Natalia Statsyuk | All-Russian research Institute of Phytopathology (original) (raw)

Papers by Natalia Statsyuk

Research paper thumbnail of Rapid and simple detection of two potato cyst nematode species by real-time multiplex PCR using preserved microarray-based test systems

Accurate diagnostics of plant pathogens is very important to prevent the spread of infection and ... more Accurate diagnostics of plant pathogens is very important to prevent the spread of infection and significant yield losses. This paper presents an easy, rapid and inexpensive system for the complex real-time multiplex PCR diagnostics of two potato cyst nematode species, Globodera rostochiensis and G. pallida, using disposable long-stored stationary PCR microarrays. Laboratory trials have demonstrated an excellent diagnostic efficiency and sensitivity of the developed test systems with the DNA detection limit equal to 1 and 10 pg for G. rostochiensis and G. pallida, respectively. The specificity of the developed primers was successfully confirmed using DNA samples of related nematode species and host plant. A small reaction volume (1.2 μl) and a special technology of lyophilisation and stabilisation of PCR reagents within microreactors significantly reduce the total time of amplification (~30 min) and the number of required manipulations. Ready-to-use microarrays with flexible architecture can be stored up to 6 months at room temperature without any loss of their detection efficiency. The developed test system has a good potential as a rapid field diagnostic method, especially in the case of a low pathogen concentration in plant tissues or soil.

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Research paper thumbnail of Phytophthora Infestans Populations from the European Part of Russia: Genotypic Structure and Metalaxyl Resistance

Journal of Plant Pathology, 2015

Populations of the potato and tomato late blight pathogen from different regions of the European ... more Populations of the potato and tomato late blight pathogen from different regions of the European part of Russia were studied for their genotypic structure and metalaxyl resistance. The majority of the studied populations showed a high genotypic diversity, i.e. strains of A1 and A2 mating types, Ia and IIa mitochondrial DNA haplotypes, and different genotypes at Pep1 and Pep2 loci were identified. The highest genotypic diversity was registered among P. infestans populations from the North Caucasus, Mariy El Republic, and the Moscow region. The obtained data confirm a high probability of crossing and hybridization within the most part of the field populations of P. infestans from the European part of Russia. Most of the studied populations included strains with high and low metalaxyl resistance, the incidence of resistant strains depending on the use of metalaxyl-containing fungicides.

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Research paper thumbnail of 10.7251/AGSY1303607S Changes in Phenotypic Characteristics of the Moscow Phytopht Hora Infestans Population in the Period of 2000 -2011

A long-term monitoring of the Moscow population of Phytophthora infestans (Mont.) De Bary, a caus... more A long-term monitoring of the Moscow population of Phytophthora infestans (Mont.) De Bary, a causal agent of the late blight disease of potato, was performed in the period from 2000 to 2011. A total of 1097 isolates has been assessed for changes in phenotyp ic characteristics such as the virulence pattern, mating type and metalaxyl resistance. A trend toward an increase in the percentage of the A2 mating type was observed in the first half of the period surveyed. During the whole period, metalaxyl-sensitive isolates remained d ominant in the population. In recent years the frequency of the virulence gene 2 began to sharply decrease, whereas the frequency of the gene 10 increased. Among rare virulence genes (genes 5, 6 and 9), the gene 9, wh ich has not been revealed in the Moscow Ph. infestans population before 2000, has been stably observed since 2006. Thus, the current Ph. infestans population of the Moscow region includes all 11 virulence genes. During the whole period of the stu...

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Research paper thumbnail of Virulence of Alternaria strains toward potato and tomato cultivars

SUMMARY Alternaria alternata is one of the causal agents of the early blight, a dangerous disease... more SUMMARY Alternaria alternata is one of the causal agents of the early blight, a dangerous disease of potato and tomato, which is common for almost all regions, where these crops are grown. In this study the virulence and aggressiveness of A. alternata isolates, obtained from the leaves and tubers of potato and leaves and fruits of tomato, has been studied on 13 potato cultivars of different maturity groups and 5 large-fruited tomato cultivars. In the case of two isolates, obtained from tomato leaves and significantly differing in their virulence, the activity of subtilisin- and tripsin- like serine proteases has been also analyzed. The performed study has revealed intraspecific differences in the virulence and aggressiveness of A. alternata toward the leaves of different potato and tomato cultivars. Some isolates successfully infected cultivars, highly resistant toward other isolates that probably evidences some potato and tomato cultivars have genes of specific resistance to A. alt...

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Research paper thumbnail of New bacitracin-resistant nisin-producing strain of Lactococcus lactis and its physiological characterization

AIMS Microbiology

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Research paper thumbnail of Assessment of the Sensitivity of Some Plant Pathogenic Fungi to 6-Demethylmevinolin, a Putative Natural Sensitizer Able to Help Overcoming the Fungicide Resistance of Plant Pathogens

Antibiotics

Agricultural fungicides contaminate the environment and promote the spread of fungicide-resistant... more Agricultural fungicides contaminate the environment and promote the spread of fungicide-resistant strains of pathogenic fungi. The enhancement of pathogen sensitivity to these pesticides using chemosensitizers allows the reducing of fungicide dosages without a decrease in their efficiency. Using Petri plate and microplate bioassays, 6-demethylmevinolin (6-DMM), a putative sensitizer of a microbial origin, was shown to affect both colony growth and conidial germination of Alternaria solani, A. alternata, Parastagonospora nodorum, Rhizoctonia solani, and four Fusarium species (F. avenaceum, F. culmorum, F. oxysporum, F. graminearum) forming a wheat root rot complex together with B. sorokiniana. Non- or marginally toxic 6-DMM concentrations suitable for sensitizing effect were determined by the probit analysis. The range of determined concentrations confirmed a possibility of using 6-DMM as a putative sensitizer for the whole complex of root rot agents, other cereal pathogens (A. alter...

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Research paper thumbnail of Evaluation of eliciting activity of peptidil prolyl cys/trans isomerase from Pseudonomas fluorescens encapsulated in sodium alginate regarding plant resistance to viral and fungal pahogens

AIMS Microbiology

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Research paper thumbnail of Development and testing of a weather-based model to determine potential yield losses caused by potato late blight and optimize fungicide application

Frontiers of Agricultural Science and Engineering

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Research paper thumbnail of Co-application of Difenoconazole with Thymol Results in Suppression of a Parastagonospora Nodorum Mutant Strain Resistant to this Triazole

KnE Life Sciences

Results of in vitro study of thymol, a natural chemosensitizer, as a potential agent for overcomi... more Results of in vitro study of thymol, a natural chemosensitizer, as a potential agent for overcoming of difenoconazole resistance of Parastagonospora nodorum causing glume and leaf blotch of wheat are first reported. The level of difenoconazole resistance of a natural mutant PNm1 strain with low sensitivity to the Dividend fungicide (a.i. difenoconazole) was determined by the cultivation of this isolate on potato dextrose agar in the presence of the fungicide at sub-lethal and lethal (in relation to the initial fungicide-sensitive strain) concentrations. A principal possibility of the thymol use to overcome resistance of P. nodorum to DMI (demethylation inhibitors) fungicides is shown. Co-application of this compound with Dividend SC, 3 % resulted in a significant reduction of resistance of the mutant strain and enhancement of its sensitivity to difenoconazole up to the level corresponding to the initial non-resistant isolate.

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Research paper thumbnail of Minimization of Fungicidal Applications Against Potato Late Blight in the North Caucasian Region: Use of the “Agrodozor” System

KnE Life Sciences

Late blight is considered to be the most devastating potato disease, which control requires appli... more Late blight is considered to be the most devastating potato disease, which control requires application of fungicides able to significantly contaminate the environment and accumulate in agricultural products. Pesticide load on potato fields can be reduced via optimization of the scheme of protective treatments. Such optimization can be performed using a mathematical simulator describing the dependence of late blight-caused yield losses on the weather data and an “Agrodozor” decision support system (DSS). The performed analysis of a situation in the regions of the North Caucasus allowed us to determine three zones differing in the disease harmfulness and characterized by potential yield losses at the level of <10, 10-20, and >20 % and the probability of disease appearance in field during 30, 77, and 90 % of seasons, respectively. Using the “Agrodozor” DSS, we calculated the optimum dates of fungicidal treatments for these zones and showed that, comparing to the commonly used ro...

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Research paper thumbnail of Potato Pathogens in Russia’s Regions: An Instrumental Survey with the Use of Real-Time PCR/RT-PCR in Matrix Format

Pathogens

Viral and bacterial diseases of potato cause significant yield loss worldwide. The current data o... more Viral and bacterial diseases of potato cause significant yield loss worldwide. The current data on the occurrence of these diseases in Russia do not provide comprehensive understanding of the phytosanitary situation. Diagnostic systems based on disposable stationary open qPCR micromatrices intended for the detection of eight viral and seven bacterial/oomycetal potato diseases have been used for wide-scale screening of target pathogens to estimate their occurrence in 11 regions of Russia and to assess suitability of the technology for high-throughput diagnostics under conditions of field laboratories. Analysis of 1025 leaf and 725 tuber samples confirmed the earlier reported data on the dominance of potato viruses Y, S, and M in most regions of European Russia, as well as relatively high incidences of Clavibacter michiganensis subsp. sepedonicus, Pectobacterium atrosepticum, and P. carotovorum subsp. carotovorum, and provided detailed information on the phytosanitary status of select...

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Research paper thumbnail of Preserved Microarrays for Simultaneous Detection and Identification of Six Fungal Potato Pathogens with the Use of Real-Time PCR in Matrix Format

Biosensors

Fungal diseases of plants are of great economic importance causing 70–80% of crop losses associat... more Fungal diseases of plants are of great economic importance causing 70–80% of crop losses associated with microbial plant pathogens. Advanced on-site disease diagnostics is very important to maximize crop productivity. In this study, diagnostic systems have been developed for simultaneous detection and identification of six fungal pathogens using 48-well microarrays (micromatrices) for qPCR. All oligonucleotide sets were tested for their specificity using 59 strains of target and non-target species. Detection limit of the developed test systems varied from 0.6 to 43.5 pg of DNA depending on target species with reproducibility within 0.3−0.7% (standard deviation). Diagnostic efficiency of test systems with stabilized and freeze-dried PCR master-mixes did not significantly differ from that of freshly prepared microarrays, though detection limit increased. Validation of test systems on 30 field samples of potato plants showed perfect correspondence with the results of morphological iden...

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Research paper thumbnail of Development of a real-time assay for detection of Synchytrium endobioticum, a quarantine pathogen of potato

Potato wart disease caused by a quarantine pathogen Synchytrium endobioticum may cause up to 50-1... more Potato wart disease caused by a quarantine pathogen Synchytrium endobioticum may cause up to 50-100% of yield losses; moreover, contaminated fields may be scheduled for 20 years. To provide an appropriate and timely control of this pathogen, reliable, simple, and robust diagnostic tools are needed. In this study we developed a test system for the real-time PCR detection and identification of S. endobioticum suitable for the use in qPCR micromatrices providing multiplex simultaneous detection of a range of pathogens. The designed primers and probe successfully tested for specificity using 10 samples of non-target fungal pathogens of potato. Detection limit of the test system was 0.1 pg of DNA per reaction. Reproducibility assay showed the standard error for Ct value did not exceed 4.5%. The further validation of the test system using culture samples of S. endobioticum of different pathotypes and infected field samples of potato should be done, and the efficient protocol for the pathogen detection in soil samples should be developed in future studies.Potato wart disease caused by a quarantine pathogen Synchytrium endobioticum may cause up to 50-100% of yield losses; moreover, contaminated fields may be scheduled for 20 years. To provide an appropriate and timely control of this pathogen, reliable, simple, and robust diagnostic tools are needed. In this study we developed a test system for the real-time PCR detection and identification of S. endobioticum suitable for the use in qPCR micromatrices providing multiplex simultaneous detection of a range of pathogens. The designed primers and probe successfully tested for specificity using 10 samples of non-target fungal pathogens of potato. Detection limit of the test system was 0.1 pg of DNA per reaction. Reproducibility assay showed the standard error for Ct value did not exceed 4.5%. The further validation of the test system using culture samples of S. endobioticum of different pathotypes and infected field samples of potato should be done, and the efficient protocol for the pathogen detection in soil s...

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Research paper thumbnail of Sensitization of plant pathogenic fungi to the tebuconazole-based commercial fungicide using some analogues of natural amino acids

Commercial fungicides provide an efficient control of fungal diseases of crops. At the same time,... more Commercial fungicides provide an efficient control of fungal diseases of crops. At the same time, plant pathogenic fungi can develop resistance to fungicides that results in epiphytoties and significant yield losses. The most common response of potato growers is more extensive use of fungicides associated with environmental pollution and a range of various health risks. Therefore, the search for approach preventing the development of pest resistance to fungicides is of great practical interest. A potentially new strategy to overcome or minimize this problem is an enhancement of pathogen sensitivity to fungicides using various chemical compounds. In this study, we examined several putative sensitizers chosen among structural analogues of some natural amino acids known for their inhibiting effect on the polyketide biosynthetic pathway, since polyketide mycotoxins play an important role in a pathogenicity of fungi. A possible synergism between the action of these compounds and a commercial fungicide Folicur on the growth of Phoma glomerata and Fusarium culmorum colonies was evaluated using the Limpel’s criterion. For all four compounds tested, a significant chemosensitizing effect was revealed for both non-fungicidal and sub-fungicidal concentrations that confirmed the initial hypothesis. The further screening of other amino acid analogues for their chemosensitizing activity is planned, as well as the study of the mechanism of their sensitizing action. Revealing of a compound providing significant chemosensitizing effect and characterized by a relatively low cost of synthesis would provide a low-cost solution for reduction of fungicidal treatments, while still providing sufficient protection even against fungicide-resistant strains of plant pathogens.Commercial fungicides provide an efficient control of fungal diseases of crops. At the same time, plant pathogenic fungi can develop resistance to fungicides that results in epiphytoties and significant yield losses. The most common response of potato growers is more extensive use of fungicides associated with environmental pollution and a range of various health risks. Therefore, the search for approach preventing the development of pest resistance to fungicides is of great practical interest. A potentially new strategy to overcome or minimize this problem is an enhancement of pathogen sensitivity to fungicides using various chemical compounds. In this study, we examined several putative sensitizers chosen among structural analogues of some natural amino acids known for their inhibiting effect on the polyketide biosynthetic pathway, since polyketide mycotoxins play an important role in a pathogenicity of fungi. A possible synergism between the action of these compounds and a commercial fungicide Folicur ...

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Research paper thumbnail of Matrix approach to the simultaneous detection of multiple potato pathogens by real-time PCR

Journal of Applied Microbiology

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Research paper thumbnail of SOME NATURAL AND SYNTHETIC COMPOUNDS INHIBITING THE BIOSYNTHESIS OF AFLATOXIN B1 AND MELANIN IN Aspergillus flavus

Sel'skokhozyaistvennaya Biologiya

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Research paper thumbnail of A new highly productive Propionibacterium acidipropionici FL-48 strain with increased resistance to propionic acid and the scaling up of its production for industrial bioreactors

Biosystems Diversity

Propionic acid bacteria, including Propionibacterium acidipropionici, are widely used in the chem... more Propionic acid bacteria, including Propionibacterium acidipropionici, are widely used in the chemical industry to produce propionic acid and also for food and feed preservation. However, the efficiency of the industrial production of these bacteria is limited by their sensitivity to high concentrations of propionic acid excreted into the cultivation medium. Therefore, the development of new biotechnological processes and strains able to overcome this limitation and to improve the profitability of the microbiological production remains a relevant problem. A new P. acidipropionici FL-48 strain characterized by an increased resistance to 10 g/L of propionic acid (the number of viable cells after 24-h cultivation reached 1.05 × 106) was developed by a two-step induced mutagenesis using UV and diethyl sulphate from the P. acidipropionici VKPM B-5723 strain. The mutant strain exceeded the parental strain in the biomass accumulation rate and the amount of produced propionic and acetic aci...

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Research paper thumbnail of Effect of Various Compounds Blocking the Colony Pigmentation on the Aflatoxin B1 Production by Aspergillus flavus

Toxins, 2016

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Research paper thumbnail of Effect of a Pre-Planting Treatment of Seed Tubers with Low-Frequency Pulse Electric Field on the Growth of Potato Plants of Different Varieties

Sel'skokhozyaistvennaya Biologiya, 2016

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Research paper thumbnail of Characterization of Russian Phytophthora infestans populations: DNA fingerprinting and SSR analysis

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Research paper thumbnail of Rapid and simple detection of two potato cyst nematode species by real-time multiplex PCR using preserved microarray-based test systems

Accurate diagnostics of plant pathogens is very important to prevent the spread of infection and ... more Accurate diagnostics of plant pathogens is very important to prevent the spread of infection and significant yield losses. This paper presents an easy, rapid and inexpensive system for the complex real-time multiplex PCR diagnostics of two potato cyst nematode species, Globodera rostochiensis and G. pallida, using disposable long-stored stationary PCR microarrays. Laboratory trials have demonstrated an excellent diagnostic efficiency and sensitivity of the developed test systems with the DNA detection limit equal to 1 and 10 pg for G. rostochiensis and G. pallida, respectively. The specificity of the developed primers was successfully confirmed using DNA samples of related nematode species and host plant. A small reaction volume (1.2 μl) and a special technology of lyophilisation and stabilisation of PCR reagents within microreactors significantly reduce the total time of amplification (~30 min) and the number of required manipulations. Ready-to-use microarrays with flexible architecture can be stored up to 6 months at room temperature without any loss of their detection efficiency. The developed test system has a good potential as a rapid field diagnostic method, especially in the case of a low pathogen concentration in plant tissues or soil.

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Research paper thumbnail of Phytophthora Infestans Populations from the European Part of Russia: Genotypic Structure and Metalaxyl Resistance

Journal of Plant Pathology, 2015

Populations of the potato and tomato late blight pathogen from different regions of the European ... more Populations of the potato and tomato late blight pathogen from different regions of the European part of Russia were studied for their genotypic structure and metalaxyl resistance. The majority of the studied populations showed a high genotypic diversity, i.e. strains of A1 and A2 mating types, Ia and IIa mitochondrial DNA haplotypes, and different genotypes at Pep1 and Pep2 loci were identified. The highest genotypic diversity was registered among P. infestans populations from the North Caucasus, Mariy El Republic, and the Moscow region. The obtained data confirm a high probability of crossing and hybridization within the most part of the field populations of P. infestans from the European part of Russia. Most of the studied populations included strains with high and low metalaxyl resistance, the incidence of resistant strains depending on the use of metalaxyl-containing fungicides.

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Research paper thumbnail of 10.7251/AGSY1303607S Changes in Phenotypic Characteristics of the Moscow Phytopht Hora Infestans Population in the Period of 2000 -2011

A long-term monitoring of the Moscow population of Phytophthora infestans (Mont.) De Bary, a caus... more A long-term monitoring of the Moscow population of Phytophthora infestans (Mont.) De Bary, a causal agent of the late blight disease of potato, was performed in the period from 2000 to 2011. A total of 1097 isolates has been assessed for changes in phenotyp ic characteristics such as the virulence pattern, mating type and metalaxyl resistance. A trend toward an increase in the percentage of the A2 mating type was observed in the first half of the period surveyed. During the whole period, metalaxyl-sensitive isolates remained d ominant in the population. In recent years the frequency of the virulence gene 2 began to sharply decrease, whereas the frequency of the gene 10 increased. Among rare virulence genes (genes 5, 6 and 9), the gene 9, wh ich has not been revealed in the Moscow Ph. infestans population before 2000, has been stably observed since 2006. Thus, the current Ph. infestans population of the Moscow region includes all 11 virulence genes. During the whole period of the stu...

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Research paper thumbnail of Virulence of Alternaria strains toward potato and tomato cultivars

SUMMARY Alternaria alternata is one of the causal agents of the early blight, a dangerous disease... more SUMMARY Alternaria alternata is one of the causal agents of the early blight, a dangerous disease of potato and tomato, which is common for almost all regions, where these crops are grown. In this study the virulence and aggressiveness of A. alternata isolates, obtained from the leaves and tubers of potato and leaves and fruits of tomato, has been studied on 13 potato cultivars of different maturity groups and 5 large-fruited tomato cultivars. In the case of two isolates, obtained from tomato leaves and significantly differing in their virulence, the activity of subtilisin- and tripsin- like serine proteases has been also analyzed. The performed study has revealed intraspecific differences in the virulence and aggressiveness of A. alternata toward the leaves of different potato and tomato cultivars. Some isolates successfully infected cultivars, highly resistant toward other isolates that probably evidences some potato and tomato cultivars have genes of specific resistance to A. alt...

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Research paper thumbnail of New bacitracin-resistant nisin-producing strain of Lactococcus lactis and its physiological characterization

AIMS Microbiology

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Research paper thumbnail of Assessment of the Sensitivity of Some Plant Pathogenic Fungi to 6-Demethylmevinolin, a Putative Natural Sensitizer Able to Help Overcoming the Fungicide Resistance of Plant Pathogens

Antibiotics

Agricultural fungicides contaminate the environment and promote the spread of fungicide-resistant... more Agricultural fungicides contaminate the environment and promote the spread of fungicide-resistant strains of pathogenic fungi. The enhancement of pathogen sensitivity to these pesticides using chemosensitizers allows the reducing of fungicide dosages without a decrease in their efficiency. Using Petri plate and microplate bioassays, 6-demethylmevinolin (6-DMM), a putative sensitizer of a microbial origin, was shown to affect both colony growth and conidial germination of Alternaria solani, A. alternata, Parastagonospora nodorum, Rhizoctonia solani, and four Fusarium species (F. avenaceum, F. culmorum, F. oxysporum, F. graminearum) forming a wheat root rot complex together with B. sorokiniana. Non- or marginally toxic 6-DMM concentrations suitable for sensitizing effect were determined by the probit analysis. The range of determined concentrations confirmed a possibility of using 6-DMM as a putative sensitizer for the whole complex of root rot agents, other cereal pathogens (A. alter...

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Research paper thumbnail of Evaluation of eliciting activity of peptidil prolyl cys/trans isomerase from Pseudonomas fluorescens encapsulated in sodium alginate regarding plant resistance to viral and fungal pahogens

AIMS Microbiology

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Research paper thumbnail of Development and testing of a weather-based model to determine potential yield losses caused by potato late blight and optimize fungicide application

Frontiers of Agricultural Science and Engineering

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Research paper thumbnail of Co-application of Difenoconazole with Thymol Results in Suppression of a Parastagonospora Nodorum Mutant Strain Resistant to this Triazole

KnE Life Sciences

Results of in vitro study of thymol, a natural chemosensitizer, as a potential agent for overcomi... more Results of in vitro study of thymol, a natural chemosensitizer, as a potential agent for overcoming of difenoconazole resistance of Parastagonospora nodorum causing glume and leaf blotch of wheat are first reported. The level of difenoconazole resistance of a natural mutant PNm1 strain with low sensitivity to the Dividend fungicide (a.i. difenoconazole) was determined by the cultivation of this isolate on potato dextrose agar in the presence of the fungicide at sub-lethal and lethal (in relation to the initial fungicide-sensitive strain) concentrations. A principal possibility of the thymol use to overcome resistance of P. nodorum to DMI (demethylation inhibitors) fungicides is shown. Co-application of this compound with Dividend SC, 3 % resulted in a significant reduction of resistance of the mutant strain and enhancement of its sensitivity to difenoconazole up to the level corresponding to the initial non-resistant isolate.

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Research paper thumbnail of Minimization of Fungicidal Applications Against Potato Late Blight in the North Caucasian Region: Use of the “Agrodozor” System

KnE Life Sciences

Late blight is considered to be the most devastating potato disease, which control requires appli... more Late blight is considered to be the most devastating potato disease, which control requires application of fungicides able to significantly contaminate the environment and accumulate in agricultural products. Pesticide load on potato fields can be reduced via optimization of the scheme of protective treatments. Such optimization can be performed using a mathematical simulator describing the dependence of late blight-caused yield losses on the weather data and an “Agrodozor” decision support system (DSS). The performed analysis of a situation in the regions of the North Caucasus allowed us to determine three zones differing in the disease harmfulness and characterized by potential yield losses at the level of <10, 10-20, and >20 % and the probability of disease appearance in field during 30, 77, and 90 % of seasons, respectively. Using the “Agrodozor” DSS, we calculated the optimum dates of fungicidal treatments for these zones and showed that, comparing to the commonly used ro...

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Research paper thumbnail of Potato Pathogens in Russia’s Regions: An Instrumental Survey with the Use of Real-Time PCR/RT-PCR in Matrix Format

Pathogens

Viral and bacterial diseases of potato cause significant yield loss worldwide. The current data o... more Viral and bacterial diseases of potato cause significant yield loss worldwide. The current data on the occurrence of these diseases in Russia do not provide comprehensive understanding of the phytosanitary situation. Diagnostic systems based on disposable stationary open qPCR micromatrices intended for the detection of eight viral and seven bacterial/oomycetal potato diseases have been used for wide-scale screening of target pathogens to estimate their occurrence in 11 regions of Russia and to assess suitability of the technology for high-throughput diagnostics under conditions of field laboratories. Analysis of 1025 leaf and 725 tuber samples confirmed the earlier reported data on the dominance of potato viruses Y, S, and M in most regions of European Russia, as well as relatively high incidences of Clavibacter michiganensis subsp. sepedonicus, Pectobacterium atrosepticum, and P. carotovorum subsp. carotovorum, and provided detailed information on the phytosanitary status of select...

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Research paper thumbnail of Preserved Microarrays for Simultaneous Detection and Identification of Six Fungal Potato Pathogens with the Use of Real-Time PCR in Matrix Format

Biosensors

Fungal diseases of plants are of great economic importance causing 70–80% of crop losses associat... more Fungal diseases of plants are of great economic importance causing 70–80% of crop losses associated with microbial plant pathogens. Advanced on-site disease diagnostics is very important to maximize crop productivity. In this study, diagnostic systems have been developed for simultaneous detection and identification of six fungal pathogens using 48-well microarrays (micromatrices) for qPCR. All oligonucleotide sets were tested for their specificity using 59 strains of target and non-target species. Detection limit of the developed test systems varied from 0.6 to 43.5 pg of DNA depending on target species with reproducibility within 0.3−0.7% (standard deviation). Diagnostic efficiency of test systems with stabilized and freeze-dried PCR master-mixes did not significantly differ from that of freshly prepared microarrays, though detection limit increased. Validation of test systems on 30 field samples of potato plants showed perfect correspondence with the results of morphological iden...

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Research paper thumbnail of Development of a real-time assay for detection of Synchytrium endobioticum, a quarantine pathogen of potato

Potato wart disease caused by a quarantine pathogen Synchytrium endobioticum may cause up to 50-1... more Potato wart disease caused by a quarantine pathogen Synchytrium endobioticum may cause up to 50-100% of yield losses; moreover, contaminated fields may be scheduled for 20 years. To provide an appropriate and timely control of this pathogen, reliable, simple, and robust diagnostic tools are needed. In this study we developed a test system for the real-time PCR detection and identification of S. endobioticum suitable for the use in qPCR micromatrices providing multiplex simultaneous detection of a range of pathogens. The designed primers and probe successfully tested for specificity using 10 samples of non-target fungal pathogens of potato. Detection limit of the test system was 0.1 pg of DNA per reaction. Reproducibility assay showed the standard error for Ct value did not exceed 4.5%. The further validation of the test system using culture samples of S. endobioticum of different pathotypes and infected field samples of potato should be done, and the efficient protocol for the pathogen detection in soil samples should be developed in future studies.Potato wart disease caused by a quarantine pathogen Synchytrium endobioticum may cause up to 50-100% of yield losses; moreover, contaminated fields may be scheduled for 20 years. To provide an appropriate and timely control of this pathogen, reliable, simple, and robust diagnostic tools are needed. In this study we developed a test system for the real-time PCR detection and identification of S. endobioticum suitable for the use in qPCR micromatrices providing multiplex simultaneous detection of a range of pathogens. The designed primers and probe successfully tested for specificity using 10 samples of non-target fungal pathogens of potato. Detection limit of the test system was 0.1 pg of DNA per reaction. Reproducibility assay showed the standard error for Ct value did not exceed 4.5%. The further validation of the test system using culture samples of S. endobioticum of different pathotypes and infected field samples of potato should be done, and the efficient protocol for the pathogen detection in soil s...

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Research paper thumbnail of Sensitization of plant pathogenic fungi to the tebuconazole-based commercial fungicide using some analogues of natural amino acids

Commercial fungicides provide an efficient control of fungal diseases of crops. At the same time,... more Commercial fungicides provide an efficient control of fungal diseases of crops. At the same time, plant pathogenic fungi can develop resistance to fungicides that results in epiphytoties and significant yield losses. The most common response of potato growers is more extensive use of fungicides associated with environmental pollution and a range of various health risks. Therefore, the search for approach preventing the development of pest resistance to fungicides is of great practical interest. A potentially new strategy to overcome or minimize this problem is an enhancement of pathogen sensitivity to fungicides using various chemical compounds. In this study, we examined several putative sensitizers chosen among structural analogues of some natural amino acids known for their inhibiting effect on the polyketide biosynthetic pathway, since polyketide mycotoxins play an important role in a pathogenicity of fungi. A possible synergism between the action of these compounds and a commercial fungicide Folicur on the growth of Phoma glomerata and Fusarium culmorum colonies was evaluated using the Limpel’s criterion. For all four compounds tested, a significant chemosensitizing effect was revealed for both non-fungicidal and sub-fungicidal concentrations that confirmed the initial hypothesis. The further screening of other amino acid analogues for their chemosensitizing activity is planned, as well as the study of the mechanism of their sensitizing action. Revealing of a compound providing significant chemosensitizing effect and characterized by a relatively low cost of synthesis would provide a low-cost solution for reduction of fungicidal treatments, while still providing sufficient protection even against fungicide-resistant strains of plant pathogens.Commercial fungicides provide an efficient control of fungal diseases of crops. At the same time, plant pathogenic fungi can develop resistance to fungicides that results in epiphytoties and significant yield losses. The most common response of potato growers is more extensive use of fungicides associated with environmental pollution and a range of various health risks. Therefore, the search for approach preventing the development of pest resistance to fungicides is of great practical interest. A potentially new strategy to overcome or minimize this problem is an enhancement of pathogen sensitivity to fungicides using various chemical compounds. In this study, we examined several putative sensitizers chosen among structural analogues of some natural amino acids known for their inhibiting effect on the polyketide biosynthetic pathway, since polyketide mycotoxins play an important role in a pathogenicity of fungi. A possible synergism between the action of these compounds and a commercial fungicide Folicur ...

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Research paper thumbnail of Matrix approach to the simultaneous detection of multiple potato pathogens by real-time PCR

Journal of Applied Microbiology

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Research paper thumbnail of SOME NATURAL AND SYNTHETIC COMPOUNDS INHIBITING THE BIOSYNTHESIS OF AFLATOXIN B1 AND MELANIN IN Aspergillus flavus

Sel'skokhozyaistvennaya Biologiya

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Research paper thumbnail of A new highly productive Propionibacterium acidipropionici FL-48 strain with increased resistance to propionic acid and the scaling up of its production for industrial bioreactors

Biosystems Diversity

Propionic acid bacteria, including Propionibacterium acidipropionici, are widely used in the chem... more Propionic acid bacteria, including Propionibacterium acidipropionici, are widely used in the chemical industry to produce propionic acid and also for food and feed preservation. However, the efficiency of the industrial production of these bacteria is limited by their sensitivity to high concentrations of propionic acid excreted into the cultivation medium. Therefore, the development of new biotechnological processes and strains able to overcome this limitation and to improve the profitability of the microbiological production remains a relevant problem. A new P. acidipropionici FL-48 strain characterized by an increased resistance to 10 g/L of propionic acid (the number of viable cells after 24-h cultivation reached 1.05 × 106) was developed by a two-step induced mutagenesis using UV and diethyl sulphate from the P. acidipropionici VKPM B-5723 strain. The mutant strain exceeded the parental strain in the biomass accumulation rate and the amount of produced propionic and acetic aci...

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Research paper thumbnail of Effect of Various Compounds Blocking the Colony Pigmentation on the Aflatoxin B1 Production by Aspergillus flavus

Toxins, 2016

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Research paper thumbnail of Effect of a Pre-Planting Treatment of Seed Tubers with Low-Frequency Pulse Electric Field on the Growth of Potato Plants of Different Varieties

Sel'skokhozyaistvennaya Biologiya, 2016

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Research paper thumbnail of Characterization of Russian Phytophthora infestans populations: DNA fingerprinting and SSR analysis

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Research paper thumbnail of Blocking of some stages of melaninogenesis can enhance aflatoxin B1 production in Aspergillus flavus

Both aflatoxin B1 (AFB1) and melanin produced by Aspergillus flavus are products of a polyketide ... more Both aflatoxin B1 (AFB1) and melanin produced by Aspergillus flavus are products of a polyketide biosynthesis. Inhibition of AFB1 biosynthesis reduces the harmfulness of A. flavus, whereas blocking of melanin biosynthesis is able to reduce the viability of this fungus. Thus, the development of a complex preparation blocking both AFB1 and melanin production in A. flavus is a point of practical interest.
As we have shown earlier, some phosphoanalogues of amino acids and peptides are able to regulate polyketide biosynthesis in phytopathogenic fungi. The purpose of this study was to screen such compounds for potential inhibitors of the AFB1 and melanin production in A. flavus. As a result, two different groups of inhibitors have been revealed:
I. Inhibitors of AFB1 biosynthesis
1. CH3(NH2)-P(S)(OH)2
2. CH3-CH(NH2)-P(S)(OH)2
3. CH3-CH(NH2)-P(O)(OC2H5)OH
4. CH3-CH(NH2)-CO-NH -CH(CH3)-P(O)(OCH3)OH
II. Inhibitors of melaninogenesis
5. CH3-CH(NH2)-P(O)(OH)2
6. NH2-CH2-CH2-CH2-ONH2
7. D,L-Ala-P(O)(CH3)OH
8. NH2-CH2-CH2-CH2-CH2-NHOH
Compounds 1-4 reduced AFB1 production by A. flavus up to 6 times as against the control, but did not influence on a colony pigmentation. Compounds 5-8 caused colony discoloration and, at the same time, significantly enhanced the AFB1 production.
It is known that AFB1 and melanin biosynthetic pathways have common initial stages and then diverge. Colony discoloration could be caused by blocking of melaninogenesis after the divergence point (see figure); due to such blocking, toxin production is increased.
The further testing of different combinations of compounds from the groups I and II will probably allow us to develop a complex preparation able to block both toxin and melanin production.

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