Identification of an A2 population of Phythophthora andina attacking tree tomato in Peru indicates a risk of sexual reproduction in this pathosystem

Tree tomato, Solanum betaceum, is an Andean fruit crop previously shown to be attacked by Phytophthora andina in Ecuador and Colombia. Blight‐like symptoms were discovered on tree tomato plants in the central highlands of Peru in 2003 and shown to be caused by P. andina. Isolates of P. andina, collected from three different plantations in Peru over a 6‐year time span (2003–2008), were compared genetically with P. andina isolates from Colombia and Ecuador to test whether the pathogen population is geographically structured in the Andes. Restriction fragment length polymorphism (RFLP), mitochondrial DNA and simple sequence repeat (SSR) genetic markers, and mating type behaviour indicated that the Peruvian P. andina population from tree tomato is genetically distinct from populations infecting tree tomato in Colombia (CO‐1) and Ecuador (EC‐3, Ia, A1), but is more similar to the population infecting solanaceous hosts of the Anarrhichomenum complex (EC‐2, Ic, A2). Such geographic substructuring within this pathogen species could result from spatial isolation. Most strikingly, in contrast to the Ecuadorian and Colombian P. andina isolates from tree tomato, the Peruvian isolates have the A2 mating type. The presence of both mating types in the Andean population of P. andina attacking tree tomato indicates a risk of sexual reproduction and the presence of long‐lasting oospores in this pathosystem.     

Large subclonal variation in Phytophthora infestans populations associated with Ecuadorian potato landraces

The population of Phytophthora infestans on potato landraces in three provinces (Carchi, Chimborazo and Loja) of Ecuador was analysed. All isolates (n = 66) were of the A1 mating type. Simple sequence repeats (SSR) were used to assess the genetic diversity of the isolates. The P. infestans isolates from the potato landraces grouped in a single clade together with reference isolates belonging to the clonal lineage EC‐1. In the 66 SSR profiles obtained, 31 multilocus genotypes were identified. The 66 isolates constituted 49 different races according to the Solanum demissum differential set ( R1 to R11). The P. infestans population was complex and virulent on 4 to 11 R genes. Analysis showed that the subclonal variation in the Ecuadorian EC‐1 clone is increasing over time and is much larger than clonal variation in lineages in the Netherlands and Nicaragua, suggesting high mutation rates and little or no selection in Ecuador.     

Phytophthora urerae sp. nov., a new clade 1c relative of the Irish famine pathogen Phytophthora infestans from South America

An unknown Phytophthora species was discovered in the central Peruvian Andes on blighted foliage of the native South American plant species Urera lacineata. Urera is a genus of native flowering shrubs in the nettle family Urticaceae. This new taxon Phytophthora urerae sp. nov. is herewith formally described based on extensive morphological analysis, phylogenetic analysis of nuclear and mitochondrial loci, and AFLP analysis. Phytophthora urerae sp. nov. is a close relative of the Irish famine pathogen, Phytophthora infestans, and only the third clade 1c taxon described from South America to date. In contrast to the clade 1c taxon Phytophthora andina, first described in South America as a hybrid, P. urerae does not appear to be a hybrid based on cloning and sequencing nuclear loci. Findings of new species in South America may provide novel insights into the origin and evolutionary history of clade 1c Phytophthora species.     

SSR assessment of Phytophthora infestans populations on tomato and potato in British gardens demonstrates high diversity but no evidence for host specialization

Phytophthora infestans populations can differ in composition as a result of host specialization on tomato and potato hosts. In Great Britain many amateur gardeners grow outdoor tomatoes but there is little or no commercial tomato production outdoors. This study analysed isolates of P. infestans from British gardens with 12 multiplexed simple sequence repeat markers that are used to monitor the disease on commercial potato crops. Samples of P. infestans from tomato hosts were collected in 3 years and from potato in 1 year from across Great Britain. Seven previously unreported clonal lineages were detected in garden populations and higher frequencies of unique clonal lineages (28–40%) were present compared with populations from British commercial potato crops reported elsewhere. Garden populations had a lower proportion (11–48% less) of the most common lineages (13_A2 and 6_A1) that together made up at least 86% of the commercial potato populations during the sampling period. Host species accounted for only 2·0% of molecular variance detected between garden potato‐ and tomato‐hosted samples. No significant difference in clonal lineage composition was found between host species in Great Britain and this could be due to the whole P. infestans population overwintering on potato. British garden populations on both hosts were much more diverse than those on commercial potato crops; this finding may be influenced by less frequent fungicide use by gardeners and a higher diversity of unsprayed susceptible potato cultivars, enabling metalaxyl‐sensitive and less aggressive genotypes to survive in gardens.     

Phytophthora hydropathica,a new pathogen identified from irrigation water,Rhododendron catawbiense and Kalmia latifolia

A new species of Phytophthora, previously referred to as taxon Dre II, is named Phytophthora hydropathica. It is heterothallic, but all isolates recovered to date are of the A1 compatibility type. Plerotic oospores are produced. Its sporangia are usually obpyriform and are nonpapillate and noncaducous. Isolates of P. hydropathica had nearly identical single‐strand conformation polymorphism (SSCP)‐based DNA fingerprints that are distinct from those of all existing species. Their closest relatives are P. parsiana and P. irrigata. This new species is able to grow at relatively high temperatures, with an optimum of 30°C and a maximum of 40°C. It was frequently isolated from irrigation water during warm summers. This species caused leaf necrosis and shoot blight of Rhododendron catawbiense and collar rot of Kalmia latifolia at two nurseries where irrigation reservoirs yielded P. hydropathica. Its potential impact on other horticultural crops is discussed.     

Genetic Structure of the Population of Phytophthora infestans Attacking Solanum ochranthum in the Highlands of Ecuador

Thirty-nine isolates of Phytophthora infestans were collected from the wild host Solanum ochranthum in the highland tropics of Ecuador and characterized with a set of phenotypic and molecular markers (mating type, metalaxyl sensitivity, the allozyme loci Gpi, and Pep, mitochondrial DNA haplotype, RFLP, and SSR), as well as for pathogenicity on various hosts. Three groups of isolates (A, B, and C) were identified based on their multilocus genotypes and variable abilities to cause disease on different hosts. Group A had a combination of alleles for the Gpi (86/100), Pep (96/100) and mtDNA (Ia) loci, as well as an RFLP fingerprint, that have not been reported for P. infestans in Ecuador, or elsewhere. Group B shares many marker characteristics with the US-1 lineage described in Ecuador on tomato, pear melon (S. muricatum), and S. caripense, but has SSR alleles not present in typical US-1 isolates. Group C for all markers tested is identical to the EC-1 lineage described on cultivated and wild potatoes in Ecuador. All isolates from S. ochranthum were able to re-infect their host of origin in the detached leaf assay; however, we did not draw clear conclusions as to the relative aggressiveness of the three groups on this host. Isolates of group A were the most specialized and were generally non-pathogenic or weakly pathogenic on all hosts other than S. ochranthum. Groups B and C infected tuber-bearing hosts, including the cultivated potato but were generally non-pathogenic on other non-tuber bearing hosts. Solanum ochranthum was infected by isolates coming from tuber-bearing Solanum hosts (i.e., the EC-1 lineage of P. infestans) and some US-1 isolates from non-tuber bearing hosts. Thus, in nature this species might be a potential reservoir of inoculum of different pathogen populations able to infect the cultivated hosts potato, tomato and pear melon (S.␣muricatum). Unlike potato and tomato in Ecuador, each of which is primarily attacked by a highly specialized pathogen population, S. ochranthum appears to harbour at least three pathogen groups of␣different genetic make-up. The unresolved issue of potential host specificity in isolates found on S.␣ochranthum could complicate efforts to use this species in tomato improvement.     

Population structure of Phytophthora infestans in Turkey reveals expansion and spread of dominant clonal lineages and virulence

Late blight, caused by the oomycete Phytophthora infestans, is considered the most important and destructive disease of potato in Turkey. In this study, characterization of 367 isolates of P. infestans obtained from the potato-growing areas of the country was carried out to evaluate the pathogen population structure over the 2017–2019 production seasons. The isolates were characterized by numerous features including mating type, in vitro mefenoxam sensitivity, simple-sequence repeat (SSR) markers, and virulence against a set of potato differential lines. Most isolates were A2 mating type (353 isolates). Also, 68% of isolates were resistant to mefenoxam; the remainder were intermediate in their sensitivity and there were no sensitive isolates. SSR-based genotypic analysis of P. infestans populations showed a low genetic diversity. The 13_A2 clonal lineage predominated with a frequency of 92.1%, followed by 34_A1 (3.3%) and 37_A2 (2.7%). Genotypes 34_A1 and 37_A2 were detected only in 2019. This is the first report of 34_A1 and 37_A2 clonal lineages causing late blight disease of potato in Turkey. The most abundant virulence type was one overcoming resistance genes R1, R2, R3, R4, R6, R7, R10, and R11. These results emphasized that the migration of individuals and the asexual generation of subclonal differences were the main factors driving the population structure of P. infestans in Turkey.     

Genotypic and phenotypic characterization of Phytophthora infestans in South Korea during 2009–2016 reveals clonal reproduction and absence of EU_13_A2 genotype

In order to better understand the Phytophthora infestans population structure in South Korea, 172 isolates were collected between 2009 and 2016 from four major potato cultivation areas. Fungicide (metalaxyl and dimethomorph) response, mating type, and microsatellite (SSR) genetic fingerprints were analysed to characterize these isolates. Ten isolates collected in Gyeongnam Province, which specializes in protected winter cultivation in polytunnels, were A2 mating type. All other isolates were A1 mating type. Overall, 42% of the isolates were resistant to metalaxyl, and 43% were sensitive. All isolates were sensitive to dimethomorph. From the SSR fingerprints, 45 distinct genotypes were identified, which could be clustered into four clonal lineages: KR_1_A1, KR_2_A2, SIB-1, and US-11. KR_1_A1 was the predominant P. infestans genotype in South Korea. KR_2_A2 was only found in Gyeongnam Province; all isolates were A2 mating type and resistant to metalaxyl. SIB-1 was dominant until 2013 but its frequency has gradually decreased in more recent years. US-11 was first found in 2014, after which its frequency has increased to become codominant with KR_1_A1. The calculated standardized index of association (I_A) suggests that the South Korean P. infestans population is undergoing clonal reproduction. When compared with populations of P. infestans from the Netherlands, it has less genetic diversity and the dominant Netherlands P. infestans genotype, EU_13_A2 (Blue_13), was not found in South Korea. Such monitoring of the pathogen population contributes to a more efficient integrated pest management-based control strategy for potato late blight control in South Korea.     

Population genetic analysis of Phytophthora infestans in northwestern China

Late blight caused by Phytophthora infestans is the most devastating disease of potato worldwide. To understand the P. infestans population structure and dynamics in northwestern China, 959 single‐lesion isolates were purified in three consecutive years (2009–2011) and were characterized for mating type, pathotype, mtDNA haplotype and molecular variation at eight SSR loci. The results showed that the distribution of mating types changed significantly over years, with self‐fertile isolates dominant in 2010 and 2011. SSR genotyping distinguished 959 isolates into 151 genotypes, and association analysis indicated that P. infestans populations in 2010 and 2011 were strictly asexual while in 2009 they showed signs of sexual reproduction. Population analysis showed that the majority of genetic variation was within P. infestans populations. Isolates sharing identical SSR genotypes were detected in distant regions, indicating that migration of P. infestans could have occurred between regions. Pathogenicity assays on a set of potato differential lines containing R1 to R11 resistance genes detected four pathotypes from 74 selected isolates, with the pathotype virulent against all 11 R genes being dominant. Three mtDNA haplotypes (Ia, IIa, IIb) were detected with Ia being dominant among 507 isolates examined. Phylogenetic analysis indicated that P. infestans populations in northwestern China are distant from European lineages including 13‐A2 (blue‐13) at the time of this survey. The results have implications for the trade of healthy seed tubers as a means of managing late blight.     

Production and Germination of Oospores of Phytophthora infestans (Mont.) de Bary in Morocco

One hundred and eight isolates of Phytophthora infestans were collected from infected potato and tomato crops in the middle-north of Morocco during 1997–2000. Pairings of these isolates with tester isolates of mating type A1 and A2 revealed that 60% of the isolates were mating type A2 (65/108) and 40% were mating type A1. After 10 days incubation at 20 °C and a 16-h photoperiod, approximately 25% and 18% of the oospores produced in-vitro germinated in potato soil extract and potato root extract, respectively. Oospores were observed in potato leaf tissues in pairings that were fertile in-vitro. Maximum production of oospores was obtained in potato leaves of cultivars that were moderately susceptible (Desirée, Nicola) after 10 days of incubation at 15 °C and a 16-h photoperiod. These results confirm the presence of P. infestans strains that are sexually compatible under Moroccan climatic conditions. Production of oospores constitutes a threat for these crops because of the occurrence of recombinants with new virulences which may be difficult to control and as a consequence survival of oospores in absence of the host plant in the soil.     

Diversity of Phytophthora infestans from Poland

A collection of 96 Polish isolates of Phytophthora infestans sampled in the years 2006, 2008 and 2009 were analysed using phenotypic and genotypic markers. Mating type, virulence, resistance to metalaxyl, mitochondrial haplotype and polymorphism at 12 simple sequence repeat (SSR) loci were determined. The majority of isolates were of the A1 mating type, mitochondrial haplotype Ia and sensitive to metalaxyl. Virulence factors against potato R genes R1, R3, R4, R7, R10 and R11 were present in most isolates. Genotyping using SSR markers revealed high genetic diversity within the Polish P. infestans population. Amongst the 96 isolates 66 unique genotypes were identified, 49 of which were observed only in single isolates. Eight isolates of the genotype 13_A2 lineage that has been reported in other parts of Europe were also found in Poland. The implications of these results are discussed.     

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.     

Population structure of Phytophthora infestans in China – geographic clusters and presence of the EU genotype Blue_13

The population structure of Phytophthora infestans in China was studied and three mitochondrial haplotypes (Ia, IIa, IIb) were observed. Genetic analysis with 10 highly informative SSR markers identified 68 different genotypes, including three dominant clonal lineages. In the Chinese P. infestans population, the genotypes were strongly clustered according to their geographic origin. One of dominant clonal lineages was genetically similar to Blue_13, a dominant clonal lineage found in Europe since 2004. This is the first report of Blue_13 outside Europe. Only one mating type (A1) was found in the northern and southeastern provinces, but in southern and northwestern China both mating types were observed. The mating type ratio and SSR allele frequencies indicate that in China the sexual cycle of P. infestans is rare. These results emphasize that the migration of asexual propagules and the generation of subclonal variation are the dominant driving factors of the population structure of P. infestans in China. They may also have implications for the role of monitoring P. infestans populations in potato late blight management strategies in China.     

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.     

Phytophthora infestans populations in central,eastern and southern African countries consist of two major clonal lineages

Limited knowledge is available on Phytophthora infestans populations in Sub‐Saharan Africa (SSA). Therefore, and in response to recent severe late blight epidemics, P. infestans isolates from potato, tomato and Petunia × hybrida from eight SSA countries were characterized. Isolates were characterized with ‘old’ markers, including mating type (176 isolates), mitochondrial DNA haplotype (mtDNA) (281 isolates), glucose‐6‐phosphate isomerase (Gpi) (70 isolates), restriction fragment length polymorphism analysis with probe RG‐57 (49 isolates), and by metalaxyl sensitivity (64 isolates). Most isolates belonged to the US‐1 genotype or its variants (US‐1.10 and US‐1.11). The exceptions were genotype KE‐1 isolates (A1 mating type, mtDNA haplotype Ia, Gpi 90/100 and unique RG‐57 genotype), identified in two fields in Kenya, which are related to genotypes previously identified in Rwanda (RW‐1 and RW‐2), Ecuador and Europe. Metalaxyl‐resistant P. infestans isolates from potato were present in all the countries except Malawi, whereas all the isolates from tomato were sensitive. Genotyping of 176 isolates with seven simple sequence repeat (SSR) markers, including locus D13 that was difficult to score, revealed 79 multilocus genotypes (MLGs) in SSA. When this locus was excluded, 35 MLGs were identified. Genetic differentiation estimates between regional populations from SAA were significant when locus D13 was either excluded (P = 0·05) or included (P = 0·007), but population differentiation was only low to moderate (F_ST = 0·044 and 0·053, respectively).     

Distribution of major clonal lineages EU_13_A2, EU_2_A1, and EU_23_A1 of Phytophthora infestans associated with potato late blight across crop seasons and regions in Algeria

Potato is one of the most important agricultural crops in Algeria and worldwide. Each year, potato late blight, caused by Phytophthora infestans, is responsible for significant damage that leads to large production losses, and is thus a direct threat to food security in Algeria. In this study, 131 isolates of P. infestans and 92 DNA fingerprints captured on FTA cards were sampled from commercial and seed production fields in three major potato production regions (western, eastern, and central) during the main-season and late-season in Algeria over six cropping seasons (2010–2016). Genotypes of P. infestans and population genetic diversity were analysed using a 17-plex simple-sequence repeat (SSR) marker assay, and the mating type of all isolates was characterized. Both mating types (A1 and A2) were found, and often occurred in the same field. Differences in mating type proportion were observed between regions and between sampling periods. Analysis with SSR markers showed the prevalence of the EU_13_A2 lineage (70%) over EU_2_A1 (16%), EU_23_A1 (10%), and 4% of unknown multilocus lineage (MLL). The EU_13_A2 showed differentiation within the group. EU_23_A1 was found mainly in late-season crops. However, the cropping region did not influence the distribution of lineages due to the dispersal of the pathogen in Algeria by seeds. Genetic structure did not reveal a clear variation in distribution of the three lineages throughout the sampling regions. These data provide important new information on the composition and change over time of P. infestans populations in Algeria and open the way for a better understanding of the local epidemiology of this important pathogen.     

Evidence for selection pressure from resistant potato genotypes but not from fungicide application within a clonal Phytophthora infestans population

Insight into pathogen population dynamics provides a key input for effective disease management of the potato late blight pathogen Phytophthora infestans. Phytophthora infestans populations vary from genetically complex to more simple with a few clonal lineages. The presence or absence of certain strains of P. infestans may impact the efficacy of fungicides or host resistance. Current evidence indicates that genetically, the Irish populations of P. infestans are relatively simple with a few clonal lineages. In this study, P. infestans populations were genetically characterized based on samples collected at the national centre for potato breeding during the period 2012–16. The dominance of clonal lineages within this P. infestans population was confirmed and the potential selection pressure of fungicide treatment (2013–15) and host resistance (2016) on this clonal P. infestans population was then investigated. It was found that fungicide products did not notably affect the genetic structure of sampled populations relative to samples from untreated control plants. In contrast, samples taken from several resistant potato genotypes were found to be more often of the EU_13_A2 lineage than those taken from control King Edward plants or potato genotypes with low resistance ratings. Resistant potato varieties Sarpo Mira and Bionica, containing characterized R genes, were found to strongly select for EU_13_A2 strains.     

Characteristics of Phytophthora infestans Isolates from Uruguay

Isolates of Phytophthora infestans were obtained from late blighted plants from several potato-growing regions of Uruguay in 1998 and 1999. Of these, 25 representative isolates (4 from 1998, 21 from 1999) from the main potato-growing areas of the country, were characterised in terms of mating type, metalaxyl resistance, allozyme genotype, mitochondrial haplotype, RG57 fingerprint (1999 isolates only) and pathotype. All isolates proved to be A2 mating type, monomorphic and homozygous at the loci coding for glucose-6-phosphate isomerase and peptidase (Gpi 100/100, Pep 100/100) and to possess mitochondrial haplotype IIa. Metalaxyl-resistant isolates constituted 92% of the total. All the 1999 isolates possessed the same RG57 fingerprint, which was that previously reported as associated with the clonal lineage BR-1 from Brazil and Bolivia, which is also A2, Gpi 100/100, Pep 100/100. Most of the isolates displayed broad-spectrum virulence and five carried virulence to 10 of the 11 R genes tested despite the absence of R genes in commercially grown potato cultivars. It was concluded that the Uruguayan P. infestans isolates resembled isolates from neighbouring South American countries, notably Brazil, and belong to the new populations of the pathogen now predominant in many countries.     

SGT1 and HSP90 are essential for age-related non-host resistance of Nicotiana benthamiana against the oomycete pathogen Phytophthora infestans

The oomycete pathogen, Phytophthora infestans, is the causal agent of potato late blight, which is one of the most destructive and economically important plant diseases. We investigated the interaction between P. infestans and Solanaceous model plant Nicotiana benthamiana. Mature N. benthamiana plants were resistant to 8 isolates of P. infestans, whereas relatively young plants were susceptible to all isolates. Analysis with virus-induced gene silencing (VIGS) indicated that NbSGT1 and NbHSP90, genes essential for the function of R proteins, are required for the resistance of N. benthamiana to P. infestans. NbSGT1 was also required for the production of reactive oxygen species (ROS), hypersensitive cell death and expression of NbEAS, a gene for phytoalexin biosynthesis, induced by INF1, a secretory protein derived from P. infestans. These results suggested that N. benthamiana possibly possesses a broad-spectrum R protein against P. infestans, which requires an SGT1/HSP90-dependent mechanism, for the recognition of a conserved molecular pattern of P. infestans.