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.     

Identification of mating types and metalaxyl resistance in North American populations ofPhytophthora infestans

The A² mating type ofPhytophthora infestans was first reported in the United States in 1990. Concurrently,P. infestans strains resistant to metalaxyl ere found in the Pacific Northwest. Collaborative surveys were undertaken during 1991–1993 to investigate the frequency of occurrence of A² mating types and metalaxyl resistant strains in populations ofP. infestans isolated from outbreaks of late blight in potato and tomato crops in North America.In vitro testing indicated that isolates from the northeastern U.S. and Atlantic Canada were primarily (52/55) metalaxyl sensitive and all were A¹ mating types. Among 85 isolates from late blight epidemics in Florida and Texas, greater than 61% were both metalaxyl resistant and A² mating type. Metalaxyl resistance and A² mating types were identified also in a few tomato isolates from North Carolina. Although the majority of 134 isolates from the Pacific Northwest (British Columbia and Washington) were metalaxyl resistant, only 2 isolates from Washington were A² mating types. Among 111 isolates from 2 sites in central Mexico, 63% and 77% were both metalaxyl resistant and A² mating types. The data indicate also a higher frequency of metalaxyl resistance in A² isolates, than in A¹ isolates, among isolates from Florida and Texas. Highest metalaxyl resistance levels were found, however, in A¹ isolates from California, where no A² isolates were recovered.     

Changes in metalaxyl resistance among glucose phosphate isomerase genotypes ofPhytophthora infestans in Canada during 1997 and 1998

A total of 930 isolates ofPhytophthora infestans collected during 1997 and 1998 from commercial potato and tomato fields, home gardens, and potato storages in Canada was analyzed for glucose phosphate-isomerase (Gpi) genotype, mating type, and resistance to metalaxyl. Both mating types and diversity in resistance to metalaxyl were detected within and amongGpi-genotypes. Pathogen populations were dominated by the 100:111:122Gpi-genotype (characteristic of US-8) in all the provinces where late blight was detected, except in 1997 in British Columbia (BC) where the 100:100:111Gpi-genotype (characteristic of US-11) predominated.Gpi-genotypes 100:111, 111:122, and 100:122 (characteristic of US-7, US-10 and US-14/US-17, respectively) were less common. Nearly all of the isolates with the 100:100:111Gpi-genotype were the A1 mating type, except for a few collected in BC in 1997. A majority of isolates with the 100:111:122Gpi-genotype were A2 except for a few found in Québec in 1998. Based on colony growth on media amended with 1,10, or 100 μg/ml metalaxyl, the 100:100:111Gpi-genotype isolates had higher levels of resistance to the fungicide in 1998 compared to 1997. In 1998, level of metalaxyl resistance was higher within the 100:100:111Gpi-genotype whereas it was lower within the 100:111: 122Gpi-genotype, when compared to 1997. BC was the only Canadian province where an increase in metalaxyl resistance within the 100:111:122Gpi-genotype was recorded between 1997 and 1998. Isolates from leaves had higher metalaxyl resistance levels than stem or tuber isolates, regardless of genotype.     

Phenotypes ofPhytophthora infestans in Nepal: mating types and metalaxyl sensitivity

Summary Potato late blight has appeared in epidemic proportions in Nepal since the mid 1990s and fungicides have been reported to be decreasingly effective in managing the disease.Phytophthora infestans isolates were collected from potato crops during 1999–2000 and analysis of 371 isolates for mating types and 270 isolates for metalaxyl sensitivity revealed the presence of both new and old populations. This is the first report on the presence of metalaxyl-resistant isolates in Nepal. The frequencies of A1 and A2 were 83 and 17%, respectively. Metalaxyl-resistant, intermediate and sensitive isolates were recorded as 10%, 12% and 78% respectively. Metalaxyl resistance was distributed in both mating types. Sites with a history of metalaxyl use had a significantly higher number of resistant and intermediate isolates ({ie337-1}, P<0.01) than sites where metalaxyl had not been used. This study confirms the changes in population structure ofP. infestans in Nepal.     

Survey ofPhytophthora infestans populations in Serbia: Mating type, metalaxyl resistance and virulence properties

Summary A total of 37 single-lesion isolates ofPhytophthora infestans were collected during 2000 from different potato growing regions in the central part of Serbia. They were paired in Al and A2 matings to determine their mating types. Of these 37 isolates, 28 were found to belong to the A2 mating type, eight to the Al type and one was found to be self-fertile (A1/A2). Using the in vitro mycelial radial growth method no metalaxyl resistantP. infestans isolate could be found. The rate of occurrence of intermediate isolates was not high, on the average 2.70%. All other isolates were sensitive to metalaxyl. There was no correlation between mating type and the intermediate metalaxyl resistance found.     

Virulence Complexity and High Levels of Fungicide Resistance Suggest Population Change of Phytophthora infestans in the Heilongjiang Province of China

Samples of Phytophthora infestans were isolated from 12 city and county fields in the main potato production areas of Heilongjiang province of China between 2004 and 2008. A total of 72 isolates were tested for virulence on a potato R gene differential set and 39 of these isolates were also analysed for their resistance to the fungicide metalaxyl. All 11 differential plants were infected by one or more of the isolates collected; the most commonly overcome differentials were R1, R3, R4, R7, R10, and R11. Most (89.7%) of the tested isolates were resistant to metalaxyl and the rest (10.3%) were sensitive. The proportion of metalaxyl-resistant isolates differed between sites (χ ²?=?25.42, df?=?10, p?<?0.005), but not between years (χ ²?=?4.63, df?=?4, p?>?0.05). Taken together and in comparison with earlier pathogen population studies, our findings suggest that the population of P. infestans in Heilongjiang is different from that existing in the province two decades ago and that at various locations within the province, the new population has a high frequency of resistance to metalaxyl.     

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.     

Characterisation of Phytophthora infestans Isolates Collected from Potato and Tomato Crops in Tunisia During 2006–2008

Severe late blight epidemics in Tunisia in recent years prompted population studies on the pathogen responsible for this disease, Phythophthora infestans. Characterisation of 165 Tunisian P. infestans isolates collected from 2006 to 2008 was performed for the mating type and mt haplotype, while subsets were analysed for metalaxyl sensitivity (n?=?65), virulence on differential set of 11 R genes of Solanum demissum (n?=?31), aggressiveness on cv. Bintje (n?=?36) and measurement of the radial growth on agar medium at three temperatures (n?=?38). Most isolates from potato and all isolates from tomato had the A1 mating type. The A2 mating type was detected in the north-east and northern areas, but not in the north-west. All the A2 mating type isolates were metalaxyl resistant and seem to be part of a new generation of the P. infestans isolates which are more aggressive, with more complex races, and tolerant to higher temperatures. The increased severity of epidemics during 2006 to 2008 can be attributed to favourable weather conditions during growing seasons, adaptation of new genotypes, widespread phenylamide resistance in potato production regions and most probably incorrect spray programmes. In contrast to the presence of complex pathotypes in two major potato crop regions (north-east and northern areas), the P. infestans population detected in the other regions and in tomato crops was still relatively simple. Compared with the situation in Europe and the American continent, or even compared with neighbouring countries such as Algeria, the genetic changes in Tunisia are still comforting and require strict management decision on late blight control to avoid the spread of new P. infestans populations from Europe or neighbouring countries.     

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.     

Mating Type, Virulence, Aggressiveness and Metalaxyl Resistance of Isolates of Phytophthora Infestans in Poland

A total of 93 single-lesion isolates of Phytophthora infestans were collected from late blight diseased potatoes grown in several locations in Poland, in the years 2002–2004. They were tested for mating type, virulence, aggressiveness and metalaxyl resistance. Sixty one percent of the isolates was of the A1 and 39% of the A2 mating type. Regarding virulence, complex races predominated. Most of the isolates were highly and moderately aggressive. Among isolates, tested together with 38 previously collected (1995–2001) ones, 14.5% was metalaxyl resistant, 3.5% intermediately resistant and 82.0% of the isolates was sensitive to metalaxyl. In addition, 14 isolates tested with three simple sequence repeat markers showed high genetic diversity. These data indicate that Polish population of P. infestans is diverse and aggressive, with potential for sexual recombination and spreading of fungicide insensitivity.     

Occurrence of the A2 mating type ofPhytophthora infestans in potato fields in the United States and Canada

Ten isolates ofP. infestans collected from blighted potato fields in the U.S. and Canada during the years 1983–1989 were examined for mating type on lima bean, oatmeal, and rye grain media. Two of the ten isolates produced oospores in less than 15 days when cultured in the presence of known A¹ mating types from the USA, Mexico, and Europe, indicating they are A² mating types. When the two A² isolates (one from Pennsylvania and one from Vancouver, B.C.) were cultured singly or with known A² isolates, no oospores formed after 6 weeks of culture. Pathogenicity tests showed that both isolates produced typical late blight symptoms on potato foliage and stems, and there appeared to be no difference in virulence between these two A² and the eight A¹ isolates. Inoculations with a mixture of A¹ + A² sporangia also produced oospores in host tissues. This is the first report of the presence of the A² mating type ofP. infestans in the U.S. and Canada.     

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.     

Monitoring metalaxyl resistance in populations ofPhytophthora infestans

Summary Samples of potato foliage infected withPhytophthora infestans were collected from the main potato growing areas in Ireland over a three-year period starting in 1981 when metalaxyl-based fungicides were no longer commercially available. Using the floating disc method, 75% of the isolates tested were found to be resistant to metalaxyl in 1981, 21% in 1982 and only 6% in 1983. The reliability of the test method and the sampling procedure to detect low levels of resistance is discussed.     

Characterization ofPhytophthora infestans from Maine during 1999 and 2000

A total of 77 isolates ofPhytophthora infestans was collected in 1999 and 93 isolates were collected in 2000 from growers’ fields, home gardens, and research plots in Maine. Genotype diversity based onglucose-6-phosphate isomerase (Gpi) allozyme analysis was greater in the collections from 2000, with six genotypes identified vs two genotypes identified in 1999. The US-8 (Gpi 100/111/122) genotype constituted 88% of the isolates collected in 1999 and 75% of those collected in 2000. The US-14 (Gpi 100/122) genotype constituted 12% and 7% of the isolates collected in 1999 and 2000, respectively. Three newGpi genotypes constituted 7% of the isolates collected from potato in northern Maine in 2000. A new genotype (A1 mating type,Gpi 100/100) that was aggressive on both tomato and potato was identified from home gardens in western Maine and constituted 11% of the isolates collected in 2000. A1, A2, and A1A2 mating types as well as self-fertile isolates were collected in both years. Self-fertility in artificial culture appeared to be relatively common in field isolatesof P. infestans. Potential origins of the new genotypes are discussed.     

Response ofColletotrichum coccodes to fungicidesin vitro

Black dot is an important disease of potato that affects all plant parts. The causal agent,Colletotrichum coccodes, is commonly found on potato tubers used for seed. Our objective was to determine thein vitro sensitivity ofC. coccodes to the fungicides maneb, thiabendazole, imazalil and CGA 173506, a phenylpyrrole. These fungicides are currently used, or are being tested for use, as potato seed treatments in the United States. All four fungicides reduced radial growth, sclerotial germination and spore germination ofC. coccodes at 10 μg/ml and higher. The fungicide concentrations that inhibited radial growth by 50% were 2 μg/ml, 5 μg/ml, 35 μg/ml, and 40 μg/ml for imazalil, CGA 173506, maneb and thiabendazole, respectively. TBZ was less inhibitory than the other three fungicides and stimulated sclerotial productionin vitro. Isolates ofC. coccodes differed in their sensitivity to maneb, thiabendazole and imazalil in radial growth, but not spore and sclerotial germination, assays. On CGA 173506, radial growth of fourC. coccodes isolates was inhibited, but these isolates produced sectors which grew normally. Germination of most sclerotia from these isolates was inhibited, but some germinated and produced normal colonies in the presence of CGA 173506. OneC. coccodes isolate was resistant to this fungicide in all assays.     

马铃薯晚疫病菌对瑞毒霉与霜脲氰的交互抗性

2008~2011年在黑龙江省共分离了41个马铃薯晚疫病菌,并测定了这些菌株对瑞毒霉和霜脲氰的敏感性和交互抗性。瑞毒霉测定结果显示6个(14.63%)菌株显示为敏感性,11个(26.83%)菌株显示为中抗,24个(58.54%)菌株显示为抗性。这说明黑龙江省分离的晚疫病菌已对瑞毒霉产生抗性。对于霜脲氰,EC50值为0.0630~2.1289μg/mL,最不敏感菌株是最敏感菌株的33.79倍,敏感性基线值为0.1879μg/mL。交互抗性分析结果显示,未产生交互抗性的菌株为73.16%,产生交互抗性菌株为26.84%。以上结果说明霜脲氰和瑞毒霉不能交叉使用。     

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.     

Characterization of <Emphasis Type="Italic">Phytophthora infestans</Emphasis> Isolates from Jersey,Channel Islands

Potato production on the island of Jersey, in the English Channel, is dominated by Jersey Royal, a selection of the early cultivar International Kidney. Jersey Royal is very susceptible to Phytophthora infestans, the cause of potato late blight, and Jersey’s climate is frequently conducive to infection. During 2004–2006, isolates of P. infestans were obtained from Jersey Royal plant samples from 41 different sites (24 crops, one infected tuber, 16 volunteers) and from five outdoor tomato crops and characterised by mating type, mtDNA haplotype, Gpi and Pep allozyme genotype, metalaxyl resistance and RG57 fingerprint. A subset of 17 isolates was additionally characterised by simple sequence repeat (SSR). All but one isolate from potato belonged to a single A1 multilocus genotype or a variant. The five isolates from tomato represented three distinct genotypes, none of which was found on potato and included one which was A2 mating type. The populations of P. infestans on potato and tomato in Jersey appear distinct, with that on potato being highly clonal.     

Comparative analysis of quantitative trait loci for foliage resistance toPhytophthora infestans in tuber-bearingSolanum species

Previously published data for potato foliage resistance to late blight were examined in a combined analysis to determine how the genetic backgrounds of the host and pathogen affect the occurrence of resistance QTLs. Data from 19 diploid populations and one tetraploid population that originated from at least 12 different tuber-bearingSolanum species were included. Comparative analysis across all populations revealed three highly active genomic regions on the distal parts of chromosomes 3,4, and 5. The region most consistently detected that conferred foliage resistance inSolanum was located on chromosome 5, near marker locus GP21. A previously identified cluster of three race-specific R-genes on chromosome 11 was not associated with polygenic resistance. Statistical examination of active QTLs indicates that congruence among QTLs is significantly affected by both genetic relatedness of mapping populations and race ofPhytophthora infestans used for resistance tests. The proportion of congruent QTLs for late blight resistance decreased from 0.52 detected in relatedSolanum populations tested with the same race ofPhytophthora infestans to 0.20 found in unrelated mapping populations tested with different races ofPhytophthora infestans. Analysis of resistance mapping tests provided statistical evidence for the occurrence of race-specific QTLs in tuber-bearingSolanum. The highly conserved genomic regions identified in the comparative analysis are likely to be good candidates for gene cloning or marker assisted selection in potato breeding programs.