Oospore formation by Phytophthora infestans in host tissue after inoculation with isolates of opposite mating type found in the Netherlands

Samenvatting Uit combinaties van A1- en A2-paringstypen vanPhytophthora infestans werden zowel bij tomaat als bij aardappel oösporen verkregen in de plant. Oösporen werden niet in de bladeren gevormd, maar werden veelvuldig aangetroffen in de epidermis van de stengels. De proeven werden uitgevoerd in de klimaatkamer; de veldsituatie moet nog onderzocht worden.     

Aggressiveness of Phytophthora infestans on detached potato leaflets in four Nordic countries

Potato fields in Denmark, Finland, Norway and Sweden were sampled for single-lesion isolates of Phytophthora infestans . The aggressiveness of the isolates was determined on detached leaflets of potato cvs Bintje (susceptible) and Matilda (moderately resistant). The aggressiveness tests were carried out in the respective home countries of the isolates, with the exception of the Danish isolates. Fifteen Danish isolates were studied in each of the other three countries, including five isolates tested in all three laboratories. Results obtained from the Danish isolates revealed substantial differences between the test laboratories for infection efficiency, lesion growth rate and sporulation capacity on detached leaflets. When the laboratory effect was taken into account, the differences in aggressiveness between the countries were generally small or inconsistent between the test cultivars and epidemiologically insignificant. By contrast, variation among isolates within countries was substantial. The magnitude of the variation depended on country and cultivar. Maximal variation for the means of the isolates was between 89 and 185 h for latent period, between 100 and 1297 sporangia mm⁻² for sporulation capacity and between nearly zero and 6 mm day⁻¹ for lesion growth rate. Typically less than 1% of sporangia were able to cause infections, except in Norway. These extraordinarily low values may be an artefact of the testing method. High variation in results between the test laboratories emphasizes the need for caution when comparing results obtained by different research groups.     

Stability of partial resistance in potato cultivars exposed to aggressive strains of Phytophthora infestans

Potato cultivars were evaluated for their resistance responses to aggressive strains of Phytophthora infestans in field and laboratory experiments. Analysis of variance revealed differential cultivar-by-isolate interactions for both foliar and tuber blight resistance. Differential responses occur as revealed by specific susceptibilities of cultivars to certain pathogen genotypes and changing rank order. In general, severity of late blight epidemics as observed in the haulms did not correlate well with foliar blight resistance ratings as presented in the National List of Recommended Potato Varieties. No significant correlation was found between tuber blight incidence under field conditions and the tuber blight rating in the National List. Also, there was no relation between the field and laboratory tuber blight resistance assessments. A significant association was demonstrated between late blight infection in the foliage and tuber blight incidence under field conditions. The presence of differential interaction, independent of R-gene-based resistance, indicates some adaptation of P. infestans to partial resistance and consequently adverse effects on the stability and durability of partial resistance to potato late blight.     

Genetic structure and pathogenicity of populations of Phytophthora infestans from organic potato crops in France, Norway, Switzerland and the United Kingdom

Genetic variation and pathogenicity of Phytophthora infestans isolates collected from organic potato crops of the susceptible cv. Bintje and the moderately resistant cv. Santé were assessed in France, Norway, and the United Kingdom in 2001 and in Switzerland in 2001 and 2002. Population structures differed considerably between the four P. infestans populations. Those from France, Switzerland and the UK were mainly clonal populations showing restricted levels of genetic diversity, whilst those from Norway were mixed A1 and A2 mating type populations with high levels of genetic diversity, suggesting periodical sexual reproduction. Isolates collected from cv. Bintje were on average more aggressive than or comparable to isolates from cv. Santé. Race complexity varied considerably between the regional P. infestans populations, with isolates from France and Switzerland showing the highest number of virulence factors. In all pathogen samples but the French, isolates collected from cv. Santé were more complex than isolates collected from cv. Bintje. No directional selection towards increased aggressiveness towards the more resistant cultivar Santé was observed. This suggests that there is no shift towards increased levels of pathogenicity in P. infestans populations following the large-scale introduction of more resistant potato varieties in organic production systems in Europe.     

French isolates of Phytophthora infestans from potato and tomato differ in phenotype and genotype

Prior to 1996, the A2 mating type of Phytophthora infestans was not detected on potato in France, but was found at one site on tomato in 1995. This finding lead to the question of the extent of differences and relationships existing between the populations of P. infestans present on each host. A collection of 76 isolates collected in France, mainly in 1996, from potato and tomato was characterised for mating type, allozyme genotype at the Gpi and Pep loci, and mitochondrial DNA haplotype; 74 of these isolates were also characterised for multilocus RFLP fingerprint, and 62 for virulence. All isolates except four showed allozyme genotypes (Gpi 90/100 or 100/100, Pep 83/100 or 100/100) and mtDNA haplotypes (Ia or IIa) characteristic of the populations introduced into Europe in the late 1970s. The four exceptions were isolates collected from tomato in Southern France in 1988-1991, which showed some characteristics of the former European populations (Gpi 86/100, Pep 92/100, mtDNA Ib). Both mating types were present among the collections from both hosts, but isolates with the A2 mating type were found on potato only in one garden crop, adjacent to tomato. Nine different RG57 fingerprints were observed, with a greater diversity among tomato isolates. Furthermore, tomato and potato collections differed markedly in the frequencies of genotypes present. Finally, tomato isolates generally had a lower virulence complexity than potato isolates. These data suggest that P. infestans populations on tomato and potato are largely separated, despite the occurrence of limited gene flow.     

Sensitivity of Phytophthora infestans to mandipropamid and the effect of enforced selection pressure in the field

Sensitivity to the new carboxylic acid amide (CAA) fungicide mandipropamid (MPD) in Phytophthora infestans was measured for isolates collected between 1989 and 2002 in Israel prior to the commercial use of MPD (baseline sensitivity, 44 isolates), and from MPD-treated (25 isolates) and untreated fields (215 isolates) in nine European countries and Israel between 2001 and 2005. All isolates were sensitive to MPD, with EC₅₀ values ranging between 0·02 and 2·98  µ g mL⁻¹. Plastic-tunnel (UK), shade-house (Israel) and field experiments (Israel) conducted during 2001–05 showed that enforced selection pressure, applied preventively or curatively, imposed by repeated sublethal (5  µ g mL⁻¹) or excessive (500–1000  µ g mL⁻¹) doses of MPD on mixed isolates of P. infestans produced no isolates resistant to the compound. The results of this study indicate that the probability of a buildup of resistant sub-populations of P. infestans to mandipropamid in the field is low.     

Sensitivity of Phytophthora infestans to flumorph: in vitro determination of baseline sensitivity and the risk of resistance

The sensitivity of 127 Phytophthora infestans isolates to flumorph was determined in 2003 and 2004. The isolates originated from two geographical regions and showed similar levels of sensitivity in both years. Baseline sensitivities were distributed as a unimodal curve with EC₅₀ values for growth of mycelia ranging from 0·1016 to 0·3228  µ g mL⁻¹, with a mean of 0·1813 (± 0·0405) µ g mL⁻¹. There was no cross-resistance between flumorph and metalaxyl. Laboratory studies were conducted to evaluate the risk of P. infestans developing resistance to flumorph. Mutants resistant to metalaxyl or flumorph were obtained by treating mycelium of wild-type isolates with ultraviolet radiation. Metalaxyl-resistant mutants were obtained with a high frequency and exhibited resistance factor values (EC₅₀ resistant/EC₅₀ sensitive phenotypes) of more than 100, while flumorph-resistant mutants were obtained at much lower frequencies and had very small resistance factors (1·5–3·2). There was cross-resistance between flumorph and dimethomorph, but not with azoxystrobin or cymoxanil. Most flumorph-resistant mutants showed decreases in hyphal growth in vitro and in sporulation both in vitro and on detached leaf tissues. These studies suggested that the risk of resistance developing was much lower for flumorph than metalaxyl. However, as P. infestans is a high-risk pathogen, appropriate precautions against resistance development should be taken.     

Population dynamics of metalaxyl-sensitive and metalaxyl-resistant isolates of Phytophthora infestans in untreated crops of potato

Each of six pairs of metalaxyl-sensitive (MS) and metalaxyl-resistant (MR) field isolates of Phytophthora infestans was inoculated as a mixture of 90% MS sporangia with 10% MR sporangia onto fungicide-free potato plants (cv. Alpha) in six walk-in plastic tunnels in each of 2 years. In both experiments, late blight killed the crops in 26-31 days. In all tunnels, the frequency of MR sporangia increased during the logarithmic phase of the epidemic to 70-85% of the population. Later, however, the frequency of M R sporangia decreased to 7-38% in six tunnels but continued to increase to 94-100% in the other six. The increased frequency of MR during the logarithmic phase was attributed to the shorter latent period and larger lesion size of MR compared to MS isolates and to the unlimited availability of uninfected host tissue. The later decrease in the frequency of MR in six tunnels was attributed to the lower sporulation capacity and the shorter infectious period of MR compared to MS isolates, and to the limitation of uninfected host tissue. The data showed that the population dynamics of P. infestans was dependent not only on fitness components of isolates, but also on the availability of uninfected host tissue.     

Effects of cropping history and origin of seed potatoes on population structure of Phytophthora infestans

The effects of origin of seed potatoes and the cropping history on the phenotypic structure of Phytophthora infestans populations was studied in northern Hessia, central Germany, from 2000 to 2002. Populations originating from fields with a history of potato cropping with only short or no rotation (old fields) were compared with populations from new fields, i.e., where no potatoes had been grown for at least 30 years and seed potatoes were either imported from breeders or produced on-farm (certified). The main goal was to determine the importance of seed potato infection in the establishment of new P. infestans populations. Isolates were characterized for mating type, virulences and rep- (repetitive extragenic palindromic) PCR fingerprints. Among a total of 639 isolates sampled from 31 sites, mating types A1 and A2 co-existed in all three years in 60–92% of the sites. Over all three years, 53 pathotypes were detected in a subsample of 272 isolates. Isolates originating from the new fields had significantly higher frequencies of the virulences v1, v2, v3, v6 and v7, indicating general effects of seed introduction into a new region. Thirty-six fingerprints were detected in a subsample of 281 isolates of which 22 were unique while four occurred in all three years and in many sites. Pathogen populations from potato fields that were grown from seed tubers of geographically different origin differed significantly based on χ ² tests. While the Nei genetic distances were less than .1 among the local populations, distances to the US lineages US-1, US-6, US-7 and US-8 ranged from .22 to .47; however, the bootstrap values were not significant. Populations from old fields were more diverse and 14 of the 22 rep-PCR types occurred there among 132 isolates tested in comparison to six in the new fields (n = 140 isolates) and two among six isolates from volunteers. The results also suggest that both sexual and asexual reproduction play a role.     

Stability of resistance to Phytophthora infestans in potato: an international evaluation

Ten institutions in nine countries joined together to test the stability of resistance of 14 potato genotypes to the oomycete pathogen Phytophthora infestans in three separate trials. Seven of the genotypes were tested in one trial involving seven locations, and all 14 were tested in two subsequent trials, each involving eight locations. Stability of resistance was tested with nonparametric tests and with an additive main effects and multiplicative interaction (AMMI) model. Overall, resistance to P. infestans was robust; resistant genotypes were consistently resistant in all locations and trials. The nonparametric analysis indicated that specific genotypes were basically stable across sites for resistance. In trial 3, the Z statistic for overall stability was significant at 0·05%, indicating a significant level of interaction across the trial, but there were no significant interactions for specific genotypes in this trial. The genotype by environment (G × E) effect of the AMMI model was highly significant in both trials, but the mean square of G × E was less than 10% of the genotype effect in each trial. The first two principal components (PCA1 and PCA2) of the AMMI analyses together explained 75 and 80% of the interaction effects in trials 2 and 3, respectively. Based on both nonparametric and AMMI analyses, Ecuador and Argentina were locations of relatively high interaction effects for both trials 2 and 3, although in Ecuador this interaction was not associated with any particular potato genotype. Other locations also had high interaction effects, but these occurred in only one trial. The genotypes Chata Blanca and, to a lesser extent, Torridon were relatively unstable in trials 2 and 3, but in the case of Torridon, resistant, this did not represent a significant loss of resistance.     

Selection and use of a mathematical model to evaluate components of resistance to Phytophthora infestans in potato

Five models of general epidemics, spatially homogeneous, were all shown to fit well to disease progress data forPhytophthora infestans on a susceptible potato cultivar. The models were: the logistic equation, the paralogistic or Vanderplank equation, two models from medical epidemiology with similar complexity, and a slightly more complex model with explicit treatment of lesion expansion. The use of the models for analysing the sensitivity of disease progress to changes in resistance components is discussed. Sensitivity analysis of the most complex model, by varying components within their range of genetic variation, indicates lesion expansion and infection efficiency as the components offering the best perspectives for resistance breeding. Improving two components simultaneously reduces disease progress slightly more than additively, but not enough to add other components to the list of breeding objectives. Pitfalls in using models for component sensitivity analysis, in the form of erroneous model initializations, are discussed, including implications for the role of components in the development of natural epidemics and in resistance breeding trials.     

Displacement of US‐1 clonal lineage by a new lineage of Phytophthora infestans on potato in Kenya and Uganda

Phytophthora infestans, the pathogen responsible for causing late blight in potato and tomato, is a rapidly evolving and highly adaptable pathogen. Following the appearance of a new clonal genotype in Kenya (KE‐1 lineage) in 2007, there was a need to document possible displacement of the US‐1 genotype by the newer lineage. Samples from Kenya (260) collected on potato and from Uganda collected on potato (134) and tomato (32) were genotyped with simple sequence repeat (SSR) and mitochondrial DNA haplotype (mtDNA) markers. Results show that, four years after its discovery, the KE‐1 lineage has completely displaced the US‐1 lineage on potato in Kenya. The KE‐1 lineage has also migrated into Uganda through the eastern part of the country that borders western Kenya. All potato samples from eastern Uganda were of the KE‐1 lineage. Moreover, samples from eight fields in western Uganda were also of the KE‐1 lineage, which is in agreement with an ongoing westward displacement. However, the new lineage has not displaced the old lineage on tomato, as all samples from tomato were US‐1. Principal coordinate analysis (PCA) scatter plots revealed clustering that indicated separation between the lineages and also between the subpopulations of the US‐1 lineage on potato and tomato.     

Population structure and host range of the potato late blight pathogen Phytophthora infestans in Peru spanning two decades

The genetic diversity of the late blight pathogen Phytophthora infestans infecting cultivated potato and alternative hosts growing in the vicinity of fields in the main potato-growing areas of the Peruvian Andes was characterized using collections from 1997–2013 as reference. The Peruvian P. infestans population, including previously collected and current isolates, consists of four clonal lineages (EC-1, US-1, PE-3 and PE-7) that belong to the A1 mating type and have been present in the country for decades. The first report of US-1 was in isolates collected between 1982 and 1986; meanwhile, EC-1 and PE-3 appeared for the first time in isolates from 1992 and PE-7 was found in 1997. The pathogen has a very broad host range among the solanaceous plants infecting cultivated potato, tomato, pear melon and several wild species. The solanaceous species growing in the vicinity of the potato fields sampled were identified and surveyed for late blight-like symptoms. Phytophthora infestans was isolated from nine wild species, including three new host species: Solanum zahlbruckneri, Solanum grandidentatum and Iochroma grandiflorum. There was no clear host specialization, but geographical substructuring was found as well as changes in the pathogen populations at the regional level. The clonal lineage EC-1, which is mostly resistant to metalaxyl, has complex virulence and contains a high level of subclonal variation, continues to dominate the population. Some multilocus genotypes of the EC-1 lineage were sampled in high frequencies and were found among the previously collected and new samples.     

Genetic diversity and host differentiation among isolates of Phytophthora infestans from cultivated potato and wild solanaceous hosts in Peru

To test the hypothesis that isolates of Phytophthora infestans attacking wild Solanaceae exhibit specialization for particular host species, 115 isolates of P. infestans were collected from cultivated potatoes, nontuber-bearing Solanum spp. of the Basarthrum section and wild tomatoes from five departments in the northern and central highlands of Peru, and characterized using several neutral markers. All isolates belonged to one of four clonal lineages described previously in Peru: EC-1, US-1, PE-3 and PE-7. There was a strong association of three lineages with host species: PE-3 was only isolated from cultivated potato, while PE-7 and US-1 were only isolated from nontuber-bearing Solanum spp. ( Basarthrum section and wild tomatoes). EC-1 was isolated from all host groups sampled. A subset ( n  = 74) of the isolates was evaluated for metalaxyl resistance. High levels of resistance were found almost exclusively in EC-1 and PE-3, while US-1 and PE-7 isolates were generally sensitive. In a detached-leaf assay for lesion diameter using five EC-1 isolates from S. caripense and seven EC-1 isolates from cultivated potato, there was a significant interaction between isolate origin and inoculated host, caused by higher aggressiveness of EC-1 from cultivated potato on its host of origin. In a comparison of EC-1 (seven isolates from cultivated potato) and US-1 (three isolates from S. caripense ), each pathogen lineage was more aggressive on its original host species, causing a highly significant interaction between isolate origin and inoculated host. Wild tomatoes and nontuber-bearing Solanum spp. harbour several pathogen lineages in Peru and could serve as reservoirs of inoculum that might contribute to epidemics on potato or tomato. Potential risks associated with the use of wild Solanum hosts as sources of resistance to P. infestans are discussed .     

Diversity of populations of Phytophthora infestans in relation to patterns of potato crop management in Latvia and Lithuania

Potato crop losses can be substantial when conditions for late blight (Phytophthora infestans) development and spread are favourable. In this study, drivers of differences between the P. infestans population structures in Latvia and Lithuania, two neighbouring countries with similar potato-growing traditions, were investigated. Genotypes of P. infestans and population genetic diversity were analysed using a 12-plex simple sequence repeat (SSR) marker assay. High genetic diversity was demonstrated in both populations, with population diversity being higher in Latvia. It would appear that local populations established from soilborne oospores early in the season are well adapted to the conditions in the region. However, somewhat greater spread and survival of local clones was detected in Lithuania, suggesting that potato cropping there is more vulnerable to clonal invasion than in Latvia. For effective disease management, current strategies should be adjusted according to the specific pathogen populations in the region, considering the reproduction and survival of the pathogen. Potato growers should implement late blight preventive measures such as longer field rotation to prevent oospore infections, especially in Latvia, and should use more disease resistant cultivars and high-quality seed potatoes.     

Activation of defence responses to Phytophthora infestans in potato by BABA

Late blight caused by Phytophthora infestans is one of the most devastating diseases of the potato crop. Resistance breeding and current fungicides are unable to control the rapidly evolving P. infestans and new control strategies are urgently needed. This study examined mechanisms of dl ‐β‐aminobutyric acid (BABA)‐induced resistance (IR) in the potato–P. infestans system. Leaves from two cultivars that differ in their degree of resistance, Bintje and Ovatio, were analysed after foliar treatment with BABA. Rapid activation of various defence responses and a significant reduction in P. infestans growth were observed in leaves treated with BABA. In the more resistant cultivar, Ovatio, the activation was both faster and stronger than in Bintje. Microscopic analysis of leaves treated with BABA revealed induction of small hypersensitive response (HR)‐like lesions surrounded by callose, as well as production of hydrogen peroxide (H₂O₂). Molecular and chemical analyses revealed soluble phenols such as arbutin and chlorogenic acid and activation of PR‐1. These results show a direct activation of defence responses in potato, rather than priming as reported for other plant species. They also show that the efficiency of BABA‐IR differs between cultivars, which highlights the importance of taking all aspects into consideration when establishing new methods for disease management.     

Severe outbreaks of late blight on potato and tomato in South India caused by recent changes in the Phytophthora infestans population

Late blight, caused by Phytophthora infestans, has emerged as the most destructive disease of potato and tomato in South India since 2008. One hundred and fifty‐seven isolates of Phytophthora infestans, 63 from potato and 94 from tomato, were collected from major potato and tomato production areas of South India between 2010 and 2012. Their phenotypic and genotypic characteristics were determined and compared with reference isolates. Isolates were characterized based on mating type, in vitro metalaxyl sensitivity, mitochondrial DNA haplotype, RG57 DNA fingerprinting patterns, SSR markers and aggressiveness on potato and tomato, in order to monitor population changes in P. infestans. All isolates were A2 mating type, metalaxyl resistant, mtDNA haplotype Ia and had RG57 and SSR fingerprints almost identical to the 13_A2 clonal lineage reported in Europe. Variation at the D13 and SSR4 loci allowed discrimination of minor variants, designated as 13_A2_3, 13_A2_3b, 13_A2_3c and 13_A2_1. A comparison of the lesion diameters caused by 157 isolates on detached leaflets of three potato and tomato cultivars showed all isolates to be equally aggressive, confirming that the same clonal population is infecting both hosts. This study demonstrates that the 13_A2 lineage was responsible for severe late blight outbreaks on potato and tomato in South India and has replaced the prior population represented by the US‐1 and other genotypes. Revised management strategies will be required to combat this destructive 13_A2 clonal lineage and monitoring of the population across other potato‐ and tomato‐growing regions of India is warranted.     

Evaluation of foliar resistance in potato to Phytophthora infestans based on an international field trial network

During the period 2000–03, local potato cultivars in Estonia, Latvia, Lithuania, Poland and Denmark were tested for foliar resistance to Phytophthora infestans (late blight) in an international field trial network. Four standard cultivars were included in the trials: Sava, Oleva, Danva and Kuras. Primary disease-assessment data were entered into a common database, and parameters from the disease progress curves were calculated and made available on interactive web pages. A regression model, using relative area under disease progress curve (RAUDPC) values for cv. Oleva as a reference, was developed for the estimation of 1–9 scale values, where 1 = most susceptible. Standard deviations for the estimated 1–9 scale values and a nonparametric rank stability analysis of RAUDPC were used to evaluate the stability of resistance of the cultivars. Overall, the results showed stability of resistance for cvs Sava, Oleva and Danva, but not for Kuras. Use of the Internet-based Web-Blight service in this study facilitated comparison of results among countries for the level and stability of resistance. The estimated 1–9 scale values were similar to, or slightly lower than, those from official cultivar lists or from the European Cultivated Potato Database, especially for the more resistant cultivars. Possible reasons for discrepancies from different sources and locations are discussed. It is concluded that RAUPDC and the derived 1–9 scale values are useful for ranking cultivars for resistance to P. infestans , but this information is not detailed enough for use in a decision support system for late blight control.     

Aggressiveness and other factors relating to displacement of populations of Phytophthora infestans in England and Wales

Mating type, in vitro sensitivity to the phenylamide fungicide metalaxyl, and mitochondrial (mtDNA) haplotype were determined in some or all of 618 isolates of Phytophthora infestans from the years between 1978 and 1995. A2 mating type occurred infrequently in most but not all years and insensitivity to metalaxyl increased over time. After 1982, the mtDNA Ib haplotype was largely replaced (except for one isolate in 1986 and one in 1995) by two new haplotypes, Ia and IIa. Type Ia was much more common than type IIa.Approximately one quarter of these isolates (165) were compared using two components of fitness associated with aggressiveness (infection frequency × number of sporangia per lesion) on detached leaves of cultivars Maris Piper, Cara and Stirling, which were chosen as exhibiting increasing levels of race non-specific resistance. Isolates were compared with three standard isolates of low, intermediate or high aggressiveness, and the data standardised for comparison between experiments. On cvs. Cara and Stirling, but not on Maris Piper, mtDNA Ia and IIa haplotypes were more aggressive than type Ib in several separate experiments. Similarly, metalaxyl sensitive phenotypes were more aggressive than insensitive phenotypes on Cara and Stirling but not on Maris Piper. The displacement of mtDNA type Ib by types Ia and IIa over this period may have been a result of the lower aggressiveness and lack of complete insensitivity to metalaxyl in type Ib isolates.