Abstract
An accurate assessment of the disease resistance status of plants to fungal pathogens is an essential requirement for the development of resistant crop plants. Many disease resistance phenotypes are partial rather than obvious immunity and are frequently scored using subjective qualitative estimates of pathogen development or plant disease symptoms. Here we report a method for the accurate comparison of total fungal biomass in plant tissues. This method, called the WAC assay, is based upon the specific binding of the plant lectin wheat germ agglutinin to fungal chitin. The assay is simple, high-throughput, and sensitive enough to discriminate between single Puccinia graminis f.sp tritici infection sites on a wheat leaf segment. It greatly lends itself to replication as large volumes of tissue can be pooled from independent experiments and assayed to provide truly representative quantification, or, alternatively, fungal growth on a single, small leaf segment can be quantified. In addition, as the assay is based upon a microscopic technique, pathogen infection sites can also be examined at high magnification prior to quantification if desired and average infection site areas are determined. Previously, we have demonstrated the application of the WAC assay for quantifying the growth of several different pathogen species in both glasshouse grown material and large-scale field plots. Details of this method are provided within.
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References
Strange RN (2005) Plant disease: a threat to global food security. Annu Rev Phytopathol 43:83–116
Ayliffe MA, Singh R, Lagudah ES (2008) Durable resistance to stem rust needed. Curr Opin Plant Biol 11:187–192
Fisher MC, Henk DA, Briggs CJ, Brownstein JS, Madoff LC, McCraw SL, Gurr SL (2012) Emerging fungal threats to animal, plant and ecosystem health. Nature 484:186–194
Gassner G, Starib W (1932) Die bestimmung der biologischen rassen de weizengelbrostes (Puccinia glumarum f.sp tritici (Schmidt.) Erikss.undHenn.) Arbeiten der Biologischen Reichsanstalt fur Land und Forstwirtschaft, Berlin, 20: 141–163
Stakman EC, Stewart DM, Loegering WQ (1962) Identification of physiologic races of Puccinia graminis var. tritici. Agricultural Research Service E617. United States Department of Agriculture, Washington, DC
Horsfall JG, Barratt RW (1945) An improved grading system for measuring plant disease. Phytopathology 35:655
Bock CH, Gottwald TR, Parker PE, Ferrandino F, Welham S, van den Bosch F, Parnell S (2010) Some consequences of using the Horsfall-Barratt scale for hypothesis testing. Phytopathology 100:1030–1041
Ayliffe M, Periyannan S, Feechan A, Dry I, Schumann U, Wang M-B, Pryor A, Lagudah E (2013) A simple method for comparing fungal biomass in infected plant tissues. Mol Plant Microbe Interact 26: 658–667
Mirelman D, Galun E, Sharon N, Lotan R (1975) Inhibition of fungal growth by wheat germ agglutinin. Nature 256:414–416
Molano J, Bowers B, Cabib E (1980) Distribution of chitin in the yeast cell wall. J Cell Biol 85:199–212
Chong J, Harder DE, Rohringer R (1985) Cytochemical studies on Puccinia graminis f. sp. tritici in a compatible wheat host. I. Walls of intercellular hyphal cells and haustorium mother cells. Can J Bot 63:1713–1724
O’Connell RJ, Ride JP (1990) Chemical detection and ultrastructural localisation of chitin in cells walls of Colletotrichum lindemuthianum. Physiol Mol Plant Pathol 37:39–53
Allen AK, Neuberger A, Sharon N (1973) The purification, composition and specificity of wheat germ agglutinin. J Biochem 131:155–162
Roth J (1978) The biological functions of the lectins, 2. Wheat germ agglutinin. In: Roth J (ed) The lectins: molecular probes in cell biology and membrane research. VEB Gustav Fischer Verlag, Jena, pp 21–23
Ayliffe M, Devilla R, Mago R, White R, Talbot M, Pryor A, Leung H (2011) Non-host resistance of rice to rust pathogens. Mol Plant Microbe Interact 24:1143–1155
Acknowledgments
The authors wish to thank the Australian Grains Research and Development Corporation for financial support, and Melanie Soliveres and Soma Chakraborty for technical support.
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Ayliffe, M. et al. (2014). Simple Quantification of In Planta Fungal Biomass. In: Birch, P., Jones, J., Bos, J. (eds) Plant-Pathogen Interactions. Methods in Molecular Biology, vol 1127. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-986-4_13
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DOI: https://doi.org/10.1007/978-1-62703-986-4_13
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