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.     

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.     

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.     

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.     

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.     

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.     

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.     

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.     

Testing for resistance to metalaxyl inPhytophthora infestans isolates from northwestern Washington

Outbreaks of late blight occurred in commercial potato fields in northwestern Washington in 1989 and reached epidemic proportions in 1990. Since most blighted fields reportedly had received 2–4 late-season applications of metalaxyl, the possibility existed that pathogenic strains ofPhytophthora infestans had developed resistance to this systemic fungicide. In testing this hypothesis,P. infestans was isolated consistently from diseased leaves, stems, and tubers of potato when small tissue pieces were surface-disinfected in 0.5% NaOCl for 3 min. Virulence of isolates was assessed on detached leaflets of eight differential (R-gene) potato genotypes. Although simple races (single and double) were recovered, there was also a high frequency of complex race combinations. When tested in metalaxyl-amended media, 81% of the 73 isolates recovered from diseased tissues were highly resistant to 10 μg metalaxyl/ml and 19% yielded intermediate resistance. Sensitive isolates were not recovered. Fifty-five percent of 40 isolates sporulated on potato tuber disks in the presence of ≥10 ug metalaxyl/ml. On leaf tissues, 85% of 20 isolates sporulated in the presence of 10 μg metalaxyl/ml, and 50% sporulated in the presence of 100 μg/ml. This is the first report ofP. infestans resistance to metalaxyl in the U.S.     

Incidence of the A2 mating type ofPhytophthora infestans on potato crops in Northern Ireland

Summary Of 250 isolates ofPhytophthora infestans obtained from Northern Ireland potato crops between 1981 and 1993, only six proved to be of the A2 mating type, the remainder being A1. The first A2 isolate was obtained from a tuber in 1987. The frequency of A2 isolates from 1987 onwards was 3%, a lower incidence than has been reported for England and Wales and the Republic of Ireland. There was no association between phenylamide resistance and mating type.     

黑龙江省和吉林省马铃薯晚疫病菌multi-locus基因型分析

2006～2008年在黑龙江省和吉林省共分离获得99株马铃薯晚疫病菌,并测定了这些菌株的交配型、瑞毒霉敏感性、同工酶基因型和mtDNA单倍型。交配型测定结果显示在吉林省和黑龙江省分离的所有菌株均为A1交配型。瑞毒霉敏感性测定结果显示敏感性菌株占14.1%,中抗菌株占7.1%,抗性菌株占78.8%,表明吉林省和黑龙江省发生的晚疫病菌已对瑞毒霉产生抗药性。黑龙江省和吉林省分离的所有菌株中发现了两种mt DNA单倍型(Ⅰa和Ⅱa)。其中,Ⅰa单倍型占11.1%,Ⅱa单倍型占88.9%。根据Gpi和Pep图谱,本试验发现了3种同工酶基因型,其中优势同工酶基因型是Gpi:100/100,Pep:100/100。根据菌株的交配型、同工酶基因型和mtDNA单倍型,共发现4种基因型,其中,multi-locus基因型A(84.9%)是黑龙江省和吉林省发现的优势基因型,但也出现了新的基因型分化。     

Impact of Phytosanitary Quality of Seed Potato and Temporal Epidemic Progress on the Phenotypic Diversity of Phytophthora infestans Populations

Altogether 365 isolates of Phytophthora infestans were sampled from potatoes propagated from seed potatoes of high (multiplied for two years in open field after meristem phase) and low (commercial certified seed multiplied for several consecutive years in open field) phytosanitary quality at different phases of epidemic progress during the growing seasons of 2001–2007 from field plots at two experimental institutes in Estonia, North-East Europe. High or low phytosanitary quality of seed potatoes had no effect on mating type ratio or response to metalaxyl in populations of P. infestans isolated from these two different groups of potato material. In contrast, the incidence of certain virulence factors, as well as the diversity of pathotypes, was very high in populations collected from potatoes propagated from low-quality seed in comparison to those from high-quality seed. The incidence of A2 mating type and fully metalaxyl sensitive strains was statistically significantly higher at the epidemic outbreak than later during epidemic progress. The incidence of most virulence factors and overall pathotype diversity were not affected by the temporal progress of the epidemic. Rare virulence factors 5 and 9 were more frequent at the outbreak of the epidemic and declined in the population during the course of epidemic.     

Occurrence of the A2 mating type ofPhytophthora infestans in India

Summary One hundred and four isolates ofPhytophthora infestans were collected during 1990–93 from different potato growing regions of India. They were paired with mating type A1 ofP. capsici to determine their mating type. All isolates from the Indo-Gangetic plains and Assam were the A1 mating type, whereas a mixed population of A1 and A2 types occurred at Shillong, Darjeeling and Shimla hills. The frequency of A2 isolates was 5–30% in the Shimla hills and 60% and 58% respectively in the Shillong and Darjeeling hills.     

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.     

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.     

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.     

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。     

Emerging and Re-emerging Potato Diseases in the Andes

Outbreaks of a new mating type (A2) of late blight (Phythopthora infestans) and the spread of bacterial wilt (Ralstonia solanacearum) were the most notable events among fungi and bacteria during the last 10 to 15 years in the Andes. The emergence of the A2 mating type of P. infestans was probably a result of seed movement from the Northern hemisphere. Black scurf caused by the fungus Rhizoctonia solani spread in Peru and Bolivia causing economic losses estimated at 5 to 10%. A high incidence of common scab on potato tubers, caused by the bacteria Streptomyces scabies was found in some localities in Bolivia. Among the emerging viruses, the NTN strains of PVY have been found in some countries including Peru. Recently, two other viruses have been found spreading in potatoes. One is coded SB26/29 and is transmitted by Russelliana solanicola, the first psyllid vector found for a plant virus. The other emerging virus is coded SB41 and is apparently also new to potatoes. It causes a severe mosaic on the affected plants and the virus is transmitted experimentally only by grafting. Some other viruses of minor importance have also been detected in potatoes.     

Characterization and Distribution of Three New Clonal Lineages of Phytophthora infestans Causing Late Blight in Wisconsin from 2009 to 2012

Phytophthora infestans causes late blight of potato and tomato, a disease that has been estimated to cost U.S. potato growers $287.8 million annually. We collected isolates of P. infestans from Wisconsin from 2009 to 2012 and determined distribution of clonal lineages and mating types and sensitivity to the systemic fungicide mefenoxam. We also sought to evaluate the current utility of an analysis of the Glucose-6-phosphate isomerase (Gpi) allozyme locus for predicting mefenoxam sensitivity with the aim of delivering timely information to growers. Overall, 143 isolates were collected from 52 locations in 20 Wisconsin counties from 2009 to 2012. Three clonal lineages, US-22, US-23, and US-24, were identified and were novel to Wisconsin and the U.S. US-22 is of the A2 mating type and sensitive to mefenoxam, with Gpi 100/122. US-23 and US-24 are of the A1 mating type and primarily intermediately sensitive to mefenoxam, with Gpi 100/100 and 100/100/111, respectively. Because of this close correlation and the unique Gpi patterns for each lineage present, we were able to predict mefenoxam sensitivity directly from samples using the allozyme assay and quickly deliver management information to growers. Both mating types were present in Wisconsin in 2009 and 2010 but were spatially separated and no evidence of sexual recombination or soil persistence was detected. The presence of new clonal lineages of P. infestans in Wisconsin indicates a need for continued close monitoring of late blight to facilitate generation of timely information for enhanced short-term and long-term late blight management.