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Applied and Environmental Microbiology, October 1998, p. 4007-4014, Vol. 64, No. 10
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.

Polymorphisms in Phytophthora infestans: Four Mitochondrial Haplotypes Are Detected after PCR Amplification of DNA from Pure Cultures or from Host Lesions

Gareth W. Griffith* and David S. Shaw

School of Biological Sciences, University of Wales, Bangor, Gwynedd, LL57 2UW, Wales, United Kingdom

Received 23 February 1998/Accepted 21 July 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.

* Corresponding author. Mailing address: Institute of Biological Sciences, University of Wales, Aberystwyth, Ceredigion SY23 3DA, Wales, United Kingdom. Phone: 44-1970-622325. Fax: 44-1970-622350. E-mail: GWG{at}aber.ac.uk.

Applied and Environmental Microbiology, October 1998, p. 4007-4014, Vol. 64, No. 10
0099-2240/98/$04.00+0
Copyright © 1998, American Society for Microbiology. All rights reserved.


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