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Dr. Alexandra Dubrovina
Laboratory of Biotechnology, Federal Scientific Center of the East Asia Terrestrial Biodiversity, Far Eastern Branch of the Russian Academy of Sciences, 690022 Vladivostok, Russia

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0 SiRNA
0 RNA interference
0 dsRNA
0 Transgenic plants
0 Foliar RNA application

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dsRNA
RNA interference
Transgenic plants
SiRNA
Foliar RNA application
Plant gene regulation

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Journal article
Published: 24 June 2021 in Plants
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Excessive ultraviolet B (UV-B) irradiation is one of the most serious threats leading to severe crop production losses. It is known that secondary metabolite biosynthesis plays an important role in plant defense and forms a protective shield against excessive UV-B irradiation. The contents of stilbenes and other plant phenolics are known to sharply increase after UV-B irradiation, but there is little direct evidence for the involvement of stilbenes and other plant phenolics in plant UV-B protection. This study showed that foliar application of trans-resveratrol (1 and 5 mM) and trans-piceid (5 mM) considerably increased tolerance to a shock of UV-B (10 min at 1800 µW cm−2 of irradiation intensity) of four-week-old Arabidopsis thaliana plants that are naturally incapable of stilbene production. Application of trans-resveratrol and trans-piceid increased the leaf survival rates by 1–2%. This stilbene-induced improvement in UV-B tolerance was higher than after foliar application of the stilbene precursors, p-coumaric and trans-cinnamic acids (only 1–3%), but less than that after treatment with octocrylene (19–24%), a widely used UV-B absorber. Plant treatment with trans-resveratrol increased expression of antioxidant and stress-inducible genes in A.thaliana plants and decreased expression of DNA repair genes. This study directly demonstrates an important positive role of stilbenes in plant tolerance to excessive UV-B irradiation, and offers a new approach for plant UV-B protection.

ACS Style

Zlata Ogneva; Vlada Volkonskaia; Alexandra Dubrovina; Andrey Suprun; Olga Aleynova; Konstantin Kiselev. Exogenous Stilbenes Improved Tolerance of Arabidopsis thaliana to a Shock of Ultraviolet B Radiation. Plants 2021, 10, 1282 .

AMA Style

Zlata Ogneva, Vlada Volkonskaia, Alexandra Dubrovina, Andrey Suprun, Olga Aleynova, Konstantin Kiselev. Exogenous Stilbenes Improved Tolerance of Arabidopsis thaliana to a Shock of Ultraviolet B Radiation. Plants. 2021; 10 (7):1282.

Chicago/Turabian Style

Zlata Ogneva; Vlada Volkonskaia; Alexandra Dubrovina; Andrey Suprun; Olga Aleynova; Konstantin Kiselev. 2021. "Exogenous Stilbenes Improved Tolerance of Arabidopsis thaliana to a Shock of Ultraviolet B Radiation." Plants 10, no. 7: 1282.

Journal article
Published: 23 June 2021 in Plants
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Plant endophytes are known to alter the profile of secondary metabolites in plant hosts. In this study, we identified the main bacterial and fungal representatives of the wild grape Vitis amurensis Rupr. microbiome and investigated a cocultivation effect of the 14 endophytes and the V. amurensis cell suspension on biomass accumulation and stilbene biosynthesis. The cocultivation of the V. amurensis cell culture with the bacteria Agrobacterium sp., Bacillus sp., and Curtobacterium sp. for 2 weeks did not significantly affect the accumulation of cell culture fresh biomass. However, it was significantly inhibited by the bacteria Erwinia sp., Pantoea sp., Pseudomonas sp., and Xanthomonas sp. and fungi Alternaria sp., Biscogniauxia sp., Cladosporium sp., Didymella sp. 2, and Fusarium sp. Cocultivation of the grapevine cell suspension with the fungi Didymella sp. 1 and Trichoderma sp. resulted in cell death. The addition of endophytic bacteria increased the total stilbene content by 2.2–5.3 times, while the addition of endophytic fungi was more effective in inducing stilbene accumulation by 2.6–16.3 times. The highest content of stilbenes in the grapevine cells cocultured with endophytic fungi was 13.63 and 13.76 mg/g of the cell dry weight (DW) after cultivation with Biscogniauxia sp. and Didymella sp. 2, respectively. The highest content of stilbenes in the grapevine cells cocultured with endophytic bacteria was 4.49 mg/g DW after cultivation with Xanthomonas sp. The increase in stilbene production was due to a significant activation of phenylalanine ammonia lyase (PAL) and stilbene synthase (STS) gene expression. We also analyzed the sensitivity of the selected endophytes to eight antibiotics, fluconazole, and trans-resveratrol. The endophytic bacteria were sensitive to gentamicin and kanamycin, while all selected fungal strains were resistant to fluconazole with the exception of Cladosporium sp. All endophytes were tolerant of trans-resveratrol. This study showed that grape endophytes stimulate the production of stilbenes in grape cell suspension, which could further contribute to the generation of a new stimulator of stilbene biosynthesis in grapevine or grape cell cultures.

ACS Style

Olga Aleynova; Andrey Suprun; Nikolay Nityagovsky; Alexandra Dubrovina; Konstantin Kiselev. The Influence of the Grapevine Bacterial and Fungal Endophytes on Biomass Accumulation and Stilbene Production by the In Vitro Cultivated Cells of Vitis amurensis Rupr. Plants 2021, 10, 1276 .

AMA Style

Olga Aleynova, Andrey Suprun, Nikolay Nityagovsky, Alexandra Dubrovina, Konstantin Kiselev. The Influence of the Grapevine Bacterial and Fungal Endophytes on Biomass Accumulation and Stilbene Production by the In Vitro Cultivated Cells of Vitis amurensis Rupr. Plants. 2021; 10 (7):1276.

Chicago/Turabian Style

Olga Aleynova; Andrey Suprun; Nikolay Nityagovsky; Alexandra Dubrovina; Konstantin Kiselev. 2021. "The Influence of the Grapevine Bacterial and Fungal Endophytes on Biomass Accumulation and Stilbene Production by the In Vitro Cultivated Cells of Vitis amurensis Rupr." Plants 10, no. 7: 1276.

Journal article
Published: 23 June 2021 in International Journal of Molecular Sciences
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Exogenous application of double-stranded RNAs (dsRNAs) and small-interfering RNAs (siRNAs) to plant surfaces has emerged as a promising method for regulation of essential genes in plant pathogens and for plant disease protection. Yet, regulation of plant endogenous genes via external RNA treatments has not been sufficiently investigated. In this study, we targeted the genes of chalcone synthase (CHS), the key enzyme in the flavonoid/anthocyanin biosynthesis pathway, and two transcriptional factors, MYBL2 and ANAC032, negatively regulating anthocyanin biosynthesis in Arabidopsis. Direct foliar application of AtCHS-specific dsRNAs and siRNAs resulted in an efficient downregulation of the AtCHS gene and suppressed anthocyanin accumulation in A. thaliana under anthocyanin biosynthesis-modulating conditions. Targeting the AtMYBL2 and AtANAC032 genes by foliar dsRNA treatments markedly reduced their mRNA levels and led to a pronounced upregulation of the AtCHS gene. The content of anthocyanins was increased after treatment with AtMYBL2-dsRNA. Laser scanning microscopy showed a passage of Cy3-labeled AtCHS-dsRNA into the A. thaliana leaf vessels, leaf parenchyma cells, and stomata, indicating the dsRNA uptake and spreading into leaf tissues and plant individual cells. Together, these data show that exogenous dsRNAs were capable of downregulating Arabidopsis genes and induced relevant biochemical changes, which may have applications in plant biotechnology and gene functional studies.

ACS Style

Konstantin Kiselev; Andrey Suprun; Olga Aleynova; Zlata Ogneva; Alexander Kalachev; Alexandra Dubrovina. External dsRNA Downregulates Anthocyanin Biosynthesis-Related Genes and Affects Anthocyanin Accumulation in Arabidopsis thaliana. International Journal of Molecular Sciences 2021, 22, 6749 .

AMA Style

Konstantin Kiselev, Andrey Suprun, Olga Aleynova, Zlata Ogneva, Alexander Kalachev, Alexandra Dubrovina. External dsRNA Downregulates Anthocyanin Biosynthesis-Related Genes and Affects Anthocyanin Accumulation in Arabidopsis thaliana. International Journal of Molecular Sciences. 2021; 22 (13):6749.

Chicago/Turabian Style

Konstantin Kiselev; Andrey Suprun; Olga Aleynova; Zlata Ogneva; Alexander Kalachev; Alexandra Dubrovina. 2021. "External dsRNA Downregulates Anthocyanin Biosynthesis-Related Genes and Affects Anthocyanin Accumulation in Arabidopsis thaliana." International Journal of Molecular Sciences 22, no. 13: 6749.

Journal article
Published: 09 April 2021 in Metabolites
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Grapes and wines represent the most important source of edible stilbenes and other phenolic metabolites, which demonstrate a wide range of valuable biological activities. However, there is no information about the profile and content of phenolic compounds in Russian wines. We firstly analyzed phenolics (stilbenes, phenolic acids, and flavonols) in some representatives of Russian wines, including eleven red and seven white Russian wines from Fanagoria, Krasnodarsky Territory. The Russian red wines contained six stilbenes (trans-resveratrol, cis-resveratrol, trans-, cis-piceid, trans-piceatannol, δ-viniferin), while the white wines contained only five stilbenes (cis-resveratrol, trans-, cis-piceid, trans-piceatannol, trans-resveratrol). More than a half of the total stilbenes in the wines (65% of all stilbenes) were presented by trans-piceid and cis-piceid, while trans-resveratrol reached 16% of all the stilbenes. The red wines also contained six phenolic acids and six flavonols, while the white wines contained six phenolic acids and only three flavonols. Myrecitin-3-O-glucoside, quercetin-3-O-glucoside, and myricetin were the major flavonols in the red wines, while dihydroquercetin-3-O-rhamnoside was the major flavonol in the white wines. The red wines contained markedly higher amounts of stilbenes, phenolic acids, and flavonols than the white wines. Thus, the data showed that young red Russian Fanagoria wines represent a rich source of phenolic compounds. The study also revealed that younger wines were more abundant in phenolics, and wine storage for six months in the dark at +10 °C led to a decrease in the total content of phenolics, primarily monomeric stilbenes and quercetin-3-O-glucoside and quercetin flavonols.

ACS Style

Andrey Suprun; Alexandra Dubrovina; Alexey Tyunin; Konstantin Kiselev. Profile of Stilbenes and Other Phenolics in Fanagoria White and Red Russian Wines. Metabolites 2021, 11, 231 .

AMA Style

Andrey Suprun, Alexandra Dubrovina, Alexey Tyunin, Konstantin Kiselev. Profile of Stilbenes and Other Phenolics in Fanagoria White and Red Russian Wines. Metabolites. 2021; 11 (4):231.

Chicago/Turabian Style

Andrey Suprun; Alexandra Dubrovina; Alexey Tyunin; Konstantin Kiselev. 2021. "Profile of Stilbenes and Other Phenolics in Fanagoria White and Red Russian Wines." Metabolites 11, no. 4: 231.

Original article
Published: 25 February 2021 in Molecular Biology Reports
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The cauliflower mosaic virus (CaMV) 35S promoter is known as the most frequently used promoter in plant biotechnology. Although it is widely considered to be a strong constitutive promoter exhibiting high transcriptional activity, the transcriptional stability of CaMV 35S has not been extensively studied. Using the model plant species Arabidopsis thaliana, this study aimed for a comprehensive expression analysis of two widely used plant transgenes, neomycin phosphotransferase II (NPTII) and enhanced green fluorescent protein (EGFP), regulated by a double CaMV 35S promoter depending on the organ type, time of day, plant age, and in response to abiotic stress conditions. Quantitative real-time PCR (qRT-PCR) analysis revealed that the NPTII and EGFP transcript levels were markedly higher in the cotyledons, young leaves, and roots than in the inflorescences, stems, and adult leaves of three independent transgenic A. thaliana lines. The expression of NPTII and EGFP varied during the day and was elevated with the plant age. Drought and cold stress considerably affected the expression of the transgenes, while heat, high salinity, and wounding had no significant effect. This study shows that transgenes driven by a common constitutive promoter can exhibit marked variations in transcriptional activity depending on plant organ, physiological conditions, and in response to abiotic stress. Therefore, to ensure high and stable transgene activity, considerable attention should be given to the transgenic plant material and incubation conditions before harvesting the plant material.

ACS Style

Konstantin V. Kiselev; Olga A. Aleynova; Zlata V. Ogneva; Andrey R. Suprun; Alexandra S. Dubrovina. 35S promoter-driven transgenes are variably expressed in different organs of Arabidopsis thaliana and in response to abiotic stress. Molecular Biology Reports 2021, 48, 2235 -2241.

AMA Style

Konstantin V. Kiselev, Olga A. Aleynova, Zlata V. Ogneva, Andrey R. Suprun, Alexandra S. Dubrovina. 35S promoter-driven transgenes are variably expressed in different organs of Arabidopsis thaliana and in response to abiotic stress. Molecular Biology Reports. 2021; 48 (3):2235-2241.

Chicago/Turabian Style

Konstantin V. Kiselev; Olga A. Aleynova; Zlata V. Ogneva; Andrey R. Suprun; Alexandra S. Dubrovina. 2021. "35S promoter-driven transgenes are variably expressed in different organs of Arabidopsis thaliana and in response to abiotic stress." Molecular Biology Reports 48, no. 3: 2235-2241.

Journal article
Published: 30 January 2021 in Plants
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: Recent studies have revealed that foliar application of double-stranded RNAs (dsRNAs) or small-interfering RNAs (siRNAs) encoding specific genes of plant pathogens triggered RNA interference (RNAi)-mediated silencing of the gene targets. However, a limited number of reports documented silencing of plant endogenes or transgenes after direct foliar RNA application. This study analyzed the importance of physiological conditions (plant age, time of day, soil moisture, high salinity, heat, and cold stresses) and different dsRNA application means (brush spreading, spraying, infiltration, inoculation, needle injection, and pipetting) for suppression of neomycin phosphotransferase II (NPTII) transgene in Arabidopsis thaliana, as transgenes are more prone to silencing. We observed a higher NPTII suppression when dsRNA was applied at late day period, being most efficient at night, which revealed a diurnal variation in dsRNA treatment efficacy. Exogenous NPTII-dsRNA considerably reduced NPTII expression in 4-week-old plants and only limited it in 2- and 6-week-old plants. In addition, a more discernible NPTII downregulation was detected under low soil moisture conditions. Treatment of adaxial and abaxial leaf surfaces by brushes, spraying, and pipetting showed a higher NPTII suppression, while infiltration and inoculation were less efficient. Thus, appropriate plant age, late time of day, low soil moisture, and optimal dsRNA application modes are important for exogenously induced gene silencing.

ACS Style

Konstantin V. Kiselev; Andrey R. Suprun; Olga A. Aleynova; Zlata V. Ogneva; Alexandra S. Dubrovina. Physiological Conditions and dsRNA Application Approaches for Exogenously induced RNA Interference in Arabidopsis thaliana. Plants 2021, 10, 264 .

AMA Style

Konstantin V. Kiselev, Andrey R. Suprun, Olga A. Aleynova, Zlata V. Ogneva, Alexandra S. Dubrovina. Physiological Conditions and dsRNA Application Approaches for Exogenously induced RNA Interference in Arabidopsis thaliana. Plants. 2021; 10 (2):264.

Chicago/Turabian Style

Konstantin V. Kiselev; Andrey R. Suprun; Olga A. Aleynova; Zlata V. Ogneva; Alexandra S. Dubrovina. 2021. "Physiological Conditions and dsRNA Application Approaches for Exogenously induced RNA Interference in Arabidopsis thaliana." Plants 10, no. 2: 264.

Journal article
Published: 26 October 2020 in International Journal of Molecular Sciences
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Calmodulin-like proteins (CMLs) represent a large family of plant calcium sensor proteins involved in the regulation of plant responses to environmental cues and developmental processes. In the present work, we identified four alternatively spliced mRNA forms of the grapevine CML21 gene that encoded proteins with distinct N-terminal regions. We studied the transcript abundance of CML21v1, CML21v2, CML21v3, and CML21v4 in wild-growing grapevine Vitis amurensis Rupr. in response to desiccation, heat, cold, high salinity, and high mannitol stress using quantitative real-time RT-PCR. The levels of all four splice variants of VaCML21 were highly induced in response to cold stress. In addition, VaCML21v1 and VaCML21v2 forms were highly modulated by all other abiotic stress treatments. Constitutive expression of VaCML21v2 and VaCML21v4 improved biomass accumulation of V. amurensis callus cell cultures under prolonged low temperature stress. Heterologous expression of the grapevine CML21v2 and VaCML21v4 splice variants in Arabidopsis improved survival rates of the transgenic plants after freezing. The VaCML21v2 overexpression enhanced activation of the cold stress-responsive marker genes AtDREB1A and AtDREB2A, while VaCML21v4 overexpression—AtCOR47, AtRD29A, AtRD29B, and AtKIN1 genes after freezing stress in the transgenic Arabidopsis. The results indicate that the grapevine CML21 gene acts as a positive regulator in the plant response to cold stress. The detected variety of CML21 transcripts and their distinct transcriptional responses suggested that this expansion of mRNA variants could contribute to the diversity of grapevine adaptive reactions.

ACS Style

Olga A. Aleynova; Konstantin V. Kiselev; Zlata V. Ogneva; Alexandra S. Dubrovina. The Grapevine Calmodulin-Like Protein Gene CML21 Is Regulated by Alternative Splicing and Involved in Abiotic Stress Response. International Journal of Molecular Sciences 2020, 21, 7939 .

AMA Style

Olga A. Aleynova, Konstantin V. Kiselev, Zlata V. Ogneva, Alexandra S. Dubrovina. The Grapevine Calmodulin-Like Protein Gene CML21 Is Regulated by Alternative Splicing and Involved in Abiotic Stress Response. International Journal of Molecular Sciences. 2020; 21 (21):7939.

Chicago/Turabian Style

Olga A. Aleynova; Konstantin V. Kiselev; Zlata V. Ogneva; Alexandra S. Dubrovina. 2020. "The Grapevine Calmodulin-Like Protein Gene CML21 Is Regulated by Alternative Splicing and Involved in Abiotic Stress Response." International Journal of Molecular Sciences 21, no. 21: 7939.

Applied genetics and molecular biotechnology
Published: 14 January 2020 in Applied Microbiology and Biotechnology
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Recent research has shown that plants can uptake long dsRNAs and dsRNA-derived siRNAs that target important genes of infecting fungi or viruses when applied on the surface of plant leaves. The external RNAs were capable of local and systemic movement inducing plant resistance against the pathogens. Few studies have been made for plant gene regulation by foliar application of RNAs. In this study, several types of ssRNA and siRNA duplexes targeting the neomycin phosphotransferase II (NPTII) transgene were in vitro-synthesized and externally applied to the leaf surface of 4-week-old transgenic Arabidopsis thaliana plants. External application of the synthetic NPTII-encoding siRNAs down-regulated NPTII transcript levels in transgenic A. thaliana 1 and 7 days post-treatment with a higher and more consistent effect being observed for siRNAs methylated at 3' ends. We also analyzed the effects of external NPTII-encoding dsRNA precursors and a dsRNA-derived heterogenous siRNA mix. Digestion of the NPTII-dsRNA to the heterogeneous siRNAs did not improve efficiency of the transgene suppression effect. Key Points• Foliar application of siRNAs down-regulated a commonly used transgene in Arabidopsis. • A more consistent effect was observed for methylated siRNAs. • The findings are important for development of plant gene regulation approaches.

ACS Style

Alexandra S. Dubrovina; Olga A. Aleynova; Andrey Suprun; Zlata V. Ogneva; Konstantin V. Kiselev. Transgene suppression in plants by foliar application of in vitro-synthesized small interfering RNAs. Applied Microbiology and Biotechnology 2020, 104, 2125 -2135.

AMA Style

Alexandra S. Dubrovina, Olga A. Aleynova, Andrey Suprun, Zlata V. Ogneva, Konstantin V. Kiselev. Transgene suppression in plants by foliar application of in vitro-synthesized small interfering RNAs. Applied Microbiology and Biotechnology. 2020; 104 (5):2125-2135.

Chicago/Turabian Style

Alexandra S. Dubrovina; Olga A. Aleynova; Andrey Suprun; Zlata V. Ogneva; Konstantin V. Kiselev. 2020. "Transgene suppression in plants by foliar application of in vitro-synthesized small interfering RNAs." Applied Microbiology and Biotechnology 104, no. 5: 2125-2135.

Review article
Published: 11 January 2020 in Biotechnology and Applied Biochemistry
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Plant stilbenes have attracted special attention, since they possess valuable health benefits and improve plant resistance to environmental stresses. Stilbenes are synthesized via the phenylpropanoid pathway, where stilbene synthase (STS, EC 2.3.1.95) directly catalyzes the formation of t‐resveratrol (monomeric stilbene). This review discusses the features of using STS genes in genetic engineering and plant biotechnology with the purpose to increase plant resistance to environmental stresses and to modify secondary metabolite production. This article is protected by copyright. All rights reserved

ACS Style

Konstantin V. Kiselev; Alexandra S. Dubrovina. Overexpression of stilbene synthase genes to modulate the properties of plants and plant cell cultures. Biotechnology and Applied Biochemistry 2020, 68, 13 -19.

AMA Style

Konstantin V. Kiselev, Alexandra S. Dubrovina. Overexpression of stilbene synthase genes to modulate the properties of plants and plant cell cultures. Biotechnology and Applied Biochemistry. 2020; 68 (1):13-19.

Chicago/Turabian Style

Konstantin V. Kiselev; Alexandra S. Dubrovina. 2020. "Overexpression of stilbene synthase genes to modulate the properties of plants and plant cell cultures." Biotechnology and Applied Biochemistry 68, no. 1: 13-19.

Journal article
Published: 13 December 2019 in Plants
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Plant calmodulins (CaMs) and calmodulin-like proteins (CMLs) are important plant Ca2+-binding proteins that sense and decode changes in the intracellular Ca2+ concentration arising in response to environmental stimuli. Protein Ca2+ sensors are presented by complex gene families in plants and perform diverse biological functions. In this study, we cloned, sequenced, and characterized three CaM and 54 CML mRNA transcripts of Vitis amurensis Rupr., a wild-growing grapevine with a remarkable stress tolerance. Using real-time quantitative RT-PCR, we analyzed transcript abundance of the identified VaCaMs and VaCMLs in response to water deficit, high salinity, high mannitol, cold and heat stresses. Expression of VaCaMs and 32 VaCMLs actively responded to the abiotic stresses and exhibited both positive and negative regulation patterns. Other VaCML members showed slight transcriptional regulation, remained essentially unresponsive or responded only after one time interval of the treatments. The substantial alterations in the VaCaM and VaCML transcript levels revealed their involvement in the adaptation of wild-growing grapevine to environmental stresses.

ACS Style

Alexandra S. Dubrovina; Olga A. Aleynova; Zlata V. Ogneva; Andrey R. Suprun; Alexey A. Ananev; Konstantin V. Kiselev. The Effect of Abiotic Stress Conditions on Expression of Calmodulin (CaM) and Calmodulin-Like (CML) Genes in Wild-Growing Grapevine Vitis amurensis. Plants 2019, 8, 602 .

AMA Style

Alexandra S. Dubrovina, Olga A. Aleynova, Zlata V. Ogneva, Andrey R. Suprun, Alexey A. Ananev, Konstantin V. Kiselev. The Effect of Abiotic Stress Conditions on Expression of Calmodulin (CaM) and Calmodulin-Like (CML) Genes in Wild-Growing Grapevine Vitis amurensis. Plants. 2019; 8 (12):602.

Chicago/Turabian Style

Alexandra S. Dubrovina; Olga A. Aleynova; Zlata V. Ogneva; Andrey R. Suprun; Alexey A. Ananev; Konstantin V. Kiselev. 2019. "The Effect of Abiotic Stress Conditions on Expression of Calmodulin (CaM) and Calmodulin-Like (CML) Genes in Wild-Growing Grapevine Vitis amurensis." Plants 8, no. 12: 602.

Review
Published: 08 May 2019 in International Journal of Molecular Sciences
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Recent investigations documented that plants can uptake and process externally applied double-stranded RNAs (dsRNAs), hairpin RNAs (hpRNAs), and small interfering RNAs (siRNAs) designed to silence important genes of plant pathogenic viruses, fungi, or insects. The exogenously applied RNAs spread locally and systemically, move into the pathogens, and induce RNA interference-mediated plant pathogen resistance. Recent findings also provided examples of plant transgene and endogene post-transcriptional down-regulation by complementary dsRNAs or siRNAs applied onto the plant surfaces. Understanding the plant perception and processing of exogenous RNAs could result in the development of novel biotechnological approaches for crop protection. This review summarizes and discusses the emerging studies reporting on exogenous RNA applications for down-regulation of essential fungal and insect genes, targeting of plant viruses, or suppression of plant transgenes and endogenes for increased resistance and changed phenotypes. We also analyze the current understanding of dsRNA uptake mechanisms and dsRNA stability in plant environments.

ACS Style

Alexandra S. Dubrovina; Konstantin V. Kiselev. Exogenous RNAs for Gene Regulation and Plant Resistance. International Journal of Molecular Sciences 2019, 20, 2282 .

AMA Style

Alexandra S. Dubrovina, Konstantin V. Kiselev. Exogenous RNAs for Gene Regulation and Plant Resistance. International Journal of Molecular Sciences. 2019; 20 (9):2282.

Chicago/Turabian Style

Alexandra S. Dubrovina; Konstantin V. Kiselev. 2019. "Exogenous RNAs for Gene Regulation and Plant Resistance." International Journal of Molecular Sciences 20, no. 9: 2282.

Journal article
Published: 29 March 2019 in International Journal of Molecular Sciences
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Recent investigations show that exogenously applied small interfering RNAs (siRNA) and long double-stranded RNA (dsRNA) precursors can be taken up and translocated in plants to induce RNA interference (RNAi) in the plant or in its fungal pathogen. The question of whether genes in the plant genome can undergo suppression as a result of exogenous RNA application on plant surface is almost unexplored. This study analyzed whether it is possible to influence transcript levels of transgenes, as more prone sequences to silencing, in Arabidopsis genome by direct exogenous application of target long dsRNAs. The data revealed that in vitro synthesized dsRNAs designed to target the gene coding regions of enhanced green fluorescent protein (EGFP) or neomycin phosphotransferase II (NPTII) suppressed their transcript levels in Arabidopsis. The fact that, simple exogenous application of polynucleotides can affect mRNA levels of plant transgenes, opens new opportunities for the development of new scientific techniques and crop improvement strategies.

ACS Style

Alexandra S. Dubrovina; Olga A. Aleynova; Alexander V. Kalachev; Andrey Suprun; Zlata V. Ogneva; Konstantin V. Kiselev. Induction of Transgene Suppression in Plants via External Application of Synthetic dsRNA. International Journal of Molecular Sciences 2019, 20, 1585 .

AMA Style

Alexandra S. Dubrovina, Olga A. Aleynova, Alexander V. Kalachev, Andrey Suprun, Zlata V. Ogneva, Konstantin V. Kiselev. Induction of Transgene Suppression in Plants via External Application of Synthetic dsRNA. International Journal of Molecular Sciences. 2019; 20 (7):1585.

Chicago/Turabian Style

Alexandra S. Dubrovina; Olga A. Aleynova; Alexander V. Kalachev; Andrey Suprun; Zlata V. Ogneva; Konstantin V. Kiselev. 2019. "Induction of Transgene Suppression in Plants via External Application of Synthetic dsRNA." International Journal of Molecular Sciences 20, no. 7: 1585.

Review
Published: 06 July 2017 in Planta
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This review analyzes the advances in understanding the natural signaling pathways and environmental factors regulating stilbene biosynthesis. We also discuss the studies reporting on stilbene content and repertoire in plants. Stilbenes, including the most-studied stilbene resveratrol, are a family of phenolic plant secondary metabolites that have been the subject of intensive research due to their valuable pharmaceutical effects and contribution to plant disease resistance. Understanding the natural mechanisms regulating stilbene biosynthesis in plants could be useful for both the development of new plant protection strategies and for commercial stilbene production. In this review, we focus on the environmental factors and cell signaling pathways regulating stilbene biosynthesis in plants and make a comparison with the regulation of flavonoid biosynthesis. This review also analyzes the recent data on stilbene biosynthetic genes and summarizes the available studies reporting on both stilbene content and stilbene composition in different plant families.

ACS Style

A. S. Dubrovina; K. V. Kiselev. Regulation of stilbene biosynthesis in plants. Planta 2017, 246, 597 -623.

AMA Style

A. S. Dubrovina, K. V. Kiselev. Regulation of stilbene biosynthesis in plants. Planta. 2017; 246 (4):597-623.

Chicago/Turabian Style

A. S. Dubrovina; K. V. Kiselev. 2017. "Regulation of stilbene biosynthesis in plants." Planta 246, no. 4: 597-623.

Original paper
Published: 19 December 2016 in Plant Growth Regulation
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Ca2+-dependent protein kinases (CDPKs or CPKs) are essential primary sensors of Ca2+ in plants and are known to play important roles in plant abiotic and biotic stress responses. Vitis amurensis is a wild grapevine species with a high level of cold and disease resistance. It has previously been shown that transcription of 10 CDPK genes of V. amurensis was elevated under salt, desiccation, high mannitol, cold, and heat stress conditions. Expression of VaCPK29 was induced under high and low temperatures, water deficit, and high mannitol stress in plant cuttings of V. amurensis. The present study revealed that the callus cell cultures of V. amurensis and soil-grown plants of Arabidopsis thaliana overexpressing VaCPK29 exhibited higher tolerance to heat and high mannitol stress in comparison with the control transformed with the empty vector. Cold, salt, and drought stress tolerance of the transgenic V. amurensis calli and A. thaliana plants was comparable to that of the controls. The stress-responsive genes AtDREB1A, AtDREB2A, AtRD29A, AtRD29B, and AtABF3 were up-regulated in the VaCPK29-overexpressing A. thaliana plants under heat stress. Taken together, the data indicate that the VaCPK29 gene may act as a positive regulator in the grapevine response to heat and osmotic stresses.

ACS Style

Alexandra S. Dubrovina; Konstantin V. Kiselev; Valeriya S. Khristenko; Olga A. Aleynova. The calcium-dependent protein kinase gene VaCPK29 is involved in grapevine responses to heat and osmotic stresses. Plant Growth Regulation 2016, 82, 79 -89.

AMA Style

Alexandra S. Dubrovina, Konstantin V. Kiselev, Valeriya S. Khristenko, Olga A. Aleynova. The calcium-dependent protein kinase gene VaCPK29 is involved in grapevine responses to heat and osmotic stresses. Plant Growth Regulation. 2016; 82 (1):79-89.

Chicago/Turabian Style

Alexandra S. Dubrovina; Konstantin V. Kiselev; Valeriya S. Khristenko; Olga A. Aleynova. 2016. "The calcium-dependent protein kinase gene VaCPK29 is involved in grapevine responses to heat and osmotic stresses." Plant Growth Regulation 82, no. 1: 79-89.

Journal article
Published: 01 November 2016 in Phytochemistry
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Stilbenes are valuable phenolic compounds that are synthesized in plants via the phenylpropanoid pathway where stilbene synthase (STS) directly catalyzes resveratrol or pinosylvin formation. Currently, there is a lack of information about the stilbene biosynthetic pathway in spruce (Picea). Resveratrol and piceatannol derivatives have been detected in the spruce bark, needles, and roots. We analyzed seasonal variation in stilbene spectrum and content in the needles of different ages of one tree of spruce Picea jezoensis. HPLC analysis revealed the presence of nine stilbenes: t- and cis-astringin, t- and cis-piceid, t- and cis-isorhapontin, and t-piceatannol were present in amounts of 0.01-6.07 mg/g of dry weight (DW), while t-isorhapontigenin and t-resveratrol were present in traces (0.001-0.312 μg/g DW). T-astringin prevailed over other stilbenoid compounds (66-86% of all stilbenes). The highest total stilbene content was detected in one-year-old needles collected in the autumn and spring (5.4-7.77 mg/g DW). We previously cloned and sequenced full-length cDNAs of the four STS transcripts (PjSTS1a, PjSTS1b, PjSTS2, and PjSTS3) of P. jezoensis. This study presents a detailed analysis of seasonal variations in PjSTS1a, 1b, 2, and 3 transcript levels in the needles of P. jezoensis of different ages using qRT-PCR. PjSTS1a and PjSTS1b transcription was higher in the needles collected in the autumn, spring, or summer than in the winter. PjSTS2 was actively transcribed in the needles of all ages collected in the winter, spring, and summer. PjSTS3 expression did not significantly change during the year and did not depend on the age of the needles. Therefore, the data show that high levels of the stilbene glucosides and PjSTS expression are present in the needles of P. jezoensis.

ACS Style

K.V. Kiselev; V.P. Grigorchuk; Z.V. Ogneva; Andrey Suprun; Alexandra Dubrovina. Stilbene biosynthesis in the needles of spruce Picea jezoensis. Phytochemistry 2016, 131, 57 -67.

AMA Style

K.V. Kiselev, V.P. Grigorchuk, Z.V. Ogneva, Andrey Suprun, Alexandra Dubrovina. Stilbene biosynthesis in the needles of spruce Picea jezoensis. Phytochemistry. 2016; 131 ():57-67.

Chicago/Turabian Style

K.V. Kiselev; V.P. Grigorchuk; Z.V. Ogneva; Andrey Suprun; Alexandra Dubrovina. 2016. "Stilbene biosynthesis in the needles of spruce Picea jezoensis." Phytochemistry 131, no. : 57-67.

Original article
Published: 29 September 2016 in Planta
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We detected and quantified six stilbenes (cis-piceid,t-piceid,t-ε-viniferin,cis-ε-viniferin,t-resveratrol, andt-δ-viniferin) in the leaves, petioles, berry skins, and seeds of wild-growingVitis amurensis. The highest content of stilbenes and expression of stilbene biosynthesis genes were in the probes collected in the autumn and after ultraviolet elicitation. Stilbenes, including the best-studied stilbene resveratrol, are known to display valuable bioactivities and protect plants against various pathogens. There is a lack of studies on stilbene quantities and spectrum combined with an analysis of the stilbene biosynthesis pathway gene expression in Vitaceae species, despite grapevine is an important source of stilbenes. This study presents an analysis of stilbene spectrum, stilbene content, and expression of stilbene biosynthesis genes both in natural conditions and after ultraviolet (UV-C) elicitation in the leaves, petioles, berry skins, and seeds of wild-growing Vitis amurensis, a highly stress-tolerant plant species. Using HPLC analysis, we detected six main stilbenes: cis-piceid (up to 0.257 mg/g of dry weight (DW) of plant material), t-piceid (up to 0.055 mg/g DW), t-ε-viniferin (up to 0.122 mg/g DW), cis-ε-viniferin (up to 0.031 mg/g DW), t-resveratrol (from 0.004 to 0.121 mg/g DW), and t-δ-viniferin (up to 0.019 mg/g DW). The stilbenes were actively synthesized in the leaves (total stilbenes 0.39 mg/g DW) and berry skins (total stilbenes 0.249 mg/g DW) of V. amurensis collected in the autumn. qRT-PCR revealed that the stilbene synthase (STS), resveratrol O-glucosyltransferase (Glu1), and polyphenol oxidase (PPO1) genes were actively expressed in the analyzed tissues. The resveratrol methyltransferase (Romt1) gene, which is known to catalyze biosynthesis of pterostilbene, was also expressed, but no pterostilbene has been detected in V. amurensis. The content of all detected stilbenes and expression of stilbene biosynthesis genes increased after UV-C treatment, except for Romt1. The data are important for understanding the stilbene biosynthesis in grapevine.

ACS Style

Konstantin V. Kiselev; Olga A. Aleynova; Valeria P. Grigorchuk; Alexandra Dubrovina. Stilbene accumulation and expression of stilbene biosynthesis pathway genes in wild grapevine Vitis amurensis Rupr. Planta 2016, 245, 151 -159.

AMA Style

Konstantin V. Kiselev, Olga A. Aleynova, Valeria P. Grigorchuk, Alexandra Dubrovina. Stilbene accumulation and expression of stilbene biosynthesis pathway genes in wild grapevine Vitis amurensis Rupr. Planta. 2016; 245 (1):151-159.

Chicago/Turabian Style

Konstantin V. Kiselev; Olga A. Aleynova; Valeria P. Grigorchuk; Alexandra Dubrovina. 2016. "Stilbene accumulation and expression of stilbene biosynthesis pathway genes in wild grapevine Vitis amurensis Rupr." Planta 245, no. 1: 151-159.

Original paper
Published: 16 May 2016 in Biologia plantarum
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Little is known about the contributions of DNA methylation/demethylation to plant aging and senescence. We used Arabidopsis thaliana to study how increasing age of an annual plant species influences DNA methylation. Based on methylation-sensitive DNA fragmentation assay, it could be concluded that aging A. thaliana was accompanied by DNA demethylation. Bisulfite sequencing reveals that cytosine methylation within the Actin2 3’ untranslated region and internal transcribed spacer with 5.8S rRNA (ITS1-5.8SrRNA-ITS2) DNA regions decreased with A. thaliana growth and aging. We show that transcription of methyltransferase genes, chromomethyltransferase AtCMT3 and methyltransferse AtMETI, significantly decreased during development and aging of the A. thaliana plants, whereas expression of demethylase genes - repressor of silencing AtROS1, demeter AtDME, and demeter-like AtDML2 and AtDML3 - increased at least at some stages of plant development. The data obtained in the present study suggest that plant DNA regions may undergo demethylation during plant aging via reduction of DNA methylation processes and activation of active DNA demethylation.

ACS Style

Z. V. Ogneva; A. S. Dubrovina; K. V. Kiselev. Age-associated alterations in DNA methylation and expression of methyltransferase and demethylase genes in Arabidopsis thaliana. Biologia plantarum 2016, 60, 628 -634.

AMA Style

Z. V. Ogneva, A. S. Dubrovina, K. V. Kiselev. Age-associated alterations in DNA methylation and expression of methyltransferase and demethylase genes in Arabidopsis thaliana. Biologia plantarum. 2016; 60 (4):628-634.

Chicago/Turabian Style

Z. V. Ogneva; A. S. Dubrovina; K. V. Kiselev. 2016. "Age-associated alterations in DNA methylation and expression of methyltransferase and demethylase genes in Arabidopsis thaliana." Biologia plantarum 60, no. 4: 628-634.

Journal article
Published: 24 February 2016 in Acta Physiologiae Plantarum
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Calcium-dependent protein kinases (CDPKs or CPKs) are important Ca2+ sensors in plants implicated in plant abiotic and biotic stress responses, e.g. in drought-induced stomata closure or pathogen-induced stilbene production. We previously found that the VaCPK9 gene of Vitis amurensis was subjected to unusual post-transcriptional processing mediated by canonical splice sites leading to production of three mRNA isoforms that lacked important catalytic kinase subdomains. We also detected numerous unusual VaCPK9 and VaCPK3a cDNAs possessing short direct repeated sequences instead of canonical splice sites and suggested that these cDNAs were false alternative transcripts generated by reverse transcriptase template switching in vitro. The present study has been attempted to determine if the full-length and spliced CDPKs play a role in the grapevine growth regulation, stilbene biosynthesis, and abiotic stress adaptation. We generated transgenic V. amurensis callus cultures overexpressing different true and false VaCPK9 and VaCPK3a transcripts under control of the CaMV 35S promoter. Overexpression of the full-length VaCPK9 and VaCPK3a transcripts slightly increased biomass accumulation but did not affect resistance to abiotic stresses and biosynthesis of resveratrol (3,5,4′-trihydroxy-trans-stilbene) in the calli. Overexpression of the true alternative VaCPK9SF1, VaCPK9SF2, and VaCPK9SF3 transcripts considerably improved growth of the transgenic calli, while overexpression of the false alternative VaCPK3aSF1, VaCPK3aSF2, and VaCPK3aSF3 transcripts did not considerably influence it. The results show that the VaCPK9 and VaCPK3a genes function in growth regulation of V. amurensis but not in stilbene biosynthesis and abiotic stress tolerance. Alternative splicing of VaCPK9 may serve to reinforce the positive effect of VaCPK9 expression on biomass accumulation.

ACS Style

Alexandra Dubrovina; O. A. Aleynova; K. V. Kiselev. Influence of overexpression of the true and false alternative transcripts of calcium-dependent protein kinase CPK9 and CPK3a genes on the growth, stress tolerance, and resveratrol content in Vitis amurensis cell cultures. Acta Physiologiae Plantarum 2016, 38, 78 .

AMA Style

Alexandra Dubrovina, O. A. Aleynova, K. V. Kiselev. Influence of overexpression of the true and false alternative transcripts of calcium-dependent protein kinase CPK9 and CPK3a genes on the growth, stress tolerance, and resveratrol content in Vitis amurensis cell cultures. Acta Physiologiae Plantarum. 2016; 38 (3):78.

Chicago/Turabian Style

Alexandra Dubrovina; O. A. Aleynova; K. V. Kiselev. 2016. "Influence of overexpression of the true and false alternative transcripts of calcium-dependent protein kinase CPK9 and CPK3a genes on the growth, stress tolerance, and resveratrol content in Vitis amurensis cell cultures." Acta Physiologiae Plantarum 38, no. 3: 78.

Original paper
Published: 09 February 2016 in Plant Cell, Tissue and Organ Culture
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Stilbenes (resveratrol, piceid, viniferins, etc.) are acting as phytoalexins and possess valuable biologically active properties. To improve stilbene production in grapevine, the effect of VaSTS1, VaSTS2, and VaSTS7 gene overexpression on stilbene production in callus cell cultures of wild grapevine Vitis amurensis Rupr. were accessed. Overexpression of VaSTS1 increased the total content of stilbenes by 1.2–2.1 times up to 0.36 mg per g of the dry weight (mg g−1 dry wt) or 0.036 % of dry wt, while overexpression of VaSTS2 and VaSTS7 increased the content of stilbenes by 1.5–3.5 and 2.5–2.8 times (up to 0.48 and 0.60 mg g−1 dry wt), respectively. P-coumaric acid (CA), a precursor of stilbenes, induced stilbene content and production in all cell lines with the highest positive effect on stilbene accumulation being observed in the STS2- and STS7-transgenic cell lines. After the CA feeding, the stilbene content increased up to 1.09–1.16 mg g−1 dry wt in the STS2- and one STS7-transgenic cell lines and only up to 0.35 and 0.40 mg g−1 dry wt in the control and STS1-transgenic cell lines, respectively. Thus, overexpression of VaSTS1, VaSTS2, and VaSTS7 genes had a positive effect on stilbene accumulation in the cells of V. amurensis cultivated in vitro. T-resveratrol and t-resveratrol glycosides were the prevalent stilbenes synthesized in the STS-transgenic calli before and after CA feeding. The results obtained are innovative for plant biotechnology, since stilbene production in plant cell cultures overexpressing a STS gene has not been studied until now.

ACS Style

O. A. Aleynova; V. P. Grigorchuk; A. S. Dubrovina; V. G. Rybin; K. V. Kiselev. Stilbene accumulation in cell cultures of Vitis amurensis Rupr. overexpressing VaSTS1, VaSTS2, and VaSTS7 genes. Plant Cell, Tissue and Organ Culture 2016, 125, 329 -339.

AMA Style

O. A. Aleynova, V. P. Grigorchuk, A. S. Dubrovina, V. G. Rybin, K. V. Kiselev. Stilbene accumulation in cell cultures of Vitis amurensis Rupr. overexpressing VaSTS1, VaSTS2, and VaSTS7 genes. Plant Cell, Tissue and Organ Culture. 2016; 125 (2):329-339.

Chicago/Turabian Style

O. A. Aleynova; V. P. Grigorchuk; A. S. Dubrovina; V. G. Rybin; K. V. Kiselev. 2016. "Stilbene accumulation in cell cultures of Vitis amurensis Rupr. overexpressing VaSTS1, VaSTS2, and VaSTS7 genes." Plant Cell, Tissue and Organ Culture 125, no. 2: 329-339.

Journal article
Published: 30 September 2015 in Plant Cell, Tissue and Organ Culture (PCTOC)
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Ca2+-dependent protein kinases (CDPKs) are known to play important roles in signal transduction in response to various abiotic stresses that limit crop yields. Vitis amurensis is a wild grape species that possesses a high adaptive potential and displays a high resistance to environmental stresses, such as low temperatures or microbial pathogens. The roles of most grape CDPKs in the adaptation to various stresses have not yet been uncovered. A recent CDPK gene expression analysis revealed that expression of VaCPK21 gene was significantly up-regulated under salt stress and, to a less degree, under high mannitol and high temperature stresses in V. amurensis plant cuttings. In this study, the effects of overexpressing the VaCPK21 gene in callus cell lines of V. amurensis and transgenic plants of A. thaliana on their responses to abiotic stresses were investigated. The VaCPK21-overexpressing cell cultures of V. amurensis, soil-grown plants of A. thaliana, and seedlings of A. thaliana exhibited higher tolerance to salt stress in comparison with the controls transformed with the “empty” vector. Heat, cold, and drought stress resistance of the transgenic V. amurensis calli and A. thaliana plants was comparable to that of the controls. Under salt stress conditions, the stress-related genes AtCOR15, AtCOR47, AtCAT1, AtCSD1, AtNHX1, AtKIN1, AtRD26, and AtRD29B demonstrated strong up-regulation in the VaCPK21-transgenic but not in the control Arabidopsis plants. Together, the results indicate that VaCPK21 may act as a positive regulator involved in the grapevine response to salt stress.

ACS Style

Alexandra S. Dubrovina; Konstantin V. Kiselev; Valeriya S. Khristenko; Olga A. Aleynova. VaCPK21, a calcium-dependent protein kinase gene of wild grapevine Vitis amurensis Rupr., is involved in grape response to salt stress. Plant Cell, Tissue and Organ Culture (PCTOC) 2015, 124, 137 -150.

AMA Style

Alexandra S. Dubrovina, Konstantin V. Kiselev, Valeriya S. Khristenko, Olga A. Aleynova. VaCPK21, a calcium-dependent protein kinase gene of wild grapevine Vitis amurensis Rupr., is involved in grape response to salt stress. Plant Cell, Tissue and Organ Culture (PCTOC). 2015; 124 (1):137-150.

Chicago/Turabian Style

Alexandra S. Dubrovina; Konstantin V. Kiselev; Valeriya S. Khristenko; Olga A. Aleynova. 2015. "VaCPK21, a calcium-dependent protein kinase gene of wild grapevine Vitis amurensis Rupr., is involved in grape response to salt stress." Plant Cell, Tissue and Organ Culture (PCTOC) 124, no. 1: 137-150.