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Late blight disease, caused by the plant pathogen Phytophthora infestans, is one of the major threats for tomato and potato crops. Monitoring the populations of P. infestans is important to determine if there are changes in the sensitivity to fungicides and host preference. In this study, microsatellite markers and mitochondrial haplotypes were used to assess the genotype of isolates of P. infestans collected from tomato and potato plants in Colombia. Furthermore, sensitivity to the three fungicides cymoxanil (penetrant fungicide), mefenoxam, and fluopicolide (systemic fungicides), and tomato–potato host preference, were evaluated. Mitochondrial haplotyping showed that isolates collected on tomato were from the genetic groups Ia and Ib, while isolates collected on potatoes belonged to group IIa. Microsatellite analyses showed that isolates from tomato form two groups, including the Ib mitochondrial haplotype (which is genetically close to the US-1 clonal lineage) and the Ia haplotype (related to the EC-3 lineage), whereas Colombian isolates from potato formed a separate group. Furthermore, differences in sensitivity to fungicides were observed. Eighty-one percent of the isolates tested were resistant to mefenoxam with an EC50 >10 μg ml−1. Forty-two percent of the isolates showed an intermediate resistance to cymoxanil. The EC50 values ranged between 1 and 10 μg ml−1. For fluopicolide, 90% of the isolates were sensitive, with EC50 −1. Host preference assays showed that potato isolates infected both host species. Thus, isolates that infect potatoes may pose a risk for tomato crops nearby.

Phytophthora infestans, the causal agent of late blight disease of potatoes, is mainly controlled by the use of fungicides. Isolates that are resistant to commonly used fungicides have been reported. Also, several studies show that originally mefenoxam-sensitive isolates acquire resistance to this fungicide when exposed to sublethal concentrations. This phenomenon, termed “mefenoxam-acquired resistance,” has been observed in different Phytophthora species and seems to be unique to mefenoxam. In this study, we aimed to elucidate the molecular mechanism mediating this type of resistance as well as a possible regulatory process behind it. A combination of computational analyses and experimental approaches was used to identify differentially expressed genes with a potential association to the phenomenon. These genes were classified into seven functional groups. Most of them seem to be associated with a pleiotropic drug resistance (PDR) phenotype, typically involved in the expulsion of diverse metabolites, drugs, or other substances out of the cell. Despite the importance of RNA Polymerase I for the constitutive resistance of P. infestans to mefenoxam, our results indicate no clear interaction between this protein and the acquisition of mefenoxam resistance. Several small non-coding RNAs were found to be differentially expressed and specifically related to genes mediating the PDR phenotype, thus suggesting a possible regulatory process. We propose a model of the molecular mechanisms acting within the cell when P. infestans acquires resistance to mefenoxam after exposed to sublethal concentrations of the fungicide. This study provides important insights into P. infestans’ cellular and regulatory functionalities.

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.

Phenotypic diversity among individuals defines the potential for evolutionary selection in a species. Phytophthora infestans epidemics are generally thought to be favored by moderate to low temperatures, but temperatures in many locations worldwide are expected to rise as a result of global climate change. Thus, we investigated variation among individuals of P. infestans for relative growth at different temperatures. Isolates of P. infestans came from three collections: (i) individual genotypes recently dominant in the United States, (ii) recently collected individuals from Central Mexico, and (iii) progeny of a recent sexual recombination event in the northeastern United States. In general, these isolates had optimal mycelial growth rates at 15 or 20°C. However, two individuals from Central Mexico grew better at higher temperatures than did most others and two individuals grew relatively less at higher temperatures than did most others. The isolates were also assessed for mefenoxam sensitivity and mating type. Each collection contained individuals of diverse sensitivities to mefenoxam and individuals of the A1 and A2 mating type. We then searched for genomic regions associated with phenotypic diversity using genotyping-by-sequencing. We found one single nucleotide polymorphism (SNP) associated with variability in mycelial growth at 20°C, two associated with variability in mycelial growth at 25°C, two associated with sensitivity to mefenoxam, and one associated with mating type. Interestingly, the SNPs associated with mefenoxam sensitivity were found in a gene-sparse region, whereas the SNPs associated with growth at the two temperatures and mating type were found both at more gene-dense regions.

Host–pathogen interactions of a new species of Phytophthora, causal agent of late blight of tree tomato (Solanum betaceum Cav.), identified as Phytophthora betacei, were investigated with four different cultivars. Thirty-six P. betacei isolates, collected from southern Colombia between 2008 and 2009, were used to inoculate common tree tomato cultivars, Común, Híbrido, Injerto, and Holandés. Data on incubation and latent periods as well as infection efficiency, lesion development, and total sporulation were collected via detached leaf assays. Significant differences in susceptibility, based on the parameters measured, were observed. Común was the most susceptible cultivar, followed by Injerto, Híbrido, and Holandés. The mean incubation period was lowest for Común at 125.6 h post-inoculation (hpi) and highest for Híbrido at 139.4 hpi. No significant differences in latent period were observed. All 36 isolates produced necrotic lesions on Común, and 33, 24, and 21 caused infection on Injerto, Híbrido, and Holandés, respectively. Two isolates were able to cause infection only on Común, and 13 isolates were able to infect all four cultivars. Infection efficiency was significantly higher for the cultivar Común, followed by Injerto, Híbrido, and Holandés. Average lesion size was larger on Común than on any other cultivar. An inverse relationship of lesion size and total sporulation was observed. Común had significantly lower total sporulation than Híbrido and Holandés, which had the smallest average lesion sizes. These data show variation in pathogenicity of P. betacei isolates, under controlled conditions, and differential susceptibility of four distinct S. betaceum cultivars.

In most locations worldwide, populations of Phytophthora infestans are asexual and are often dominated by one or a few clonal lineages. The dominance in a population by any of these clonal lineages can be ephemeral. Studies in the USA and in the United Kingdom illustrate that dominance by any one clonal lineage may range from a year to two to fifteen years. In the eastern USA, lineage US22 dominated in 2009 and 2010. However, from 1994 to 2009, US8 was consistently dominant on potatoes throughout most of the continental USA. Migrations of exotic clonal lineages contribute substantially to changes in the population composition in most locations worldwide. Because these exotic strains can have “novel” phenotypic characteristics, they may not respond to traditional disease management techniques and thus may cause considerable damage which can lead to significant human suffering. Thus, while the disease persists in a location, the genotypes of P. infestans may change dramatically.

Phytophthora infestans is the causal agent of late blight disease of potatoes and tomatoes. This disease causes devastating economic losses each year, and control is mainly achieved by the use of fungicides. Unfortunately, populations of P. infestans resistant to fungicides have been documented. Furthermore, studies have reported that sensitive isolates to the phenylamide fungicide, mefenoxam, become less sensitive in vitro after a single passage through sublethal concentrations of fungicide-amended medium. The first objective of this study was to investigate if isolates of P. infestans are capable of acquiring resistance to two additional systemic fungicides, fluopicolide (benzamide) and cymoxanil (cyanoacetamide-oxime). In contrast to the situation with mefenoxam, exposure of isolates to sublethal concentrations of fluopicolide and cymoxanil did not induce reduced sensitivity to these two fungicides. The second objective was to assess if reduced sensitivity to mefenoxam could occur in naturally sensitive isolates of other Phytophthora species and of Phytopythium sp., another oomycete plant pathogen. All Phytophthora spp. assessed (P. infestans, P. betacei, and P. pseudocryptogea) as well as Phytopythium sp. acquired resistance to mefenoxam after previous exposure through medium containing 1 µg ml−1 of mefenoxam. Interestingly, isolate 66 of Phytopythium sp. and the isolate of P. pseudocryptogea tested do not seem to be acquiring resistance to mefenoxam after exposure to medium containing 5 µg ml−1 of this fungicide. The tested isolates of P. palmivora and P. cinnamomi were extremely sensitive to mefenoxam, and thus it was not possible to perform a second transfer to access acquisition of resistance to this fungicide.

Sporangia of Phytophthora infestans from pure cultures on agar plates are typically used in lab studies, whereas sporangia from leaflet lesions drive natural infections and epidemics. Multiple assays were performed to determine if sporangia from these two sources are equivalent. Sporangia from plate cultures showed much lower rates of indirect germination and produced much less disease in field and moist-chamber tests. This difference in aggressiveness was observed whether the sporangia had been previously incubated at 4°C (to induce indirect germination) or at 21°C (to prevent indirect germination). Furthermore, lesions caused by sporangia from plates produced much less sporulation. RNA-Seq analysis revealed that thousands of the >17,000 P. infestans genes with a RPKM (reads per kilobase of exon model per million mapped reads) >1 were differentially expressed in sporangia obtained from plate cultures of two independent field isolates compared with sporangia of those isolates from leaflet lesions. Among the significant differentially expressed genes (DEGs), putative RxLR effectors were overrepresented, with almost half of the 355 effectors with RPKM >1 being up- or downregulated. DEGs of both isolates include nine flagellar-associated genes, and all were down-regulated in plate sporangia. Ten elicitin genes were also detected as DEGs in both isolates, and nine (including INF1) were up-regulated in plate sporangia. These results corroborate previous observations that sporangia produced from plates and leaflets sometimes yield different experimental results and suggest hypotheses for potential mechanisms. We caution that use of plate sporangia in assays may not always produce results reflective of natural infections and epidemics.

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. Scientific and Technological Research Council of Turkey (TUBITAK)Turkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [116O329]; Bolu Abant Izzet Baysal University, Scientific Research Projects Coordination Unit [2017.10.06.1155] Scientific and Technological Research Council of Turkey (TUBITAK), Grant/Award Number: 116O329; Bolu Abant Izzet Baysal University, Scientific Research Projects Coordination Unit, Grant/Award Number: 2017.10.06.1155 WOS:000613660000001 2-s2.0-85100106589

Host-pathogen interactions of a new species of

Over the past few years, symptoms akin to late blight disease have been reported on a variety of crop plants in South America. Despite the economic importance of these crops, the causal agents of the diseases belonging to the genus Phytophthora have not been completely characterized. In this study, a new Phytophthora species was described in Colombia from tree tomato (Solanum betaceum), a semi-domesticated fruit grown in northern South America. Comprehensive phylogenetic, morphological, population genetic analyses, and infection assays to characterize this new species, were conducted. All data support the description of the new species, Phytophthora betacei sp. nov. Phylogenetic analyses suggest that this new species belongs to clade 1c of the genus Phytophthora and is a close relative of the potato late blight pathogen, P. infestans. Furthermore, it appeared as the sister group of the P. andina strains collected from wild Solanaceae (clonal lineage EC-2). Analyses of morphological and physiological characters as well as host specificity showed high support for the differentiation of these species. Based on these results, a complete description of the new species is provided and the species boundaries within Phytophthora clade 1c in northern South America are discussed.

Precision agriculture has emerged as a revolutionary technology, which transforms farming related data into useful information for agricultural decision-making. This paper compares precision farming technology with calendar-based approach in scheduling fungicide applications to manage potato (Solanum tuberosum L.) late blight disease. Three fungicide scheduling strategies were evaluated: calendar-based strategy, BlightPro decision support system based strategy (DSS-based strategy), and unsprayed control. Using results from 14 yr of computer simulation experiments for 59 locations in the United States, we constructed distributions of net return to all costs excluding fungicide cost and application cost per 0.41 ha (net return per 0.41 ha) for the calendar-based and DSS-based strategies at each location. These distributions were then compared using three risk management methods: stochastic dominance, stochastic dominance with respect to a function, and stochastic efficiency with respect to a function. The DSS-based strategy was identified as the most effective approach to manage late blight in terms of disease suppression, net return per 0.41 ha, and risk-adjusted net return. Results indicate that the DSS-based strategy is the preferred method to schedule fungicide applications. Under high disease pressure circumstances, the economic benefits to potato growers of adopting the precision agriculture technology ranged from US$30 to $573 per 0.41 ha. For risk neutral individuals, who are concerned about the difference between average net return per 0.41 ha, the benefits ranged from $30 to $305 per 0.41 ha. Except for growers raising the moderately resistant potato cultivars, more risk averse individuals tended to benefit more from adopting the precision agriculture technology, with benefits ranging from $38 to $573 per 0.41 ha.

Phytophthora infestans is the causal agent of potato and tomato late blight. This pathogen, which caused the Irish potato famine, is of profound historical significance and still poses a major threat in today's agroecosystems. Research on late blight epidemics usually focuses on pathogen virulence, host resistance, environmental factors and fungicide resistance. In this study, we examined the effect of PiRV-2, an RNA virus harbored by some P. infestans isolates, on its host. Comparing isogenic isolates with or without the virus demonstrated that the virus stimulated sporangia production in P. infestans. Transcriptome analysis suggested that it achieved sporulation stimulation likely through down-regulation of ammonium and amino acid intake in P. infestans. Survey of a limited P. infestans collection found PiRV-2 presence in most strains in the US-8 lineage, a very successful clonal lineage of P. infestans in North America. We suggest that PiRV-2 may affect the ecological fitness of P. infestans and thus could contribute to late blight epidemiology.

A marker database was compiled for isolates of the potato and tomato late blight pathogen, Phytophthora infestans, originating from 41 locations which include 31 countries plus 10 regions within Mexico. Presently, the database contains information on 1,776 isolates for one or more of the following markers: restriction fragment length polymorphism (RFLP) “fingerprint” consisting of 23 bands; mating type; dilocus allozyme genotype; mitochondrial DNA haplotype; sensitivity to the fungicide metalaxyl; and virulence. In the database, 305 entries have unique RFLP fingerprints and 258 entries have unique multilocus genotypes based on RFLP fingerprint, dilocus allozyme genotype, and mating type. A nomenclature is described for naming multilocus genotypes based on the International Organization for Standardization (ISO) two-letter country code and a unique number. Forty-two previously published multilocus genotypes are represented in the database with references to publications. As a result of compilation of the database, seven new genotypes were identified and named. Cluster analysis of genotypes from clonally propagated populations worldwide generally confirmed a previously published classification of “old” and “new” genotypes. Genotypes from geographically distant countries were frequently clustered, and several old and new genotypes were found in two or more distant countries. The cluster analysis also demonstrated that A2 genotypes from Argentina differed from all others. The database is available via the Internet, and thus can serve as a resource for Phytophthora workers worldwide.

Dorrance, A. E., Inglis, D. A., Derie, M. L., Brown, C. R., Goodwin, S. B., Fry, W. E., and Deahl, K. L. 1999. Characterization of Phytophthora infestans populations in western Washington. Plant Dis. 83:423-428. The first detection in the United States of isolates of Phytophthora infestans having metalaxyl insensitivity and complex pathotypes occurred in western Washington during the early 1990s. To determine the genetic structure of the current population in western Washington, a total of 115 isolates of P. infestans were obtained during 1996 from infected tubers or foliage of potato, tomato, nightshade, and bittersweet thr oughout the region. An additional 45 isolates were collected from a single field. Based on mating type, metalaxyl-insensitivity, and molecular markers (allozymes of glucose-6-phosphate isomerase, peptidase, and RG57 DNA fingerprint), all of the isolates were A1 mating type and had the US-11 multilocus genotype. Analyses of an additional 120 isolates collected during 1997 from potato, tomato, and nightshade were performed. As in 1996, US-11 was the predominant genotype detected on all three hosts. However, three additional A2 mating type genotypes were also detected: US-7, US-8, and US-14. These three genotypes represent the first A2 mating type isolates detected in western Washington. Most of a subset of 60 isolates infected 4 to 7 of the 10 potato differentials tested. This included 90% of the isolates collected in 1996 (all US-11), plus 72% of the US-11 and 100% of the US-8 and US14 isolates collected during 1997. Virulence phenotypes in this region are complex even without the selection pressure of R-genes in the local commercial cultivars. The apparent increase in genetic variation observed in populations of P. infestans in western Washington from 1996 to 1997 most likely occurred by migration rather than by sexual recombination.

Phytophthora infestans isolates (n = 26) collected in the Columbia Basin of Oregon and Washington in 1993, which had been characterized previously for mating type, metalaxyl sensitivity, and alleles at the glucose-6-phosphate isomerase locus, were analyzed for nuclear restriction fragment length polymorphism (RFLP) bands detected by probe RG57 and mitochondrial haplotype. Analyses involving the larger set of markers indicated that this group of isolates satisfied expectations of a sexual progeny: they contained much greater genetic diversity than has been reported for most other epidemic populations of P. infestans in the United States and Canada (16 unique multilocus genotypes); both mating types were present in proximity; all possible combinations of alleles occurred at many pairs of polymorphic loci; and two distinct mitochondrial haplotypes were distributed among the isolates. An in vitro laboratory cross involving the putative parents (US-6 and US-7) as parental strains produced progeny with the same general characteristics as the field isolates. Among the field progeny were two genotypes, US-11 and US-16, that had been described previously but from subsequent and largely clonal collections. Isolates obtained from tomatoes (n = 40) and potatoes (n = 7) in 24 counties in California in 1998 were analyzed as described above, and all except one US-8 isolate from potatoes were of the US-11 clonal lineage, consistent with the hypothesis that the US-11 lineage is an especially fit clonal lineage that has survived over time and can dominate pathogen populations over a large area. We conclude that the 1993 Columbia Basin collection represents a sexual progeny that generated the US-11 lineage, and that this lineage is particularly fit when tomatoes are part of the agroecosystem.

Oospores of Phytophthora infestans produced in vitro and in planta, from a cross between US-17 and US-8 genotypes, were exposed to a variety of environments and their survival was assessed. Additionally, the pathogenic characteristics of some resultant progeny isolates were assessed. Viability of oospores as measured by plasmolysis declined slightly over a period of 18 months whether they were stored in water at 4°C, in soil at 18°C, or in soil under natural field conditions. In comparison, viability as measured by germination was lower overall but appeared to increase after storage in soil. Oospores produced in planta were buried in the field in the fall of 1998, and were capable of infecting both tomato and potato leaflets when recovered in May 1999. Single oospore progeny (n = 53) from the in vitro cross were analyzed individually for genetic and pathogenicity characteristics. All 53 progeny tested for restriction fragment length polymorphisms with probe RG57 were hybrids. All but one progeny produced sporulating lesions on detached potato or tomato leaflets in growth chamber tests, but most lesions were smaller and developed more slowly than those produced by either parental isolate. In a further test of pathogenicity, under field conditions, none of a subset of 10 A2 progeny was capable of initiating a detectable epidemic in small plots of either potatoes or tomatoes.

Oospore formation by Phytophthora infestans in nature has been detected on potato leaflets in central Mexico (1), but there are no reports of oospore formation on tubers. A severe late blight epidemic occurred in Calimaya, Mexico, in fields where potato cv. Alpha was planted during the summer of 2000. Yield was reduced despite numerous applications of fungicide. Four hundred potato tubers left in the field were collected from the upper 10 cm of soil and examined for late blight symptoms. Tubers with soft and dry rot symptoms were observed, but symptoms of pink rot (Phytophthora erythroseptica) were not found. Four percent of the tubers showed late blight symptoms. Sections of 10 tubers with late blight symptoms were air-dried for 2 weeks in the laboratory and homogenized with a mortar and pestle. Glycerol was added to the homogenized tissue and observed microscopically. Aplerotic oospores (10 to 15 oospores per tuber) with amphyginous antheridia typical of P. infestans were observed. P. mirabilis morphologically similar to P. infestans is present in the area but it does not infect potato tubers. The number of oospores observed in our tuber sample was much lower than the number reported on leaflets (>1,000 oospores per leaflet) in the Toluca Valley. Low numbers of oospores have been reported on tubers artificially inoculated with P. infestans under field conditions (2). Infected tubers left in the field may act as a source of primary inoculum. To our knowledge, this is the first report of oospores of P. infestans found on tubers in Mexico under natural field conditions. References: (1) M. E. Gallegly and J. Galindo. Phytopathology 48:274, 1958. (2) A. Levin et al. Phytopathology 91:579, 2001.

The population of Phytophthora infestans in Brazil was first characterized 12 years ago. In this research, isolates of P. infestans from potato (n = 184) and tomato (n = 267) collected in southern and southeastern Brazil were characterized to provide more detailed analysis of the current structure of the population. All 451 isolates were analyzed for mating type, and subsets of the isolates were analyzed for allozymes, restriction fragment length polymorphism fingerprint, mtDNA haplotypes, and metalaxyl resistance. Tomato isolates were all of A1 mating type, mtDNA Ib, and US-1 genotype or some variant within this clonal lineage. Of the potato isolates, 82% were A2 mating type, mtDNA IIa, BR-1 genotype, which is a new lineage of P. infestans. All A2 isolates were found on potato, whereas 91% of the A1 isolates were from tomato. A1 and A2 isolates were never found in the same field. The frequency of resistance to metalaxyl was higher in isolates from tomato (55%) than in isolates from potato (38%). After more than a decade of coexistence of isolates of the A1 and A2 mating types, the population was highly clonal, dominated by the BR-1 and US-1 clonal lineages.

Factors affecting the management of petunia and tomato late blight, caused by Phytophthora infestans, under greenhouse conditions were investigated. Late blight-infected petunias (Petunia × hybrida) and tomatoes (Lycopersicon esculentum) each produced sporangia that were dispersed throughout the greenhouse via air currents. Infected petunias produced and released fewer sporangia than infected tomatoes, but infected petunias released sporangia two times longer. Surface-directed irrigation reduced disease incidence compared with overhead irrigation that wetted the foliage. The fungicides dimethomorph-mancozeb, fosetyl-Al, azoxystrobin, and dipo-tassium phosphonate/phosphate suppressed late blight development, as did the plant defense activator acibenzolar-S-methyl. All products were applied twice at 7-day intervals. The other plant defense activator (harpin protein) and the bioantagonists (Trichoderma harzianum, Glio-cladium virens, and Bacillus subtilis) were ineffective at the rates tested.

To determine if the desert tomato, Lycopersicon pennellii, possesses resistance to late blight, caused by Phytophthora infestans, two plant populations were analyzed. Resistance was identified through assessments of disease progress in an F2 mapping population (L. esculentum × L. pennellii) and in a series of introgression lines (L. pennellii into L. esculentum). Levels of resistance varied widely among individuals within each population. However, the response of individuals to different strains of P. infestans was consistent. In the mapping population, a quantitative trait locus (QTL) was detected near marker T1556 on chromosome 6. This QTL accounted for 25% of the phenotypic variance in the population. The occurrence of this QTL was confirmed from analysis of the introgression lines (ILs), where IL 6-2 (containing marker T1556) was the most resistant IL in 2002 and the second most resistant IL in 2001. The identification of an additional QTL for resistance to late blight in tomato will aid in the development of durable resistance to this devastating disease.

Isolates of Phytophthora infestans (n = 178) were collected in 2002 to 2009 from the eastern United States, Midwestern United States, and eastern Canada. Multilocus genotypes were defined using allozyme genotyping, and DNA fingerprinting with the RG-57 probe. Several previously described and three new mulitilocus genotypes were detected. The US-8 genotype was found commonly on commercial potato crops but not on tomato. US-20 was found on tomato in North Carolina from 2002 through 2007 and in Florida in 2005. US-21 was found on tomato in North Carolina in 2005 and Florida in 2006 and 2007. US-22 was detected on tomato in 2007 in Tennessee and New York and became widespread in 2009. US-22 was found in 12 states on tomato and potato and was spread on tomato transplants. This genotype accounted for about 60% of all the isolates genotyped. The US-23 genotype was found in Maryland, Virginia, Pennsylvania, and Delaware on both tomato and potato in 2009. The US-24 genotype was found only in North Dakota in 2009. A1 and A2 mating types were found in close proximity on potato and tomato crops in Pennsylvania and Virginia; therefore, the possibility of sexual reproduction should be monitored. Whereas most individuals of US-8 and US-20 were resistant to mefenoxam, US-21 appeared to be intermediately sensitive, and isolates of US-22, US-23, and US-24 were largely sensitive to mefenoxam. On the basis of sequence analysis of the ras gene, these latter three genotypes appear to have been derived from a common ancestor. Further field and laboratory studies are underway using simple sequence repeat genotyping to monitor current changes in the population structure of P. infestans causing late blight in North America.

The tomato late blight pandemic of 2009 made late blight into a household term in much of the eastern United States. Many home gardeners and many organic producers lost most if not all of their tomato crop, and their experiences were reported in the mainstream press. Some CSAs (Community Supported Agriculture) could not provide tomatoes to their members. In response, many questions emerged: How did it happen? What was unusual about this event compared to previous late blight epidemics? What is the current situation in 2012 and what can be done? It's easiest to answer these questions, and to understand the recent epidemics of late blight, if one knows a bit of the history of the disease and the biology of the causal agent, Phytophthora infestans.

Cooke, David/0000-0002-9154-7954 WOS: 000509100200009 Potato late blight, caused by Phytophthora infestans, has been the most damaging disease in Turkey since 2010. In this study, 127 isolates of P. infestans were obtained from the main growing areas of Turkey between 2015 and 2017. Their phenotypic and genotypic features were revealed and presented with those of reference isolates. These isolates were categorized by their mating type, in vitro mefenoxam sensitivity, mtDNA haplotype, RG57 DNA fingerprinting patterns, simple sequence repeat (SSR) markers, and aggressiveness on a set of potato differential lines. All isolates were of the A2 mating type and mtDNA haplotype Ia, were resistant to mefenoxam, and had RG57 and SSR fingerprints similar to the 13_A2 clonal lineage reported in Europe. This is the first report of 13_A2 in Turkey. Virulence abilities against potato resistance (R) genes R1, R2, R3, R4, R6, R7, R10, and R11 were observed in most of the isolates. The mating type ratios and SSR marker analysis indicate that in Turkey, the sexual reproduction of P. infestans is limited. These results underline that the movement of asexual individuals and the generation of sub-clonal difference are the factors driving the population structure of P. infestans in Turkey. TUBITAKTurkiye Bilimsel ve Teknolojik Arastirma Kurumu (TUBITAK) [116O329] This research was supported by TUBITAK (grant 116O329).

Phytophthora infestans, a pathogenic oomycete that is the causal agent of potato and tomato late blight, has devastating effects worldwide. The genetic composition of P. infestans populations in Canada has changed considerably over the last few years, with the appearance of several new genotypes showing different mating types and sensitivity to the fungicide metalaxyl. Genetic markers allowing for a rapid assessment of genotypes from small amounts of biological material would be beneficial for the early detection and control of this pathogen throughout Canada. Mining of the P. infestans genome revealed several regions containing single-nucleotide polymorphisms (SNP) within both nuclear genes and flanking sequences of microsatellite loci. Allele-specific oligonucleotide polymerase chain reaction (ASO-PCR) assays were developed from 14 of the 50 SNP found by sequencing. Nine optimized ASO-PCR assays were validated using a blind test comprising P. infestans and other Phytophthora spp. The assays revealed diagnostic profiles unique to each of the five dominant genotypes present in Canada. The markers developed in this study can be used with environmental samples such as infected leaves, and will contribute to the genomic toolbox available to assess the genetic diversity of P. infestans at the intraspecific level. For late blight management, early warning about P. infestans genotypes present in potato and tomato fields will help growers select the most appropriate fungicides and application strategies.

The objective of this study was to evaluate the utility of the BlightPro decision support system (DSS) for late blight management using computer simulation and field tests. Three fungicide schedules were evaluated: (i) calendar-based (weekly) applications, (ii) applications according to the DSS, or (iii) no fungicide. Simulation experiments utilized 14 years of weather data from 59 locations in potato-producing states. In situations with unfavorable weather for late blight, the DSS recommended fewer fungicide applications with no loss of disease suppression; and, in situations of very favorable weather for late blight, the DSS recommended more fungicide applications but with improved disease suppression. Field evaluation was conducted in 2010, 2011, 2012, and 2013. All experiments involved at least two cultivars with different levels of resistance. DSS-guided and weekly scheduled fungicide treatments were successful at protecting against late blight in all field experiments. As expected, DSS-guided schedules were influenced by prevailing weather (observed and forecast) and host resistance and resulted in schedules that maintained or improved disease suppression and average fungicide use efficiency relative to calendar-based applications. The DSS provides an interactive system that helps users maximize the efficiency of their crop protection strategy by enabling well-informed decisions.

Phytophthora infestans is the causal agent of potato and tomato late blight. In this study, we obtained the complete genome sequence of a novel RNA virus from this plant pathogen, tentatively named “Phytophthora infestans RNA virus 2” (PiRV-2). The PiRV-2 genome is 11,170 nt in length and lacks a polyA tail. It contains a single large open reading frame (ORF) with short 5’ and 3’ untranslated regions. The ORF is predicted to encode a polyprotein of 3710 aa (calculated molecular weight, 410.94 kDa). This virus lacks significant similarity to any other known viruses, even in the conserved RNA-dependent RNA polymerase region. Phylogenetic analysis demonstrated that it did not cluster with any known virus group. We conclude that PiRV-2 belongs to a new virus family yet to be described. This virus was found to be faithfully transmitted through asexual reproduction.

Phytophthora infestansis the causal agent of potato and tomato late blight. In this study, we characterized a novel RNA virus, Phytophthora infestans RNA virus 2 (PiRV-2). The PiRV-2 genome is 11,170 nt and lacks a polyA tail. It contains a single large open reading frame (ORF) with short 5’- and 3’-untranslated regions. The ORF is predicted to encode a polyprotein of 3710 aa (calculated molecular weight 410.94 kDa). This virus lacks significant similarity to any other known viruses, even in the conserved RNA-dependent RNA polymerase region. Comparing isogenic strains with or without the virus demonstrated that the virus stimulated sporangia production inP. infestansand appeared to enhance its virulence. Transcriptome analysis revealed that it achieved sporulation stimulation likely through down-regulation of ammonium and amino acid intake inP. infestans. This virus was faithfully transmitted through asexual reproduction. Survey of PiRV-2 presence in aP. infestanscollection found it in most strains in the US-8 lineage, a very successful clonal lineage ofP. infestansin North America. We suggest that PiRV-2 may have contributed to its success, raising the intriguing possibility that a potentially hypervirulent virus may contribute to late blight epidemics.Author SummaryPotato late blight, the notorious plant disease behind the Irish Potato Famine, continues to pose a serious threat to potato and tomato production worldwide. While most studies on late blight epidemics focuses on pathogen virulence, host resistance, environmental factors and fungicide resistance, we present evidence in this study that a virus infecting the causal agent,Phytophthora infestans, may have played a role. We characterized a novel RNA virus, Phytophthora infestans RNA virus 2 (PiRV-2) and examined its effects on its host. By comparing identicalP. infestansstrains except with or without the virus, we found that PiRV-2 stimulated sporulation ofP. infestans(a critical factor in late blight epidemics) and increased its virulence. We also profiled gene expression in these strains and identified potential molecular mechanisms through which PiRV-2 asserted its sporulation stimulation effect. In a survey of PiRV-2 presence in aP. infestanscollection, we found PiRV-2 in most isolates of the US-8 clonal lineage, a very successfull ineage that dominated potato fields in North America for several decades. We suggest that PiRV-2 may have contributed to its success. Our findings raise the intriguing possibility that a potentially hypervirulent virus may contribute to late blight epidemics.

Precision agriculture technology can transform farming related data into useful information, which may lead to more efficient usage of agricultural resources and increase sustainability. This paper compares precision agriculture technology with traditional practices in scheduling fungicide application so as to manage late blight disease in tomato production. The following three fungicide scheduling strategies were evaluated: a calendar-based strategy, the BlightPro Decision Support System based strategy (DSS-based strategy), and a strategy that does not involve fungicide application. The data from field trials and computer simulation experiments were used to construct distributions of the net return per acre for the calendar-based and the DSS-based strategies. These distributions were then compared using three standard approaches to ranking risky alternatives, namely: stochastic dominance, stochastic dominance with respect to a function, and stochastic efficiency with respect to a function. Assuming no yield differences between the calendar-based and the DSS-based strategies, the calendar-based strategy was preferred for highly late blight susceptible cultivars, and the DSS-based strategy was preferred for moderately susceptible and moderately resistant cultivars. Assuming no yield differences, the value of the BlightPro Decision Support System ranged from &minus, $28 to $48 per acre. With the yield improvement for the DSS-based strategy included, the DSS-based strategy was preferred for the cultivars in all of the disease-resistance categories with the value ranging from $496 to $1714 per acre.

In many parts of the world the damaging potato late blight pathogen, Phytophthora infestans, is spread as a succession of clonal lineages. The discrimination of genetic diversity within such evolving populations provides insights into the processes generating novel lineages and the pathways and drivers of pathogen evolution and dissemination at local and global scales. This knowledge, in turn, helps optimise management practices. Here we combine two key methods for dissecting mitochondrial and nuclear diversity and resolve intra and inter-lineage diversity of over 100 P. infestans isolates representative of key clonal lineages found globally. A novel set of PCR primers that amplify five target regions are provided for mitochondrial DNA sequence analysis. These five loci increased the number of mtDNA haplotypes resolved from four with the PCR RFLP method to 37 (17, 6, 8 and 4 for Ia, Ib, IIa, and IIb haplotypes, respectively, plus 2 Herb-1 haplotypes). As with the PCR RFLP method, two main lineages, I and II were defined. Group I contained 25 mtDNA haplotypes that grouped broadly according to the Ia and Ib types and resolved several sub-clades amongst the global sample. Group II comprised two distinct clusters with four haplotypes corresponding to the RFLP type IIb and eight haplotypes resolved within type IIa. The 12-plex SSR assay revealed 90 multilocus genotypes providing accurate discrimination of dominant clonal lineages and other genetically diverse isolates. Some association of genetic diversity and geographic region of contemporary isolates was observed; US and Mexican isolates formed a loose grouping, distinct from isolates from Europe, South America and other regions. Diversity within clonal lineages was observed that varied according to the age of the clone. In combination, these fine-scale nuclear and maternally inherited mitochondrial markers enabled a greater level of discrimination among isolates than previously available and provided complementary perspectives on evolutionary questions relating to the diversity, phylogeography and the origins and spread of clonal lineages of P. infestans.

A web-based DSS for potato and tomato late blight management has been developed.Disease dynamics are predicted based on weather, crop, and management information.The web-tool enables well-informed decisions regarding fungicide use.The BlightPro DSS can help users improve the efficiency of their disease management. A web-based decision support system (DSS) for potato and tomato late blight management has been developed which links several models into a system that enables prediction of disease dynamics based on weather conditions, crop information, and management tactics. Growers identify the location of their production unit of interest (latitude and longitude of field) and the system automatically obtains observed weather data from the nearest available weather station, and location-specific forecast weather data from the National Weather Service - National Digital Forecast Database. The DSS uses these weather data along with crop and management information to drive disease forecasting systems and a validated mechanistic model of the disease to generate location-specific management recommendations for fungicide application. An integrated alert system allows users to receive notification of upcoming critical thresholds via e-mail or text message. This system provides producers, consultants, researchers, and educators with a tool to obtain management recommendations, evaluate disease management scenarios, explore comparative epidemiology, or function as a teaching aid. In field and computer simulation experiments, DSS-guided schedules were influenced by prevailing weather and host resistance and resulted in schedules that improved the efficiency of fungicide use and also reduced variance in disease suppression when compared to a weekly spray schedule. In situations with unfavorable weather, the DSS recommended fewer fungicide applications with no loss of disease suppression. In situations of very favorable weather, the DSS recommended more fungicide applications but with improved disease suppression. The DSS provides an interactive system that helps users maximize the efficiency of their crop protection strategy by enabling well-informed decisions.

Phytophthora infestans has been a named pathogen for well over 150 years and yet it continues to “emerge”, with thousands of articles published each year on it and the late blight disease that it causes. This review explores five attributes of this oomycete pathogen that maintain this constant attention. First, the historical tragedy associated with this disease (Irish potato famine) causes many people to be fascinated with the pathogen. Current technology now enables investigators to answer some questions of historical significance. Second, the devastation caused by the pathogen continues to appear in surprising new locations or with surprising new intensity. Third, populations of P. infestans worldwide are in flux, with changes that have major implications to disease management. Fourth, the genomics revolution has enabled investigators to make tremendous progress in terms of understanding the molecular biology (especially the pathogenicity) of P. infestans. Fifth, there remain many compelling unanswered questions.

Summary Hemibiotrophic plant pathogens, such as the oomycete Phytophthora infestans, employ a biphasic infection strategy, initially behaving as biotrophs, where minimal symptoms are exhibited by the plant, and subsequently as necrotrophs, feeding on dead plant tissue. The regulation of this transition and the breadth of molecular mechanisms that modulate plant defences are not well understood, although effector proteins secreted by the pathogen are thought to play a key role. We examined the transcriptional dynamics of P. infestans in a compatible interaction with its host tomato (Solanum lycopersicum) at three infection stages: biotrophy; the transition from biotrophy to necrotrophy; and necrotrophy. The expression data suggest a tight temporal regulation of many pathways associated with the suppression of plant defence mechanisms and pathogenicity, including the induction of putative cytoplasmic and apoplastic effectors. Twelve of these were experimentally evaluated to determine their ability to suppress necrosis caused by the P. infestans necrosis-inducing protein PiNPP1.1 in Nicotiana benthamiana. Four effectors suppressed necrosis, suggesting that they might prolong the biotrophic phase. This study suggests that a complex regulation of effector expression modulates the outcome of the interaction.

Summary The infection of plants by hemibiotrophic pathogens involves a complex and highly regulated transition from an initial biotrophic, asymptomatic stage to a later necrotrophic state, characterized by cell death. Little is known about how this transition is regulated, and there are conflicting views regarding the significance of the plant hormones jasmonic acid (JA) and salicylic acid (SA) in the different phases of infection. To provide a broad view of the hemibiotrophic infection process from the plant perspective, we surveyed the transcriptome of tomato (Solanum lycopersicum) during a compatible interaction with the hemibiotrophic oomycete Phytophthora infestans during three infection stages: biotrophic, the transition from biotrophy to necrotrophy, and the necrotrophic phase. Nearly 10 000 genes corresponding to proteins in approximately 400 biochemical pathways showed differential transcript abundance during the three infection stages, revealing a major reorganization of plant metabolism, including major changes in source–sink relations, as well as secondary metabolites. In addition, more than 100 putative resistance genes and pattern recognition receptor genes were induced, and both JA and SA levels and associated signalling pathways showed dynamic changes during the infection time course. The biotrophic phase was characterized by the induction of many defence systems, which were either insufficient, evaded or suppressed by the pathogen.

Phytophthora infestans causes potato late blight, an important and costly disease of potato and tomato crops. Seven clonal lineages of P. infestans identified recently in the United States were tested for baseline sensitivity to six oomycete-targeted fungicides. A subset of the dominant lineages (n = 45) collected between 2004 and 2012 was tested in vitro on media amended with a range of concentrations of either azoxystrobin, cyazofamid, cymoxanil, fluopicolide, mandipropamid, or mefenoxam. Dose-response curves and values for the effective concentration at which 50% of growth was suppressed were calculated for each isolate. The US-8 and US-11 clonal lineages were insensitive to mefenoxam while the US-20, US-21, US-22, US-23, and US-24 clonal lineages were sensitive to mefenoxam. Insensitivity to azoxystrobin, cyazofamid, cymoxanil, fluopicolide, or mandipropamid was not detected within any lineage. Thus, current U.S. populations of P. infestans remained sensitive to mefenoxam during the displacement of the US-22 lineage by US-23 over the past 5 years.

The systemic fungicide mefenoxam has been important in the control of late blight disease caused by Phytophthora infestans. This phenylamide fungicide has a negative effect on the synthesis of ribosomal RNA; however, the genetic basis for inherited field resistance is still not completely clear. We recently observed that a sensitive isolate became tolerant after a single passage on mefenoxam-containing medium. Further analyses revealed that all sensitive isolates tested (in three diverse genotypes) acquired this resistance equally quickly. In contrast, isolates that were “resistant” to mefenoxam in the initial assessment (stably resistant) did not increase in resistance upon further exposure. However, there appeared to be a cost associated with acquired resistance in the initially sensitive isolates, in that isolates with acquired resistance grew more slowly on mefenoxam-free medium than did the same isolates that had never been exposed to mefenoxam. The acquired resistance of the sensitive isolates declined slightly with subsequent culturing on medium free of mefenoxam. To investigate the mechanism of acquired resistance, we employed strand-specific RNA sequencing. Many differentially expressed genes were genotype specific, but one set of genes was differentially expressed in all genotypes. Among these were several genes (a phospholipase “Pi-PLD-like-3,” two ATP-binding cassette superfamily [ABC] transporters, and a mannitol dehydrogenase) that were up-regulated and whose function might contribute to a resistance phenotype.

Oomycetes form a deep lineage of eukaryotic organisms that includes a large number of plant pathogens which threaten natural and managed ecosystems. We undertook a survey to query the community for their ranking of plant-pathogenic oomycete species based on scientific and economic importance. In total, we received 263 votes from 62 scientists in 15 countries for a total of 33 species. The Top 10 species and their ranking are: (1) Phytophthora infestans; (2, tied) Hyaloperonospora arabidopsidis; (2, tied) Phytophthora ramorum; (4) Phytophthora sojae; (5) Phytophthora capsici; (6) Plasmopara viticola; (7) Phytophthora cinnamomi; (8, tied) Phytophthora parasitica; (8, tied) Pythium ultimum; and (10) Albugo candida. This article provides an introduction to these 10 taxa and a snapshot of current research. We hope that the list will serve as a benchmark for future trends in oomycete research.

During the summers of 2012 and 2013, 39 tomato (Solanum lycopersicum) lines or varieties were evaluated for resistance to late blight in three separate field trials. In each trial, late blight was caused by field isolates of Phytophthora infestans clonal lineage US-23. Varieties with the late blight resistance genes Ph-1, Ph-2, Ph-3, and Ph-2 + Ph-3 were included, along with several heirloom varieties with grower-reported resistance and varieties with no known resistance. All six varieties with Ph-2 + Ph-3, along with NC25P, which is homozygous for Ph-3 only, showed a high level of resistance. Plum Regal F1, which is heterozygous for Ph-3 only, showed moderate resistance. Legend, the only variety with Ph-2 alone, also showed moderate resistance. Three heirloom varieties, Matt's Wild Cherry, Lemon Drop, and Mr. Stripey, showed a high level of resistance comparable with that of varieties with Ph-2 + Ph-3. New Yorker, possessing Ph-1 only, showed no resistance. Indeterminate varieties had significantly less disease than determinate varieties in two of the three trials. Overall, this study suggests that tomato varieties with both Ph-2 and Ph-3 can be used to effectively manage late blight caused by P. infestans clonal lineage US-23. Varieties possessing only Ph-2, or heterozygous for Ph-3, were better protected than those without any late blight resistance but might still require supplemental fungicide applications, while the variety that was homozygous for Ph-3 was highly resistant. Several heirloom varieties were also highly resistant, and the unknown mechanism of their resistance warrants further research. Finally, the plasticity observed in United States P. infestans populations over the past several decades necessitates continued monitoring for genetic changes within P. infestans that could lead to the breakdown of resistance reported here.

The population structure of the Phytophthora infestans populations that caused the recent 2013–14 late blight epidemic in eastern India (EI) and northeastern India (NEI) was examined. The data provide new baseline information for populations of P. infestans in India. A migrant European 13_A2 genotype was responsible for the 2013–14 epidemic, replacing the existing populations. Mutations have generated substantial sub-clonal variation with 24 multi-locus genotypes (MLGs) found, of which 19 were unique variants not yet reported elsewhere globally. Samples from West Bengal were the most diverse and grouped alongside MLGs found in Europe, the UK and from neighbouring Bangladesh but were not linked directly to most samples from south India. The pathogen population was broadly more aggressive on potato than on tomato and resistant to the fungicide metalaxyl. Pathogen population diversity was higher in regions around the international borders with Bangladesh and Nepal. Overall, the multiple shared MLGs suggested genetic contributions from UK and Europe in addition to a sub-structure based on the geographical location within India. Our data indicate the need for improved phytosanitary procedures and continuous surveillance to prevent the further introduction of aggressive lineages of P. infestans into the country.

Phytophthora infestans, a hemibiotrophic and heterothallic oomycete pathogen, attacks a wide variety of crop plants belonging to the Solanaceae family. In 2007, late blight symptoms were observed and confirmed on Physalis peruviana in fields during a late blight survey in Colombia. Subsequent to these observations, no further symptoms have been detected in the field. This prompted an interest in investigating the interaction between P. infestans and P. peruviana using histological and molecular assays, as well as characterizing defence reactions produced on P. peruviana leaves. During this interaction, callose depositions, accumulation of hydrogen peroxide, activation of pathogenesis-related proteins (β-1,3 glucanase and peroxidase), induction of the expression of genes associated with plant defences (glucanase A and carbonic anhydrase) as well as cell death, showed that P. peruviana has active defences against P. infestans strains present in Colombia.

Phytophthora infestans is a destructive plant pathogen best known for causing the disease that triggered the Irish potato famine and remains the most costly potato pathogen to manage worldwide. Identification of P. infestan's elusive center of origin is critical to understanding the mechanisms of repeated global emergence of this pathogen. There are two competing theories, placing the origin in either South America or in central Mexico, both of which are centers of diversity of Solanum host plants. To test these competing hypotheses, we conducted detailed phylogeographic and approximate Bayesian computation analyses, which are suitable approaches to unraveling complex demographic histories. Our analyses used microsatellite markers and sequences of four nuclear genes sampled from populations in the Andes, Mexico, and elsewhere. To infer the ancestral state, we included the closest known relatives Phytophthora phaseoli, Phytophthora mirabilis, and Phytophthora ipomoeae, as well as the interspecific hybrid Phytophthora andina. We did not find support for an Andean origin of P. infestans; rather, the sequence data suggest a Mexican origin. Our findings support the hypothesis that populations found in the Andes are descendants of the Mexican populations and reconcile previous findings of ancestral variation in the Andes. Although centers of origin are well documented as centers of evolution and diversity for numerous crop plants, the number of plant pathogens with a known geographic origin are limited. This work has important implications for our understanding of the coevolution of hosts and pathogens, as well as the harnessing of plant disease resistance to manage late blight.

The oomycete Phytophthora infestans, causal agent of the tomato and potato late blight, generates important economic and environmental losses worldwide. As current control strategies are becoming less effective, there is a need for studies on oomycete metabolism to help identify promising and more effective targets for chemical control. The pyrimidine pathways are attractive metabolic targets to combat tumors, virus and parasitic diseases but have not yet been studied in Phytophthora. Pyrimidines are involved in several critical cellular processes and play structural, metabolic and regulatory functions. Here, we used genomic and transcriptomic information to survey the pyrimidine metabolism during the P. infestans life cycle. After assessing the putative gene machinery for pyrimidine salvage and de novo synthesis, we inferred genealogies for each enzymatic domain in the latter pathway, which displayed a mosaic origin. The last two enzymes of the pathway, orotate phosphoribosyltransferase and orotidine-5-monophosphate decarboxylase, are fused in a multi-domain enzyme and are duplicated in some P. infestans strains. Two splice variants of the third gene (dihydroorotase) were identified, one of them encoding a premature stop codon generating a non-functional truncated protein. Relative expression profiles of pyrimidine biosynthesis genes were evaluated by qRT-PCR during infection in Solanum phureja. The third and fifth genes involved in this pathway showed high up-regulation during biotrophic stages and down-regulation during necrotrophy, whereas the uracil phosphoribosyl transferase gene involved in pyrimidine salvage showed the inverse behavior. These findings suggest the importance of de novo pyrimidine biosynthesis during the fast replicative early infection stages and highlight the dynamics of the metabolism associated with the hemibiotrophic life style of pathogen.

Over the past few years, symptoms akin to late blight disease have been reported on a variety of crop plants in South America. Despite the economic importance of these crops, the causal agents of the diseases belonging to the genusPhytophthorahave not been completely characterized. In this study, we used an integrative approach that leveraged morphological, ecological, and genetic approaches to explore cryptic speciation withinP. infestans sensu lato. We described a newPhytophthoraspecies collected in Colombia from tree tomato (Solanum betaceum), a semi-domesticated fruit. All morphological traits and population genetic analyses, using microsatellite data and a reduced representation of single nucleotide polymorphism (SNP) data, support the description of the new species,Phytophthora betaceisp. nov. We have demonstrated that ecological differences are important in the persistence ofP. infestansandP. betaceias genetically isolated units across an overlapping area in the northern Andes.

Phytophthora infestans, the causal agent of late blight disease, has been reported in the United States and Canada since the mid-nineteenth century. Due to the lack of or very limited sexual reproduction, the populations of P. infestans in the United States are primarily reproducing asexually and, thus, show a simple genetic structure. The emergence of new clonal lineages of P. infestans (US-22, US-23, and US-24) responsible for the late blight epidemics in the northeastern region of the United States in the summers of 2009 and 2010 stimulated an investigation into phenotypic traits associated with these genotypes. Mating type, differences in sensitivity to mefenoxam, differences in pathogenicity on potato and tomato, and differences in rate of germination were studied for clonal lineages US-8, US-22, US-23, and US-24. Both A1 and A2 mating types were detected. Lineages US-22, US-23, and US-24 were generally sensitive to mefenoxam while US-8 was resistant. US-8 and US-24 were primarily pathogenic on potato while US-22 and US-23 were pathogenic on both potato and tomato. Indirect germination was favored at lower temperatures (5 and 10°C) whereas direct germination, though uncommon, was favored at higher temperatures (20 and 25°C). Sporangia of US-24 released zoospores more rapidly than did sporangia of US-22 and US-23. The association of characteristic phenotypic traits with genotype enables the prediction of phenotypic traits from rapid genotypic analyses for improved disease management.

One of many uses of fluorescent proteins in plant pathology is their constitutive expression in a pathogen in order to facilitate microscopic visualization of host-pathogen interactions. However, if such transformants are to be useful, it is important that they be similar to wild type isolates in their ability to cause disease. The vegetable pathogen Phytophthora capsici was transformed to stably and constitutively express genes for either a green (pgfp) or red (tdTomato) fluorescent protein. All transformants fluoresced in all life stages, but varied in their intensity and contained one, two, or five copies of pgfp or tdTomato, as determined by Southern analysis. One transformant labeled with green fluorescent protein had reduced growth on artificial medium, produced smaller lesions on detached pepper fruit, and was reduced in virulence on pepper seedlings, compared to the wild type isolate. For these reasons, it is unsuitable for use in studies of host-pathogen interactions. Based on their intense fluorescence and similarity to the wild type isolate in growth and virulence, the other four transformants will be useful in future microscopy studies of interactions between P. capsici and its various hosts.

Induced resistance by chemicals such as acibenzolar-S-methyl -ASM (commercialized as Actigard by Syngenta Inc) mimics the biological activation of systemic acquired resistance (SAR). ASM takes the place of salicylic acid (SA) in the SAR signal pathway inducing the same molecular markers and range of resistance. The goal of our work was to understand the downstream molecular events by which ASM confers resistance to Phytophthora infestans in tomatoes. To accomplish this goal we assayed gene expression in ASM-treated plants using a microarray with more than 12,000 tomato ESTs. As many as 300 genes were responsive to ASM. Of these, 117 were detected in most of the biological replications. Basal defense associated genes as well as SAR and disease resistance genes (R-like) involved in induced resistance and effector-triggered immunity were highly expressed. We attempted to determine the phenotype of 13 of these genes by virus induced gene silencing (VIGS). These 13 genes were selected on the basis of previous implication in plant defense response and by reliability of induction by ASM. VIGS was partially successful for three of the 13 genes, but this partial silencing did not lead to a significant reduction in the effect of ASM. The ethylene pathway was also activated in response to ASM, but a tomato mutant not responsive to ethylene remained responsive to ASM. It seems most likely that the ASM effect is complex and polygenic, depending on the effect of several genes.