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.     

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.     

Application of a PCR-based test for detection of potato late blight and pink rot in tubers

A polymerase chain reaction (PCR)-based test for potato late blight (Phytophthora infestans) and pink rot (P.erythroseptica, P. nicotianae) diseases has been developed for use with potato tuber tissue. Primers based on sequence analysis of the ITS2 region of ribosomal DNA of late blight and pink rot pathogens were utilized in PCR assays of inoculated tubers and tubers harvested from plots known to have late blight and/or pink rot. Assays of artificially inoculated Kennebec and Russet Burbank tubers revealed thatP. infestans was detected by PCR as early as 72 h after inoculation and in the absence of visible symptoms. Much higher detection frequencies were obtained by PCR compared with plating on selective medium or placement of tissue in moist chambers. Tubers from plots known to have late blight and/or pink rot were tested using the PCR assay. Assay of late blight lesions showed ca. 80% recovery for late blight-infected tubers from the field. Results indicate that the PCR assay provides a rapid and accurate test for diagnosis of late blight and pink rot in potato tubers.     

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.     

Relationship between resistance to late blight in potato foliage and tubers of cultivars and breeding selections with different resistance levels

Due to changes in the pathogen population, late blight of potatoes caused byPhytophthora infestans has become a more difficult disease to manage and there is a need for new potato cultivars with greater and more stable disease resistance. Field studies are required to identify and characterize host resistance to late blight in both foliage and tubers but, as epidemics vary from year to year, combining multi-year results can be difficult. Residual maximum likelihood (REML) was successfully used to analyze data which demonstrated good correlations between foliar and tuber responses to late blight (US-1 genotype). In addition, studies on post-harvest inoculated tubers provided more reliable disease responses than field studies, and the disease resistance with this method correlated well with foliar disease responses. This tuber test would be a useful method of pre-screening selections for disease response prior to testing in the field.     

The Pathosystem Solanum tuberosum L.-Phytophthora infestans (Mont.) de Bary in Chapingo,Mexico. Expected,Observed, and Simulated

The 1:1 ratio of the Phytophthora infestans A1 and A2 mating types favors the sexually derived genetic diversity of the late blight pathogen of potatoes, which is widespread in the central highlands of México. This ratio guarantees the successful infection of this pathogen, even in resistant potato hosts. However, wild Solanum species present in the region serve as alternative hosts to the pathogen. Knowledge about the external factors that influence the dynamics of this disease facilitates the assessment and selection for genetically resistant potato cultivars to late blight, in addition to providing the capacity to predict epidemics. This work aimed to assess the expected, observed, and simulated progress of natural P. infestans infection of potatoes during two epiphytotic field seasons (2009 and 2010) at Chapingo, México. Using 8-years of weather datasets, six ideal situations were predicted with four to eight infection cycles of 6 to 12 h each. In comparison to the predictions, the observed effect of the area under the disease progress curve, and its components (AUDPC, RAUDPC, RaRAUDPC), was highly significant, with a low coefficient of variation among the potato cultivars used in the study. In conclusion, we confirm that the LATEBLIGHT-LB2004 model is useful for simulating and predicting late blight epidemics based on the weather conditions of Chapingo, except for the magnitude of the relative humidity threshold variable (RH_threshold), which requires calibration for each cultivar.     

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.     

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.     

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.     

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.     

Resistance to late blight and soft rot in six potato progenies and glycoalkaloid contents in the tubers

Glycoalkaloids are anti-nutritional compounds commonly found in wildSolanum species used as resistance sources to major potato pathogens. It is therefore important for breeding purposes to know whether selecting for resistance using such species necessarily selects also for high glycoalkaloid contents in the tubers. To test this hypothesis, we used six partial progenies from crosses betweenSolanum tuberosum and accessions ofS. andigena, S. berthaultii, S. phureja, andS. vernei to investigate the possible correlation between resistance toPhytophthora infestans and/or toErwinia carotovora subsp.atroseptica and the concentration of glycoalkaloids in tubers. Concentrations of α-solanine and α-chaconine in the tubers segregated in each progeny, as did resistance to each pathogen. Some, but not all, clones from each progeny showed hypersensitive reactions to the isolate ofP. infestans used. Furthermore, clones within each progeny also differed for components of partial resistance toP. infestans, suggesting that all four wild species could be used as sources of both race-specific and partial resistance to late blight. With the exception of low, but statistically significant, correlations between concentration of α-solanine and two late blight resistance components (incubation period and spore production per unit lesion area) in progenies derived fromS. vernei, and despite a trend towards higher glycoalkaloid concentrations in the tubers of the clones most resistant to soft rot within progenies derived fromS. berthaultii andS. vernei, no consistent relationship between resistance to either disease and concentrations of α-solanine and/or α-chaconine was observed. These results indicate that neither race-specific nor partial resistance to late blight and soft rot in the accessions used as progenitors of resistance depend on high solanine or chaconine concentrations. These resistance sources could thus prove useful in breeding programs for improved behaviour againstP. infestans and/orE. carotovora.     

Indications of soil borne inoculum ofPhytophthora infestans

Summary In this paper we present observations made during three years in a potato field in Sweden that indicated the presence of soil borne inoculum ofPhytophthora infestans. In 1994, a fungicide trial against late blight was left in the field with various degrees of late blight infection. In 1995, cereals was grown in the field. In 1996, certified potato seed tubers were planted and blight appeared early in the season in a pattern corresponding to the heavily infected areas in 1994. No volunteer potato plants were found in 1995 or 1996. Both mating types A1 and A2 were isolated in 1996 and 1997 and oospores were found in leaves, stems as well as in stolons.     

Epidemiology and Integrated Control of Potato Late Blight in Europe

Phytophthora infestans, the causal agent of late blight, is a major threat to potato production in northwestern Europe. Before 1980, the worldwide population of P. infestans outside Mexico appeared to be asexual and to consist of a single clonal lineage of A1 mating type characterized by a single genotype. It is widely believed that new strains migrated into Europe in 1976 and that this led to subsequent population changes including the introduction of the A2 mating type. The population characteristics of recently collected isolates in NW Europe show a diverse population including both mating types, sexual reproduction and oospores, although differences are observed between regions. Although it is difficult to find direct evidence that new strains are more aggressive, there are several indications from experiments and field epidemics that the aggressiveness of P. infestans has increased in the past 20?years. The relative importance of the different primary inoculum sources and specific measures for reducing their role, such as covering dumps with plastic and preventing seed tubers from becoming infected, is described for the different regions. In NW Europe, varieties with greater resistance tend not to be grown on a large scale. From the grower??s perspective, the savings in fungicide input that can be achieved with these varieties are not compensated by the higher (perceived) risk of blight. Fungicides play a crucial role in the integrated control of late blight. The spray strategies in NW Europe and a table of the specific attributes of the most important fungicides in Europe are presented. The development and use of decision support systems (DSSs) in NW Europe are described. In The Netherlands, it is estimated that almost 40% of potato growers use recommendations based on commercially available DSS. In the Nordic countries, a new DSS concept with a fixed 7-day spray interval and a variable dose rate is being tested. In the UK, commercially available DSSs are used for c. 8% of the area. The validity of Smith Periods for the new population of P. infestans in the UK is currently being evaluated.     

Phenotypic stability of resistance to late blight in potato clones evaluated at eight sites in the United States

Changes in the fungal pathogenPhytophthora infestans in the United States pose a significant threat to potato production. Sources of resistance to these new genotypes of P.infestans need to be identified for potato breeders to have parental materials for crossing, and the phenotypic stability of late blight resistance in these potato clones needs to be determined. Sixteen potato clones which reportedly have some resistance to late blight were evaluated at eight locations: Florida (FL), Maine (ME), Michigan (MI), Minnesota (MN), North Dakota (ND), New York (NY), Pennsylvania (PA) and Wisconsin (WI) in 1996. Percent infected foliage was recorded at approximately weekly intervals following the onset of the disease at each location. Area under the disease progress curve (AUDPC) was calculated. Clones were ranked for mean AUDPC within location and the nonparametric stability statistics, mean absolute rank differences and variance of the ranks, were analyzed for phenotypic stability. Neither of these statistics was significant, indicating a lack of genotype x environment interaction on the rankings of these clones across locations in 1996. The four clones with lowest AUDPC scores were U.S. clones AWN86514-2, B0692-4, B0718-3 and B0767-2. These clones should be useful parental materials for breeders seeking to incorporate genes for late blight resistance into potatoes.     

The germplasm release of B0718-3 and B0767-2: Two late blight resistant potato clones

Two potato breeding selections were released by the United States Department of Agriculture, Agricultural Research Service on November 14, 1996, as germplasm that is highly resistant to current populations ofPhytophthora infestans (Mont.) deBary, the causal organism of potato late blight: B0718-3 and B0767-2. These two clones have been evaluated for their reaction toP. infestans in replicated plots at Presque Isle, Maine since 1990. B0718-3 is a netted white-skinned selection with good yields but low specific gravity. It is both male and female fertile and crosses readily with tetraploid Tuberosum. B0767-2 is a russet-skinned selection that is low yielding and low in specific gravity. It has not been successfully crossed; however, it may have some potential use in somatic hybridizations. Neither B0718-3 nor B0767-2 process into acceptable colored chips. Genetic material of this release has been deposited in the National Plant Germplasm System where it is available for research purposes, including development and commercialization of new cultivars.     

Induction of systemic resistance in potato (Solanum tuberosum L.) plants to late blight by local treatment withPhytophthora infestans (Mont.) de Bary,Phytophthora cryptogea Pethyb. & laff. or dipotassium phosphate

Summary Induced systemic resistance to late blight was found in the potato cultivar Bintje which has no major or minor resistance genes to late blight. The prerequisite was a local inoculation with 5×10³ zoospores/mo, ofPhytophthora infestans race 1, 3, 4, 7, 8, a local inoculation with 2×10³ zoospores/ml ofPhytophthora cryptogea isolated from wheat or local treatment with 50–100 mM solution of K₂HPO₄. Potato plants used in the experiments were propagated in vitro from nodal cuttings. Protection caused by the different inductions was 30 to 70% (assessed as a reduced number of lesions/plant and reduced necrotic tissue/leaf or plant) and tended to positively influence yield and dry matter of the haulm. The induced protection in the potato plant was better in young developing leaves than in fully expanded leaves.     

Impact of Agronomic Strategies (Seed Tuber Pre-sprouting,Cultivar Choice) to Control Late Blight (<Emphasis Type="Italic">Phytophthora infestans</Emphasis>) on Tuber Growth and Yield in Organic Potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) Crops

In order to optimize potato crop management in organic farming systems, knowledge of crop growth processes in relation to limitations and reductions by nitrogen (N) and disease is necessary. This paper examines the effect of different prevention measures (seed tuber pre-sprouting, choice of cultivars: resistance to Phytophthora infestans; earliness of tuber initiation) against disease-related constraints on yields, depending on the N-mediated growing conditions of organic potato crops. Under conditions of a relatively high N supply, accelerating the early development and tuber initiation of potato crops by seed tuber pre-sprouting (yield increase of c. 18–23%) or the selective choice of cultivars with an earlier tuber initiation (yield increase of c. 0–21%) are the most effective strategies in combating late blight. They bring forward the crop development c. 7–10 days, escaping the negative impact of the fungus on tuber growth. Under conditions of a relatively low N supply, these strategies do not affect final tuber yield. The reason for the differences of the effect depending on N supply of the crops is, that the lower the N supply, the shorter the period of time over which tuber growth takes place, independent of whether P. infestans becomes severe or not. Tuber growth of organic crops low in N has mostly ceased by the time late blight becomes potentially important for limiting yield, with the consequence that preventative measures against P. infestans are meaningless. Surprisingly, the level of cultivar haulm resistance to late blight did not affect tuber yields in years with an early and strong late blight development. Probably, the positive effect of a longer resistance to the fungus (c. 1 week) was counterbalanced by a later tuber initiation (also c. 1 week). In years with a moderate late blight incidence, less susceptible cultivars were able to produce higher tuber yields on fields with a lower N availability (c. 17–20%), probably due to higher N use efficiency and a better match of N mineralization and N uptake. Under conditions of a relatively high N supply, the tested cultivars with a higher haulm resistance to late blight were not able to improve tuber yields.     

Foliar reaction toPhytophthora infestans in inoculated potato field trials in Michigan

Late blight, caused byPhytophthora infestans, is the most important disease of potato worldwide and foliar resistance is an important component of managing late blight in the field. The objective of this research was to identify germplasm for use in breeding cultivars with foliar resistance toP. infestans. More than 500 clones were tested from 1997 to 2002 in inoculated (US8 genotype) field experiments conducted at the Michigan State University Muck Soils Research Farm in Bath, Michigan. All of the current commercial cultivars tested were classified as susceptible toP. infestans. The most resistant clones were A90586-11, AWN86514-2, B0718-3, Jacqueline Lee (MSG274-3), MSI152-A, MSJ307-2, MSJ317-1, MSJ453-4Y, MSJ456-2, MSJ456-4, MSJ461-1, MSK101-2, MSK128-1, NY121, LBR8, LBR9, Tollocan, and Torridon. Some of these resistant selections were from crosses with B0718-3, Jacqueline Lee, and Tollocan suggesting that the resistance toP. infestans was transmissible. These resistant clones will provide the opportunity to breed late-blight-resistant cultivars from a diverse pool of cultivated germplasm. Consistent foliar reaction toP. infestans over years suggested that the Michigan State University Muck Soils Research Farm is a valuable location for North American breeders to assess the reaction of potato germplasm to the US8 genotype of late blight.     

Changes in the distribution of phenylamide-resistant strains ofPhytophthora infestans (Mont.) de Bary in Ireland 1985–89

Summary Phenylamide-based fungicides were reintroduced to the Irish market in 1985. Mixtures of phenylamides and mancozeb were recommended for no more than 3 sprays in potato late blight control programmes with the last systemic spray to be applied not later than 31 July. Samples of potato foliage infected withP. infestans were collected from the main potato growing areas in Ireland over a five-year period starting in 1985. Using the floating disc method, 11% of the crops sampled in 1985 were found to have phenylamide-resistant strains ofP. infestans present. The number of crops with resistant strains increased rapidly up to 1988 when 83% of the crops sampled had resistance. Factors affecting the distribution of resistance and the efficacy of a phenylamide-based spray programme are discussed.