Polymorphisms in Phytophthora infestans: four mitochondrial haplotypes are detected after PCR amplification of DNA from pure cultures or from host lesions

• Published in: Applied and Environmental Microbiology Vol. 64; no. 10; pp. 4007 - 4014
• Main Authors: Griffith, G.W. (University of Wales, Wales, UK.), Shaw, D.S
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01.10.1998
• Subjects: ADN; Biological and medical sciences; DNA; Fundamental and applied biological sciences. Psychology; Fungal plant pathogens; LYCOPERSICON ESCULENTUM; Microbiology; MITOCHONDRIA; MITOCHONDRIE; MITOCONDRIA; Mycology; Pathogenicity, host-agent relations, miscellaneous strains, epidemiology; PCR; Phytopathology. Animal pests. Plant and forest protection; PHYTOPHTHORA INFESTANS; POLYMERASE CHAIN REACTION; RFLP; SOLANUM TUBEROSUM; Variation, races, biotypes, parasitic specialization, genetics; Plant pathogen; Solanum tuberosum; Phycomycetes; Primer; Mitochondrial DNA; Identification; Method; Fungi; Polymerase chain reaction; Restriction fragment length polymorphism; Dicotyledones; Angiospermae; Lycopersicon esculentum; Phytophthora infestans; Spermatophyta; Solanaceae; Haplotype; Thallophyta
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/aem.64.10.4007-4014.1998

Four pairs of primers were designed for PCR amplification of known polymorphic regions of the mitochondrial genome of Phytophthora infestans. Digestion of the amplified products with restriction enzymes allows identification of previously identified haplotypes. Product P2 cut with MspI uniquely identifies haplotypes Ib and IIa, while types Ia and IIb are differentiated by digestion of product P4 with EcoRI. Digestion of products P1 and P3 gave results similar to that with digestion of P4, but amplification of these products was less robust. Thus, all four common haplotypes are identified by amplifying and digesting products P2 and P4. Identification of haplotypes was also possible from DNA extracted directly from small, late-blight lesions on both tomato and potato leaves, making isolation of the fungus unnecessary. A rapid and efficient method of monitoring changes in the pathogen population is facilitated. These PCR primers were also useful for differentiating other Phytophthora species