Host–pathogen interaction between Phytophthora infestans and Solanum nigrum, S. villosum, and S. scabrum

Potato and tomato are the two major hosts for Phytophthora infestans causing late blight. The susceptibility of leaves and whole plants of Solanum nigrum, S. villosum, and S. scabrum to infection by P. infestans was tested under laboratory conditions. Out of 39 plants representing 38 different S. nigrum accessions, 16 were highly resistant (seven accessions did not show any symptoms of infection, nine were highly resistant showing necrotic lesions in the place of infection), and 23 plants of S. nigrum were colonized by, at least, 1 of the 2 isolates of P. infestans (17 accessions were infected with two P. infestans isolates, and 6 accessions showed different reactions depending on the isolate used for inoculation). Three accessions of S. villosum, and one accession of S. scabrum were tested and did not show any symptoms of infection. The majority of S. nigrum accessions infected by P. infestans in a detached leaf assay were also infected in the whole plant assay. The reaction of field- and greenhouse-grown plants to inoculation with P. infestans in detached leaf assays was similar, but in some cases leaves from field-grown plants reacted as resistant in comparison with the leaves from greenhouse-grown plants, which were susceptible.     

Activity against plant pathogenic fungi of phomalactone isolated from Nigrospora sphaerica

Phomalactone, 5,6‐dihydro‐5‐hydroxy‐6‐prop‐2‐enyl‐2H‐pyran‐2‐one, produced by the fungus Nigrospora sphaerica, was tested in vitro against nine plant pathogenic fungi, and specifically inhibited the mycelial growth of Phytophthora infestans, with an MIC value of 2.5 mg litre^?1. Its inhibitory activities against sporangium and zoospore germination of P infestans were similar to those against Phytophthora capsici. In vivo, at 100 and 500 mg litre^?1, it reduced the development of tomato late blight caused by P infestans. © 2001 Society of Chemical Industry     

The insecticidal and anticholinesterase activities of O,O-diethyl O-4-(2,2-disubstituted vinyl)phenyl phosphates and phosphorothioates

A series of O,O-diethyl O-4-(2,2-disubstituted-vinyl)phenyl phosphorothioates and phosphates was synthesised and examined for anticholinesterase activity to housefly and bovine erythrocyte ChE, and also for their toxicity to Musca domestica and Triatoma infestans. The synthesised phosphates were potent anticholinesterases and stronger inhibitors of housefly acetylcholinesterase than bovine erythrocyte acetyl-cholinesterase. Satisfactory correlation was obtained between the anticholinesterase activity and the Hammett σ^? constants against both enzymes, but it was not significantly improved by introducing a term for the Hansch π constant. The phosphorothioates showed poor and variable toxicity to M. domestica. On T. infestans, the only compound that showed some toxicity was the di(acetyl)-substituted product [O-4-(2-acetyl-3-oxobut-1-enyl)phenyl O,O-diethyl phosphorothioate]. For comparative studies, some commercial products were also assayed on T. infestans.     

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.     

Inheritance of resistance in <Emphasis Type="Italic">Solanum nigrum</Emphasis> to <Emphasis Type="Italic">Phytophthora infestans</Emphasis>

Solanum nigrum, black nightshade, is a wild non-tuber bearing hexaploid species with a high level of resistance to Phytophthora infestans (Colon et al. 1993), the causal agent of potato late blight, the most devastating disease in potato production. However, the genetic mode of resistance in S. nigrum is still poorly understood. In the present study, two S. nigrum accessions, 984750019 (N19) and #13, resistant (R) and susceptible (S), respectively, to three different isolates of P. infestans, were sexually crossed. The various kinds of progeny including F1, F2, F3, and backcross populations (BC₁; F1 × S), as well as two populations produced by self-pollinating the R parent and S parent, were each screened for susceptibility to P. infestans isolate MP 324 using detached leaf assays. Fifty seedling plant individuals of the F1 progeny were each resistant to this specific isolate, similarly to the seedling plants resulting from self-pollination of the resistant R parent. Thirty seedling plants obtained from self-pollination of the S parent were susceptible. Among a total of 180 F2 plants, the segregation ratio between resistant and susceptible plants was approximately 3: 1. Among the 66 seedling plants of the BC₁ progeny originating from crossing an F1 plant with the susceptible S parent, there were 26 susceptible and 40 resistant plants to P. infestans. The segregation patterns obtained indicated monogenic dominant inheritance of resistance to P. infestans isolate MP 324 in S. nigrum acc. 984750019. This gene, conferring resistance to P. infestans, may be useful for the transformation of potato cultivars susceptible to late blight.     

Biological Activity of a Furanyl Anti-Juvenile Hormonal Compound on Triatomine Larvae

Applied topically to larvae of Rhodnius prolixus, Triatoma infestans, Triatoma Vitticeps, Panstrongylus herrera and Panstrongylus megistus a novel, synthetic, furan-containing anti-juvenile hormonal compound, 2-(2-ethoxyethoxy)ethyl furfuryl ether (EEFE) induced a variety of biological responses attributable to an enforced cessation of juvenile hormone secretion, including precocious metamorphosis into diminutive adultoids and retardation of molting. The highest percentage of precocious adultoids was obtained when EEFE was applied prior to feeding. In decreasing order of sensitivity to this anti-juvenile hormone (AJH) were T. infestans, R. prolixus, P. herrera, P. megistus and T. vitticeps. The ED₅₀ of EEFE in R. prolixus compared well with that of the botanically derived AJH, precocene II, but T. infestans was significantly more sensitive to the furanyl antihormone than to precocene II. The significance of these findings is discussed in terms of the complexity of expression of the anti-juvenile hormonal activity during the post-embryonic development of triatomines.     

Development of a quantitative real‐time PCR assay for Phytophthora infestans and its applicability to leaf,tuber and soil samples

A sensitive real‐time polymerase chain reaction (PCR) assay was developed for the quantification of Phytophthora infestans, the cause of foliar and tuber late blight in potato. A primer pair (PinfTQF/PinfTQR) and a fluorogenic probe (PinfTQPR) were designed to perform a quantitative assay for the detection of P. infestans in leaves, tubers and soils. The assay was shown to be specific to P. infestans and the very closely taxonomically related non‐potato pathogen species P. mirabilis, P. phaseoli and P. ipomoea, but did not detect the potato pathogens P. erythroseptica and P. nicotianae. The assay was able to reliably detect P. infestans DNA at 100 fg per reaction and was effective in quantifying P. infestans in infected leaf tissue from 24 h after inoculation and also in infected symptomless tubers and diseased tubers. Attempts to detect oospores of P. infestans in naturally and artificially infested soil samples are described and compared with baiting tests and previous literature. It was not possible to detect oospores in soil samples due to problems with DNA extraction from the oospores themselves. However, the assay was shown to detect even very low levels of asexual inoculum (sporangia and mycelium) in soil. This work assembles all the necessary features of a quantitative P. infestans assay, which have previously been somewhat disparate: the sensitivity, specificity and quantitation are fully validated, the assay is shown to work in common applications in leaf and tuber tissue and the problems with P. infestans oospore detection are explored and tested experimentally.     

Phosphite compounds reduce disease severity in potato seed tubers and foliage

Phosphites (Phi) are alkali metal salts of phosphorous acid, with the ability to protect plants against different pathogens. In this research, the effect of Phi applied to potato plants on severity of three important potato diseases in Argentina was assessed. Seed tubers and foliage of potato cvs Shepody and Kennebec were treated with Phi to assess effects on resistance against Phytophthora infestans, Fusarium solani and Rhizoctonia solani. Protection resulting from Phi treatment in seed tubers was high against P. infestans, intermediate against F. solani, and low against R. solani. In addition, seed tubers treated with calcium or potassium phosphites (CaPhi and KPhi, respectively) at 1% of commercial product emerged earlier than untreated ones. When Phi were foliarly applied two or four times at different doses, high levels of protection against P. infestans were achieved in both cultivars. Higher protection was observed in Kennebec when CaPhi was applied, while in Shepody this was true for KPhi. Expression of β-1,3-glucanases was induced at different times after treatment but no correlation between β-1,3-glucanases expression and foliar protection level was found. On the other hand, Phi positive protection effects did not produce negative effects in plant growth. Leaves from CaPhi-treated plants showed a darker green colour than leaves from control plants; also an increase in Rubisco protein and a delay in crop senescence was observed.     

Involvement of Iron and Ferritin in the Potato–Phytophthora infestans Interaction

This work shows that the infection of potato (Solanum tuberosum) detached leaves by the late blight pathogen Phytophthora infestans, was drastically reduced by adding deferoxamine, an exogenous iron chelator. Reactive oxygen species in leaves inoculated with P. infestans were also reduced after adding deferoxamine. A leaf ferritin cDNA fragment was obtained by PCR and used as probe for screening a tuber cDNA library. A cDNA (named StF1) encoding the iron-storing potato ferritin was cloned. StF1 is 915 bp in length and has an open reading frame of 230 amino acids that contains the information for the mature 28 kDa subunit of potato ferritin. StF1 was used as probe in northern blot hybridizations to analyze expression of the ferritin gene. In leaves, ferritin mRNA accumulated in response to pathogen attack. In tubers, ferritin mRNA increased upon treatment with the elicitor eicosapentaenoic acid. These results suggest that iron plays a role in the potato-P. infestans interaction.     

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.     

Inhibition by aluminum of mycelial growth and of sporangial production and germination inPhytophthora infestans

Mycelial growth on clarified V8 agar of the potato late blight pathogenPhytophthora infestans was inhibited when either aluminum chloride (AlCl₃, 6 H₂O) or aluminum sulfate (Al₂(SO₄)₃, 18 H₂O) was added to the culture medium at concentrations of 2.5–100 mg.l^–1 Al³⁺. Toxicity of Al³⁺ varied among the fiveP. infestans isolates tested, but toxicity of sulfate and chloride salts was similar for a given isolate. Overall sporangial production was affected in all five isolates by both Al³⁺ forms. Al³⁺ also decreased sporangial germination at concentrations equal to or greater than 10 mg.l^–1 in two isolates. These data support the hypothesis of aluminum toxicity as a major factor in soil suppressiveness toP. infestans.     

Dominance of a single clonal lineage in the Phytophthora infestans population from northern Shaanxi,China revealed by genetic and phenotypic diversity analysis

Late blight caused by Phytophthora infestans is the most serious disease of potato worldwide. To understand the P. infestans population structure in northern Shaanxi, an emerging potato production region in China, 125 single‐lesion isolates were randomly collected from farmers' fields in 2009 and characterized phenotypically and genotypically. A mating type assay showed that 94 isolates were A1 mating type. Virulence determination of selected isolates on a set of differential potato lines containing R1 to R11, respectively, showed the presence of two pathotypes, of which the pathotype lacking avirulence genes Avr3, Avr4 and Avr10 was dominant. Isolates lacking all avirulence factors Avr1 to Avr11 were detected but at lower frequency (13·6%). Analysis for mtDNA haplotype showed all 61 examined isolates were IIa. A total of seven multilocus genotypes were distinguished among 125 isolates, as determined with seven polymorphic microsatellite markers. The genotype SG‐1 was dominant in the population with a frequency of 75·2% and was present throughout the region. Analysis of the phenotypic and genotypic structures of P. infestans populations indicated strict clonal reproduction of the pathogen and suggested that sexual reproduction probably does not occur. Potential implications for disease management are discussed.     

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.     

BABA effects on the behaviour of potato cultivars infected by <Emphasis Type="Italic">Phytophthora infestans</Emphasis> and <Emphasis Type="Italic">Fusarium solani</Emphasis>

Since most plants possess resistance mechanisms which can be induced upon pre-treatment with a variety of chemical compounds, the use of β-aminobutyric acid (BABA) as a defence inducer without reported toxic effect on the environment was studied. The aim of this work was to analyse the effectiveness of BABA to induce resistance against Phytophthora infestans and Fusarium solani in potato cultivars differing in their level of resistance to late blight. The behaviour of some components of biochemical mechanisms by which BABA increases resistance against P. infestans, as well as the effect of BABA on the activity of a potential pathogenic factor of F. solani, were studied. Plants with four applications of BABA throughout the crop cycle produced tubers more resistant to P. infestans and F. solani than non-treated plants. In addition, tuber slices from treated plants, inoculated with P. infestans, showed an increase in phenol and phytoalexin content. The aspartyl protease StAP1 accumulation was also higher in tubers obtained from treated plants and inoculated with P. infestans. This result was observed only in the more resistant potato cv. Pampeana, early after infection. In the potato–F. solani interaction, infected tubers coming from BABA-treated plants showed minor fungal proteolytic activity than infected, non-treated ones. For potato cvs Pampeana and Bintje, the BABA treatment improved the yield of harvested tubers. The number of tubers per plant and total weight of harvested tubers was greater for those obtained from treated plants with two early or four applications of BABA. The results show that the BABA treatment increases the resistance of potatoes but the degree of increase depends on the original level of resistance present in each cultivar.     

Control of potato late blight using controlled-release granules containing ofurace

In an attempt to reduce or eliminate the need for spraying to control potato late blight caused by Phytophthora infestans, investigations were made on the use of controlled-release granules containing the systemic fungicide ofurace. The granules when mixed with sand and buried in soil released the fungitoxicant for about 100 days. When the granules were applied in furrows at planting, protection of potato plants (cv. Maris Bard), assessed by inoculation of detached leaflets with P. infestans, lasted for between 85 and 100 days. Application of the granules when the plants were earthed up 48 days after planting did not result in better late-season protection, possibly due to poorer uptake of the fungitoxicant by the plants at this time. Chemical analysis of leaves from plants that had received in-furrow treatment indicated that ofurace at 0.2–0.5 μg g^?1 fresh weight was needed to confer protection from late blight.     

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.     

Sensitivity profiles of Mycosphaerella graminicola and Phytophthora infestans populations to different classes of fungicides

Prior to the use of fungicides, the baseline sensitivity of individuals in a pathogen population may already differ by a factor of 10 to 100 between the least and the most sensitive isolates. In Mycosphaerella graminicola populations, this factor, measured in vitro, was 5 to 20 for both the strobilurin analogue azoxystrobin (baseline) and the triazole cyproconazole which has been in use for several years. In Phytophthora infestans populations, this factor, measured in a leaf disc assay, was about 100 for azoxystrobin (baseline), up to 1000 for the cyanoacetamide cymoxanil and >10000 for the phenylamide oxadixyl; both of the latter have been used for many years. In M. graminicola, cross-sensitivity was present between all azole fungicides for the majority of the isolates, whereas no correlation was found between triazoles and azoxystrobin. Despite the existence of cross-sensitivity between azoles, ‘box-and-whiskers’ plots revealed large variations in the sensitivity profiles of some triazoles; isolates resistant to triazoles have not been detected in M. graminicola populations. In P. infestans populations, the proportion of the phenylamide-resistant sub-population increased during the season more rapidly in treated than in untreated fields, but it was low at the beginning of the next season in all fields. During disease epidemics, the fitness of phenylamide-resistant P. infestans isolates, as characterised by lesion size, was higher than that of the sensitive isolates, but after the overwintering period, the recovery of resistant isolates was apparently lower. The presence of both A1 and A2 mating types of P. infestans in European populations, although at different frequencies, allows sexual recombination and increased genetic diversity, affecting sensitivity and fitness. Such mixed populations can still be adequately controlled by using sound anti-resistance strategies. ©1997 SCI     

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.