Abstract: An important biological event in phytopathogens of the genus Phytophthora is sexual reproduction, which is conducted by two mating types, A1 and A2. A factor known as hormone a1 is secreted by the A1 mating type and induces the formation of sexual spores (oospores) in the A2 mating type. Here we describe the asymmetric synthesis and assignment of the absolute configuration of hormone a1 by oospore-inducing assays of the synthesized isomers.
Abstract: A method was elaborated to isolate oospores of Plasmopara halstedii from tissue of its host, Helianthus annuus. Isolated oospores were studied microscopically and germination was documented with respect to the time course and the mode of germination. Formation of primary sporangia was similarly observed in oospores, harvested from 4 to 6-week-old systemically infected
sunflower plants, grown under constant conditions at 160C, as well as from field plants, harvested late in the season. Pretreatment of oospores with cold temperatures, previously assumed to stimulate the rate and to accelerate the speed of oospore germination, did not result in such effects. Germination usually occurred within 10-30 days of incubation at a highly variable rate of about 1 to 17%) average 6.7% in deionized water.
Abstract: The ability of the late blight pathogen Phytophthora infestans to form oospores in leaves of seven potato cultivars was examined at different incubation temperatures under controlled environmental conditions and under field conditions. At 10°C, the oospore formation in three intermediate-resistant cultivars all differed significantly from each other (P < 0.05), with the lowest amount formed in cv. Asterix. This latter cultivar did not form oospores at any other temperature. Under field conditions oospores were formed abundantly in a naturally infected field. A significant date by cultivar interaction showed that P. infestans increased the oospore formation in foliage by time in
cvs Columbo, Hertha and Matilda, whereas no significant differences between dates were found for other cultivars. The genetic structure of P. infestans in the naturally infected field plot, where oospores formed abundantly, was studied by using amplified fragment length polymorphism and a high genetic diversity was revealed. Oospore germination from two Scandinavian (A1 and A2) P. infestans isolates was stimulated in visible light and in 1 : 2 and 1 : 10 soil extract. The effect of light and nutrients on oosporogenesis is discussed.
Abstract: Oospore viability, oospore germinability, and phenotypic variation among zoospore and hyphal tip derivatives of the standard and natural variant types of the hybrid alder Phytophthoras were investigated. Oospore viability in the standard hybrid, estimated by the tetrazolium bromide method, was low (» 31–36%). No germination was observed in more than 4000 oospores, although germination did occur in the Phytophthora cactorum, Phytophthora citricola and Phytophthora cambivora controls. This is consistent with the known meiotic irregularities in this hybrid. Mean oospore viabilities in the natural variants were significantly different (p < 0.001), ranging from approximately 24% in the UK variant to approximately 75% in the Dutch variant. Again, no oospore germination was observed. Zoospore and hyphal tip derivatives of the standard hybrid and of the Swedish and Dutch variants resembled the parent isolate in phenotype. The derivatives of the German and UK variants, however, often differed from the parent type. Those of the UK variant were extremely and continuously variable in colony patterns, growth rates, temperature–growth relationships and fertility levels. Although these results do not support the view that the natural variants arise as genetic breakdown products of the standard hybrid, this possibility cannot yet be excluded. The apparent inviability of the oospores suggests that the mycelium and zoospores are mainly responsible for the survival and spread of the alder Phytophthoras in the field.
Abstract: To determine the potential of sexual reproduction among host-adapted populations of Phytophthora infestans sensu lato in Ecuador, 13 A1 isolates belonging to clonal lineages US-1, EC-1 and EC-3, and 11 A2 isolates belonging to the clonal lineage EC-2, were paired on agar plates to induce crossing. In the first experiment, six A1 isolates (three US-1, two EC-1 and one EC-3) were each crossed with three A2 isolates (total = 18 crosses). Matings involving isolates of the EC-1 lineage produced more oospores of healthy appearance than did matings with isolates of US-1 or EC-3. In the second experiment, the oospores of 35 crosses (21 EC-1 × EC-2; 10 US-1 × EC-2; four EC-3 × EC-2) were dispersed on water agar to assess oospore germination. Overall, germination percentages were low. Only one cross produced enough progeny for evaluation. Twenty-three single-oospore offspring were isolated and evaluated for mating type; electrophoretic patterns of glucose-6-phosphate isomerase (Gpi) and peptidase (Pep) alloenzyme loci; mitochondrial DNA haplotype; and genomic DNA fingerprint. Multilocus genotype data indicated that all 23 isolates resulted from meiotic recombination. Four progeny with homothallic phenotype appeared to be unstable heterokaryons. Markers at several loci segregated according to simple Mendelian expectations for a diploid organism, but the ratios of three RFLP loci and the Pep locus were not consistent with Mendelian expectations. All progeny were nonpathogenic on hosts of the parental genotypes. Reduced mating success and reduced pathogenic fitness of progeny appear to be postmating mechanisms of reproductive isolation in populations of P. infestans sensu lato in Ecuador.
Abstract: Pink rot of potato, most commonly caused by Phytophthora erythroseptica, is a major field and post-harvest problem in southern Idaho, USA, particularly since 1998 when isolates resistant to the phenylamide fungicide metalaxyl-M (mefenoxam) were detected. Isolates of P. erythroseptica were collected from infected tubers in 2001 and 2002 from six Idaho counties and tested for resistance to metalaxyl-M on amended agar. Metalaxyl-M resistant (MR) and metalaxyl-M-sensitive (MS) isolates were identified in six counties; 160 isolates were highly resistant, seven moderately resistant and 57 sensitive to metalaxyl-M with mean EC50 values of 182, 23 and 0·5 mg L-1 ai metalaxyl-M, respectively. Mycelial growth rates and oospore production in agar were assessed for 20 MS and 20 MR isolates at 10, 15, 20, 25 and 30°C. Growth rates of MR isolates were between 2·5 and 3·1 times greater (P < 0·05) than those of MS isolates at 10, 15, 20 and 25°C, and oospore production was between 6·8 and 20·5 times greater (P < 0·0001) for MR than for MS isolates at the same temperatures. Colony growth in V8 broth at 18°C was greater for MR than MS isolates (P < 0·0032). However, zoospore production at 18°C was greater for MS than for MR isolates (P < 0·0109), and zoospore production mM-1 of colony circumference was also greater for MS than for MR isolates, 14 191 and 9959, respectively (P = 0·0109).Sexual reproduction of MR isolates in nature may be greater than MS isolates, but MS isolates may be more asexually fit based on the fitness parameters studied.
Abstract: Nine isolates of known oospore mycoparasites comprised of six actinomycetes {Actinoplanes missouriensis, A. philippinensis, A. utahensis, Amorphosporangium auranticolor, Ampullariella regularis, Spirillospora albida) and three fungi {Acremonium sp., Humicola fuscoatra, Vertidllium
chlamydosporium) were tested in the greenhouse for their ability to suppress or delay the onset of crown rot of pepper caused by Phytophthora capsici. Vertidllium chlamydosporium applied as a root dip increased the number of healthy plants by more than 100% when peppers were transplanted into soil artificially infested with oospores of Phytophthora capsici, but not when peppers were transplanted into soil naturally infested with P. capsici. The other mycoparasites were ineffective in the greenhouse. All the mycoparasites tested parasitized oospores of P. capsici in vitro.
Abstract: Fifty single oospore progeny were established from an in vitro mating of A1 and A2 mating type isolates of Phytophthora cinnamomi from South Africa. Forty-nine progeny were identified as F1 hybrids using seven random amplified polymorphic DNA (RAPD) primers, and one was a selfed isolate of the A1 mating type parent. Among the hybrid progeny, 24 and 25 were A1 and A2 mating type, respectively. Aggressiveness of progeny and parental isolates
was assessed on 1-year-old seedlings of Eucalyptus smithii. The mean aggressiveness of hybrid oosporic isolates, expressed as lesion length, was significantly (P=0.0001) lower than that of the parental isolates. No significant difference in aggressiveness of A1 and A2 mating type F1 hybrid isolates was observed. This is the first report demonstrating sexual recombination in vitro in P. cinnamomi.
Abstract: Nineteen crosses were carried out in vitro between seven and nine isolates of Phytophthora infestans originating from potato and tomato, respectively. Oospores were produced abundantly in all but two crosses, but oospore germination was generally low (a few per cent) and depended on the combination of parental isolates. The highest fertility in F1 progeny
was observed when at least one parental isolate originated from tomato, and the lowest in crosses of isolates from potato; half the crosses were not fertile. Forty-three F1 and 51 F2 single-oospore progeny of a selected cross, along with the 16 field isolates, were analysed phenotypically and with molecular markers. Phenotypic characterization included mating
type; sensitivity to the phenylamide fungicide metalaxyl-M; specific virulence on potato R-gene differentials; and aggressiveness (infected leaf area) on potato and tomato leaf discs (host preference). Isolates and progeny were also assessed for mitochondrial DNA haplotype with RFLP–PCR (restriction fragment length polymorphism–polymerase chain reaction), and characterized with AFLP (amplified fragment length polymorphism) and SSR (simple sequence repeats,
microsatellites). Parental isolates were sensitive and resistant to metalaxyl-M, whereas all F1 were intermediate phenotypes. In the
F2 progeny, the majority of isolates (43 of 51) were intermediate in sensitivity and four each were sensitive and resistant to metalaxyl-M, respectively. In both F1 and F2 progeny, four isolates emerged through selfing. The A1 : A2 ratio was 25 : 18 in F1 and 24 : 21 (plus six self-fertiles) among the F2 progeny. Many F1 progeny isolates were highly aggressive on both hosts, but 15 and 23% of isolates preferred tomato and potato, respectively. Among F2 progeny, few highly aggressive isolates were recorded and a quarter of isolates lost pathogenicity almost completely. Isolates preferring tomato increased, and those preferring potato significantly decreased in the F2 progeny. Inheritance of mitochondrial haplotype in F1 progeny was uniparental and mostly (25 of 27) from one parent only. Six and four different SSR genotypes
were identified in F1 and F2 progeny, respectively, of which two were identical to the parents. The two microsatellite loci,4B and 4G, segregated in the ratios 15 : 22 : 2 and 22 : 17 in F1 and 24 : 17 and 26 : 15 in F2 progeny, respectively, while the majority of AFLP markers segregated in either a 1 : 0, 1 : 1, 3 : 1 or 1 : 2: 1 ratio. There was no obvious association between AFLP and SSR genotypes, nor between genotypic and phenotypic traits.
Abstract: When oospores of Phytophthora cactorum from 30-day-old culture were
treated with 0.25% KMnO4 for 20 min and incubated at 24°C under light for 10 days, 65-75% germinated on water agar and water agarose but only 1-21% germinated on V-8 agar and S-l-L agar. Water agarose was selected because germinated oospores formed restricted colonies on this medium that could be isolated easily. KMnO4 treatment killed sporangia, chlamydospores and mycelial fragments present in oospore suspensions. Under the above conditions, approximately 44% of oospores from 10-day-old culture germinated and the optimum germination rate of about 75% was obtained when oospores reached about 20 days old.
Abstract