Detection of fungal pathogens using the polymerase chain reactio

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

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536 231, 536 243, 435 912, C12Q 168, C07H 2102, C07H 2104, C12P 1934

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059552743

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BRIEF SUMMARY
FIELD OF THE INVENTION

The present invention relates to the use of species-specific primers in polymerase chain reaction assays for the detection of fungal pathogens. The use of these primers enables the detection of specific isolates of fungal pathogens and the monitoring of disease development in plant populations.


BACKGROUND OF THE INVENTION

Diseases in plants cause considerable crop loss from year to year resulting both in economic deprivation to farmers and additionally in many parts of the world to shortfalls in the nutritional provision for local populations. The widespread use of fungicides has provided considerable security against plant pathogen attack. However, despite $1 billion worth of expenditure on fungicides, worldwide crop losses amounted to approximately 10% of crop value in 1981 (James, 1981; Seed Sci. & Technol. 9: 679-685).
The severity of the destructive process of disease depends on the aggressiveness of the pathogen and the response of the host. One aim of most plant breeding programs is to increase the resistance of host plants to disease. Typically, different races of pathogens interact with different varieties of the same crop species differentially, and many sources of host resistance only protect against specific pathogen races. Furthermore, some pathogen races show early signs of disease symptoms, but cause little damage to the crop. Jones and Clifford (1983; Cereal Diseases, John Wiley) report that virulent forms of the pathogen are expected to emerge in the pathogen population in response to the introduction of resistance into host cultivars and that it is therefore necessary to monitor pathogen populations. In addition, there are several documented cases of the evolution of fungal strains which are resistant to particular fungicides. As early as 1981, Fletcher and Wolfe (1981; Proc. 1981 Brit. Crop Prot. Conf.) contended that 24% of the powdery mildew populations from spring barley, and 53% from winter barley showed considerable variation in response to the fungicide triadimenol and that the distribution of these populations varied between varieties with the most susceptible variety also giving the highest incidence of less susceptible types. Similar variation in the sensitivity of fungi to fungicides has been documented for wheat mildew (also to triadimenol), Botrytis (to benomyl), Pyrenophora (to organomercury), Pseudocercosporella (to MBC-type fungicides) and Mycosphaerella fijiensis to triazoles to mention just a few (Jones and Clifford; Cereal Diseases, John Wiley, 1983).
Cereal species are grown world-wide and represent a major fraction of world food production. Although yield loss is caused by many pathogens, the necrotizing pathogens Septoria and Pseudocercosporella are particularly important in the major cereal growing areas of Europe and North America (Jones and Clifford; Cereal Diseases, John Wiley, 1983). In particular, the differential symptomology caused by different isolates and species of these fungi make the accurate predictive determination of potential disease loss difficult. Consequently, the availability of improved diagnostic techniques for the rapid and accurate identification of specific pathogens will be of considerable use to field pathologists.
Four Septoria species parasitize the small grain species. Septoria tritici is the causative agent of leaf blotch and is virulent on wheat but also parasitizes triticale and rye. It typically causes leaf necrosis. Septoria nodorum is the causative agent of glume blotch and is parasitic on wheat, triticale, rye and barley and although mainly restricted to glumes is also found on leaf blades and sheaths. Septoria avenae is parasitic on oats, wheat and triticale and Septoria passerinii is restricted to barley. Septoria diseases occur in all wheat growing areas at economically important levels. Different Septoria diseases frequently occur concurrently within fields and on individual plants, where the disease symptoms may be collectively referred to as the "Septoria complex". Typically, the most commonly fou

REFERENCES:
patent: 4683195 (1987-07-01), Mullis et al.
patent: 4683202 (1987-07-01), Mullis
patent: 5324632 (1994-06-01), Weisburg et al.
patent: 5447848 (1995-09-01), Barns et al.
patent: 5585238 (1996-12-01), Ligon et al.
Ursi et al., APMIS 100: 635-639 (1992).
The Stratagene Catalog, p. 39 (1988).
Johanson et al. Use of PCR for detection of Mycosphaerella fijiensis and M. Musicola", the causal agents of Sigatoka leaf spots in banana and plantain", Mycol. Res., 97:670-674 (1993).
Nazar, R.N., et al, "Potential use of PCR-amplified ribosomal intergeneic sequences in the detection and differentiation of verticillium wilt pathogens", Physiol. and Molec. Plant Pathol., 39:1-11 (1991).
Poupard et al., "Molecular characterization of Pseudocerosporella herpotrichoides isolates by amplification of ribosomal DNA internal transcribed spaces", Plant Pathology, 42: 873-881 (1993).
Schesser et al., "Use of Polymerase Chain Reaction To Detect the Take-All Fungus, Gaeumannomyces graminis, in Infected Wheat Plants", Applied and Environ. Microbiol., 57(2):553-556 (1991).
Stratagene Catalog, 1988, p.39.
Tisserat et al., "Selective Amplification of rDNA Internal Transcribed Spacer Regions to Detect Opiosphaerella korrae and O. herpotricha", Phytopathology, 84(5): 478-482 (1994).
White, T.J., et al., "Amplification and Direct Sequencing of Fungal Ribosomal RNA Genes for Phylogenetics", In: PCR Protocols; Academic Press Inc., pp. 315-322 (1990).
Xue et al., "Pathotype identification of Leptosphaeria maculans with PCR and oligonucleotide primrers from ribosomal internal transribed spacer sequences", Physiological and Molecular Plant Pathology, 41: 179-188 (1992).
International Search Report Jul. 20, 1995.
GenBank Accession No. UO4237, computer printout, Jan. 3, 1994.
Gene characterization kits. Stratagene Catalog, 1988.

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