Impact of seed potatoes infected withPhytophthora infestans (US-1 or US-8 genotypes) on crop growth and disease risk

As the surface area with late blight symptoms of seed tubers increased, the incidence of plant emergence decreased. This relationship was more dramatic with the US-8 genotype than the US-1 genotype ofhytophthora infestans, causal agent of late blight. However, when seed tubers were inoculated with both US-1 and US-8 isolates, the plant growth responses were generally intermediate to those obtained with the two genotypes alone. More resistant cultivars (eg. Dorita, Island Sunshine, and Sebago) had greater plant emergence and less seed rot than more susceptible cultivars (eg. Green Mountain and Shepody) but the response differences between resistant and susceptible cultivars was less clear with US-8 as most cultivars had 100% seed rot. Seed handling treatments affected crop growth and disease risk; as the inoculum levels and in some cases, the pre-planting storage period increased, plant emergence decreased generally due to greater seed rot incidences. Of the one registered seed treatment fungicide and several non-registered and/or experimental fungicides tested, only two significantly improved plant emergence. During three years of study, “diseased” seed treatments generally led to seed rot except in 1996 when one plant in one experimental plot of infected seed had subsequent spread of the disease to the foliage. Furthermore, within one week, a field epidemic developed outwardly from the single infected plant due to the occurrence of favourable weather conditions. Further research opportunities are discussed relative to the impact of infected seed on crop growth and disease risk     

Impact of Different US Genotypes of <Emphasis Type="Italic">Phytophthora infestans</Emphasis> on Potato Seed Tuber Rot and Plant Emergence in a Range of Cultivars and Advanced Breeding Lines

Seed pieces of different potato cultivars and advanced breeding lines (ABLs) from north central US breeding programmes were inoculated with different genotypes of Phytophthora infestans (US-1, US-1.7, US-8, US-11 and US-14). The effect of these genotypes of P. infestans on seed piece rot severity after re-storage was assessed using an image analysis technique. P. infestans genotypes demonstrated variable ability to cause seed piece rot and to reduce plant emergence measured as final plant stand (%) and the relative area under the plant emergence curve (RAUEPC). The US-8 genotype of P. infestans was the most aggressive genotype, as indicated by tuber rot severity across all cultivars/ABLs tested, followed by US-14 in both years. The US-1, US-1.7 and US-11 genotypes were the least aggressive, causing only moderate seed piece rotting across cultivars/ABLs tested. Similar trends were observed in two field experiments, where the US-8 and US-14 genotypes delayed or reduced emergence. Values of final plant stand (%) and RAUEPC demonstrated that the cultivars/ABLs Atlantic, MSJ453-4Y and Torridon were the least susceptible across all P. infestans genotypes. In both experiments cv. Pike was the most susceptible. Other cultivars/ABLs demonstrated variable responses to different genotypes of P. infestans. No symptoms of P. infestans were observed on emerged plants up to 60 days after planting. The variability of susceptibility of tubers to different genotypes of P. infestans has implications for plant breeding efforts in that the major emphasis in the past has been to breed for foliar resistance, with limited emphasis on the reaction of the tuber. Results from this study suggest that highly aggressive genotypes of P. infestans such as US-8 may lead to severe tuber rotting and deterioration of tubers before emergence, but despite this observation the US-8 genotype is still predominant in North America.     

Changes in race structure of Canadian populations ofPhytophthora infestans based on specific virulence to selected potato clones

Summary A collection of 80 isolates ofPhytophthora infestans, representing multilocus genotypes of the pathogen found in Canada from 1994 to 1996, was screened on a series of nine differential potato host cultivars to determine the physiological race of the isolates. A total of 28 pathotypes were found among the 11 genotypes present in the collection. There was a significant increase in the complexity of pathotypes from 1994 to 1996 which reflected the displacement of the US-1 genotype (mean of 2.2 host differentials infected) by the US-8 genotype (mean of 8.1 host differentials infected). Eighteen of the US-8 isolates could overcome all nine of the resistance genes tested. Virulence genes which overcame host resistance conferred by R8 were the rarest in the populations studied. The predominance of complex populations ofP. infestans in Canada implies that the introduction of cultivars with specific R genes for resistance would be an ineffective control measure. AAFC Contribution No. 901     

US-8 and US-11 genotypes ofPhytophthora infestans from potato and tomato respond differently to commercial fungicides

Isolates ofPhytophthora infestans collected in Canada in 1997 from both potatoes and tomatoes, were tested on potato leaf discs for their response to an equal active ingredient concentration (10 μg a.i./mL) of the following commercial fungicides: Acrobat MZ (Dimethomorph and Mancozeb), Ridomil Gold (metalaxyl-m), Dithane (Mancozeb), Curzate (Cymoxanil), Bravo (Chlorothalonil), and Tattoo C (Propamocarb and Chlorothalonil). Relative percent leaf infection values, estimated on fungicide-treated vs fungicide-free leaf discs, were compared among isolates from the US-8 and US-11 genotypes isolated from the two host plants. Based on an equal concentration of each fungicide’s active ingredients, variations in relative percent leaf infection were recorded between US-8 and US-11 genotypes, and between potato and tomato isolates within each genotype. Bravo and Tattoo C used with similar active ingredients concentrations were the most inhibitory to all groups of isolates. Dithane and Ridomil Gold provided uniform low inhibition againstP. infestans when tested on potato leaf discs. The different behavior ofP. infestans isolates from potato vs tomato suggests that management of late blight in these two important crops must take such differences into consideration. In particular, the nature and concentration of the fungicides to be applied must take into account any information available about genotypes present on each crop.     

Variability in responses of US-8 and US-11 genotypes of potato and tomato isolates ofPhytophthora infestans to commercial fungicidesin vitro

Isolates ofPhytophthora infestans collected in Canada from potato and tomato plants from 1994 to 1998 were tested for their responsein vitro to equal active ingredient concentrations (1, 2.5, and 5 μg a.i/mL) of six commercial fungicides: Acrobat MZ (dimethomorph and mancozeb), Ridomil Gold (metalaxyl-m), Dithane (mancozeb), Curzate (cymoxanil), Bravo (chlorothalonil), and Tattoo C (propamocarb and chlorothalonil). Relative mycelial growth of isolates estimated on fungicideamended vs fungicide-free media was compared among isolates of the US-8 and US-11 genotypes from the two host plants. The effects of these fungicides on spore germination of isolates from the two genotypes were also estimated. Mycelial growth of US-8 and US-11 isolates was most affected by Bravo and Tattoo C, followed by Acrobat, and then Curzate and Dithane. Ridomil Gold was not effective in inhibitingin vitro growth of US-11 isolates at the concentrations tested. Variations in sensitivity to different fungicides among isolates of the same genotype collected in different years were observed. For example, based on 2.5 and 5 μg a.i./mL, potato/US-11 isolates collected from 1995 to 1997 showed increased sensitivity to Curzate and Ridomil Gold, while decreased sensitivity was recorded with Dithane. For US-8 isolates, responses to the fungicides varied according to fungicide and year. Variations were generally not significant for Acrobat, Curzate, Bravo, and Tattoo C over time. Conversely, potato/US-8 isolates from 1995 and 1998 were less sensitive to Dithane and Ridomil Gold than those from 1994, 1996, and 1997.     

Variation in tuber pathogenicity ofPhytophthora infestans in the Netherlands

Summary Variation in aggressiveness to tubers among isolates ofPhytophthora infestans sampled from three potato growing regions in the Netherlands was compared. Variation in the ability to infect tubers of cv. Bintje was found between isolates of each of the three regional populations. The most aggressive isolate of the old population matched the average level of the new population in its ability to infect tubers. As a consequence, the commonly used reference isolate VK 6C can no longer be considered to be representative of the present population ofP. infestans. Therefore it is recommended that testing tuber resistance for the official list of potato cultivars with this isolate should be discontinued. Tuber infection and subsequent spread of the fungus in the tuber tissues were not found to be correlated. The components of tuber pathogenicity studied were not correlated to pathogenicity factors in the foliage, as measured under growth chamber conditions.     

Differential resistance to tuber late blight in potato cultivars without R-genes

Summary In 1983 and 1984 there were reports in Norway of more severe late blight infection than usual on tubers of cv. Pimpernel, a cultivar without R-genes known to be somewhat resistant to both haulm and tuber infection. Results from routine tests confirmed earlier experience that cv. Pimpernel shows stable resistance to tuber late blight, although the ranking of cultivars may vary between years. In experiments during three harvest seasons tubers of different cultivars were inoculated with isolates ofP. infestans obtained from stocks of cv. Pimpernel with severe tuber late blight and with other isolates. In these experiments the isolates from cv. Pimpernel caused more rot on tubers of Pimpernel than did other isolates.     

Relationships between pathotypes and RAPDs, Gpi-allozyme patterns, mating types, and resistance to metalaxyl ofPhytophthora infestans in Canada in 1997

Relationships between pathotypes and other markers such as mating types, metalaxyl resistance, Gpi-allozyme and random amplified polymorphic DNA (RAPD) patterns, were investigated among 33 isolates ofPhytophthora infestans obtained from both potato and tomato across Canada in 1997. Pathogenicity testings, based on virulence against potato differentials carrying single-resistance genes and the size of lesions on leaves, revealed higher pathogenic diversity in 1997 than in the few past years, with more pathotypes and more race-complex isolates. A higher pathogenic diversity was observed within A2 (US-8) than within A1 (US-11) isolates. There was no correlation between the level of resistance to metalaxyl of an isolate and its pathogenicity. Nevertheless, when grouped into metalaxyl-sensitive (MS), -intermediate (MI), and -resistant (MR) classes, both average pathotype complexity and average lesion size were lowest for MS group. RAPD data revealed a good distinction between A1 (US-11) and A2 (US-8) mating type groups, and between isolates from British Columbia and those from other provinces. The detection in the last few years of both A1 and A2 mating types in Canada and the occurrence in 1997 of some isolates sharing different characteristics of both of these two groups suggest that sexual reproduction may have occurred locally.     

Assessment of mating types and resistance to metalaxyl of Canadian populations ofPhytophthora infestans in 1997

The distribution of A1 and A2 mating types, and of metalaxyl-sensitive (MS), metalaxyl-intermediate (MI), and metalaxyl-resistant (MR) strains ofPhytophthora infestans, the causal pathogen of late blight, were determined in Canada in 1997. Potato plants or tubers and tomato plants or fruits, suspected to have late blight infection, were received from seven Canadian provinces in 1997, and were assessed for the presence ofP. infestans. Among 220 samples suspected to have late blight,P. infestans was recovered from 132 samples (60%) received from British Columbia (BC), Manitoba, New Brunswick, Prince Edward Island, and Quebec. From the 132 samples, 364 isolates were obtained and tested for their mating type and response to metalaxyl. Populations ofP. infestans within Canadian provinces were homogeneous for mating types, except within BC, where populations comprised both A1 and A2 mating types both within fields and sometimes within samples. The A2 mating type was found in all provinces where late blight was present. The A1 mating type was found only in Manitoba (1 isolate) and in BC (38 isolates). Unlike the old A1 isolates, those found in BC in 1997 were highly resistant to metalaxyl. These A1 isolates are even more resistant than A2 isolates. Based on assays with 10 ug.ml^?1 of metalaxyl, 14% of all tested isolates were MS, 69.8% were MI and 16.2% were MR. In addition, all provinces contained MS, MI, and MR isolates. No specific mating type or metalaxyl sensitivity level were specific to tomato or potato.     

Influence of storage temperature on rate of potato tuber tissue infection caused byPhytophthora infestans (mont.) de bary estimated by digital image analysis

Summary Potato tubers were inoculated with two biotypes ofPhytophthora infestans then stored at 3,7, 10 and 15°C. Image analysis quantified average reflective intensity (ARI) of diseased tissue from cut surfaces of sample tubers. Tuber tissue infection and infection rate were measured by calculating Mean ARI of samples. Average tuber tissue infection and infection rate was minimal at 3°C (P.i.-US8 orP.i.-US1). Tuber tissue infection increased at temperatures >3°C, from 220 Mean ARI seven days after inoculation (dai) to 190–150 Mean ARI 50 dai (depending on cultivar and biotype ofP. infestans). Rate of tuber tissue infection caused byP.i.-US1 at 7°C was about zero in cv. Snowden but greater than −0.2 ARI day⁻¹ (cvs Russet Burbank and Superior). Rate of late blight infection in tuber tissue generally increased with temperature from −0.2 ARI day⁻¹ (at 7°C) in all cultivars to a maximum of −0.8 ARI day⁻¹ (10°C).     

Sporulation ofPhytophthora infestans (Mont.) de Bary on the surface of diseased potatoes and tuber to tuber spread of infection during handling

Summary Laboratory experiments and observations in commercial stores showed that viable sporangia ofPhytophthora infestans can be produced on infected potato tubers of different cultivars. Infection was also found to spread from diseased to healthy tubers during handling.     

The infection of potatoes byPhytophthora infestans during simulated washing and its control using a hot water treatment

Summary A single blighted tuber released on average 1.39 mg ofP. infestans mycelia and sporangia during simulated washing. In subsequent experiments up to 100% of tubers were infected when healthy tubers were washed in a suspension ofP. infestans equivalent to a concentration of 1.65 μg ml⁻¹, i.e. that calculated to be in a commercial washing plant after washing a potato stock with approximately 1% of tubers infected with blight for 4 hours. Immature tubers were more prone to infection during washing than mature tubers. Damaging tubers, irrespective of their maturity, increased the incidence of tuber infection. The viability ofP. infestans isolates was significantly reduced byin vitro immersion in water at 44°C for 5 minutes. The infection of daughter tubers dipped in a suspension ofP. infestans for 3 minutes was prevented when the suspension temperature was 44°C. There was no indication of tuber damage at this temperature.     

Production of phytoalexins, glycoalkaloids and phenolics in leaves and tubers of potato cultivars with different degrees of field resistance after infection withPhytophthora infestans

Summary The kinetics of accumulation of phytoalexins, glycoalkaloids and phenolics was studied in two potato cultivars differing in their degrees of field resistance when infected withPhytophthora infestans. Tuber slices and leaves of cvs Pampeana INTA (high degree of field resistance, free of R genes) and Bintje (susceptible) were infected with race C (complex race 1, 3, 5, 7, 11) ofPhytophthora infestans. Phytoalexins and phenolics accumulated in tuber and leaf tissues which had been inoculated. The levels of these compounds in the susceptible cv. Bintje were relatively low and similar to those found before inoculation. Leaves of cv. Pampeana INTA had a very high glycoalkaloid content, suggesting that glycoalkaloids may play a role in protection of leaves against the fungus. However, we could find no correlation between resistance and glycoalkaloid content of tubers. Our results suggest a major role of phytoalexins, phenolics and glycoalkaloids in the complex mechanisms of field resistance.     

The effect of the moisture level of potting compost on the susceptibility of glasshouse-grown tubers to late blight

Summary The effect of the moisture level of potting compost at harvest on the susceptibility of tubers of three cultivars to late blight was investigated in the glasshouse in three successive years. Three moisture regimes (dry, moist, wet) were imposed at flowering time, the tubers harvested 3 weeks later and immediately dipped in a zoospore suspension ofPhytophthora infestans. Tubers from dry compost were significantly more susceptible than those from wet or moist compost, which showed similar levels of susceptibility.     

Comparison of different culture media on the mycelial growth, sporangia and oospore production ofPhytophthora infestans

The purpose of this study was to evaluate various culture media, with or without β-sitosterol, on mycelial growth, sporangia and oospore production by isolates ofPhytophthora infestans. Nine media were compared to rye agar (RA) which is frequently used to cultureP. infestans. Poor fungal growth across most isolates was observed on soybean agar, carrot agar and carrot agar with sterols relative to RA. Only clarified V8 juice containing β-sitosterol (CLV8S) resulted in significantly more sporangia production compared to RA. All other media performed similar to the RA. No significant differences were found in the sizes (length and width) of sporangia produced among the isolates tested on various media. Oospore production was greatest whenP. infestans was cultured on carrot agar with β-sitosterol (CAS) or CLV8S when compared to RA. Responses of isolate matings varied somewhat with an A2 isolate from Nova Scotia consistently producing fewer oospores in matings with A1 isolates compared to the other A1/A2 mating combinations. Rye agar and oat meal agar with or without β-sitosterol as well as clarified V8 juice agar with β-sitosterol are useful forin vitro growth of mycelium.     

Differences in tuber rot development for North American clones ofPhytophthora infestans

The relative aggressiveness ofPhytophthora infestons clones in potato tubers was compared in three trials using 7 to 24 isolates of 2 to 4 clones. Visible rot developed slowly at 13C with isolates of the US-1 genotype, the only significant clone found in North America prior to 1979, but substantially faster with most isolates of the newer clonal genotypes US-6, US-7 and US-8. Certain US-7 isolates were similar to US-1, and US-6 isolates also had a broad range of aggressiveness. Secondary infection byFusarium sp. increased rot development in many instances, but this effect was not clone-related. Differences in rot development may affect potato storage or late blight disease transmission.     

Novel genotypes ofPhytophthora infestans in Canada during 1994 and 1995

Populations ofPhytophthora infestans, causing late blight of potato, changed significantly in Canada in the mid-1990s. The US-1 genotype (A1, metalaxylsensitive [MS]) was displaced by novel genotypes (mainly US-8) in much of the country in 1994 and 1995. Many of the new genotypes were insensitive to metalaxyl and were of the A2 mating type, although metalaxyl-insensitive (MI) Al isolates of the gll (or US-11) genotype were found in British Columbia. A total of 14 unique multilocus genotypes were described over the two years of the study based on mating type (Al or A2), metalaxyl sensitivity (MS or MI), allozyme banding patterns (Gpi andPep), and random amplified polymorphic DNA (RAPD) markers. Six of these genotypes could be distinguished by allozyme banding patterns at theGpi locus alone; RAPD analysis revealed additional variation within allozyme groupings and defined additional genotypes. Among 82 isolates collected from various Canadian provinces in 1994, US-1 and US-8 constituted 30% and 49% of isolates, respectively; other genotypes occurred in minor frequency. Among 50 isolates collected in 1995, US-1 and US-8 constituted 10% and 74%, respectively. Isolates of gll (US-11) were found in British Columbia in 1995. Diversity of genotypes was greater in 1994 (11 genotypes) than in 1995 (six genotypes). We hypothesize that sexual reproduction may have been responsible for generating some of the genetic variability inP. infestans populations, during a time period in which A2 isolates first migrated into many provinces in Canada.     

Identification of late blight, Colorado potato beetle, and blackleg resistance in three Mexican and two South American wild 2x (1EBN)Solanum species

Wild potatoes are important sources of genes for resistance to disease and insect pests. A collection of wild Mexican and South AmericanSolarium species from the US potato Genebank was evaluated under laboratory and/or field conditions for their reaction to late blight (Phytophthora infestans), Colorado potato beetle (CPB,Leptinotarsa decemlineata Say), and blackleg (Erwinia carotovora subsp.atroseptica (van Hall) Dye) in order to identify individual genotypes with multiple resistance genes. Late blight inoculations using aggressive isolates (US-8/A2 and US-11/A1 mating types) of P.infestans revealed a wide range of variation for resistance between and within the accessions of the wild species tested. For late blight, susceptible as well as moderately to highly resistant genotypes were observed in all the species tested. However, at least one accession from the three Mexican and one South American wild diploid species tested showed a relatively uniform high level of resistance toP. infestans. These includedS. bulbocastanum, S. pinnatisectum, S. cardiophyllum, andS. circaeifolium. Two accessions from South American speciesS. commersonii were highly susceptible to late blight. For the Colorado potato beetle test, only one species,S. pinnatisectum appeared uniformly resistant to CPB under field conditions. Results of screening for blackleg resistance showed that there were major differences between genotypes in the wild species. Accessions ofS. circaeifolium PI 498119 andS. bulbocastanum PI 243504 were identified as having significantly higher blackleg resistance than cultivated potato and the other wild species tested. However, genotypes from these two accessions were more susceptible to late blight and CPB. Characterization of theP. infestans isolate P1801C.16 used for late blight evaluation and multi-locus isolate tests using US-8/A2 and US-11/A1 races revealed that the resistance inS. pinnatisectum genotypes tested corresponded to a race-non-specific genetic system, which was different from any existing R genes.Solanum pin-natisectum genotypes with both high levels of late blight and CPB resistance as well as blackleg resistance genotypes identified in the present study represent a diverse gene pool that may be useful for development of new potato cultivars with multiple disease and insect resistance. The potential utilization of these valuable sources for improvement of cultivated potato is discussed.     

Reaction toPhytophthora infestans of potato genotypes with race-specific resistance

Summary Phytophthora infestans isolates collected in Poland in 1987–1995 were evaluated on detached leaflets of a differential series consisting of potato genotypes possessing resistance genes R1–R11. On the same differentials was evaluated over three years a highly virulentP. infestans isolate MP 245. Isolate MP 245 and those from natural populations ofP. infestans were seldom or inconsistently pathogenic to R5, R8 and R9. When leaflets of the differentials were infected with MP 245 the lesions were often small and sporulation weak. A statistically significant interaction was found between years and pathogenicity of MP 245 to individual differentials. Attempts to train the isolate MP 245 for pathogenicity to R9 were unsuccessful. The variation in specific resistance of potato genotypes may complicate the evaluation of their resistance toP. infestans, as well as the evaluation of the virulence spectrum of natural fungus populations. This variation may be due to changes in specific pathogenicity of the fungus or in specific resistance of the potato. Some differentials have a possibly useful resistance toP. infestans.     

Detecting resistance to the root-knot nematodesMeloidogyne hapla andM. chitwoodi in potato and wildSolanum spp.

Summary Various methods of screening for resistance to root-knot nematodes were compared and evaluated. Seedling populations ofSolanum spp., grown in clay pots and plastic tubes with silver sand and inoculated with juveniles ofMeloidogyne chitwoodi andM. hapla, showed large differences in the number of egg masses on roots 7 weeks after inoculation. The differences were reproducible when re-testing was done with cuttings and plants from tubers. No resistance toMeloidogyne spp. was observed with ten potato cultivars when grown in clay pots, plastic tobes or closed containers. Plants from tubers in growth pouches developed a large two-dimensional root system, and after inoculation with juveniles the infection process could be observed over 8 weeks. A method of infecting potato tuber tissue withMeloidogyne is described, using tuber slices in Petri dishes as a potential screening test for tuber resistance.