Search Results (17)

Phytophthora infestans is the causal agent of late blight in potato. The occurrence of P. infestans with both A1 and A2 mating types in the field may result in sexual reproduction and the generation of recombinant strains. Such strains with new combinations of traits can be highly aggressive, resistant to fungicides, and can make the disease difficult to control in the field. Metalaxyl-resistant isolates are now more prevalent in potato fields. Understanding the genetic structure and rapid identification of mating types and metalaxyl response of P. infestans in the field is a prerequisite for effective late blight disease monitoring and management. Molecular and phenotypic assays involving molecular and phenotypic markers such as mating types and metalaxyl response are typically conducted separately in the studies of the genotypic and phenotypic diversity of P. infestans. As a result, there is a pressing need to reduce the experimental workload and more efficiently assess the aggressiveness of different strains. We think that employing genetic markers to not only estimate genotypic diversity but also to identify the mating type and fungicide response using machine learning techniques can guide and speed up the decision-making process in late blight disease management, especially when the mating type and metalaxyl resistance data are not available. This technique can also be applied to determine these phenotypic traits for dead isolates. In this study, over 600 P. infestans isolates from different populations—Estonia, Pskov region, and Poland—were classified for mating types and metalaxyl response using machine learning techniques based on simple sequence repeat (SSR) markers. For both traits, random forest and the support vector machine demonstrated good accuracy of over 70%, compared to the decision tree and artificial neural network models whose accuracy was lower. There were also associations (p < 0.05) between the traits and some of the alleles detected, but machine learning prediction techniques based on multilocus SSR genotypes offered better prediction accuracy. Full article

Late blight, caused by the oomycete Phytophthora infestans, is one of the most important diseases affecting tomato and potato production worldwide. In Thailand, the disease is widespread in the north and northeast, especially in the Chiang-Mai and Tak provinces. The mating type, metalaxyl sensitivity, mitochondrial DNA (mtDNA) haplotype, RG57 fingerprinting, and microsatellite were used to characterize the P. infestans populations. The study revealed that the P. infestans of tomato isolates in Thailand are of the same lineage as those from 1994 until 2002. The clonal lineages that were found in the potato populations have changed since 1994. The changes in P. infestans isolates in the potato populations have likely been the result of the import of seed potatoes to Thailand. Furthermore, the P. infestans populations in potatoes show resistance to metalaxyl, whereas those from tomato isolates show sensitivity to fungicides. The reasons for the different responses can be attributed to (i) the use of metalaxyl, (ii) the host preferences of P. infestans, and (iii) the migration of new genotypes from infected potato seeds. Full article

The high incidence of fungal resistance to commercial fungicides and the negative effects of chemical fungicides on the environment and human health necessitate the development of novel biofungicides for the efficient management of fungal diseases. This study aims to greenly synthesize iron oxide nanoparticles (IONPs) using the aqueous extract of Laurus nobilis leaves and characterize these nanoparticles using various physicochemical techniques. The biogenic IONPs were tested against two pathogenic strains of Alternaria alternata and compared to the metalaxyl–mancozeb fungicide. The food poisoning technique was used to assess the antifungal efficacy of the greenly synthesized IONPs and the commercial metalaxyl–mancozeb fungicide against the tested pathogenic A. alternata strains. The biogenic IONPs showed a higher antifungal efficiency against the A. alternata OR236467 and A. alternata OR236468 strains at concentrations of 800 ppm compared to metalaxyl– mancozeb fungicide, with relative growth inhibition percentages of 75.89 and 60.63%, respectively. The commercial metalaxyl–mancozeb fungicide (800 ppm) showed growth inhibition percentages of 72.23 and 58.54% against the same strains. The biogenic IONPs also showed potential antioxidant activities against 1,1-diphenyl-2-picrylhydrazyl (DPPH) radicals, with DPPH inhibition percentages of 34.61% to 83.27%. In conclusion, the biogenic IONPs derived from L. nobilis leaves have the potential to be employed as biofungicides for the effective control of fungal phytopathogens, reducing reliance on harmful chemical fungicides. Full article

Mal secco, caused by Plenodomus tracheiphilus, is an economically important fungal vascular disease in citrus-growing countries of the Mediterranean basin. Preventing fungal infections usually requires a high number of copper treatments but European legislation imposes the minimization of their accumulation in soil. In our study, biological control agents (BCAs) and a plant resistance inducer (PRI), tested in four different experiments on citrus seedlings under controlled conditions, have resulted in promising strategies to control mal secco disease. Foliar (Experiment I) and soil (Experiment II) applications of two formulations of Bacillus amyloliquefaciens strain D747 (Amylo-X^® LC and Amylo-X^® WG) provided similar performances in reducing the disease amount (incidence and symptoms severity) over time compared to the untreated control, whereas copper hydroxide (Kocide Opti^®) used as standard was the most effective treatment over time. In the third experiment, Pythium oligandrum strain M1 (Polyversum^®) and Trichoderma asperellum strain ICC012 + Trichoderma gamsii strain ICC080 (Remedier^®) were able to reduce disease incidence and symptoms severity compared to the untreated control. Remedier^® provided the best performances in reducing the disease amount, whereas the Polyversum^® application was the least effective treatment over time. The effectiveness of the Trichoderma spp. formulation in reducing P. tracheiphilus infections did not significantly differ from the standard copper compound (Kocide Opti^®). Comprehensively, in the last experiment (IV), acibenzolar-S-methyl (ASM) alone and in mixture with metalaxyl-M proved as effective as B. amyloliquefaciens strain FZB24, with no dose–response relationships observed. These findings provide important insight for the integrated management of mal secco disease. Full article

Potato (Solanum tuberosum L.) and tomato (S. lycopersicum L.) are the most economically important vegetable crops in Egypt and worldwide. The winter crop in Egypt is particularly prone to late blight caused by Phytophthora infestans. A total of 152 P. infestans isolates were isolated from the 2013, 2014, 2016 and 2018 winter crops with 82 isolates from potato, 69 from tomato and one isolate from eggplant (S. melongena L.). All isolates belonged to the A1 mating type with no evidence of A2 or self-fertile strains. The majority of isolates (53%) were sensitive to metalaxyl, 32% were intermediate and 15% were resistant. Variation in aggressiveness between three P. infestans isolates EG-005 (13_A2) and EG-276 (23_A1) from potato, and EG-237 (23_A1) from eggplant was determined on tuber slices and leaflets of 10 potato cultivars. The eggplant isolate EG-237 showed higher sporulation capacity compared with the other tested isolates and was able to infect potato (Lady Rosetta cv) and tomato (Super Strain B cv). The simple sequence repeat (SSR) genotyping data showed that in contrast to our previous work (3-year period 2010–12) in which the proportion of 13_A2 lineage was 35%, all isolates belonged to the 23_A1 lineage. There was no evidence for the existence of the A2 mating type or 13_A2 lineage even in the destroyed field crops of some cultivars (Cara, Bellini and Valor) that had been reported as resistant to 23_A1. The data have been submitted into the Euroblight database to allow temporal and spatial genetic diversity to be examined in comparison with other regional P. infestans populations. The AVR2 and AVR2-like RXLR effector genes were amplified and sequenced. In the avirulent AVR2 gene, only one heterozygous SNP was detected at position 31 in the N terminus in six isolates out of eleven, whereas two heterozygous SNPs were detected at position 29 in the N-terminus and ninety-two in the C- terminus of the AVR2-like gene. This suggests that changes in the previously reported virulence profile of 23_A1 are not related to commercial cultivars carrying the R2 gene. In addition, this is the first report of P. infestans on eggplant in Egypt. Full article

Phytophthora, a blight of pigeonpea caused by Phytophthora cajani, has been significantly increasing in major pigeonpea production regions of India. Limited information on infection with this pathogen and its epidemiology, as well as a lack of adequate resistant cultivars, is hampering the management of Phytophthora blight significantly. Therefore, five fungicides, viz., metiram + dimethomorph, cymoxanil + mancozeb, famoxadone + cymoxanil, mancozeb, and metalaxyl-M + mancozeb, were evaluated against P. cajani under control conditions to control zoospore induction, as well as the infection of zoospores, at the seedling stage. The half-maximal effective concentration (EC50) of fungicides for mycelial inhibition was calculated. The lowest EC50 was recorded in metiram + dimethomorph (0.17 µg/mL), followed by the metalaxyl-M + mancozeb (2.49 µg/mL) and cymoxanil + mancozeb (8.23 µg/mL) fungicides. The formation of the sporangium and zoospores was most significantly affected by metalaxyl-M + mancozeb, followed by metiram + dimethomorph and cymoxanil + mancozeb, in terms of sporangia viability and zoospore germination and encystment. Further, under glasshouse conditions, different fungicide application methods (e.g., seed-treatment; soil-drench; foliar-spray, either singly or in combinations) were evaluated with fungicides on susceptible (ICP 7119) moderately resistant pigeonpea (ICPL 99010, ICPL 20135 and ICPL 99048) cultivars. The seed-treatment + soil-drench, soil-drench + foliar-spray, and soil-drench of fungicide application methods were found to be effective in controlling the Phytophthora blight, at p < 0.001. A combination of the seed-treatment + soil-drench and soil-drench + foliar-spray methods, using metalaxyl-M + mancozeb or metiram + dimethomorph fungicides on moderately resistant cultivars (ICPL 99010), has a synergistic effect on the ability to control the Phytophthora blight at the seedling stage. Full article

Phytophthora cactorum is considered an important plant pathogen which is causing major damage to strawberry plants worldwide. In the current study, the ability of the active ingredients of seven different fungicides, azoxystrobin, cymoxanil, dimethomorph, fenamidone, fluopicolide, metalaxyl and propamocarb, to suppress the mycelial growth, sporangial formation and zoospore release of P. cactorum isolates, was tested. The variation in resistance against various fungicides was found among the isolates. The active ingredients are also unequally efficient against different life stages of P. cactorum, which is probably associated with their different modes of action. A significant level of resistance was recorded against metalaxyl and dimethomorph; however, these were totally inefficient against the zoospore release, while azoxystrobin did not inhibit mycelial growth. The only fungicide efficient against all three P. cactorum life stages tested was fluopicolide, although the calculated resistance factor gives evidence of the rise of resistance in the majority of isolates even against this fungicide. Significant differences were found between responses to fungicides of isolates from strawberry and from other host species. Based on the Mahalanobis distances calculated in the discriminant analysis comprising all of the assays performed, the similarities among isolates were estimated. Full article

Long-term site-specific studies describing changes in the phenotypic variability of Phytophthora infestans populations allow quantitative predictions of pathogen spread and possible outbreaks of epidemics, and provide key input for regional resistance breeding programs. Late blight samples were collected from potato (Solanum tuberosum) breeding fields in Estonia during a twelve-year study period between 2001 and 2014. In total, 207 isolates were assessed for mating type and 235 isolates for metalaxyl resistance and 251 isolates for virulence factors. The frequency of mating types strongly fluctuated across the years, whereas the later period of 2010–2014 was dominated by the A2 mating. Despite fluctuations, both mating types were recorded in the same fields in most years, indicating sustained sexual reproduction of P. infestans with oospore production. Metalaxyl-resistant and intermediately resistant strains dominated in the first years of study, but with the progression of the study, metalaxyl-sensitive isolates became dominant, reaching up to 88%. Racial diversity, characterized by normalized Shannon diversity index decreased in time, varying from 1.00 in 2003 to 0.43 in 2013. The frequency of several virulence factors changed in a time-dependent manner, with R2 increasing and R6, R8, and R9 decreasing in time. Potato cultivar resistance background did not influence the frequency of P. infestans mating type, response to metalaxyl, and racial diversity. However, the diversity index decreased in time among isolates collected from resistant and susceptible cultivars, and remained at a high level in moderately resistant cultivars. These data demonstrate major time-dependent changes in racial diversity, fungicide resistance, and virulence factors in P. infestans, consistent with alterations in the control strategies and popularity of potato cultivars with different resistance. Full article

There is limited understanding of the genetic variability in Phytophthora infestans in the major potato cultivation region of north-western Russia, where potato is grown primarily by small households with limited chemical treatment of late blight. In this study, the mating type, sensitivity to metalaxyl, and genotype and population genetic diversity (based on 12 simple sequence repeat (SSR) markers) of 238 isolates of P. infestans from the Pskov region during the years 2010–2013 were characterized. The aim was to examine the population structure, phenotypic and genotypic diversity, and the prevalent reproductive mode of P. infestans, as well as the influence of the location, time, and agricultural management practices on the pathogen population. The frequency of the A2 mating was stable over the four seasons and ranged from 33 to 48% of the sampled population. Both mating types occurred simultaneously in 90% of studied fields, suggesting the presence of sexual reproduction and oospore production in P. infestans in the Pskov region. Metalaxyl-sensitive isolates prevailed in all four years (72%), however, significantly fewer sensitive isolates were found in samples from large-scale conventional fields. A total of 50 alleles were detected in the 141 P. infestans isolates analyzed for genetic diversity. Amongst the 83 SSR multilocus genotypes (MLGs) detected, 65% were unique and the number of MLGs varied between locations from 3 to 20. These results, together with the high genotypic diversity observed in all the locations and the lack of significance of linkage disequilibrium, suggest that sexual recombination is likely responsible for the unique MLGs and the high genetic diversity found in the Pskov region population, resembling those of north-eastern European populations. Full article

Late blight disease of potato and tomato, caused by Phytophthora infestans, results in serious losses to Egyptian and global potato and tomato production. To understand the structure and dynamics of the Egyptian population of P. infestans, 205 isolates were collected from potato and tomato plants during three growing seasons in 2010–2012. The characterization was achieved by mating-type assay, metalaxyl sensitivity assay, and virulence pattern. Additionally, genotyping of 85 Egyptian isolates and 15 reference UK isolates was performed using 12 highly informative microsatellite (SSR) markers and five effector (RxLR) genes. Mating-type testing showed that 58% (118 of 205) of the isolates belonged to mating type A1, 35% (71 isolates) to mating type A2, and the rest 8% (16 isolates) were self-fertile. The phenotype of metalaxyl response was represented as 45% resistant, 43% sensitive, and 12% as intermediate. Structure analysis grouped the 85 identified genotypes into two main clonal lineages. The first clonal lineage comprised 21 isolates belonging to A2 mating type and 8 self-fertile isolates. This clonal lineage was identified as Blue_13 or EU_13_A2. The second main clonal lineage comprised 55 isolates and was identified as EU_23_A1. A single isolate with a novel SSR genotype that formed a distinct genetic grouping was also identified. The effector sequencing showed good correspondence with the virulence data and highlighted differences in the presence and absence of loci as well as nucleotide polymorphism that affect gene function. This study indicated a changing population of P. infestans in Egypt and discusses the findings in the context of late blight management. Full article

Baby leaf wild rocket cropping systems feeding the high convenience salad chain are prone to a set of disease agents that require management measures compatible with the sustainability-own features of the ready-to-eat food segment. In this light, bio-based disease resistance inducers able to elicit the plant’s defense mechanism(s) against a wide-spectrum of pathogens are proposed as safe and effective remedies as alternatives to synthetic fungicides, to be, however, implemented under practical field applications. Hyperspectral-based proximal sensing was applied here to detect plant reflectance response to treatment of wild rocket beds with Trichoderma atroviride strain TA35, laminarin-based Vacciplant^®, and Saccharomyces cerevisiae strain LAS117 cell wall extract-based Romeo^®, compared to a local standard approach including synthetic fungicides (i.e., cyprodinil, fludioxonil, mandipropamid, and metalaxyl-m) and a not-treated control. Variability of the spectral information acquired in VIS–NIR–SWIR regions per treatment was explained by three principal components associated with foliar absorption of water, structural characteristics of the vegetation, and the ecophysiological plant status. Therefore, the following model-based statistical approach returned the interpretation of the inducers’ performances at field scale consistent with their putative biological effects. The study stated that compost and laminarin-based treatments were the highest crop impacting ones, resulting in enhanced water intake and in stress-related pigment adjustment, respectively. Whereas plants under the conventional chemical management proved to be in better vigor and health status than the untreated control. Full article

Thailand is the leading producer and exporter of durians worldwide. Serious diseases in durians include root rot, stem rot, and fruit rot, which are caused by Phytophthora palmivora, P. nicotianae, and Pythium cucurbitacearum, respectively. Thai farmers have applied fungicides for more than 20 years to control rot, but it remains difficult to control. Thus, the monitoring of fungicide-resistance development in pathogens is important for disease management. Pathogens were isolated from naturally infected durians between 2016 and 2017 in southern Thailand. The sequences of the internal transcribed spacer (ITS) and 5.8S regions of rDNA were used for the identification of their species. Seventeen out of twenty isolates were confirmed to be P. palmivora. All the isolates were tested for mycelium-growth sensitivity to metalaxyl, azoxystrobin, and dimethomorph. The results showed that nine isolates were resistant to metalaxyl with the 50% effective concentration (EC₅₀) higher than 100 mg L⁻¹. By contrast, all the isolates were sensitive to both azoxystrobin and dimethomorph, with EC₅₀ < 1 mg L⁻¹. Metalaxyl-resistant isolates were not controlled (−25.6% to 22.2%) by the treatment of the detached leaves of ‘Monthong’ durian with 100 mg L⁻¹ metalaxyl prior to inoculation, but all the metalaxyl-sensitive and moderately metalaxyl-resistant isolates were better controlled (33.0% to 62.6%). These results clearly indicate that metalaxyl-resistant strains are present in the populations of P. palmivora in Thailand. Full article

Metalaxyl is an effective approach to control Phytophthora palmivora infection in durian plantation. However, inappropriate metalaxyl usage may increase production cost, pathogen with fungicide resistance, and environmental toxicity. This study established and validated a simple and reproducible procedure to measure metalaxyl concentration in the durian leaf using HPLC. Linearity of the detection ranged from 1–100 µg/mL. The limits of detection (LOD) and quantification (LOQ) were 0.27 and 0.91 µg/mL, respectively. The extraction method gave recovery rates ranging from 88% to 103%. Durian seedlings were treated with 4 g/L metalaxyl either by foliar spray or soil drench. The highest metalaxyl accumulation in durian leaf was found between 6–24 h after treatment and persisted above its effective concentration at least 60 days after foliar application. The dissipation pattern fit to a first-order kinetics equation showed a half-life of 16.50 days. Soil drenching led to eight times higher metalaxyl concentrations in plants than foliar spraying and caused plant death within 15 days after application. These results suggest that foliar spraying of 4 g/L metalaxyl or soil drenching at a lower concentration every two months is sufficient in controlling P. palmivora infection in durian seedlings. Full article

Metalaxyl is one of the main fungicides used to control pepper blight caused by Phytophthora capsici. Metalaxyl resistance of P. capsici, caused by the long-term intense use of this fungicide, has become one of the most serious challenges facing pest management. To reveal the potential resistance mechanism of P. capsici to fungicide metalaxyl, a metalaxyl-resistant mutant strain SD1-9 was obtained under laboratory conditions. The pathogenicity test showed that mutant strain SD1-9 had different pathogenicity to different host plants with or without the treatment of metalaxyl compared with that of the wild type SD1. Comparative transcriptome sequencing of mutant strain SD1-9 and wild type SD1 led to the identification of 3845 differentially expressed genes, among them, 517 genes were upregulated, while 3328 genes were down-regulated in SD1-9 compared to that in the SD1. The expression levels of 10 genes were further verified by real-time RT-PCR. KEGG analysis showed that the differentially expressed genes were enriched in the peroxisome, endocytosis, alanine and tyrosine metabolism. The expression of the candidate gene XLOC_020226 during 10 life history stages was further studied, the results showed that expression level reached a maximum at the zoospores stage and basically showed a gradually increasing trend with increasing infection time in pepper leaves in SD1-9 strain, while its expression gradually increased in the SD1 strain throughout the 10 stages, indicated that XLOC_020226 may be related to the growth and pathogenicity of P. capsici. In summary, transcriptome analysis of plant pathogen P. capsici strains with different metalaxyl resistance not only provided database of the genes involved in the metalaxyl resistance of P. capsici, but also allowed us to gain novel insights into the potential resistance mechanism of P. capsici to metalaxyl in peppers. Full article

Phytophthora cinnamomi is a phytopathogen that causes extensive damage in different crops, and therefore, produces important economic losses all around the world. Chemical fungicides are a key factor for the control of this disease. However, ecological and environmental considerations, as well as the appearance of strains that are resistant to commercial fungicides, have prompted the quest for new antifungal agents which are of low ecological impact. In this work, a series of new 2-allylphenol derivatives was synthesized, and their structures were confirmed by FT-IR, NMR, and MS. Some of the synthesized compounds, more specifically nitro derivatives, exhibit strong growth inhibition of P. cinnamomi with EC₅₀ as low as 10.0 µg/mL. This level of activity is similar to that exhibited by METALAXYL MZ 58 WP, a commonly-used commercial fungicide; therefore, these compounds might be of agricultural interest due to their potential use as fungicides against P. cinnamomi. The results indicate that this activity depends on the chemical structures of the 2-allylphenol derivatives, and that it is strongly enhanced in molecules where nitro and hydroxyl groups adopt a -para configuration. These effects are discussed in terms of the electronic distribution of the aromatic ring induced by substituent groups. Full article