Expanding populations are often less genetically diverse at their margins than at the centre of a species' range. Established, older populations of the chestnut blight fungus, Cryphonectria parasitica, are more variable for vegetative compatibility (vc) types than in expanding populations in southeastern Europe where C. parasitica has colonized relatively recently. To test whether vc types represent clones, we genotyped 373 isolates of C. parasitica from southern Italy, Romania, Bulgaria, Macedonia, Greece and Turkey using 11 sequence-characterized amplified region (SCAR) markers. Ten SCAR loci and six vegetative incompatibility (vic) loci were polymorphic in these samples. These populations are clonal by all criteria tested: (i) among 373 isolates, we found only eight multilocus haplotypes, and the same haplotypes were found in multiple countries, sometimes separated in time by as much as 12 years; (ii) the number of haplotypes observed was significantly less than expected under random mating; (iii) populations are in linkage disequilibrium; (iv) the two sets of independent markers, SCARs and vc types, are highly correlated; and (v) sexual structures of C. parasitica were found only in Bulgaria and Romania. One mating type (MAT-1) was found in 98% of the isolates sampled. In contrast, a population in northern Italy, in the central part of the range in Europe, had 12 multilocus haplotypes among 19 isolates. The spread of a few clones could be the result either of founder effect and restricted migration, or these clones have greater fitness than others and spread because they are better adapted to conditions in southeastern Europe.
The genus Cuphea (Lythraceae) includes approximately 250 species of annual, evergreen perennials and short shrubs native to Central and South America. During the springs of 2003 and 2004, 10% of the nursery stock of approximately 12,000 potted cigar-flowers (C. ignea A. DC) grown in a screenhouse at a commercial ornamental nursery near Piedimonte Etneo, Sicily, had symptoms of wilt, defoliation, and rapid collapse of the entire plant. These foliar symptoms were associated with a reduced root system, browning of the collar, and dark brown discolored roots. A Phytophthora species was consistently recovered by plating small pieces of rotted roots of symptomatic plants onto selective medium (3); pure cultures were obtained by single-hypha transfers. On potato dextrose-agar (PDA), cardinal temperatures for growth were 10 to 35°C and the optimum was 28 to 30°C. Sporangiophores were umbellate or in a close monoclasial sympodium and mean dimensions of sporangia were 52 × 26 mm, with a mean length/width ratio of 2:1. Sporangia produced on V8 juice agar (VJA) were ellipsoid, fusiform, or limoniform with a tapered base. They were papillate, occasionally bipapillate, caducous, with a long pedicel (as much as 150 μm). All isolates were mating type A1 determined by pairing with A2 reference isolates of P. palmivora (Butl.) Butl. and P. nicotianae Breda de Haan. Oogonia with amphigynous antheridia were formed on VJA after 10 to 15 days at 24°C in the dark. Occasionally, 10 of 15 isolates formed small chlamydospores on VJA. Electrophoretic patterns of total mycelial proteins and four isozymes (acid and alkaline phosphatase, esterase, and malate dehydrogenase) on polyacrylamide slab gels (3) of all Cuphea isolates were very similar to those of reference isolates of P. tropicalis M. Aragaki & J. Y. Uchida from Convolvulus cneorum L. (IMI 391714) and Rhamnus alaternus L., respectively. In addition, the Cuphea isolates were clearly distinct from reference isolates of other species including P. capsici Leon., P. citricola Sawada, P. citrophthora (R. E. Smith & E. H. Smith) Leon., P. nicotianae, and P. palmivora. On the basis of morphological cultural characters and the electrophoretic phenotype, the isolates were identified as P. tropicalis. Internal transcribed spacer (ITS) regions of rDNA sequences (2) confirmed the identification. Koch's postulates were fulfilled by testing three cigar-flower isolates, including isolate IMI 391709, on 10 6-month-old potted cuttings of Cuphea inoculated by applying a 10-ml zoospore suspension (2 × 104 zoospores/ml) to the crowns, incubated for 24 h at 100% relative humidity, and maintained in the greenhouse at 20 to 24°C. After 10 days, crowns and stems were brown and all plants wilted within 20 days. Ten control plants treated with water remained healthy. P. tropicalis was reisolated from infected tissues. The test was repeated with similar results. In Europe, P. tropicalis has been reported on Cyclamen persicum Mill. in Germany (4) and C. cneorum and R. alaternus in Italy (1), indicating a broad host range and spreading in ornamental nurseries. References: (1) S. O. Cacciola et al. Boll. Acc. Gioenia Sci. Nat. 31:57, 1999. (2) S. O. Cacciola et al. For. Snow Landsc. Res. 76:387, 2001. (3) D. C. Erwin and O. K. Ribeiro. Pages 39–41, 138–139 in: Phytophthora Diseases Worldwide. The American Phytopathological Society, St. Paul MN. 1996. (4) W. W. P. Gerlach and A. Schubert. Plant Dis. 85:334, 2001.
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