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A Simple Method for Comparing Fungal Biomass in Infected Plant Tissues

June 2013 , Volume 26 , Number  6
Pages  658 - 667

Michael Ayliffe,1 Sambasivam K. Periyannan,1 Angela Feechan,2 Ian Dry,2 Ulrike Schumann,1 Ming-Bo Wang,1 Anthony Pryor,1 and Evans Lagudah1

1CSIRO Plant Industry, Box 1600, Clunies Ross Street, Canberra, ACT, 2601, Australia; 2CSIRO Plant Industry, Box 350, Glen Osmond, SA, 5064, Australia

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Accepted 7 February 2013.

Plant phenotypes resistant and susceptible to fungal pathogens are usually scored using qualitative, subjective methods that are based upon disease symptoms or by an estimation of the amount of visible fungal growth. Given that plant resistance genes often confer partial resistance to fungal pathogens, a simple, sensitive, nonsubjective quantitative method for measuring pathogen growth would be highly advantageous. This report describes an in planta quantitative assay for fungal biomass based upon detection of chitin using wheat germ agglutinin conjugated to a fluorophore. Using this assay, the growth of wheat rust pathogens on wheat was assayed and the additivity of several adult plant and seedling resistance genes to Puccinia striiformis, P. graminis, and P. triticina was assayed on both glasshouse- and field-grown material. The assay can discriminate between individual rust pustules on a leaf segment or, alternatively, compare fungal growth on field plots. The quantification of Erysiphe necator (powdery mildew) growth on Vitis vinifera (grapevine) is also demonstrated, with resistant and susceptible cultivars readily distinguished. Given that chitin is a major cell wall component of many plant fungal pathogens, this robust assay will enable simple and accurate measurement of biomass accumulation in many plant–fungus interactions.

© 2013 The American Phytopathological Society