RESUMO
In a study of the Phytophthora infestans population in Taiwan, samples with symptoms typical of late blight were collected from field crops in an important potato- (Solanum tuberosum) and tomato-(Lycopersicon esculentum) production area in the central highlands region. Isolates were obtained by surface disinfecting leaf sections and plating them onto antibiotic-amended rye A agar (1). After subculturing, the pathogen was confirmed as P. infestans on the basis of morphological characters (2). Mating type was determined by co-inoculating unamended rye agar plates with mycelial plugs of the test isolate and a reference P. infestans isolate of either the A1 or A2 mating type (four plates per test isolate, two with different A1, and two with different A2 reference isolates). After incubation (15°C darkness, 7 to 14 days), plates were examined microscopically for the presence of oospores where the colonies interacted. In 2004, one isolate of 200 tested, and in 2006, one isolate of 102 tested, produced oospores only with A1 reference isolates and were concluded to be A2 mating type. In vitro testing showed the two A2 isolates were metalaxyl-resistant (ED50 values >100 mg of metalaxyl per liter on rye grain agar), which is typical of recent P. infestans isolates from potato and tomato in this area (2). Twenty-one single-sporangial isolates from each of the two A2 strains were tested for mating type against two different A1 isolates of P. infestans and confirmed as A2. These isolates were characterized using the techniques described by Deahl et al. (1) and had the allozyme genotype 100/100/111, 100/100 at the loci coding for glucose-6-phosphate isomerase and peptidase, respectively, and were mitochondrial haplotype IIb. This multi-locus genotype is characteristic of recent P. infestans isolates from tomato and potato in Taiwan, but all previous such isolates were A1 mating type and attributed to the US-11 clonal lineage (1). When evaluated on differential hosts, both A2 isolates were tomato race PH-1 and complex potato race R 0,1,2,3,4,7,9,11. RG57 fingerprinting showed that the A2 isolates had fingerprints identical to each other and to A1 P. infestans isolates of the US-11 clonal lineage from tomato in Taiwan (101 011 100 100 110 101 011 001 1). Koch's postulates were completed and the two A2 isolates were found to be highly aggressive on cultivars of potato and tomato. To our knowledge, this is the first report of A2 mating type strains of P. infestans in the field in Taiwan, but currently, their incidence is very low (<1%). One crop from which an A2 isolate was obtained also yielded an A1 isolate, while A1 isolates were obtained from crops in the vicinity of the other. The concurrent presence of the two mating types of P. infestans poses a risk of sexual reproduction and oospore formation in tomato or potato in Taiwan. References: (1) K. L. Deahl et al. Pest Manag. Sci. 58:951, 2002. (2). D. C. Erwin and O. K. Ribeiro, Page 346 in: Phytophthora Diseases Worldwide. The American Phytopathological Society. St. Paul, MN, 1996.
RESUMO
There is only one published record of natural infection of black nightshade (Solanum nigrum L.) by Phytophthora infestans (Mont.) de Bary in England (3) and none from Wales. In August 2001, brown, necrotic leaf lesions with pale green margins were found on black nightshade weeds in a potato trial naturally infected with P. infestans at Henfaes Research Centre, University of Wales, Bangor. Although the plants were low growing with large, succulent leaves 4 to 5 cm long instead of having a more erect habit and smaller leaves, their identity was confirmed as S. nigrum; their atypical appearance may relate to the known phenotypic plasticity of this species (4). Infected leaflets incubated in moist chambers produced sporangia typical of P. infestans, and zoospores were released after chilling in water. Five isolates obtained from leaf fragments had growth on rye agar that was indistinguishable from that of P. infestans from potato. Detached leaflets of S. nigrum and S. tuberosum cv. Green Mountain inoculated with the S. nigrum isolates developed sporulating lesions under high humidity in 7 to 10 days; uninoculated controls remained symptomless. Inoculation of attached leaves of 10 potted S. nigrum plants resulted in seven plants developing necrotic lesions with a few sporangia 10 to 14 days later; sporulation developed mainly on lower leaves of plants that were older or had senesced. The remaining plants developed necrotic lesions with no sporulation, and P. infestans was reisolated from sporulating and nonsporulating lesions. All isolates were A1 mating type, metalaxyl-sensitive, and mitochondrial haplotype IIa, which are characteristics found commonly in isolates of P. infestans from potato in Wales (1). Single-sporangial isolates from each isolate were homozygous for glucose-6-phosphate isomerase and peptidase (Gpi 100/100, Pep 100/100). RG57 fingerprint analysis further established that all five black nightshade isolates were identical to each other and to some local P. infestans isolates from potato. P. infestans in Wales belongs to the new population (1), which may infect a wider host-range than the old US-1 clonal lineage. However, infected black nightshade was only found after late blight was widespread in potato fields. In subsequent years at the same site, weeds of S. nigrum have remained noninfected despite high levels of late blight pressure on adjacent potato plots. There is no evidence to suggest that this species acts as an overwintering host in Wales since it is an annual and lacks frost resistance. Field infection of S. nigrum by P. infestans has recently been reported in the Netherlands (2). Our observations confirm the potential of P. infestans to infect another solanaceous plant species. Alternative hosts may interfere with current disease control strategies because infected weeds would escape fungicide application and could serve as reservoirs of inoculum throughout the growing season. References: (1) J. P. Day and R. C. Shattock. Eur. J. Plant Pathol. 103:379, 1997. (2) W. G. Flier et al. Plant Pathol. 52:595, 2003. (3) J. M. Hirst and O. J. Steadman. Ann. Appl. Biol. 48:489, 1960. (4) B. S. Rogers and A. G. Ogg Jr. Page 30 in: Biology of Weeds of the Solanum Nigrum Complex (Solanum Section Solanum) in North America. USDA Publication ARM-W-23, 1981.
RESUMO
Control of Helminthosporium solani, the cause of silver scurf in potato tubers, has been impaired by selection of benzimidazole-resistant strains as a result of repeated use of the fungicide thiabendazole. Identification of thiabendazole-resistant strains of H. solani by conventional techniques takes several weeks. Primers designed from conserved regions of the fungal beta-tubulin gene were used to PCR amplify and sequence a portion of the gene. A point mutation was detected at codon 198 in thiabendazole-resistant isolates causing a change in the amino acid sequence from glutamic acid to alanine or glutamine. Species-specific PCR primers designed to amplify this region were used in conjunction with a restriction endonuclease to cause cleavage in sensitive isolates only and thus provide a rapid diagnostic test to differentiate field isolates.
