Личная информация

1975—1980 — Биофизик, Институт физиологии им. И. С. Бериташвили, Тбилиси, Грузия (СССР)R
1980—1982 — Молекулярный биолог, Институт биоорганической химии им. М. М. Шемякина РАН, Москва
1982—1986 — Старший научный сотрудник (мол. Биология), Институт, НИИ Сельскохозяйственной биотехнологии ВАСХНИЛ, Москва
1986—2005 — Заведующий Лабораторией молекулярной вирусологии Института Сельскохозяйственной биотехнологии ВАСХНИЛ, Москва
1987—1992 — Заместитель директора по науке Института сельскохозяйственной биотехнологии РАСХН, Москва
1993 — Приглашённый профессор Университета Северной Каролины, США
2000—2005 — Заместитель директора по науке Института сельскохозяйственной биотехнологии РАСХН, Москва
2005–н. в. — Заведующий Отделом молекулярной биологии и биотехнологии растений, Заведующий Лабораторией молекулярной диагностики, Заведующий Отделом международных научных связей Института биоорганической химии им. М. М. Шемякина и Ю. А. Овчинникова РАН, Москва

Образование

Избранные публикации

1. Tsybulsky D.A., Kvach M.V., Ryazantsev D.Y., Aparin I.O., Stakheev A.A., Prokhorenko I.A., Martynenko Y.V., Gontarev S.V., Formanovsky A.A., Zatsepin T.S., Shmanai V.V., Korshun V.A., Zavriev S.K. (2016). Molecular beacons with JOE dye: Influence of linker and 3' couple quencher. Mol. Cell. Probes 30 (5), 285–290 [+]

   Molecular beacons carrying JOE dye (4',5'-dichloro-2',7'-dimethoxy-6-carboxyfluorescein) on a rigid or flexible linker and one or two BHQ1 quenchers have been prepared and tested in real-time PCR using Fusarium avenaceum elongation factor 1α DNA template. The probes were different in their structures (loop size and stem length), linkers for dye attachment (6-aminohexanol or trans-4-aminocyclohexanol), quencher composition (single and double BHQ1) to elucidate the influence of all these features. Fluorogenic properties of the probes were studied and compared to those of FAM (fluorescein)-based probes. All the factors - stem length, JOE vs FAM, rigid vs flexible linker, single vs double quencher - appeared to play a considerable role in the probe's fluorescent properties and determine the usability of the probe at two different temperatures of fluorescence detection (55°С and 64°С).

   ID:1582
2. Stakheev A.A., Khairulina D.R., Zavriev S.K. (2016). Four-locus phylogeny of Fusarium avenaceum and related species and their species-specific identification based on partial phosphate permease gene sequences. International Journal of Food Microbiology 225, 27–37 [+]

   The fungus Fusarium avenaceum and its closest relatives are responsible for contamination of agricultural plants and their products by mycotoxins such as enniatins and moniliformin. Precise identification of mycotoxin producers is necessary for estimation of the accumulation risk of those compounds and for preventing the consumption of highly contaminated products. Nucleic acids amplification-based techniques proved to be the most rapid and reliable approach for pathogen diagnostics and identification. In this study partial phosphate permease gene (PHO) sequences were determined for Fusarium avenaceum (including one isolate identified as F. arthrosporioides), F. tricinctum, F. acuminatum and F. torulosum. Phylogenetic analysis of 40 isolates of those species from different climates and geographical regions of Russia and some neighboring countries based on sequences of PHO, translation elongation factor 1 alpha (TEF1α), beta-tubulin (β-TUB), enniatin synthetase (Esyn1) genes and combined data set demonstrated that the PHO gene possesses the highest rate of variability among them and can be considered as an informative marker for phylogenetic studies of these species. According to the combined data set phylogeny, the isolates of each species formed clusters with a high bootstrap support. Analysis of PHO sequences revealed a high intraspecific variability of F. avenaceum: there were 5 independent clusters on the dendrogram, including one cluster which was closer to F. torulosum than to other F. avenaceum isolates. Variable sites in PHO sequences have been used for the design of species-specific primers and a fluorescent hydrolysis probe. The specificity of the assay was shown for DNA samples extracted from 68 isolates of 23 Fusarium species. Quantitative PCR approach was applied to estimate the contamination rate of 17 naturally infected oat and barley samples, previously characterized by microbiological procedures.

   ID:1613
3. Vasilchenko A.S., Yuryev M., Ryazantsev D.Y., Zavriev S.K., Feofanov A.V., Grishin E.V., Rogozhin E.A. (2016). Studying of cellular interaction of hairpin-like peptide EcAMP1 from barnyard grass (Echinochloa crusgalli L.) seeds with plant pathogenic fungus Fusarium solani using microscopy techniques. Scanning , [+]

   An interaction of recombinant hairpin-like cationic peptide EcAMP1 with conidia of plant pathogenic fungus Fusarium solani at the cellular level was studied by a combination of microscopic methods. EcAMP1 is from barnyard grass (Echinochloa crusgalli L.), and obtained by heterologous expression in Escherichia coli system. As a result, a direct relationship between hyphal growth inhibition and increasing active peptide concentration, time of incubation and fungal physiological condition has been determined. Dynamics of accumulation and redistribution of the peptide studied on fungal cellular cover and inside the conidia cells has been shown. The dynamics are dependent on time of coupling, as well as, a dissimilarity of EcAMP1 binding with cover of fungal conidia and its stepwise accumulation and diffuse localization in the cytoplasm. Correlation between structural disruption of fungal conidia and the presence of morphological changes has also been found. The correlation was found under the influence of peptide high concentrations at concentrations above 32 μM. The results indicate the presence of a binding of EcAMP1 with the surface of fungal conidia, thus, demonstrating a main specificity for its antifungal action at the cellular level. These results, however, cannot exclude the existence of attendant EcAMP1 action based on its intracellular localization on some specific targets. SCANNING 9999:1-8, 2016. © 2016 Wiley Periodicals, Inc.

   ID:1624
4. Shcherbakova L.A., Odintsova T.I., Stakheev A.A., Fravel D.R., Zavriev S.K. (2016). Identification of a Novel Small Cysteine-Rich Protein in the Fraction from the Biocontrol Fusarium oxysporum Strain CS-20 that Mitigates Fusarium Wilt Symptoms and Triggers Defense Responses in Tomato. Front Plant Sci 6, 1207 [+]

   The biocontrol effect of the non-pathogenic Fusarium oxysporum strain CS-20 against the tomato wilt pathogen F. oxysporum f. sp. lycopersici (FOL) has been previously reported to be primarily plant-mediated. This study shows that CS-20 produces proteins, which elicit defense responses in tomato plants. Three protein-containing fractions were isolated from CS-20 biomass using size exclusion chromatography. Exposure of seedling roots to one of these fractions prior to inoculation with pathogenic FOL strains significantly reduced wilt severity. This fraction initiated an ion exchange response in cultured tomato cells resulting in a reversible alteration of extracellular pH; increased tomato chitinase activity, and induced systemic resistance by enhancing PR-1 expression in tomato leaves. Two other protein fractions were inactive in seedling protection. The main polypeptide (designated CS20EP), which was specifically present in the defense-inducing fraction and was not detected in inactive protein fractions, was identified. The nucleotide sequence encoding this protein was determined, and its complete amino acid sequence was deduced from direct Edman degradation (25 N-terminal amino acid residues) and DNA sequencing. The CS20EP was found to be a small basic cysteine-rich protein with a pI of 9.87 and 23.43% of hydrophobic amino acid residues. BLAST search in the NCBI database showed that the protein is new; however, it displays 48% sequence similarity with a hypothetical protein FGSG_10784 from F. graminearum strain PH-1. The contribution of CS20EP to elicitation of tomato defense responses resulting in wilt mitigating is discussed.

   ID:1427
5. Morozov S.Y., Miliytina I.A., Bobrova V.K., Ryazantsev D.Y., Erokhina T.N., Zavriev S.K., Agranovsky A.A., Solovyev A.G., Troitsky A.V. (2015). Structural evolution of the 4/1 genes and proteins in non-vascular and lower vascular plants. Biochimie , [+]

   The 4/1 protein of unknown function is encoded by a single-copy gene in most higher plants. The 4/1 protein of Nicotiana tabacum (Nt-4/1 protein) has been shown to be alpha-helical and predominantly expressed in conductive tissues. Here, we report the analysis of 4/1 genes and the encoded proteins of lower land plants. Sequences of a number of 4/1 genes from liverworts, lycophytes, ferns and gymnosperms were determined and analyzed together with sequences available in databases. Most of the vascular plants were found to encode Magnoliophyta-like 4/1 proteins exhibiting previously described gene structure and protein properties. Identification of the 4/1-like proteins in hornworts, liverworts and charophyte algae (sister lineage to all land plants) but not in mosses suggests that 4/1 proteins are likely important for plant development but not required for a primary metabolic function of plant cell.

   ID:1343
6. Beyshova I.S., Chuzhebaeva G.D., Aubakirov M.Z.h., Kozhmuhametova A.S., Stakheev A.A., Ryazantsev D.Y., Zavriev S.K., Oleynik A.T. (2015). Development of PCR diagnosis of pathogenic fungi of the genus Septoria affecting cereal crops in Northern Kazakhstan. . Biosciences Biotechnology Asia 12 (2), 1321 [+]

   Traditional methods of complex diagnostics, including the selection of species in pure culture, selection environments and cultivation conditions, obtaining monoporosa isolates and microscopy are long, time-consuming and ineffective. Immune-enzyme analysis method takes a few hours, however, developed on the basis of the methodology of species-specific, predictive and quantification S. Tritici and S. nodorum in the leaves and seeds of wheat insensitive do not always provide a clear identification of these species. The introduction of express methods of diagnostics of Septoria leaf blotch on the basis of the polymerase chain reaction (PCR) in modifications of Real-Time lets quickly and accurately diagnose the disease. A system of highly specific sensitive PCR diagnostics of pathogenic fungi Septoria tritici and Stagonospora nodorum, causing diseases of cereals in Northern Kazakhstan is developed. The primers and probes are designed based on partial sequences of the internal transcribed spacer of ribosomal DNA (ITS) provides a quick and accurate identification of pathogens Septoria by quantitative PCR without the risk of contamination of the work area with amplification products. Effectiveness of the test-system was also demonstrated in samples of total DNA isolated from the infected herbarium material.

   ID:1426
7. Ryazantsev D.Y., Kvach M.V., Tsybulsky D.A., Prokhorenko I.A., Stepanova I.A., Martynenko Y.V., Gontarev S.V., Shmanai V.V., Zavriev S.K., Korshun V.A. (2014). Design of molecular beacons: 3' couple quenchers improve fluorogenic properties of a probe in real-time PCR assay. Analyst 139 (11), 2867–72 [+]

   Convenient preparation of fluorogenic hairpin DNA probes (molecular beacons) carrying a pair of FAM fluorophores (located close to 5'-terminus of the probe) or a pair of BHQ1 quenchers on 3'-terminus (with (BHQ1)2 or BHQ1-BHQ1 composition) is reported. These probes were used for the first time in a real-time PCR assay and showed considerable improvements in fluorogenic properties (the total fluorescence increase or signal-to-background ratio) in assay conditions vs. conventional one-FAM-one-BHQ1 molecular beacon probes as well as vs. hydrolyzable one-FAM-one-BHQ1 TaqMan probes. At the same time, such multiple modifications of the probe do not influence its Cq (a fractional PCR cycle used for quantification). The probe MB14 containing a BHQ1-BHQ1 pair showed a PCR fluorescence/background value of 9.6 which is more than two times higher than that of a regular probe MB2 (4.6). This study demonstrates prospects for the design of highly fluorogenic molecular beacon probes suitable for quantitative real-time PCR and for other potential applications (e.g. intracellular RNA detection and SNP/mutation analysis).

   ID:1019
8. Kapustin D.V., Prostyakova A.I., Alexeev Y.I., Varlamov D.A., Zubov V.P., Zavriev S.K. (2014). High-throughput Method of One-Step DNA Isolation for PCR Diagnostics of Mycobacterium tuberculosis. Acta Naturae 6 (2), 48–52 [+]

   The efficiency of one-step and multi-step protocols of DNA isolation from lysed sputum samples containing the Mycobacterium tuberculosis complex has been compared. DNA was isolated using spin-cartridges containing a special silica-based sorbent modified with fluoroplast and polyaniline, or using an automated isolation system. One-step isolation using the obtained sorbent has been shown to ensure a significantly lower DNA loss and higher sensitivity in the PCR detection of Mycobacterium tuberculosis as compared to a system based on sorption and desorption of nucleic acids during the isolation.

   ID:1424
9. Рязанцев Д.Ю., Дробязина П.Е., Хлгатян С.В., Завриев С.К., Свирщевская Е.В. (2014). Экспрессия аллергенов клещей домашней пыли Der f 1 и Der f 2 в листьях Nicotiana benthamiana. Биоорг. хим. 40 (4), . 468–478 ID:1157
10. Rogozhin E.A., Ryazantsev D.Y., Grishin E.V., Egorov T.A., Zavriev S.K. (2012). Defense peptides from barnyard grass (Echinochloa crusgalli L.) seeds. Peptides 38 (1), 33–40 [+]

    A number of defense polypeptides from latent seeds of weed cereal barnyard grass (Echinochloa crusgalli L.) has been isolated and characterized using an acidic extraction and high performance liquid chromatography methods in combination with MALDI-TOF mass spectrometry and Edman sequencing. Members of three antimicrobial peptide families and two protease inhibitor families were found to be localized in barnyard grass seeds. Their biological activity concerning to Gram-Positive and Gram-Negative phytopathogenic bacteria, as well as oomycete Phytophthora infestans, has been investigated. Diversity of barnyard grass defense peptides is a significant factor that provides a resistance of E. crusgalli seeds to germination and latent phases.

    ID:805
11. Ryazantsev D.Y., Tsybulsky D.A., Prokhorenko I.A., Kvach M.V., Martynenko Y.V., Philipchenko P.M., Shmanai V.V., Korshun V.A., Zavriev S.K. (2012). Two-dye and one- or two-quencher DNA probes for real-time PCR assay: synthesis and comparison with a TaqMan™ probe. Analytical and bioanalytical chemistry 404 (1), 59–68 [+]

    A typical TaqMan™ real-time PCR probe contains a 5'-fluorescent dye and a 3'-quencher. In the course of the amplification, the probe is degraded starting from the 5'-end, thus releasing fluorescent dye. Some fluorophores (including fluorescein) are known to be prone to self-quenching when located near each other. This work is aimed at studying dye-dye and dye-quencher interactions in multiply modified DNA probes. Twenty-one fluorogenic probes containing one and two fluoresceins (FAM), or a FAM-JOE pair, and one or two BHQ1 quenchers were synthesized using non-nucleoside reagents and "click chemistry" post-modification on solid phase and in solution. The probes were tested in real-time PCR using an ~300-bp-long natural DNA fragment as a template. The structural prerequisites for lowering the probe background fluorescence and increasing the end-plateau fluorescence intensity were evaluated and discussed.

    ID:753
12. Ryazantsev D.Y.u., Petrova E., Kalinina N.A., Valyakina T.I., Grishin E.V., Zavriev S.K. (2012). Application of supramolecular DNA-streptavidin complexes for ultrasensitive detection of several toxins by immuno-PCR. 14. Global J. Anal. Chem. 3 (17), [+]
    ID:650
13. Стахеев А.А., Рязанцев Д.Ю., Завриев С.К. (2011). Выявление новых генетических маркеров для таксономической характеристики и идентификации грибов рода Fusarium. Биоорг. хим. 37, 662–671 [+]
    ID:649
14. Stakheev A.A., Ryazantsev D.Y.u., Gagkaeva T.Y.u., Zavriev S.K. (2010). PCR detection of Fusarium fungi with similar profiles of the produced mycotoxins. Food Control 22 (3-4), 462–468 [+]
    ID:648
15. Рязанцев Д.Ю., Абрамов Д.Д., Завриев С.К. (2009). Диагностика карантинных фитопатогенов методом ПЦР в формате FLASH. Сельскохозяйственная биология  (3), 114–117 ID:161
16. Рязанцев Д.Ю., Завриев С.К. (2009). Эффективный метод диагностики и идентификации вирусных патогенов картофеля. Мол. биол. 43 (3), 558–567 ID:160
17. Lukhovitskaia N.I., Solov'eva A.G., Koshkina T.E., Zavriev S.K., Morozov S.I.u. (2009). [Interaction of cysteine-rich protein of Carlavirus with plant defense system]. Mol. Biol. (Mosk.) 39 (5), 896–904 [+]

    Viruses of genus Carlavirus encode a small cysteine-rich protein (CRP) of unknown function. To investigate the role of CRP of carlavirus chrysanthemum virus B (CVB), a recombinant potato virus X (PVX) genome was constructed, which carried the CVB CRP gene. Expression of CVB CRP in the PVX genetic background drastically changed the PVX symptom phenotype in N. benthamiana. Instead of symptomless infection and mild mosaic, which are characteristic of PVX in this plant host, the recombinant virus expressing CVB CRP induced formation of necrotic local lesions on inoculated leaves and necrosis of the apical leaves. In N. tabacum, the infection pattern depended on the host genotype: the recombinant PVX was able to spread systemically only in N gene-carrying plants. In agroinfiltration-mediated transient expression assay, CVB CRP did not exhibit the properties of avirulence factor in N. benthamiana and was unable to suppress post-transcriptional gene silencing. Thus, CVB CRP is the viral pathogenicity determinant controlling the virus interaction with plant hosts in a manner which depends on plant defense mediated by resistance genes such as the N gene.

    ID:164
18. Abramova S.L., Riazantsev D.Y.u., Voinova T.M., Zavriev S.K. (2009). [Diagnostics of phytopathogen fungi Septoria tritici and Stagonospora nodorum by fluorescent amplification-based specific hybridization (FLASH) PCR]. Bioorg. Khim. 34 (1), 107–13 [+]
    ID:646
19. Riazantsev D.Y.u., Abramova S.L., Evstratova S.V., Gagkaeva T.Y.u., Zavriev S.K. (2009). [FLASH-PCR diagnostics of toxigenic fungi of the genus Fusarium]. Bioorg. Khim. 34 (6), 799–807 [+]
    ID:647
20. Абрамова С.Л., Рязанцев Д.Ю., Воинова Т.М., Завриев С.К. (2008). Диагностика фитопатогенных грибов Septoria tritice и Stragonaspora nodorum методом FLASH–ПЦР. Биоорг. хим. 34, 107–113 ID:158
21. Рязанцев Д.Ю., Абрамова С.Л., Евстратова С.В., Гагкаева Т.Ю., Завриев С.К. (2008). Диагностика токсиногенных грибов рода Fusarium методом FLASH-PCR. Биоорг. хим. 35, 799–807 ID:29
22. Кокарев Н.В., Кошкина Т.Е., Рязанцев Д.Ю., Завриев С.К. (2007). Влияние дефектной РНК вируса крапчатости ежи сборной на накопление вирусного капсидного белка в растениях пшеницы. Доклады РАСХН  (3б), 13–15 ID:157
23. Mitioushkina T.Y.u., Dolgov S.V., Zavriev S.K., Kharchenko P.N. (2006). Molecular biology approach for improving chrysanthemum resistance to virus B. Acta Horticulturae. Acta Horticulturae 772, 327–332 ID:166
24. Вишниченко В.К., Рязанцев Д.Ю., Завриев С.К. (2005). Экспрессия капсидного белка Х вируса шалота в различных органах растений Allium cepa var. Aggregatum. Сельскохозяйственная биология  (1), 104–109 ID:155
25. Adams M.J., Antoniw J.F., Bar-Joseph M., Brunt A.A., Candresse T., Foster G.D., Martelli G.P., Milne R.G., Zavriev S.K., Fauquet C.M. (2004). The new plant virus family Flexiviridae and assessment of molecular criteria for species demarcation. Arch. Virol. 149 (5), 1045–60 [+]

    The new plant virus family Flexiviridae is described. The family is named because its members have flexuous virions and it includes the existing genera Allexivirus, Capillovirus, Carlavirus, Foveavirus, Potexvirus, Trichovirus and Vitivirus, plus the new genus Mandarivirus together with some related viruses not assigned to any genus. The family is justified from phylogenetic analyses of the polymerase and coat protein (CP) sequences. To help to define suitable molecular criteria for demarcation of species, a complete set of pairwise comparisons was made using the nucleotide (nt) and amino acid (aa) sequences of each fully-sequenced gene from every available accession in the family. Based on the distributions and on inspection of the data, it was concluded that, as a general rule, distinct species have less than ca. 72% identical nt or 80% identical aa between their entire CP or replication protein genes.

    ID:163
26. Кошкина Т.Е., Баранова Е.Н., Завриев С.К. (2003). Точечная мутация в гене белка оболочки влияет на дальний транспорт вируса табачной мозаики. Мол. биол. 37, 742–748 ID:154
27. Завриев С.К., Вишниченко В.К., Келдыш М.А. (2002). Обнаружение аллексивирусов в составе вирусных комплексов, поражающих декоративные луковичные культуры. Доклады РАСХН  (1), 11–13 ID:152
28. Maroon C.J.M., Zavriev S. (2002). PCR-BASED TESTS FOR THE DETECTION OF TOBAMOVIRUSES AND CARLAVIRUSES. Acta Horticulturae 598, 117–122 [+]

    Routine testing of ornamental plants involves ELISA screens that include a number of specific tests to detect the presence of a virus belonging to a particular group. This approach has been desirable for a number of reasons: cost-effectiveness, simplicity and speed. Despite these advantages, the screens can become quite complicated due to the number of specific tests that need to be conducted. Furthermore, with certain host species, non-specific tissue reaction is not at all uncommon thus prompting the use of more than one ELISA format for a particular test. As an alternative to our ELISA screens, we have developed two PCR group tests that respectively detect members of the tobamovirus and the carlavirus groups. These tests are coupled with reverse transcription using random hexamers or oligo d(T)16 primer, respectively. For the PCR part, primers are designed based on the conserved regions of the viral genome and are used under optimized conditions for the amplification of the target sequences. Using our tobamovirus group PCR test, we successfully detected CGMMV, KGMMV, ORSV, PMMoV, RMV, SHMV, TMV and ToMV from leaves, stems, fruits and seeds of infected plants. Our carlavirus group PCR test could detect all carlaviruses examined: BBScV, CLV, CVB, KLV, LSV, PotLV, PVM, PVS and SLV.

    ID:25
29. Вишниченко В.К., Стельмащик В.Я., Завриев С.К. (2002). 42K белок Х вируса шалота участвует в формировании вирусных частиц. Мол. биол. 36, 1080–1084 ID:26
30. Кошкина Т.Е., Новиков В.К., Завриев С.К. (2002). Исследование биологических свойств и структуры генома изолята К3 казахского штамма вируса табачной мозаики. Доклады РАСХН  (3), 14–15 ID:153
31. Vishnichenko V.K., Zavriev S.K. (2001). Detection of infectious viral particles in plant protoplasts inoculated with transcripts of full-length shallot virus X cDNA. Arch. Virol. 146 (6), 1213–7 [+]

    Flexible filamentous shallot virus X (ShVX) particles were detected in extracts of Beta vulgaris protoplasts inoculated with transcripts from a full-length ShVX cDNA. Extracts from ShVX-infected protoplast were infectious for ShVX-healthy shallot seedlings. Western blot analysis of inoculated plants revealed the accumulation of the ShVX coat protein, while electron microscopy confirmed the presence of ShVX virions. The results suggest that the in vitro RNA transcripts from full-length ShVX cDNA give rise to infectious viral particles.

    ID:24