho, S.V.S., Tan, K.-C., Koh, G., Gummer, J., Oliver, R.P., Trengove, R.D. & Solomon, P.S. (2010) The transcription factor StuA regulates central carbon metabolism, mycotoxin production, and effector gene expression in the wheat pathogen Stagonospora nodorum. Eukaryot. Cell 9, 1100–1108.
Halket, J.M. & Zaikin, V.G. (2003) Review: Derivatization in mass spectrometry-1. Silylation. Eur. J. Mass Spectrom. 9, 1–21.
di Mavungu, J.D., Monbaliu, S., Scippo, M.L., Maghuin-Rogister, G., Schneider, Y.J., Larondelle, Y., Callebaut, A., Robbens, J., C., v.P. & S., d.S. (2009) LC-MS/MS multi-analyte method for mycotoxin determination in food supplements. Food Addit. Contam. A 26, 885–895.
Ren, Y., Zhang, Y., Shao, S., Cai, Z., Feng, L., Pan, H. & Wang, Z. Simultaneous determination of multi-component mycotoxin contaminants in foods and feeds by ultra-performance liquid chromatography tandem mass spectrometry. J. Chromatogr. A 1143, 48–64.
Senyuva, H.Z., Gilbert, J. & Ozturkoglu, S. (2008) Rapid analysis of fungal cultures and dried figs for secondary metabolites by LC/TOF-MS. Anal. Chim. Acta 617, 97–106.
Spanjer, M.C., Rensen, P.M. & Scholten, J.M. (2008) LC-MS/MS multi-method for mycotoxins after single extraction, with validation data for peanut, pistachio, wheat, maize, cornflakes, raisins and figs. Food Addit. Contam. A 25, 472–489.
Sulyok, M., Krska, R. & Schuhmacher, R. (2007) A liquid chromatography/tandem mass spectrometric multi-mycotoxin method for the quantification of 87 analytes and its application to semi-quantitative screening of moldy food samples. Anal. Bioanal. Chem. 389, 1505–1523.
Vishwanath, V., Sulyok, M., Labuda, R., Bicker, W. & Krska, R. (2009) Simultaneous determination of 186 fungal and bacterial metabolites in indoor matrices by liquid chromatography/tandem mass spectrometry. Anal. Bioanal. Chem. 5, 1355–1372.
de Koker, T.H., Mozuch, M.D., Cullen, D., Gaskell, J. & Kersten, P.J. (2004) Isolation and purification of pyranose 2-oxidase from Phanerochaete chrysosporium and characterization of gene structure and regulation. Appl. Environ. Microbiol. 70, 5794–5800.
Donker, H.C.W. & Braaksma, A. (1997) Changes in metabolite concentrations detected by 13C-NMR in the senescing mushroom (Agaricus bisporus). Postharvest Biol. Tec. 10, 127–134.
Ceccaroli, P., Saltarelli, R., Cesari, P., Pierleoni, R., Sacconi, C., Vallorani, L., Rubini, P., Stocchi, V. & Martin, F. (2003) Carbohydrate and amino acid metabolism in Tuber borchii mycelium during glucose utilization: a 13C NMR study. Fungal Genet. and Biol. 39, 168–175.
Martin, F., Ramstedt, M., Söderhäll, K. & Canet, D. (1988) Carbohydrate and amino acid metabolism in the ectomycorrhizal ascomycete Sphaerosporella brunnea during glucose utilization. Plant Physiol. 86, 935–940.
Ramstedt, M., Martin, F. & Söderhäll, K. (1989) Mannitol metabolism in the ectomycorrhizal basidiomycete Piloderma croceum during glucose utilization. A 13C-NMR study. Agric. Ecosys. Environ. 28, 409–414.
Thomas, G.H. & Baxter, R.L. (1987) Analysis of mutational lesions of acetate metabolism in Neurospora crassa by 13C nuclear magnetic resonance. J. of Bacteriol. 169, 359–366.
Yoshida, M., Murai, T. & Moriya, S. (1984) 13C NMR spectra of plant pathogenic fungi. Agric. Biol. Chem. 48, 909–914.
Jobic, C., Boisson, A.M., Gout, E., Rascle, C., Fèvre, M., Cotton, P. & Bligny, R. (2007) Metabolic processes and carbon nutrient exchanges between host and pathogen sustain the disease development during sunflower infection by Sclerotinia sclerotiorum. Planta 226, 251–265.
Martin, F., Boiffin, V.V. & Pfeffer, P.E. (1998) Carbohydrate and amino acid metabolism in the Eucalyptus globulus-Pisolithus tinctorius ectomycorrhiza during glucose utilization. Plant Physiol. 118, 627–635.
Martin, F., Canet, D. & Marchal, J.P. (1985) 13C nuclear magnetic resonance study of mannitol cycle and trehalose synthesis during glucose utilization by the ectomycorrhizal ascomycete Cenococcum graniforme. Plant Physiol. 77, 499–502.
Martin, F., Canet, D., Marchal, J.-P. & Brondeau, J. (1984) In vivo natural-abundance 13C nuclear magnetic resonance studies of living ectomycorrhizal fungi : observation of fatty acids in Cenococcum graniforme and Hebeloma crustuliniforme. Plant Physiol. 75, 151–153.
Shachar-Hill, Y., Pfeffer, P.E., Douds, D., Osman, S.F., Doner, L.W. & Ratcliffe, R.G. (1995) Partitioning of Intermediary carbon metabolism in vesicular-arbuscular mycorrhizal leek. Plant Physiol. 108, 7–15.
Bago, B., Pfeffer, P.E., Douds, D.D., Jr., Brouillette, J., Bécard, G. & Shachar-Hill, Y. (1999) Carbon metabolism in spores of the arbuscular mycorrhizal fungus Glomus intraradices as revealed by nuclear magnetic resonance spectroscopy. Plant Physiol. 121, 263–272.
Forgue, P., Halouska, S., Werth, M., Xu, K., Harris, S. & Powers, R. (2006) NMR metabolic profiling of Aspergillus nidulans to monitor drug and protein activity. J. Proteome Res. 5, 1916–1923.
Wishart, D.S. & Sykes, B.D. (1994) Chemical shifts as a tool for structure determination. Methods Enzymol. 239, 363–392.
Kanani, H., Chrysanthopoulos, P.K. & Klapa, M.I. (2008) Standardizing GC-MS metabolomics. J. Chromatogr. B 871, 191–201.
Indarti, E., Majid, M.I.A., Hashim, R. & Chong, A. (2005) Direct FAME synthesis for rapid total lipid analysis from fish oil and cod liver oil. J. Food Compos. Anal. 18, 161–170.
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Gummer, J.P.A. et al. (2012). Metabolomics Protocols for Filamentous Fungi. 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_15
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DOI: https://doi.org/10.1007/978-1-61779-501-5_15
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