Differentiation of Phytophthora infestans Sporangia from Other Airborne Biological Particles by Flow Cytometry

ABSTRACT The ability of two different flow cytometers, the Microcyte (Optoflow) and the PAS-III (Partec), to differentiate sporangia of the late-blight pathogen Phytophthora infestans from other potential airborne particles was compared. With the PAS-III, light scatter and intrinsic fluorescence parameters could be used to differentiate sporangia from conidia of Alternaria or Botrytis spp., rust urediniospores, and pollen of grasses and plantain. Differentiation between P. infestans sporangia and powdery mildew conidia was not possible by these two methods but, when combined with analytical rules evolved by genetic programming methods, could be achieved after staining with the fluorescent brightener Calcofluor white M2R. The potential application of these techniques to the prediction of late-blight epiphytotics in the field is discussed.

[1]  K. Westerdijk,et al.  Proceedings of the workshop on the European network for development of an integrated control strategy of potato late blight : Munich, Germany, 6-10 September 2000 , 1997 .

[2]  D B Kell,et al.  Genetic programming:  a novel method for the quantitative analysis of pyrolysis mass spectral data. , 1997, Analytical chemistry.

[3]  D B Kell,et al.  Genetic programming as an analytical tool for non-linear dielectric spectroscopy. , 1999, Bioelectrochemistry and bioenergetics.

[4]  Una-May O'Reilly,et al.  Genetic Programming II: Automatic Discovery of Reusable Programs. , 1994, Artificial Life.

[5]  J. Jinks,et al.  Spontaneous variability of single isolates of Phytophthora infestans , 1970 .

[6]  David W. Galbraith,et al.  Analysis of Higher Plants by Flow Cytometry and Cell Sorting , 1989 .

[7]  D. Kell,et al.  Fluorescent brighteners: novel stains for the flow cytometric analysis of microorganisms. , 1997, Cytometry.

[8]  Douglas B. Kell,et al.  Genomic computing: explanatory modelling for functional genomics , 2000, GECCO.

[9]  G. Nebe-von Caron,et al.  Current and future applications of flow cytometry in aquatic microbiology. , 2000, FEMS microbiology reviews.

[10]  John R. Koza.  Genetic Programming III - Darwinian Invention and Problem Solving , 1999, Evolutionary Computation.

[11]  John R. Koza,et al.  Genetic Programming III - Darwinian Invention and Problem Solving , 1999, Evolutionary Computation.

[12]  Nikolay I. Nikolaev,et al.  Genetic Programming and Data Structures: Genetic Programming+Data Structures=Automatic Programming , 2001, Softw. Focus.

[13]  R. J. Gilbert,et al.  The detection of caffeine in a variety of beverages using Curie-point pyrolysis mass spectrometry and genetic programming , 1999 .

[14]  J. M. Hirst,et al.  The summer air-spora at Rothamsted in 1952. , 1957, Journal of general microbiology.

[15]  John R. Koza,et al.  Genetic Programming III: Darwinian Invention & Problem Solving , 1999 .

[16]  D. Deere,et al.  Go with the flow – use of flow cytometry in environmental microbiology , 1997 .

[17]  H M Davey,et al.  Flow cytometric analysis of microorganisms. , 2000, Methods.

[18]  O. C. Blair,et al.  Practical Flow Cytometry , 1985, The Yale Journal of Biology and Medicine.

[19]  John R. Koza,et al.  Genetic programming - on the programming of computers by means of natural selection , 1993, Complex adaptive systems.

[20]  P. H. Tsao,et al.  Phytophthora: Its Biology, Taxonomy, Ecology, and Pathology , 1988 .

[21]  A. Cunningham.  Analysis of Microalgae and Cyanobacteria by Flow Cytometry , 1993 .

[22]  J. Jinks,et al.  Spontaneous variability of single isolates of Phytophthora infestans. I. Cultural variation , 1968 .

[23]  William B. Langdon,et al.  Genetic Programming and Data Structures: Genetic Programming + Data Structures = Automatic Programming! , 1998 .

[24]  B. Kleinhenz,et al.  Results of validation trials of Phytophthora DSS in Europe in 2000 , 2001 .

[25]  R. Allman.  Characterisation of fungal spores using flow cytometry , 1992 .

[26]  Wolfgang Banzhaf,et al.  Genetic Programming: An Introduction , 1997 .

[27]  T. Cavalier-smith,et al.  A revised six‐kingdom system of life , 1998, Biological reviews of the Cambridge Philosophical Society.

[28]  David Lloyd,et al.  Flow Cytometry in Microbiology , 1993, Springer London.

[29]  D. Kell,et al.  On the Determination of the Size of Microbial Cells Using Flow Cytometry , 1993 .

[30]  D. Kell,et al.  The deconvolution of pyrolysis mass spectra using genetic programming: application to the identification of some Eubacterium species. , 1998, FEMS microbiology letters.

[31]  G. Poste,et al.  Flow Cytometry: Present and Future , 1985, Bio/Technology.

[32]  S. Bartnicki-García,et al.  Cell wall chemistry, morphogenesis, and taxonomy of fungi. , 1968, Annual review of microbiology.

[33]  S. B. Goodwin,et al.  Increased genotypic diversity via migration and possible occurrence of sexual reproduction of Phytophthora infestans in Poland , 1994 .

[34]  Douglas B. Kell,et al.  Explanatory Analysis of the Metabolome Using Genetic Programming of Simple, Interpretable Rules , 2000, Genetic Programming and Evolvable Machines.

[35]  Lynne Boddy,et al.  Identification of Phytoplankton from Flow Cytometry Data by Using Radial Basis Function Neural Networks , 1999, Applied and Environmental Microbiology.

[36]  D. Kell,et al.  Flow cytometry and cell sorting of heterogeneous microbial populations: the importance of single-cell analyses. , 1996, Microbiological reviews.

[37]  M. Melamed,et al.  Size and refractive index dependence of simple forward angle scattering measurements in a flow system using sharply-focused illumination. , 1977, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[38]  G. A. Forbes,et al.  Estimating the global severity of potato late blight with GIS‐linked disease forecast models , 2000 .

[39]  Douglas B. Kell,et al.  GMP — good modelling practice: an essential component of good manufacturing practice , 1995 .

[40]  G. Tarran,et al.  Flow Cytometry at Sea , 1993 .