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  • Author or Editor: T. Érsek x
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The mini review gives the list of 39 novel Phytophthora species and 2 species hybrids identified and described by various authors following the publication of the handbook of Erwin and Ribeiro (1996). Based upon original reports, sources and locations of isolations, morpho-physiological features and phylogenetic status of each species nova , as compared to previously known species are also summarized. Ultimately, a list of taxa to be formally described outlines prospects of further research.

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Species hybrids were created via fusion of zoospores of two morpho­logically distinct species, P. infestans and P. nicotianae. Sixteen putative hybrid isolates were recovered that expressed differential drug resistance of each parent. Repetitive DNA of P. nicotiane was detected readily in all of these isolates by hybridization with the species-specific DNA probe, pPP33A. DNA of P. infestans was detected in only two putative hybrid isolates using PCR and primer pair ITS3 and PINF2. The two true hybrids were more similar to P. nicotianae than to P. infestans on the basis of pathogenic, morphological and molecular evidence. Additionally, hybrids expressed modified host ranges compared to parental species. Fusion of zoospores or hyphae may contribute to formation of such hybrids, particularly in the case of heterothallic species in which the joint occurrence of compatible mating types is rare. Zoospore fusion may prove useful as a tool to study hybridization, pathogenesis, and sources of natural diversity of species.

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Twenty-seven isolates of Phytophthora infestans collected in Hungary in 2001 were tested for mating type, response to metalaxyl, isozyme geno­type at glucose-6-phosphate isomerase (Gpi) and peptidase A (Pep) loci and nuclear DNA fingerprints with probe RG57. The ratios of the mating types A1 to A2 were 5:6 and 9:7 among isolates from potato and tomato, respectively. Seventeen isolates were sensitive to metalaxyl, 1 isolate responded intermediately and 9 isolates were resistant. No novel combi­nations of isozyme alleles were found; all isolates were Gpi 100/100, and genotypes at the Pep locus were 96/96 (63%), 83/96 (11%) and 100/100 (26%). In contrast, all of the 22 RG57 fingerprints exhibited patterns that have not been reported in Hungary before. On the basis of combined traits, 22 multilocus genotypes, unnoted elsewhere in Europe, were con­struct­ed among the 27 isolates analysed. These results indicate that varia­bi­lity in the Hungarian P. infestans populations is likely due to local events (asexual and sexual interactions) rather than migration from other countries.

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Phytophthora alni is a species hybrid that causes a destructive root and collar rot disease of alders throughout Europe. Its subspecies, P. alni subsp. alni (Paa), P. alni subsp. uniformis (Pau) and P. alni subsp. multiformis (Pam) can be distinguished on the basis of phenotypic and genotypic traits. In this study, we report evidence of an unusual genomic combination of two subspecies occurring in two P. alni isolates from Hungary. These isolates, which had previously been identified as Paa using hybrid-specific PCR primers and morphological traits, exhibited a mitochondrial DNA restriction pattern identical to that of Pau. However, RAPD patterns and isozyme profiles of nuclear genes encoding glucose-phosphate isomerase (Gpi) and malate dehydrogenase (Mdh) of the two atypical isolates were identical to those found in all Paa isolates. Isozyme analysis also revealed a novel allele at the putative Mdh-1 locus in Paa and Pam isolates. The atypical Paa isolates have likely emerged as a result of hybridization events in the P. alni population between Paa and Pau .

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In December 2012 then in the following winter season, the occurrence of whitish mycelial coat was observed on the collar of 3- to 6-m high Bucida buceras trees grown in hydrocultures to decorate a spacious indoor community space in Vienna. (This plant [shown in Fig. 1] belongs to Combretaceae, Myrtales and commonly named black olive tree, bullet tree, gregorywood and oxhorn bucida.) The mycelium-infested area of the bark appeared to be water-soaked. Near the surface of the potting mix (earth ball embedded in clay pebbles), the roots were also covered with whitish mycelia (Fig. 2). Over the winter season when the indoor temperature increased from 20 °C to 25 °C, these symptoms were unnoticeable. Regardless of the season, the rhizosphere contained numbers of sclerotia, dark-grey, globose and 8–12 mm in diameter that occasionally developed rhizomorph-like mycelial cords.

Direct plating of mycelium fragments from the bark and sclerotia from the rhizosphere onto potato dextrose agar amended with ampicillin (500 mg/l) eventually yielded pure fungal cultures of similar characteristics. Cultures routinely incubated in the dark developed white and submerged colonies with sparse aerial mycelia. The fungus grew well between 10 °C and 25 °C, and failed to grow at either 5 °C or 32 °C. The optimal growth was measured at 20 °C with an average radial growth rate of 11 mm per day. After 10 to 12 days, a ring of sclerotia begun to develop near the edge of the colonies; they turned dark grey and sized 3–8 mm. Rather misleadingly, neither conidia, nor sexual spores were observed in these cultures. However, when the fungus was cultured in natural light under laboratory conditions at 25 °C, a completely different colony pattern was observed; it was cottony, greyish then dark grey, and produced abundant hyaline conidia borne on grey, branching tree-like conidiophores. Conidia were one-celled and egg-shaped, and their dimensions fell in the range of 9.89–14.63 (11.48±0.31) µm×7.05–10.05 (8.31±0.20) µm. These features concurred with those characterising the polyphagous grey mould fungus Botryotinia fuckeliana (anamorph: Botrytis cinerea) (Elad et al., 2007). The ITS1/ITS2 including the 5.8S subunit of rDNA of one of the isolates were amplified with primers ITS1-F/ITS4, then the PCR products were sequenced. The ITS sequence determined in this way was identical to known sequences of B. fuckeliana strains, e.g. that of CBS 131.28 (GenBank accession number: KF859918), the type material of Botrytis cinerea f. lini, DAOM 231372 (GenBank accession number: KF859924) and so on.

Pathogenicity tests resulted in rapidly (within 2 weeks) developing disease symptoms around the site of wound inoculation with a 5-mm-diametre mycelial agar plug: fruit rot on apple and lemon in the laboratory, and sunken lesions on stems of hydrocultured ornamental plants such as the herbaceous Monstera deliciosa and the woody Dracena marginata. To fulfill Koch’s postulates, the fungus was re-isolated from symptomatic apple fruit, and was found to exhibit the afore-mentioned morpho-physiological characteristics.

Inoculation test on Bucida was not performed because of the costly risk i.e., the sale price of the trees is € 3 to 10 thousand. Consequently, the actual sensitivity of Bucida to grey mould remains uncertain, so much the more because this plant species has not been recorded as a host of the pathogen or other important parasitic fungi in natural (subtropical) environment (e.g. Whelburg et al., 1975). To our knowledge, this report is the first description of Botryotinia fuckeliana on Bucida buceras. In addition to the fact that periodic emergence of fungal mycelia on the trunk impairs the tree’s aesthetic appearance, the sclerotia resting in the potting mix may cause more serious problems in the long term. However, it cannot be precluded that the elevated indoor temperature reduces disease progression and thus the economic importance of the pathogen on this plant.

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A total of 123 isolates of Phytophthora infestans were recovered from 43 commercial fields and allotments of potato and tomato in Hungary. Isolates were characterised for mating type, response to metalaxyl, isozyme genotype at glucose-6-phosphate isomerase (Gpi) and peptidase A (Pep) loci, nuclear DNA fingerprint with probe RG57 and mitochondrial haplotype. Both mating types were detected in 8 potato and 3 tomato fields. The frequency of A1 and A2 were 47.5% and 52.5% on potato and 60.5% and 39.5% on tomato. More than sixty per cent of the whole population were resistant (31.1%) or responded intermediately (30.2%) to the fungicide metalaxyl in vitro. All isolates were monomorphic at the Gpi locus (100/100), but four allele combinations 100/100 (33%), 96/100 (28%), 96/96 (31%) and 83/96 (8%) were found at the Pep locus. Genotype Pep 83/96 is rare in Europe and all isolates with this allele combination were of A1 mating type and Ia mitochondrial haplotype. As in other European populations, only two of the four mitochondrial haplotypes (Ia, 32.5% and IIa, 67.5%) were detected. Diversity of RG57 fingerprints was high. Of the 79 different fingerprints found 67 were not detected previously.

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