Abstract
The association of plants with endosymbiotic micro-organisms poses a particular challenge to metabolomics studies. The presence of endosymbionts can alter metabolic profiles of plant tissues by introducing non-plant metabolites such as fungal specific alkaloids, and by metabolic interactions between the two organisms. An accurate quantification of the endosymbiont and its metabolites is therefore critical for studies of interactions between the two symbionts and the environment.
Here, we describe methods that allow the quantification of the ryegrass Neotyphodium lolii fungal endosymbiont and major alkaloids in its host plant Lolium perenne. Fungal concentrations were quantified in total genomic DNA (gDNA) isolated from infected plant tissues by quantitative PCR (qPCR) using primers specific for chitinase A from N. lolii. To quantify the fungal alkaloids, we describe LC-MS based methods which provide coverage of a wide range of alkaloids of the indolediterpene and ergot alkaloid classes, together with peramine.
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References
Leuchtmann, A. (1992) Systematics, distribution, and host specificity of grass endophytes. Nat. Toxins 1, 150–162.
Schardl, C., Leuchtmann, L.A., and Spiering, M.J. (2004) Symbiosis of grasses with seedborne fungal endophytes. Ann. Rev. Plant Biol. 55, 315–340.
Bush, L.P., Wilkinson, H.H., and Schardl, C.L. (1997) Bioprotective alkaloids of grass-fungal endophyte symbiosis. Plant Physiol. 114, 1–7.
Gatenby, W.A., Munday-Finch, S.C., Wilkins, A.L., and Miles, C.O. (1996). Terpendole M, a novel indole-diterpenoid isolated from Lolium perenne infected with the endophytic fungus Neotyphodium lolii. J. Agric. Food Chem., 47, 1092–1097.
Panaccione, D.G., Tapper, B.A., Lane, G.A., Davies, E. and Fraser. K. (2003) Biochemical outcome of blocking the ergot alkaloid pathway of a grass endophyte. J. Agric. Food Chem. 51, 6429–6437.
Spiering, M.J., Lane, G.A., Christensen, M.J., and Schmid, J. (2005) Distribution of the fungal endophyte Neotyphodium lolii is not a major determinant of the distribution of fungal alkaloids in Lolium perenne plants. Phytochemistry 66, 195–202.
Rasmussen, S., Parsons, A.J., Bassett, S., Christensen, M.J., Hume, D.E., Johnson, L.J., Johnson, R.D., Simpson, W.R., Stacke, C., Voisey, C.,R., Xue, H., and Newman, J.A. (2007) High nitrogen supply and carbohydrate content reduce fungal endophyte and alkaloid concentration in Lolium perenne. New Phytol. 173, 787–797.
Cao, M., Koulman, A., Johnson, L.J., Lane, G.A., and Rasmussen, S. (2008) Advanced data-mining strategies for the analysis of direct-infusion ion trap mass spectrometry data from the association of perennial ryegrass with its endophytic fungus, Neotyphodium lolii. Plant Physiol. 146, 1501–1514.
Rasmussen, S., Parsons, A.J., Fraser, K., Xue, H. and Newman, J.A. (2008) Metabolic profiles of Lolium perenne are differentially affected by nitrogen supply, carbohydrate content, and fungal endophyte infection. Plant Physiol. 146, 1440–1453.
Koulman A, Lane GA, Christensen MJ, Fraser K, Tapper BA. (2006). Peramine and other fungal alkaloids are exuded in the guttation fluid of endophyte-infected grasses. Phytochemistry 68, 355–360.
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Rasmussen, S., Lane, G.A., Mace, W., Parsons, A.J., Fraser, K., Xue, H. (2011). The Use of Genomics and Metabolomics Methods to Quantify Fungal Endosymbionts and Alkaloids in Grasses. In: Hardy, N., Hall, R. (eds) Plant Metabolomics. Methods in Molecular Biology, vol 860. Humana Press. https://doi.org/10.1007/978-1-61779-594-7_14
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DOI: https://doi.org/10.1007/978-1-61779-594-7_14
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