Phytophthora Infestans Isolates Research Articles

A machine learning algorithm for the automatic classification of Phytophthora infestans genotypes into clonal lineages

AbstractPremiseThe prompt categorization of Phytophthora infestans isolates into described clonal lineages is a key tool for the management of its associated disease, potato late blight. New isolates of this pathogen are currently classified by comparing their microsatellite genotypes with characterized clonal lineages, but an automated classification tool would greatly improve this process. Here, we developed a flexible machine learning–based classifier for P. infestans genotypes.MethodsThe performance of different machine learning algorithms in classifying P. infestans genotypes into its clonal lineages was preliminarily evaluated with decreasing amounts of training data. The four best algorithms were then evaluated using all collected genotypes.ResultsmlpML, cforest, nnet, and AdaBag performed best in the preliminary test, correctly classifying almost 100% of the genotypes. AdaBag performed significantly better than the others when tested using the complete data set (Tukey HSD P < 0.001). This algorithm was then implemented in a web application for the automated classification of P. infestans genotypes, which is freely available at https://github.com/cpatarroyo/genotypeclas.DiscussionWe developed a gradient boosting–based tool to automatically classify P. infestans genotypes into its clonal lineages. This could become a valuable resource for the prompt identification of clonal lineages spreading into new regions.

First report of different mating types of Phytophthora infestans isolates from potatoes in Cameroon and their control using Thevetia peruviana (Pers.) K. Schum. seeds extracts analyzed by GC–MS

First report of different mating types of Phytophthora infestans isolates from potatoes in Cameroon and their control using Thevetia peruviana (Pers.) K. Schum. seeds extracts analyzed by GC–MS

Fitness difference between two synonymous mutations of Phytophthora infestansATP6 gene

BackgroundSequence variation produced by mutation provides the ultimate source of natural selection for species adaptation. Unlike nonsynonymous mutation, synonymous mutations are generally considered to be selectively neutral but accumulating evidence suggests they also contribute to species adaptation by regulating the flow of genetic information and the development of functional traits. In this study, we analysed sequence characteristics of ATP6, a housekeeping gene from 139 Phytophthora infestans isolates, and compared the fitness components including metabolic rate, temperature sensitivity, aggressiveness, and fungicide tolerance among synonymous mutations.ResultsWe found that the housekeeping gene exhibited low genetic variation and was represented by two major synonymous mutants at similar frequency (0.496 and 0.468, respectively). The two synonymous mutants were generated by a single nucleotide substitution but differed significantly in fitness as well as temperature-mediated spatial distribution and expression. The synonymous mutant ending in AT was more common in cold regions and was more expressed at lower experimental temperature than the synonymous mutant ending in GC and vice versa.ConclusionOur results are consistent with the argument that synonymous mutations can modulate the adaptive evolution of species including pathogens and have important implications for sustainable disease management, especially under climate change.

The Variability of Phytophthora infestans Isolates Collected from Estonian Islands in the Baltic Sea.

Knowledge of a pathogen's genetic variability and population structure is of great importance to effective disease management. In this study, 193 isolates of Phytophthora infestans collected from three Estonian islands were characterized over 3 years using simple sequence repeat (SSR) marker data complemented by information on their mating type and resistance to metalaxyl. In combination with SSR marker data from samples in the neighboring Pskov region of Northwest Russia, the impact of regional and landscape structure on the level of genetic exchange was also examined. Among the 111 P. infestans isolates from Estonian islands, 49 alleles were detected among 12 SSR loci, and 59 SSR multilocus genotypes were found, of which 64% were unique. The genetic variation was higher among years than that among islands, as revealed by the analysis of molecular variance. The frequency of metalaxyl-resistant isolates increased from 9% in 2012 to 30% in 2014, and metalaxyl resistance was most frequent among A1 isolates. The test for isolation by distance among the studied regions was not significant, and coupled with the absence of genetic differentiation, the result revealed gene flow and the absence of local adaptation. The data are consistent with a sexual population in which diversity is driven by an annual germination of soilborne oospores. The absence of shared genotypes over the years has important implications when it comes to the management of diseases. Such population diversity can make it difficult to predict the nature of the outbreak in the coming year as the genetic makeup is different for each year.

Mitochondrial Genome Contributes to the Thermal Adaptation of the Oomycete Phytophthora infestans.

As a vital element of climate change, elevated temperatures resulting from global warming present new challenges to natural and agricultural sustainability, such as ecological disease management. Mitochondria regulate the energy production of cells in responding to environmental fluctuation, but studying their contribution to the thermal adaptation of species is limited. This knowledge is needed to predict future disease epidemiology for ecology conservation and food security. Spatial distributions of the mitochondrial genome (mtDNA) in 405 Phytophthora infestans isolates originating from 15 locations were characterized. The contribution of MtDNA to thermal adaptation was evaluated by comparative analysis of mtDNA frequency and intrinsic growth rate, relative population differentiation in nuclear and mtDNA, and associations of mtDNA distribution with local geography climate conditions. Significant variation in frequency, intrinsic growth rate, and spatial distribution was detected in mtDNA. Population differentiation in mtDNA was significantly higher than that in the nuclear genome, and spatial distribution of mtDNA was strongly associated with local climatic conditions and geographic parameters, particularly air temperature, suggesting natural selection caused by a local temperature is the main driver of the adaptation. Dominant mtDNA grew faster than the less frequent mtDNA. Our results provide useful insights into the evolution of pathogens under global warming. Given its important role in biological functions and adaptation to local air temperature, mtDNA intervention has become an increasing necessity for future disease management. To secure ecological integrity and food production under global warming, a synergistic study on the interactive effect of changing temperature on various components of biological and ecological functions of mitochondria in an evolutionary frame is urgently needed.

Fitness of homothallic and heterothallic isolates of Phytophthora infestans from Mexico

Phytophthora infestans is an important pathogen of potato and tomato worldwide. Knowledge of the fitness components (FCs), such as pathogenicity, virulence, latent period, lesion area and sporulation of isolates with different mating types from the same region, is important for improved understanding of the evolutionary potential of P. infestans and its short- and long-term management. The FCs and a Compound Biological Fitness Index (CBFI) of 12 strains of P. infestans collected from the central area of Mexico, with different mating types (A1, A2 and self-fertile), were compared under controlled conditions in growth chambers. All P. infestans strains were pathogenic, indicating that the conditions of the bioassays were adequate for disease development. FCs varied significantly among and within the three mating types, indicating that each group maintains substantial levels of genetic variation among and within isolates. The self-fertile group was significantly more fit in all FCs and CBFI, followed by groups A2 and A1. These results indicate that self-fertile strains are the most aggressive and have the greatest evolutionary potential in central Mexico. All FCs showed significant correlations with one another, indicating that these parameters can be used to estimate the biological fitness of P. infestans. These results provide fundamental information for the integrated management of this pathogen in central Mexico, as well as for understanding its evolutionary potential.

Phenotypic Stability and Correlation for Late Blight Resistance in Advanced Potato Clones Under Field and Controlled Conditions

AbstractLate blight (LB) is the main potato disease worldwide and one of the most important ways to control it is the use of resistant varieties. Twenty-two potato clones from the B3 breeding population developed by the International Potato Center with high resistance to the disease and two susceptible controls were inoculated with four Peruvian complex isolates (POX67, PPA61, PLL69, and PPI112) of Phytophthora infestans, with complex virulence on potato. Whole plant inoculation assays were carried out under greenhouse and humid chamber conditions in Lima, Peru, and data obtained were correlated with data from field assays carried out in Oxapampa (Pasco), a CIP breeding site in the Peruvian rain forest. High significant correlations (α = 0.01) were found in the resistance to LB shown by potato clones, the values of the correlations under greenhouse conditions between the isolates POX67, PPA61, and PLL69 with the resistance in the field were r = 0.93, 0.92 and 0.80, respectively and under humid chamber conditions were r = 0.94, 0.93 and 0.94, respectively. Moderate correlations were found between resistance in the field and in the greenhouse (r = 0.69) and the field and in humid chamber conditions (r = 0.77) for inoculations with PPI112 isolate. The twenty-two clones tested in this study showed phenotypic stability for LB resistance according to non-parametric analysis.

Study of Phytophthora infestans Mont. de Bary isolates in the planting of potatoes

Relevance. One of the most common diseases of potatoes and other nightshade family species is late blight caused by a pathogenic oomycete of the Phytophthora infestans (Mont.) de Bary. At least 100 species of phytophthora have been described in nature, affecting a wide range of plant species. The phytophthora population is heterogeneous and is represented by races, as well as different types of mating. This leads to a rapid adaptation of the pathogen and the emergence of new, more aggressive, and resistant races. Phytophthora is a parasite, the damage from which cannot be avoided within the organic farming framework. Therefore, it is particularly important to know the pathogenesis and racial composition of phytophthora in each individual region of Solanaceae cultivation.Research methodology. Differentiation and collection of material from the natural population were carried out using potato varieties with known R-genes in the genome. Isolation and introduction into the culture were carried out from leaves with the dampening chambers method, followed by cultivation on nutrient media. The pathogen was identified by microscopic analysis. Culture filtrates were obtained on the liquid nutritious medium, followed by liquid filtration and autoclaving. Phytotoxic activity was determined by the effect on the seedlings of the nightshade, grass, and pea families by the standard method. Molecular genetic analysis of the isolates was carried out by ISSR analysis; the primer, amplification mixture, and temperature profile of the reaction were selected according to the literature data; the calculation of genetic characteristics was carried out using POPGENE software packages.Results. Samples of seven Phytophthora infestans isolates were collected and introduced into culture. As a result of in vitro cultivation, morphological differences were revealed, expressed in the structure and color of the mycelium, the shape of the colonies, the nature of sporulation, the color of the reverse, and the medium under the colonies. The genetic differences of the natural phytophthora material introduced into the culture, collected from potato varieties with single resistance genes (R1, R3, R4), were revealed. Differences in the phytotoxic activity of the studied isolates' cultural filtrates were revealed. The isolated isolates demonstrate differentiation at the phenotypic, genetic and physiological levels, which allows us to speak about their belonging to races.

Genetic mechanism, baseline sensitivity and risk of resistance to oxathiapiprolin in oomycetes.

Oxathiapiprolin is a piperidinyl thiazole isoxazoline fungicide discovered by DuPont and commercialized by Corteva Agriscience. It acts by inhibiting a novel fungal target, an oxysterol binding protein (OSBP), and is intrinsically highly active against oomycetes including grape downy mildew (Plasmopara viticola) and potato late blight (Phytophthora infestans). Because the fungicide acts at a single site there is a need to determine the risk of resistance development. Oxathiapiprolin controlled European Plasmopara viticola and Phytophthora infestans isolates at very low concentrations with half maximal effective concentration (EC50 ) values ranging from 0.001 to 0.0264 mg L-1 and 0.001 to 0.03 mg L-1 , respectively. Laboratory mutagenesis studies performed with Phytophthora capsici using ultraviolet (UV) irradiation generated mutants with reduced sensitivity to oxathiapiprolin. All resistant mutants had a base pair change in the OSBP gene that resulted in an amino acid change. Most common substitutions were S768Y, G770V, G839W and L863W. Isolates of Plasmopara viticola and Phytophthora infestans with reduced sensitivity were also detected in field trial sites where oxathiapiprolin had been applied repeatedly each season over several consecutive years. The risk of oxathiapiprolin resistance development in Plasmopara viticola and Phytophthora infestans is medium to high and strict resistance management measures are required. Over-exposure of target populations to single-site fungicides during product development should be avoided.

Late Blight Resistance Evaluation and Genome-Wide Assessment of Genetic Diversity in Wild and Cultivated Potato Species

Late blight, caused by the oomycete Phytophthora infestans, is the most devastating disease in potato-producing regions of the world. Cultivation of resistant varieties is the most effective and environmentally friendly way to control potato late blight disease, and identification of germplasms with late blight resistance and clarification their genetic relationship would promote the development of the resistant varieties. In this study, a diverse population of 189 genotypes with potential late blight resistance, consisting of 20 wild species and cultivated Solanum tuberosum Andigenum group and Chilotanum group, was screened for the presence of late blight resistance by performing challenge inoculation with four Phytophthora infestans isolates including one 13_A2 isolate, CN152. Ten elite resources with broad-spectrum resistance and 127 with isolate-specific resistance against P. infestans were identified. To improve the available gene pool for future potato breeding programs, the population was genotyped using 30 simple sequence repeat (SSR) markers covering the entire potato genome. A total of 173 alleles were detected with an average of 5.77 alleles per locus. Structure analysis discriminated the 189 potato genotypes into five populations based on taxonomic classification and genetic origin with some deviations. There was no obvious clustering by country of origin, ploidy level, EBN (endosperm balance number) value, or nuclear clade. Analysis of molecular variance showed 10.08% genetic variation existed among populations. The genetic differentiation (Fst) ranged from 0.0937 to 0.1764, and the nucleotide diversity (π) was 0.2269 across populations with the range from 0.1942 to 0.2489. Further genotyping of 20K SNP array confirmed the classification of SSRs and could uncover the genetic relationships of Solanum germplasms. Our results indicate that there exits abundant genetic variation in wild and cultivated potato germplasms, while the cultivated S. tuberosum Chilotanum group has lower genetic diversity. The phenotypic and genetic information obtained in this study provide a useful guide for hybrid combination and resistance introgression from wild gene pool into cultivated species for cultivar improvement, as well as for germplasm conservation efforts and resistance gene mining.

Virulence of Phytophthora infestans isolates from potato in Spain

The oomycete Phytophthora infestans is responsible for the disease known as late blight in potato and tomato. It is the plant pathogen that has caused the greatest impact on humankind so far and, despite all the studies that have been made, it remains the most important in this crop. In Spain during the last years a greater severity of the disease has been observed in both, potato and tomato, probably due to genetic changes in pathogen populations described recently. The aim of this study was the characterization of the physiological strains of 52 isolates of P. infestans obtained in different potato-growing areas in Spain. For this purpose, inoculations on detached leaves were performed in order to determine compatibility or incompatibility reactions. A total of 17 physiological races were found. The less frequent virulence factors were Avr5 and Avr8. By studying the epidemiology of the pathogen, a specific breeding program for late blight resistance can be implemented.

Multiple Mechanisms Drive the Evolutionary Adaptation of Phytophthora infestans Effector Avr1 to Host Resistance.

Effectors, a group of small proteins secreted by pathogens, play a central role in antagonistic interactions between plant hosts and pathogens. The evolution of effector genes threatens plant disease management and sustainable food production, but population genetic analyses to understand evolutionary mechanisms of effector genes are limited compared to molecular and functional studies. Here we investigated the evolution of the Avr1 effector gene from 111 Phytophthora infestans isolates collected from six areas covering three potato cropping regions in China using a population genetic approach. High genetic variation of the effector gene resulted from diverse mechanisms including base substitution, pre-termination, intragenic recombination and diversifying selection. Nearly 80% of the 111 sequences had a point mutation in the 512th nucleotide (T512G), which generated a pre-termination stop codon truncating 38 amino acids in the C-terminal, suggesting that the C-terminal may not be essential to ecological and biological functions of P. infestans. A significant correlation between the frequency of Avr1 sequences with the pre-termination and annual mean temperature in the collection sites suggests that thermal heterogeneity might be one of contributors to the diversifying selection, although biological and biochemical mechanisms of the likely thermal adaptation are not known currently. Our results highlight the risk of rapid adaptation of P. infestans and possibly other pathogens as well to host resistance, and the application of eco-evolutionary principles is necessary for sustainable disease management in agricultural ecosystems.

Spatial modeling increases accuracy of selection for Phytophthora infestans‐resistant tomato genotypes

AbstractAt initial breeding stages, using a replicated check design is a viable alternative to reduce experimental field area as well as financial and operational costs. In this situation, spatial modeling could act to increase prediction accuracy of plant genotypic values. The objectives of this study were to demonstrate how spatially adjusted models can be used to reduce experimental error and how to compare statistical models in order to identify the best model for accurate genotype selection. For this purpose, we assessed 200 F3:4 tomato families for their resistance to Phytophthora infestans isolates. NC1CELBR, NC25P, and the cultivar Santa Clara were used as checks. Under field conditions, plants were inoculated with P. infestans isolates and scored according to their level of disease severity. Nine statistical models were adjusted to estimate family genotypic values. The selection of the fittest model was based on residual variance values, accuracy, Akaike and Bayesian information criteria, and the maximum likelihood ratio test. We observed spatial patterns within the experimental field area. Spatial modeling decreased error, which is indicated by the better experimental variation distribution. Residual variance decreased, while genotypic variance increased ∼10% when spatial analysis was used. Spatial analysis improved selection accuracy by 19% compared with the traditional analysis. Therefore, we recommend incorporating spatial modeling into data analysis in breeding trials for disease resistance because it can provide higher gains from selection compared with traditional modeling approaches, depending on the experimental condition.

Global multi-environment resistance QTL for foliar late blight resistance in tetraploid potato with tropical adaptation.

The identification of environmentally stable and globally predictable resistance to potato late blight is challenged by the clonal and polyploid nature of the crop and the rapid evolution of the pathogen. A diversity panel of tetraploid potato germplasm bred for multiple resistance and quality traits was genotyped by genotyping by sequencing (GBS) and evaluated for late blight resistance in three countries where the International Potato Center (CIP) has established breeding work. Health-indexed, in vitro plants of 380 clones and varieties were distributed from CIP headquarters and tuber seed was produced centrally in Peru, China, and Ethiopia. Phenotypes were recorded following field exposure to local isolates of Phytophthora infestans. QTL explaining resistance in four experiments conducted across the three countries were identified in chromosome IX, and environment-specific QTL were found in chromosomes III, V, and X. Different genetic models were evaluated for prediction ability to identify best performing germplasm in each and all environments. The best prediction ability (0.868) was identified with the genomic best linear unbiased predictors (GBLUPs) when using the diploid marker data and QTL-linked markers as fixed effects. Genotypes with high levels of resistance in all environments were identified from the B3, LBHT, and B3-LTVR populations. The results show that many of the advanced clones bred in Peru for high levels of late blight resistance maintain their resistance in Ethiopia and China, suggesting that the centralized selection strategy has been largely successful.

Evidence for a synergistic effect of post-translational modifications and genomic composition of eEF-1α on the adaptation of Phytophthora infestans.

Genetic variation plays a fundamental role in pathogen's adaptation to environmental stresses. Pathogens with low genetic variation tend to survive and proliferate more poorly due to their lack of genotypic/phenotypic polymorphisms in responding to fluctuating environments. Evolutionary theory hypothesizes that the adaptive disadvantage of genes with low genomic variation can be compensated for structural diversity of proteins through post‐translation modification (PTM) but this theory is rarely tested experimentally and its implication to sustainable disease management is hardly discussed. In this study, we analyzed nucleotide characteristics of eukaryotic translation elongation factor‐1α (eEF‐lα) gene from 165 Phytophthora infestans isolates and the physical and chemical properties of its derived proteins. We found a low sequence variation of eEF‐lα protein, possibly attributable to purifying selection and a lack of intra‐genic recombination rather than reduced mutation. In the only two isoforms detected by the study, the major one accounted for >95% of the pathogen collection and displayed a significantly higher fitness than the minor one. High lysine representation enhances the opportunity of the eEF‐1α protein to be methylated and the absence of disulfide bonds is consistent with the structural prediction showing that many disordered regions are existed in the protein. Methylation, structural disordering, and possibly other PTMs ensure the ability of the protein to modify its functions during biological, cellular and biochemical processes, and compensate for its adaptive disadvantage caused by sequence conservation. Our results indicate that PTMs may function synergistically with nucleotide codes to regulate the adaptive landscape of eEF‐1α, possibly as well as other housekeeping genes, in P. infestans. Compensatory evolution between pre‐ and post‐translational phase in eEF‐1α could enable pathogens quickly adapting to disease management strategies while efficiently maintaining critical roles of the protein playing in biological, cellular, and biochemical activities. Implications of these results to sustainable plant disease management are discussed.

Mating Type and Aggressiveness of Phytophthora infestans (Mont.) de Bary in Potato-Growing Areas of Punjab, Pakistan, 2017\u20132018 and Identification of Genotype 13_A2 in 2019\u20132020

Samples of blighted potato leaves were collected from fields in six potato-growing districts of the Punjab Province of Pakistan in 2017–2018. A total of 149 isolates of Phytophthora infestans were obtained from six potato cultivars (Asterix, Barsenna, Burana, Caroda, Mazika, Sante). Of these isolates, 73% were A1 mating type, 23% were A2 mating type and 4% were self-fertile. Both mating types of P. infestans occurred in all six districts sampled, but in every case, the A1 mating type predominated. The foliar aggressiveness of 104 of these isolates (weakly pathogenic isolates were excluded) was tested on detached leaflets of potato cv. Caroda, and a composite aggressiveness index (CAI) calculated from lesion area (measured after 10 days), latent period and infection frequency was used to compare isolates. There was no difference in CAI between isolates obtained from different districts or cultivars. The A2 mating type isolates had significantly greater CAIs than A1 isolates but this does not imply a genetic linkage nor a general association between mating type and aggressiveness. It may be that the A2 isolates belonged to an aggressive clonal lineage such as 13_A2 that has been reported in other countries in the region. While it was not possible to test the isolates collected in 2017–2018, genotyping of samples collected in 2019–2020 showed the widespread occurrence of the EU_13_A2 clonal lineage in the same districts of Pakistan and supported this hypothesis. This is the first report of EU_13_A2 from Pakistan. The implications for potato late blight management in the Punjab are discussed.

Appearance and diversity of Phytophthora infestans in Oman in 2013–2015

The purpose of this study was to characterize Phytophthora infestans isolates obtained from diseased tomato tissue in Oman from 2013 to 2015. Late blight caused severe epidemics on tomatoes in Oman in 2013–2014 and 2014–2015. Prior to 2013, late blight had not previously been reported from Oman. Isolates collected from diseased tomato plants in commercial production fields in Oman were assayed for a series of traits. All fifty-five isolates tested for mating type were determined to be A1. A subset of nine isolates analysed for allozymes, sensitivity to mefenoxam, DNA fingerprint as detected with random genome probe fifty-seven (RG57), and microsatellite genotypes revealed a diverse population. A range of sensitivity to mefenoxam was observed and one multi-locus genotype (MLG) was detected in four of the nine isolates. The A2 strain Blue_13, which was recently identified in India and thought to be a possible source of the pathogen, was not detected.

Improved Genome Sequence and Gene Annotation Resource for the Potato Late Blight Pathogen Phytophthora infestans.

Phytophthora infestans is a devastating pathogen causing potato late blight (Solanum tuberosum). Here we report the sequencing, assembly and genome annotation for two Phytophthora infestans isolates sampled in Republic of Korea. Genome sequencing was carried out using long read (Oxford Nanopore) and short read (Illumina Nextseq) sequencing technologies that significantly improved the contiguity and quality of P. infestans genome assembly. Our resources would help researchers better understand the molecular mechanisms by which P. infestans causes late blight disease in the future.

The Histological, Effectoromic, and Transcriptomic Analyses of Solanum pinnatisectum Reveal an Upregulation of Multiple NBS-LRR Genes Suppressing Phytophthora infestans Infection.

Utilization of disease resistance components from wild potatoes is a promising and sustainable approach to control Phytophthora blight. Here, we combined avirulence (Avr) genes screen with RNA-seq analysis to discover the potential mechanism of resistance in Mexican wild potato species, Solanum pinnatisectum. Histological characterization displayed that hyphal expansion was significantly restricted in epidermal cells and mesophyll cell death was predominant, indicating that a typical defense response was initiated in S. pinnatisectum. Inoculation of S. pinnatisectum with diverse Phytophthora infestans isolates showed distinct resistance patterns, suggesting that S. pinnatisectum has complex genetic resistance to most of the prevalent races of P. infestans in northwestern China. Further analysis by Avr gene screens and comparative transcriptomic profiling revealed the presence and upregulation of multiple plant NBS-LRR genes corresponding to biotic stresses. Six NBS-LRR alleles of R1, R2, R3a, R3b, R4, and Rpi-smira2 were detected, and over 60% of the 112 detected NLR proteins were significantly induced in S. pinnatisectum. On the contrary, despite the expression of the Rpi-blb1, Rpi-vnt1, and Rpi-smira1 alleles, fewer NLR proteins were expressed in susceptible Solanum cardophyllum. Thus, the enriched NLR genes in S. pinnatisectum make it an ideal genetic resource for the discovery and deployment of resistance genes for potato breeding.

Analysis of the lineage of Phytophthora infestans isolates using mating type assay, traditional markers, and next generation sequencing technologies.

Phytophthora infestans (Mont.) de Bary, a hemibiotrophic oomycete, has caused severe epidemics of late blight in tomato and potato crops around the world since the Irish Potato Famine in the 1840s. Breeding of late blight resistant cultivars is one of the most effective strategies to overcome this disruptive disease. However, P. infestans is able to break down host resistance and acquire resistance to various fungicides, possibly because of the existence of high genetic variability among P. infestans isolates via sexual and asexual reproduction. Therefore, to manage this disease, it is important to understand the genetic divergence of P. infestans isolates. In this study, we analyzed the genomes of P. infestans isolates collected from Egypt and Japan using various molecular approaches including the mating type assay and genotyping simple sequence repeats, mitochondria DNA, and effector genes. We also analyzed genome-wide single nucleotide polymorphisms using double-digest restriction-site associated DNA sequencing and whole genome resequencing (WGRS). The isolates were classified adequately using high-resolution genome-wide approaches. Moreover, these analyses revealed new clusters of P. infestans isolates in the Egyptian population. Monitoring the genetic divergence of P. infestans isolates as well as breeding of resistant cultivars would facilitate the elimination of the late blight disease.