Journal articles: 'Grapevine cell culture'

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Sák, Martin, Ivana Dokupilová, Daniel Mihálik, Jana Lakatošová, Marcela Gubišová, and Ján Kraic. "Elicitation Phenolic Compounds in Cell Culture of Vitis vinifera L. by Phaeomoniella chlamydospora." Nova Biotechnologica et Chimica 13, no. 2 (December 1, 2014): 162–71. http://dx.doi.org/10.1515/nbec-2015-0006.

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Abstract The in vitro cell cultures of Vitis vinifera L. cv. St. Laurent were treated with two elicitors - synthetic methyl jasmonate and natural, prepared from grapevine plant infected with the Phaeomoniella chlamydospora, the agent causing the Esca disease of grapevine. Efficiency of phenolic compounds production after elicitation of cell culture was analysed immediately after treatment (15 min, 30 min, 60 min) and later (after 24, 48, and 72 hours). The cell growth and content of phenolic compounds (+)-catechin, (-)-epicatechin, p-coumaric acid, syringaldehyde, rutin, vanillic acid, and trans-resveratrol were analysed in cultivated cells as well as in cultivation medium. Pch-treatment increased production of total polyphenols the most significantly 15 min after the elicitation and in optimal time was 2.86 times higher than in nonelicited culture and 1.44 times higher than in MeJa induced cell culture.

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Čarná, Mária, Vladimír Repka, and Ernest Šturdí. "Proteomic Insight Into the Molecular Principles of Grapevine Habituation." Agriculture (Polnohospodárstvo) 57, no. 4 (December 1, 2011): 129–36. http://dx.doi.org/10.2478/v10207-011-0013-0.

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Proteomic Insight Into the Molecular Principles of Grapevine Habituation Two-dimensional gel electrophoresis coupled to protein microarray analysis was used to examine, for the first time, the molecular mechanisms of grapevine (Vitis vinifera L., cv. Limberger) habituation. The examination of 2-D maps derived from control and habituated cell culture revealed the presence of 55 protein spots displaying a differential expression pattern. These facts have provide a molecular evidence suggesting that the habituated cells can be used as a model for study of cell differentiation and plant defense mechanisms. Cell death, extra-cellular alkalinization and expression of genes responsible for the formation of the defense-related proteins were analyzed in suspension cultures with hormonal autonomy (habituation). Results obtained using habituated grapevine cells compared with non-habituated cells were different and strongly depended on the concentration of elicitor applied.

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Carvalho, Ana, Christina Crisóstomo, Fernanda Leal, and José Lima-Brito. "Morphological and Cytogenetic Responses of In Vitro-Grown Grapevine (Vitis vinifera L.) Plants from “Touriga Franca”, “Touriga Nacional” and “Viosinho” Varieties Under Water Stress." Stresses 4, no. 4 (October 24, 2024): 685–98. http://dx.doi.org/10.3390/stresses4040044.

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According to the climate projections, drought will increase in frequency and severity. Since water stress (WS) impacts a grapevine’s physiology and yield negatively, the evaluation and selection of tolerant genotypes are needed. To analyse the WS effects on the morphology and cell division of three grapevines (Vitis vinifera L.) varieties, “Touriga Franca” (TF), “Touriga Nacional” (TN) and “Viosinho” (VS), in vitro-grown plants were exposed to 10% polyethylene glycol 6000 (PEG) (−0.4 MPa) or 20% PEG (−0.8 MPa), incorporated in the culture medium, for four weeks. Control plants were kept in culture media without PEG. The VS and TN plants showed the highest mean numbers of nodes, shoots and leaves and average mitotic indexes under 20% PEG. The TF and TN plants showed the lowest frequencies of mitotic anomalies under 10% PEG. The VS plant growth was less affected by WS, but TF and TN presented more regular mitosis under moderate WS. Globally, in vitro culture constitutes a cost-effective experimental system for studying grapevine responses to WS and the preliminary selection of resilient genotypes. These approaches could be applied to study plant responses to other abiotic stresses based on additional evaluation techniques (e.g., transcriptional analyses or genome-wide association studies).

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Aleynova, Olga A., Andrey R. Suprun, Nikolay N. Nityagovsky, Alexandra S. Dubrovina, and Konstantin V. 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 10, no. 7 (June 23, 2021): 1276. http://dx.doi.org/10.3390/plants10071276.

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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.

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Hurtado-Gaitán, Elías, Susana Sellés-Marchart, James Hartwell, Maria José Martínez-Esteso, and Roque Bru-Martínez. "Down-Regulation of Phosphoenolpyruvate Carboxylase Kinase in Grapevine Cell Cultures and Leaves Is Linked to Enhanced Resveratrol Biosynthesis." Biomolecules 11, no. 11 (November 5, 2021): 1641. http://dx.doi.org/10.3390/biom11111641.

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In grapevine, trans-Resveratrol (tR) is produced as a defence mechanism against stress or infection. tR is also considered to be important for human health, which increases its interest to the scientific community. Transcriptomic analysis in grapevine cell cultures treated with the defence response elicitor methyl-β-cyclodextrin (CD) revealed that both copies of PHOSPHOENOLPYRUVATE CARBOXYLASE KINASE (PPCK) were down-regulated significantly. A role for PPCK in the defence response pathway has not been proposed previously. We therefore analysed the control of PPCK transcript levels in grapevine cell cultures and leaves elicited with CD. Moreover, phosphoenolpyruvate carboxylase (PPC), stilbene synthase (STS), and the transcription factors MYB14 and WRKY24, which are involved in the activation of STS transcription, were also analysed by RT-qPCR. The results revealed that under CD elicitation conditions PPCK down-regulation, increased stilbene production and loss of PPC activity occurs in both tissues. Moreover, STS transcripts were co-induced with MYB14 and WRKY24 in cell cultures and leaves. These genes have not previously been reported to respond to CD in grape leaves. Our findings thus support the hypothesis that PPCK is involved in diverting metabolism towards stilbene biosynthesis, both for in vitro cell culture and whole leaves. We thus provide new evidence for PEP being redirected between primary and secondary metabolism to support tR production and the stress response.

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Zhao, Liang, Shuangmei You, Hui Zou, and Xin Guan. "Transcriptome Analysis and Cell Morphology of Vitis rupestris Cells to Botryosphaeria Dieback Pathogen Diplodia seriata." Genes 12, no. 2 (January 27, 2021): 179. http://dx.doi.org/10.3390/genes12020179.

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Diplodia seriata, one of the major causal agents of Botryosphaeria dieback, spreads worldwide, causing cankers, leaf spots and fruit black rot in grapevine. Vitis rupestris is an American wild grapevine widely used for resistance and rootstock breeding and was found to be highly resistant to Botryosphaeria dieback. The defense responses of V. rupestris to D. seriata 98.1 were analyzed by RNA-seq in this study. There were 1365 differentially expressed genes (DEGs) annotated with Gene Ontology (GO) and enriched by the Kyoto Encyclopedia of Genes and Genomes (KEGG) database. The DEGs could be allocated to the flavonoid biosynthesis pathway and the plant–pathogen interaction pathway. Among them, 53 DEGs were transcription factors (TFs). The expression levels of 12 genes were further verified by real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). The aggregation of proteins on the plasma membrane, formation variations in the cytoskeleton and plasmodesmata and hormone regulations revealed a declined physiological status in V. rupestris suspension cells after incubation with the culture filtrates of D. seriata 98.1. This study provides insights into the molecular mechanisms in grapevine cells’ response to D. seriata 98.1, which will be valuable for the control of Botryosphaeria dieback.

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Sák, Martin, Ivana Dokupilová, Šarlota Kaňuková, Michaela Mrkvová, Daniel Mihálik, Pavol Hauptvogel, and Ján Kraic. "Biotic and Abiotic Elicitors of Stilbenes Production in Vitis vinifera L. Cell Culture." Plants 10, no. 3 (March 5, 2021): 490. http://dx.doi.org/10.3390/plants10030490.

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The in vitro cell cultures derived from the grapevine (Vitis vinifera L.) have been used for the production of stilbenes treated with different biotic and abiotic elicitors. The red-grape cultivar Váh has been elicited by natural cellulose from Trichoderma viride, the cell wall homogenate from Fusarium oxysporum and synthetic jasmonates. The sodium-orthovanadate, known as an inhibitor of hypersensitive necrotic response in treated plant cells able to enhance production and release of secondary metabolite into the cultivation medium, was used as an abiotic elicitor. Growth of cells and the content of phenolic compounds trans-resveratrol, trans-piceid, δ-viniferin, and ɛ-viniferin, were analyzed in grapevine cells treated by individual elicitors. The highest accumulation of analyzed individual stilbenes, except of trans-piceid has been observed after treatment with the cell wall homogenate from F. oxysporum. Maximum production of trans-resveratrol, δ- and ɛ-viniferins was triggered by treatment with cellulase from T. viride. The accumulation of trans-piceid in cell cultures elicited by this cellulase revealed exactly the opposite effect, with almost three times higher production of trans-resveratrol than that of trans-piceid. This study suggested that both used fungal elicitors can enhance production more effectively than commonly used jasmonates.

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Brehm, Ilka, Regina Preisig-Müller, and Helmut Kindl. "Grapevine Protoplasts as a Transient Expression System for Comparison of Stilbene Synthase Genes Containing cGMP-Responsive Promoter Elements." Zeitschrift für Naturforschung C 54, no. 3-4 (April 1, 1999): 220–29. http://dx.doi.org/10.1515/znc-1999-3-412.

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Abstract A method for preparing elicitor-responsive protoplasts from grapevine cells kept in suspension culture was established. The protoplasts were employed in order to perform transient gene expression experiments produced by externally added plasmids. Using the gene coding for bacterial β-glucuronidase as the reporter gene, the transient expression under the control of various promoters of stilbene synthase genes were analyzed. The elicitor-responsiveness of promoters from grapevine genes and heterologous promoters were assayed: the grapevine stilbene synthase gene VST-1 and pine stilbene synthase genes PST-1, PST-2 and PST-3. Compared to the expression effected by the cauliflower mosaic virus 35S RNA-promoter, the stilbene synthase promoters caused a 2-5-fold increase in GUS-activity. Incubation of transformed protoplasts with fungal cell wall further stimulated the stilbene synthase promoters but not the 35S RNA-promoter. An even more pronounced differentiation between the promoters was observed when cGMP was included in the transient expression assays. Instead of treating transformed protoplasts with fungal cell wall we administered simultaneously cGMP and the plasmid to be tested. The cGMP-responsive increase was (a) specific concerning the nucleotide applied, (b) characteristic of grapevine protoplasts, and (c) not seen with shortened promoter-GUS constructs or GUS under the control of the 35S RNA-promoter. The highest cGMP-dependent reponse to stress was shown by the promoter of the grapevine stilbene synthase gene VST-1.

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Fujita, Keiko, Yoshinao Aoki, and Shunji Suzuki. "Antidiabetic effects of novel cell culture established from grapevine, Vitis vinifera cv. Koshu." Cytotechnology 70, no. 3 (March 15, 2018): 993–99. http://dx.doi.org/10.1007/s10616-018-0203-y.

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Unknown, Unknown. "Elicitor applications to cell suspension culture for production of phenolic compounds in grapevine." Tarım Bilimleri Dergisi 22, no. 1 (2016): 42–53. http://dx.doi.org/10.1501/tarimbil_0000001366.

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Busato, Isabella, Oriana Bertaiola, Silvio Tundo, Chiara Guarnerio, Marco Lucchetta, Luca Sella, Giovanna Pressi, and Francesco Favaron. "A Phytocomplex Obtained from Salvia officinalis by Cell Culture Technology Effectively Controls the Grapevine Downy Mildew Pathogen Plasmopara viticola." Plants 11, no. 20 (October 11, 2022): 2675. http://dx.doi.org/10.3390/plants11202675.

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The negative impact of using conventional fungicides in plant disease protection has increased the interest in safer alternatives such as plant secondary metabolites, generally having a better toxicological profile. However, cultivation conditions and plant material strongly affect the quality and quantity of secondary metabolites obtained from field grown plants, limiting the standardization needed for industrial production. Plant cell culture technology can provide highly homogeneous biomasses with specific chemical characteristics. A phytocomplex with high rosmarinic acid content (10.12% w/w) was obtained from a selected cell line of Salvia officinalis and was tested against the grapevine downy mildew pathogen, Plasmopara viticola. Grapevine leaf discs were sprayed with the phytocomplex at 5 g/L and then inoculated with P. viticola sporangia. Sporulation level on each disc was assessed after 7 days with an image processing software. The phytocomplex reduced by 95% the sporulation level compared to the control and was also more effective than rosmarinic acid alone, used at the same concentration found in the phytocomplex. Persistence of the phytocomplex was also assessed: when applied 5 days before inoculation, it reduced by 90% the sporulation level compared to the control. These results highlight the possibility to take advantage of cell culture techniques to produce safer pesticides with high quality standards.

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Vera-Urbina, Juan Carlos, Susana Sellés-Marchart, Ascensión Martínez-Márquez, María José Martínez-Esteso, María Angeles Pedreño, Jaime Morante-Carriel, and Roque Bru-Martínez. "Factors Affecting the Bioproduction of Resveratrol by Grapevine Cell Cultures under Elicitation." Biomolecules 13, no. 10 (October 16, 2023): 1529. http://dx.doi.org/10.3390/biom13101529.

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Here we present a study of the characterization and optimization of the production of trans-Resveratrol (t-R) in grape (Vitis vinifera cv. Gamay) cell cultures elicited with methyl jasmonate (MeJA) and dimethyl-β-cyclodextrin (DIMEB). The aim of this study was to determine the influence of a number of factors of the grapevine cell culture on t-R production level in 250 mL shaken flasks that would enable the better control of this bioproduction system when it is upscaled to a 2 L stirred bioreactor. The factors included the optimal growth phase for elicitation, the concentration of elicitors and of biomass, the order of addition of elicitors, and the illumination regime and ageing of cells. We found out that the optimal biomass density for the production of t-R was 19% (w/v) with an optimal ratio of 0.5 g DIMEB/g biomass. The most productive concentrations of the elicitors tested were 50 mM DIMEB and 100 µM MeJA, reaching maximum values of 4.18 mg·mL−1 and 16.3 mg·g biomass−1 of t-R concentration and specific production, respectively. We found that the order of elicitor addition matters since, as compared with the simultaneous addition of both elicitors, the addition of MeJA 48 h before DIMEB results in ca. 40% less t-R production, whilst there is no significant difference when MeJA is added 48 h after DIMEB. Upon upscaling, the better conditions tested for t-R production were aeration at 1.7 vol/vol/min without agitation, 24 °C, and 30 g·L−1 sucrose, achieving production rates similar to those obtained in shaken flasks.

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Donnez, David, Kyung-Hee Kim, Sandrine Antoine, Alexandra Conreux, Vincenzo De Luca, Philippe Jeandet, Christophe Clément, and Eric Courot. "Bioproduction of resveratrol and viniferins by an elicited grapevine cell culture in a 2L stirred bioreactor." Process Biochemistry 46, no. 5 (May 2011): 1056–62. http://dx.doi.org/10.1016/j.procbio.2011.01.019.

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Gray, D. J., Z. T. Li, D. L. Hopkins, M. Dutt, S. A. Dhekney, M. M. Van Aman, J. Tattersall, and K. T. Kelley. "Transgenic Grapevines Resistant to Pierce's Disease." HortScience 40, no. 4 (July 2005): 1104D—1105. http://dx.doi.org/10.21273/hortsci.40.4.1104d.

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Pierce's disease (PD), caused by the xylem-limited bacterium Xylella fastidiosa, is endemic to the coastal plain of the southeastern United States. Although native southern grapevines are tolerant to X. fastidiosa, all varieties of Vitisvinifera grown in the region will succumb to PD. Genetic transformation to add disease resistance genes, while not disturbing desirable phenotypic characters, holds promise for expanding the southeastern U.S. grape industry by allowing use of established fruit and wine varieties. We utilize embryogenic cell cultures and Agrobacterium strain EHA105 to refine transformation systems for Vitis species and hybrids. V. vinifera`Thompson Seedless' is employed as a model variety to test various transgenes for disease resistance, since as many as 150 independent transgenic plant lines routinely are produced from 1 g of embryogenic culture material. Transgenic plants are stringently screened for PD resistance in greenhouses by mechanical inoculation with X. fastidiosa. Transgenic plants are compared with both susceptible and resistant control plants by assessing typical PD symptom development and by assaying bacterial populations in xylem sap over time. Using these procedures, nine putative PD resistance genes have been inserted into grapevine and over 900 unique transgenic lines have been evaluated. A range of susceptible-to-resistant responses has been catalogued. Thus far, the best construct for PD resistance contains a grape codon-optimized hybrid lytic peptide gene in a high-performance bi-directional 35S promoter complex. Certain transgenic plant lines containing this construct exhibit better resistance than that of resistant control vines.

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Pathirana, Ranjith, and Francesco Carimi. "Studies on Improving the Efficiency of Somatic Embryogenesis in Grapevine (Vitis vinifera L.) and Optimising Ethyl Methanesulfonate Treatment for Mutation Induction." Plants 12, no. 24 (December 11, 2023): 4126. http://dx.doi.org/10.3390/plants12244126.

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Somatic embryogenesis (SE) has many applications in grapevine biotechnology including micropropagation, eradicating viral infections from infected cultivars, mass production of hypocotyl explants for micrografting, as a continuous source for haploid and doubled haploid plants, and for germplasm conservation. It is so far the only pathway for the genetic modification of grapevines through transformation. The single-cell origin of somatic embryos makes them an ideal explant for mutation breeding as the resulting mutants will be chimera-free. In the present research, two combinations of plant growth regulators and different explants from flower buds at two stages of maturity were tested in regard to the efficiency of callusing and embryo formation from the callus produced in three white grape cultivars. Also, the treatment of somatic embryos with the chemical mutagen ethyl methanesulfonate (EMS) was optimised. Medium 2339 supplemented with β-naphthoxyacetic acid (5 μM) and 6-benzylaminopurine (BAP—9.0 μM) produced significantly more calluses than medium 2337 supplemented with 2,4-dichlorophenoxyacetic acid (4.5 µM) and BAP (8.9 µM) in all explants. The calluses produced on medium 2337 were harder and more granular and produced more SEs. Although the stage of the maturity of floral bud did not have a significant effect on the callusing of the explants, calluses produced from immature floral bud explants in the premeiotic stage produced significantly more SEs than those from more mature floral buds. Overall, immature ovaries and cut floral buds exposing the cut ends of filaments, style, etc., tested for the first time in grapevine SE, produced the highest percentage of embryogenic calluses. It is much more efficient to cut the floral bud and culture than previously reported explants such as anthers, ovaries, stigmas and styles during the short flowering period when the immature flower buds are available. When the somatic embryos of the three cultivars were incubated for one hour with 0.1% EMS, their germination was reduced by 50%; an ideal treatment considered to obtain a high frequency of mutations for screening. Our research findings will facilitate more efficient SE induction in grapevines and inducing mutations for improving individual traits without altering the genetic background of the cultivar.

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Park, Su Hyun, Yu Jeong Jeong, Sung-Chul Park, Soyoung Kim, Yong-Goo Kim, Gilok Shin, Hyung Jae Jeong, et al. "Highly Efficient Bioconversion of trans-Resveratrol to δ-Viniferin Using Conditioned Medium of Grapevine Callus Suspension Cultures." International Journal of Molecular Sciences 23, no. 8 (April 15, 2022): 4403. http://dx.doi.org/10.3390/ijms23084403.

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δ-Viniferin is a resveratrol dimer that possesses potent antioxidant properties and has attracted attention as an ingredient for cosmetic and nutraceutical products. Enzymatic bioconversion and plant callus and cell suspension cultures can be used to produce stilbenes such as resveratrol and viniferin. Here, δ-viniferin was produced by bioconversion from trans-resveratrol using conditioned medium (CM) of grapevine (Vitis labruscana) callus suspension cultures. The CM converted trans-resveratrol to δ-viniferin immediately after addition of hydrogen peroxide (H2O2). Peroxidase activity and bioconversion efficiency in CM increased with increasing culture time. Optimized δ-viniferin production conditions were determined regarding H2O2 concentration, incubation time, temperature, and pH. Maximum bioconversion efficiency reached 64% under the optimized conditions (pH 6.0, 60 °C, 30 min incubation time, 6.8 mM H2O2). In addition, in vitro bioconversion of trans-resveratrol was investigated using CM of different callus suspension cultures, showing that addition of trans-resveratrol and H2O2 to the CM led to production of δ-viniferin via extracellular peroxidase-mediated oxidative coupling of two molecules of trans-resveratrol. We thus propose a simple and low-cost method of δ-viniferin production from trans-resveratrol using CM of plant callus suspension cultures, which may constitute an alternative approach for in vitro bioconversion of valuable molecules.

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Katsirdakis, K. C., and K. A. Roubelakis-Angelakis. "Modified culture conditions for increased viability and cell wall synthesis in grapevine (Vitis vinifera L. cv. Sultanina) leaf protoplasts." Plant Cell, Tissue and Organ Culture 28, no. 3 (March 1992): 255–60. http://dx.doi.org/10.1007/bf00036121.

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Salvatore, Maria Michela, Carina Félix, Fernanda Lima, Vanessa Ferreira, Ana Sofia Duarte, Francesco Salvatore, Artur Alves, Ana Cristina Esteves, and Anna Andolfi. "Effect of γ-Aminobutyric Acid (GABA) on the Metabolome of Two Strains of Lasiodiplodia theobromae Isolated from Grapevine." Molecules 25, no. 17 (August 23, 2020): 3833. http://dx.doi.org/10.3390/molecules25173833.

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The effect of γ-aminobutyric acid (GABA) on the metabolome of two strains of Lasiodiplodia theobromae isolated from grapevine that hold a different degree of virulence to the host plant (LA-SOL3 (more virulent), LA-SV1 (less virulent)) was investigated. The culture filtrates and crude extracts from the two strains grown in the presence and absence of 10 mM of GABA were tested for phytotoxicity on tomato plant cuttings and leaves, respectively. Considering the opportunistic nature of this fungus for humans, crude extracts were also tested for cytotoxicity on mammalian cell lines. We found that culture filtrates and crude extracts have a decreased toxicity in the presence of GABA. Metabolomic analysis, conducted on both strains at both growth conditions, revealed the production of several compounds, such as indole-3-carboxylic acid (ICA, which is the main compound produced by L. theobromae), 3-indolecarboxyaldehyde, (3R,4S)-botryodiplodin, (R)-mellein. Finally, data demonstrate that GABA both induces a decrease in the amount of ICA, and a diversification of the metabolites produced by L. theobromae.

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SCAGLIUSI, SANDRA M. M., JORGE VEGA, and HUGO KUNIYUKI. "Cytopathology of callus cells infected with grapevine leafroll-associated virus 3." Fitopatologia Brasileira 27, no. 4 (July 2002): 384–88. http://dx.doi.org/10.1590/s0100-41582002000400008.

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The cytopathology of grapevine (Vitis spp.) callus tissue infected with Grapevine leafroll-associated virus 3 (GLRaV-3), genus Vitivirus was studied in order to investigate the usefulness of callus cultures to study grapevine leafroll-associated viruses. Ultrathin sections were made from in vitro callus obtained from stems and shoots of GLRaV-3 infected grapevine plants. Callus was composed of two types of tissue. Translucent, soft callus was formed and composed of large loosely arranged cells, containing big vacuoles and a thin layer of cytoplasm. Other parts of the callus were brown-coloured and composed of small compactly arranged cells, which showed flexuous and rod-shaped closterovirus-like particles, with 10-12 nm in diameter, at higher magnifications. Groups of vesicles formed by a single membrane were also observed, with sizes ranging from 50-200 nm, containing fine fibrillar material, also typical of closterovirus infections. Virus concentration was monitored by Immunosorbent electron microscopy (ISEM) tests, which showed that in vitro culture of callus tissue from grapevine infected plants, could be used to study the GLRaV viruses through many successive generations, despite the decline in virus concentration after repeated transfers. No virus particles were observed in callus tissue obtained from healthy grapevines.

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Calder�n, A. A., J. M. Zapata, and A. Ros Barcel�. "Differential expression of a cell wall-localized peroxidase isoenzyme capable of oxidizing 4-hydroxystilbenes during the cell culture of grapevine (Vitis vinifera cv. Airen and Monastrell)." Plant Cell, Tissue and Organ Culture 37, no. 2 (May 1994): 121–27. http://dx.doi.org/10.1007/bf00043605.

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Andreolli, Marco, Giacomo Zapparoli, Silvia Lampis, Chiara Santi, Elisa Angelini, and Nadia Bertazzon. "In Vivo Endophytic, Rhizospheric and Epiphytic Colonization of Vitis vinifera by the Plant-Growth Promoting and Antifungal Strain Pseudomonas protegens MP12." Microorganisms 9, no. 2 (January 23, 2021): 234. http://dx.doi.org/10.3390/microorganisms9020234.

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An evaluation was conducted of the colonization of Pseudomonas protegens MP12, a plant-growth promoting and antagonistic strain, inoculated in vine plants during a standard process of grapevine nursery propagation. Three in vivo inoculation protocols (endophytic, rhizospheric, and epiphytic) were implemented and monitored by means of both culture-dependent and independent techniques. Endophytic treatment resulted in the colonization of the bacterium inside the vine cuttings, which spread to young leaves during the forcing period. Microscopy analysis performed on transformed dsRed-tagged P. protegens MP12 cells confirmed the bacterium’s ability to penetrate the inner part of the roots. However, endophytic MP12 strain was no longer detected once the plant materials had been placed in the vine nursery field. The bacterium also displayed an ability to colonize the rhizosphere and, when the plants were uprooted at the end of the vegetative season, its persistence was confirmed. Epiphytic inoculation, performed by foliar spraying of cell suspension, was effective in controlling artificially-induced Botrytis cinerea infection in detached leaves. The success of rhizospheric and leaf colonization in vine plants suggests potential for the future exploitation of P. protegens MP12 as biofertilizer and biopesticide. Further investigation is required into the stability of the bacterium’s colonization of vine plants under real-world conditions in vineyards.

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Gan, Han Ming, Lucas Dailey, Peter Wengert, Nigel Halliday, Paul Williams, Andre Hudson, and Michael A. Savka. "Quorum sensing signals of the grapevine crown gall bacterium, Novosphingobium sp. Rr2-17: use of inducible expression and polymeric resin to sequester acyl-homoserine lactones." PeerJ 12 (December 20, 2024): e18657. https://doi.org/10.7717/peerj.18657.

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Background A grapevine crown gall tumor strain, Novosphingobium sp. strain Rr2-17 was previously reported to accumulate copious amounts of diverse quorum sensing signals during growth. Genome sequencing identified a single luxI homolog in strain Rr2-17, suggesting that it may encode for a AHL synthase with broad substrate range, pending functional validation. The exact identity of the complete suite of AHLs formed by novIspR1 is largely unknown. Methods This study validates the function of novIspR1 through inducible expression in Escherichia coli and in the wild-type parental strain Rr2-17. We further enhanced the capture of acyl homoserine lactone (AHL) signals produced by novIspR1 using polymeric resin XAD-16 and separated the AHLs by one- and two-dimensional thin layer chromatography followed by detection using AHL-dependent whole cell biosensor strains. Lastly, the complete number of AHLs produced by novIspR1 in our system was identified by LC-MS/MS analyses. Results The single LuxI homolog of N. sp. Rr2-17, NovIspR1, is able to produce up to eleven different AHL signals, including AHLs: C8-, C10-, C12-, C14-homoserine lactone (HSL) as well as AHLs with OH substitutions at the third carbon and includes 3-OH-C6-, 3-OH-C8-, 3-OH-C10-, 3-OH-C12- and 3-OH-C14-HSL. The most abundant AHL produced was identified as 3-OH-C8-HSL and isopropyl-D-1-thiogalactopyranoside (IPTG) induction of novIspR1 expression in wild type parental Rr2-17 strain increased its concentration by 6.8-fold when compared to the same strain with the vector only control plasmid. Similar increases were identified with the next two most abundant AHLs, 3-OH-C10- and unsubstituted C8-HSL. The presence of 2% w/v of XAD-16 resin in the growth culture bound 99.3 percent of the major AHL (3-OH-C8-HSL) produced by IPTG-induced overexpression of novIspR1 in Rr2-17 strain. This study significantly adds to our understanding of the AHL class of quorum sensing system in a grapevine crown gall tumor associated Novosphingobium sp. Rr2-17 strain. The identity of nine AHL signals produced by this bacterium will provide a framework to identify the specific function(s) of the AHL-mediated quorum-sensing associated genes in this bacterium.

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González-Ramos, Daniel, A. Muñoz, Anne Ortiz-Julien, Antonio Tomás Palacios, José María Heras, and Ramon González. "A Saccharomyces cerevisiae wine yeast strain overproducing mannoproteins selected through classical genetic methods." OENO One 44, no. 4 (December 31, 2010): 243. http://dx.doi.org/10.20870/oeno-one.2010.44.4.1475.

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Aims: Developing, by classical genetic methods, new wine yeast strains showing improved release of mannoproteins during wine fermentation, as well as suitable selection procedures for this purpose. These strains would be useful to improve quality characters associated to wine mannoprotein content.Methods and results: UV mutagenesis was used for genetic improvement of the industrial wine yeast strain ADY1. Cell wall-related phenotypes were used as primary selection criteria; an additional screening procedure was developed based on the detection of the released mannoproteins by hybridization with peroxidase-labeled Concanavalin A. Mannoprotein overproduction was assessed in laboratory media as well as in grapevine juice. One mutant strain, renamed HPS, was selected using these criteria. HPS showed increased mannoprotein release in different culture media, including natural must. Moreover, white wines fermented with this improved strain were less susceptible to protein haze than equivalent wines fermented with the original ADY1 strain. Red wines fermented with the mutant strain were also polysaccharide-enriched as compared to the original one.Conclusion: No clear correlation between a specific cell wall-related phenotype, or a combination of them, and improved release of polysaccharides by yeast random mutants could be established, and not all strains identified by in vitro assays as mannoprotein overproducing mutants were found positive for mannoprotein release in industrial conditions. Nevertheless, UV mutagenesis, combined with Concanavalin A detection, seems to be a viable way to improve mannoprotein release by industrial wine yeast strains.Significance and impact of the study: This study is one of the few recent reports on genetic improvement of wine yeast strains by non-recombinant genetic tools. It shows that mannoprotein release can be genetically improved and, for the first time, describes a successful selection procedure for such a complex character. These strains are potentially useful for the improvement of mannoprotein-related characters of white and red wines.

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Kiselev, Konstantin V., Zlata V. Ogneva, Olga A. Aleynova, Andrey R. Suprun, Alexey A. Ananev, Nikolay N. Nityagovsky, and Alexandra S. Dubrovina. "Influence of the 135 bp Intron on Stilbene Synthase VaSTS11 Transgene Expression in Cell Cultures of Grapevine and Different Plant Generations of Arabidopsis thaliana." Horticulturae 9, no. 4 (April 20, 2023): 513. http://dx.doi.org/10.3390/horticulturae9040513.

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Modern plant biotechnology often faces the problem of obtaining a stable and powerful vector for gene overexpression. It is known that introns carry different regulatory elements whose effects on transgene expression have been poorly studied. To study the effect of an intron on transgene expression, the stilbene synthase 11 (VaSTS11) gene of grapevine Vitis amurensis Rupr. was selected and overexpressed in grapevine callus cell cultures and several plant generations of Arabidopsis thaliana as two forms, intronless VaSTS11c and intron-containing VaSTS11d. The STS genes play an important role in the biosynthesis of stilbenes, valuable plant secondary metabolites. VaSTS11d contained two exons and one intron, while VaSTS11c contained only two exons, which corresponded to the mature transcript. It has been shown that the intron-containing VaSTS11d was better expressed in several generations of transgenic A. thaliana than VaSTS11c and also exhibited a lower level of cytosine methylation. As a result, the content of stilbenes in the VaSTS11d-transgenic plants was much higher than in the VaSTS11c-transgenic plants. Similarly, the best efficiency in increasing the content of stilbenes was also observed in grapevine cell cultures overexpressing the intron-containing VaSTS11d transcript. Thus, the results indicate that an intron sequence with regulatory elements can have a strong positive effect on both transgene expression level and its biological functions in plants and plant cell cultures.

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Ferri, Maura, Laura Righetti, and Annalisa Tassoni. "Increasing sucrose concentrations promote phenylpropanoid biosynthesis in grapevine cell cultures." Journal of Plant Physiology 168, no. 3 (February 2011): 189–95. http://dx.doi.org/10.1016/j.jplph.2010.06.027.

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Kiselev, Konstantin V., Olga A. Aleynova, Zlata V. Ogneva, Andrey R. Suprun, Alexey A. Ananev, Nikolay N. Nityagovsky, Alina A. Dneprovskaya, Alina A. Beresh, and Alexandra S. Dubrovina. "The Effect of Stress Hormones, Ultraviolet C, and Stilbene Precursors on Expression of Calcineurin B-like Protein (CBL) and CBL-Interacting Protein Kinase (CIPK) Genes in Cell Cultures and Leaves of Vitis amurensis Rupr." Plants 12, no. 7 (April 5, 2023): 1562. http://dx.doi.org/10.3390/plants12071562.

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Calcium serves as a crucial messenger in plant stress adaptation and developmental processes. Plants encode several multigene families of calcium sensor proteins with diverse functions in plant growth and stress responses. Several studies indicated that some calcium sensors may be involved in the regulation of secondary metabolite production in plant cells. The present study aimed to investigate expression of calcineurin B-like proteins (CBL) and CBL-interacting protein kinase (CIPK) in response to conditions inducting biosynthesis of stilbenes in grapevine. We investigated CBL and CIPK gene expression in wild-growing grapevine Vitis amurensis Rupr., known as a rich stilbene source, in response to the application of stilbene biosynthesis-inducing conditions, including application of stress hormones (salicylic acid or SA, methyl jasmonate or MeJA), phenolic precursors (p-coumaric acids or CA), and ultraviolet irradiation (UV-C). The influence of these effectors on the levels of 13 VaCBL and 27 VaCIPK mRNA transcripts as well as on stilbene production was analyzed by quantitative real-time RT-PCR in the leaves and cell cultures of V. amurensis. The data revealed that VaCBL4-1 expression considerably increased after UV-C treatment in both grapevine cell cultures and leaves. The expression of VaCIPK31, 41-1, and 41-2 also increased, but this increase was mostly detected in cell cultures of V. amurensis. At the same time, expression of most VaCBL and VaCIPK genes was markedly down-regulated both in leaves and cell cultures of V. amurensis, which may indicate that the CBLs and CIPKs are involved in negative regulation of stilbene accumulation (VaCBL8, 10a-2, 10a-4, 11, 12, VaCIPK3, 9-1, 9-2, 12, 21-1, 21-2, 33, 34, 35, 36, 37, 39, 40, 41-3, 41-4). The results obtained provide new information of CBL and CIPK implication in the regulation of plant secondary metabolism in response to stress hormones, metabolite precursors, and UV-C irradiation.

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Dubceac, Marcela, Evghenii Haustov, and Victor Bondarciuc. "Implementarea metodei PCR pentru identificarea tulpinilor patogene Allorhizobium Vitis ce provoacă cancerul bacterian al viței-de-vie." Agricultural Science, no. 1 (July 5, 2024): 47–54. http://dx.doi.org/10.55505/sa.2024.1.05.

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In the process of obtaining grapevine clones free from viral diseases and bacterial cancer, there are often cases where protoclones, seemingly healthy, well-developed, and with high yields, are found to be latently infected with bacterial cancer. This disease is most caused by agrobacteria from the genus Allorhizobium vitis and sometimes by Agrobacterium tumefaciens, and it is one of the most destructive diseases for grapevines from an economic perspective. The bacterial pathogen exists systemically in grapevines and spreads through infected propagation material. In the present study the Triplex End-Point PCR method was used for accurate and rapid identification of pathogenic strains of agrobacteria. One-year-old mature vines from 6 varieties and forms of grapevines were taken for the research. Microbiological testing on Roy & Sasser semiselective medium showed that five of the six plants tested were positive for Agrobacterium spp. infection. The positive tested samples were subjected to PCR testing. The presence of PehA gene, which is used to identify Allorhizobium vitis and to differentiate it from Agrobacterium tumefaciens, was confirmed in all tested samples. The virF and virD regions associated with the production of nopalins, octopins and vitopns were also detected. The test results suggested that the method allows for the reliable detection of pathogenic cultures carrying the tumor-inducing plasmid. The obtained results contribute to the development of strategies to combat bacterial cancer infections and improve phytosanitary selection practices for grapevine propagation material. În procesul de obținere a clonelor de viță de vie libere de boli virale și de cancer bacterian, adesea se observă cazuri în care plantele, aparent sănătoase, bine dezvoltate și cu o producție ridicată, se dovedesc a fi infectate latent cu cancer bacterian. Această boală este cauzată cel mai des de agrobacterii din genul Allorhizobium vitis și uneori de Agrobacterium tumefaciens și este una dintre cele mai distructive boli, din perspectivă economică, pentru vița de vie. Patogenul bacterian există sistemic în vița de vie și se răspândește prin materialul de propagare infectat. În prezentul studiu a fost utilizată metoda PCR Triplex End-Point pentru identificarea precisă și rapidă a tulpinilor patogene de agrobacterii. Pentru realizarea cercetărilor au fost prelevate vițe mature de un an de la 6 soiuri și forme de viță de vie. Testarea microbiologică pe mediul semiselectiv Roy & Sasser a demonstrat că cinci dintre cele 6 plante testate au prezentat rezultate pozitive pentru infecția cu Agrobacterium spp. Loturile testate pozitiv au fost transferate la etapa investigației prin metoda PCR. În toate probele testate a fost confirmață prezența genei PehA (466 bp), care servește pentru identificarea bacteriei Allorhizobium vitis și pentru diferențierea ei de Agrobacterium tumefaciens. De asemenea s-au detectat regiunile virF (382 bp) și virD (320 bp) asociate cu producția de nopaline, octopine și vitopine, ce servesc drept sursă de nutrienți pentru bacterii. Rezultatele testărilor au sugerat că metoda permite detectarea fiabilă a culturilor patogene purtătoare de plasmida inductoare de tumori. Rezultatele obținute contribuie la dezvoltarea strategiilor de combatere a infecțiilor cu cancer bacterian și la îmbunătățirea practicilor de selecție fitosanitară a materialului de multiplicare viticol.

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Aleynova, Olga A., Konstantin V. Kiselev, Zlata V. Ogneva, and 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 21, no. 21 (October 26, 2020): 7939. http://dx.doi.org/10.3390/ijms21217939.

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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.

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Aleynova, Olga A., Andrey R. Suprun, Alexey A. Ananev, Nikolay N. Nityagovsky, Zlata V. Ogneva, Alexandra S. Dubrovina, and Konstantin V. Kiselev. "Effect of Calmodulin-like Gene (CML) Overexpression on Stilbene Biosynthesis in Cell Cultures of Vitis amurensis Rupr." Plants 11, no. 2 (January 10, 2022): 171. http://dx.doi.org/10.3390/plants11020171.

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Stilbenes are plant phenolics known to rapidly accumulate in grapevine and other plants in response to injury or pathogen attack and to exhibit a great variety of healing beneficial effects. It has previously been shown that several calmodulin-like protein (CML) genes were highly up-regulated in cell cultures of wild-growing grapevine Vitis amurensis Rupr. in response to stilbene-modulating conditions, such as stress hormones, UV-C, and stilbene precursors. Both CML functions and stilbene biosynthesis regulation are still poorly understood. In this study, we investigated the effect of overexpression of five VaCML genes on stilbene and biomass accumulation in the transformed cell cultures of V. amurensis. We obtained 16 transgenic cell lines transformed with the VaCML52, VaCML65, VaCML86, VaCML93, and VaCML95 genes (3–4 independent lines per gene) under the control of the double CaMV 35S promoter. HPLC-MS analysis showed that overexpression of the VaCML65 led to a considerable and consistent increase in the content of stilbenes of 3.8–23.7 times in all transformed lines in comparison with the control calli, while biomass accumulation was not affected. Transformation of the V. amurensis cells with other analyzed VaCML genes did not lead to a consistent and considerable effect on stilbene biosynthesis in the cell lines. The results indicate that the VaCML65 gene is implicated in the signaling pathway regulating stilbene biosynthesis as a strong positive regulator and can be useful in viticulture and winemaking for obtaining grape cultivars with a high content of stilbenes and stress resistance.

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Jeandet, Philippe, Christophe Clément, Léo-Paul Tisserant, Jérôme Crouzet, and Éric Courot. "Use of grapevine cell cultures for the production of phytostilbenes of cosmetic interest." Comptes Rendus Chimie 19, no. 9 (September 2016): 1062–70. http://dx.doi.org/10.1016/j.crci.2016.02.013.

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Aleynova, Olga A., Konstantin V. Kiselev, Andrey R. Suprun, Alexey A. Ananev, and Alexandra S. Dubrovina. "Involvement of the Calmodulin-like Protein Gene VaCML92 in Grapevine Abiotic Stress Response and Stilbene Production." International Journal of Molecular Sciences 24, no. 21 (October 31, 2023): 15827. http://dx.doi.org/10.3390/ijms242115827.

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Calmodulin-like proteins (CMLs) are an important family of plant calcium sensor proteins that sense and decode changes in the intracellular calcium concentration in response to environmental and developmental stimuli. Nonetheless, the specific functions of individual CML family members remain largely unknown. This study aims to explore the role of the Vitis amurensis VaCML92 gene in the development of its high stress resistance and the production of stilbenes. The expression of VaCML92 was sharply induced in V. amurensis cuttings after cold stress. The VaCML92 gene was cloned and its role in the abiotic stress responses and stilbene production in grapevine was further investigated. The VaCML92-overexpressing callus cell cultures of V. amurensis and soil-grown plants of Arabidopsis thaliana exhibited enhanced tolerance to cold stress and, to a lesser extent, to the drought, while their tolerance to heat stress and high salinity was not affected. In addition, the overexpression of VaCML92 increased stilbene production in the V. amurensis cell cultures by 7.8–8.7-fold. Taken together, the data indicate that the VaCML92 gene is involved as a strong positive regulator in the rapid response to cold stress, the induction of cold stress resistance and in stilbene production in wild grapevine.

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Repka, V., and I. Baumgartnerová. "Grapevine habituation: Understanding of factors that contribute to neoplastic transformation and somaclonal variation." Acta Agronomica Hungarica 56, no. 4 (December 1, 2008): 399–408. http://dx.doi.org/10.1556/aagr.56.2008.4.4.

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Two-dimensional gel electrophoresis coupled to protein microarray analysis was used to examine for the first time the molecular mechanisms of grapevine habituation ( Vitis vinifera L., cv. Limberger) at both the proteome and the interactome level. The examination of 2-D maps derived from control and habituated cell cultures revealed the presence of 55 protein spots displaying a differential expression pattern. Using computational prediction methods, fundamental differences were found between eukaryotic interactomes. It was confirmed that all the predicted protein family interactomes (the full set of protein family interactions within a proteome) of six species are scale-free networks, and that they share a small core network comprising 16 protein families related to indispensable cellular functions predominantly involved in pathogenesis, apoptosis and plant tumorigenesis. There is molecular evidence suggesting that grapevine cells which have become habituated for one or more essential factors originated from heritable alterations in the pattern of gene expression and that they can, therefore, be used as a model for the study of cell differentiation and/or neoplastic transformation.

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Thomas, Pious, and Christopher M. M. Franco. "Intracellular Bacteria in Plants: Elucidation of Abundant and Diverse Cytoplasmic Bacteria in Healthy Plant Cells Using In Vitro Cell and Callus Cultures." Microorganisms 9, no. 2 (January 28, 2021): 269. http://dx.doi.org/10.3390/microorganisms9020269.

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This study was initiated to assess whether the supposedly axenic plant cell cultures harbored any cultivation-recalcitrant endophytic bacteria (CREB). Adopting live-cell imaging with bright-field, fluorescent and confocal microscopy and bacterial 16S-rRNA gene taxonomic profiling, we report the cytoplasmic association of abundant and diverse CREBs in long-term actively maintained callus and cell suspension cultures of different plant species. Preliminary bright-field live-cell imaging on grape cell cultures showed abundant intracellular motile micro-particles resembling bacteria, which proved uncultivable on enriched media. Bacterial probing employing DNA stains, transmission electron microscopy, and Eubacterial FISH indicated abundant and diverse cytoplasmic bacteria. Observations on long-term maintained/freshly established callus stocks of different plant species—grapevine, barley, tobacco, Arabidopsis, and medicinal species—indicated intracellular bacteria as a common phenomenon apparently originating from field shoot tissues.Cultivation-independent 16S rRNA gene V3/V3–V4 amplicon profiling on 40-year-old grape cell/callus tissues revealed a high bacterial diversity (>250 genera), predominantly Proteobacteria, succeeded by Firmicutes, Actinobacteria, Bacteriodetes, Planctomycetes, and 20 other phyla, including several candidate phyla. PICRUSt analysis revealed diverse functional roles for the bacterial microbiome, majorly metabolic pathways. Thus, we unearth the widespread association of cultivation-recalcitrant intracellular bacteria “Cytobacts” inhabiting healthy plant cells, sharing a dynamic mutualistic association with cell hosts.

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Bru, Roque, Susana Sellés, Juan Casado-Vela, Sarai Belchí-Navarro, and Maria Angeles Pedreño. "Modified Cyclodextrins Are Chemically Defined Glucan Inducers of Defense Responses in Grapevine Cell Cultures." Journal of Agricultural and Food Chemistry 54, no. 1 (January 2006): 65–71. http://dx.doi.org/10.1021/jf051485j.

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RANGEL-MONTOYA, Edelweiss A., Philippe E. ROLSHAUSEN, and Rufina HERNANDEZ-MARTINEZ. "Unravelling the colonization mechanism of Lasiodiplodia brasiliensis in grapevine plants." Phytopathologia Mediterranea 60, no. 2 (May 12, 2023): 135–49. http://dx.doi.org/10.36253/phyto-14198.

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Botryosphaeriaceae cause the degenerative disease Botryosphaeria dieback in many woody hosts, including grapevine. These pathogens penetrate host plants through pruning wounds, and colonize vascular tissues causing necrotic lesions, cankers, and eventually plant death. Colonization processes by Botryosphaeriaceae and their interactions with their hosts has been understudied. The colonization mechanisms were examined for Lasiodiplodia brasiliensis, a common pathogen causing Botryosphaeria dieback in Mexican vineyards. Lasiodiplodia brasiliensis MXBCL28 was inoculated onto grapevine ‘Cabernet Sauvignon’ plants, and after 2 months, infected tissues were observed with microscopy using histological techniques. Lasiodiplodia brasiliensis was also cultured on different carbon sources representing cell walls and non-structural plant components, to complement histology data. The host responded to wounding by developing xylem vessel occlusions with tyloses and deposition of suberin in cambium and ray parenchyma. Infection response also included deposition of suberin in pith tissues, reinforcement of cell walls with phenolic compounds, and lignin deposition in xylem vessels and ray parenchyma. The pathogen could overcome host compartmentalization mechanisms and colonize wood tissue causing extensive necrosis. The fungus was visualized in host cambium, vascular bundles, xylem vessels, and pith, and infected tissues were depleted in starch in the ray parenchyma. Cellulose, hemicellulose, and lignin in cell walls were also degraded, supporting in vitro data.

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Ananev, Alexey A., Andrey R. Suprun, Olga A. Aleynova, Nikolay N. Nityagovsky, Zlata V. Ogneva, Alexandra S. Dubrovina, and Konstantin V. Kiselev. "Effect of VaMyb40 and VaMyb60 Overexpression on Stilbene Biosynthesis in Cell Cultures of Grapevine Vitis amurensis Rupr." Plants 11, no. 15 (July 24, 2022): 1916. http://dx.doi.org/10.3390/plants11151916.

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Stilbenes are plant defense compounds known to rapidly accumulate in grapevine and some other plant species in response to microbial infection and several abiotic stresses. Stilbenes have attracted considerable attention due to valuable biological effects with multi-spectrum therapeutic application. However, there is a lack of information on natural signaling pathways and transcription factors regulating stilbene biosynthesis. It has been previously shown that MYB R2R3 transcription factor genes VaMyb40 and VaMyb60 were up-regulated in cell cultures of wild-growing grapevine Vitis amurensis Rupr. in response to UV irradiation. In this study, the effects of VaMyb40 or VaMyb60 overexpression in cell cultures of V. amurensis on their capability to produce stilbenes were investigated. Overexpression of the VaMyb60 gene led to a considerable increase in the content of stilbenes in three independently transformed transgenic lines in 5.9–13.9 times, while overexpression of the VaMyb40 gene also increased the content of stilbenes, although to a lesser extent (in 3.4–4.0 times) in comparison with stilbene levels in the empty vector-transformed calli. Stilbene content and stilbene production in the VaMyb60-transgenic calli reached 18.8 mg/g of dry weight (DW) and 150.8 mg/L, respectively. Using HPLC analysis, we detected eight individual stilbenes: t-resveratrol diglucoside, t-piceid, t-resveratrol, ε-viniferin, δ-viniferin, cis-resveratrol, cis-piceid, t-piceatannol. T-resveratrol prevailed over other stilbenoid compounds (53.1–89.5% of all stilbenes) in the VaMyb-overexpressing cell cultures. Moreover, the VaMyb40- and VaMyb60-transformed calli were capable of producing anthocyanins up to 0.035 mg/g DW, while the control calli did not produce anthocyanins. These findings show that the VaMyb40 and VaMyb60 genes positively regulate the stilbene biosynthesis as strong positive transcription regulators and can be used in biotechnological applications for stilbene production or high-quality viticulture and winemaking.

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Filippi, Antonio, Marco Zancani, Elisa Petrussa, and Enrico Braidot. "Caspase-3-like activity and proteasome degradation in grapevine suspension cell cultures undergoing silver-induced programmed cell death." Journal of Plant Physiology 233 (February 2019): 42–51. http://dx.doi.org/10.1016/j.jplph.2018.12.003.

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Martínez-Márquez, Ascensión, Jaime A. Morante-Carriel, Karla Ramírez-Estrada, Rosa M. Cusidó, Javier Palazon, and Roque Bru-Martínez. "Production of highly bioactive resveratrol analogues pterostilbene and piceatannol in metabolically engineered grapevine cell cultures." Plant Biotechnology Journal 14, no. 9 (March 7, 2016): 1813–25. http://dx.doi.org/10.1111/pbi.12539.

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Repka, V. "Elicitor-Stimulated Induction of Defense Mechanisms and Defense Gene Activation in Grapevine Cell Suspension Cultures." Biologia plantarum 44, no. 4 (December 1, 2001): 555–65. http://dx.doi.org/10.1023/a:1013742703929.

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Ben-Amar, Anis, Samia Daldoul, Dorsaf Allel, Goetz Reustle, and Ahmed Mliki. "Reliable encapsulation-based cryopreservation protocol for safe storage and recovery of grapevine embryogenic cell cultures." Scientia Horticulturae 157 (June 2013): 32–38. http://dx.doi.org/10.1016/j.scienta.2013.04.005.

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Coutos-Thevenot, Pierre, Isabelle Goebel-Tourand, Marie-Claude Mauro, Jean-Pierre Jouanneau, Michel Boulay, Alain Deloire, and Jean Guern. "Somatic embryogenesis from grapevine cells. I-Improvement of embryo development by changes in culture conditions." Plant Cell, Tissue and Organ Culture 29, no. 2 (May 1992): 125–33. http://dx.doi.org/10.1007/bf00033617.

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Miklos, E., A. Bérczi, and L. Erdei. "Plasmalemma isolation from cultured cells and roots of grapevine by aqueous two phase partition." Plant Science 66, no. 1 (January 1990): 73–80. http://dx.doi.org/10.1016/0168-9452(90)90171-j.

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Martínez-Márquez, Ascensión, Jaime A. Morante-Carriel, Javier Palazon, and Roque Bru-Martínez. "Rosa hybrida orcinol O-methyl transferase-mediated production of pterostilbene in metabolically engineered grapevine cell cultures." New Biotechnology 42 (May 2018): 62–70. http://dx.doi.org/10.1016/j.nbt.2018.02.011.

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Almagro, Lorena, Alicia De Gea-Abellán, María Isabel Rodríguez-López, Estrella Núñez-Delicado, José Antonio Gabaldón, and María Angeles Pedreño. "A Smart Strategy to Improve t-Resveratrol Production in Grapevine Cells Treated with Cyclodextrin Polymers Coated with Magnetic Nanoparticles." Polymers 12, no. 4 (April 24, 2020): 991. http://dx.doi.org/10.3390/polym12040991.

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One of the most successfully procedures used to increase the production of t-resveratrol in Vitis vinifera suspension-cultured cells is the application of cyclodextrins (CDs) and methyl jasmonate (MJ) as elicitors. In particular, β-CDs are characterized by their chemical structure which makes them special, not only by acting as elicitors, but also because they are compounds capable of trapping high added-value hydrophobic molecules such as t-resveratrol. However, the use of β-CDs as elicitors increases the production costs of this compound, making their industrial exploitation economically unfeasible. Therefore, the development of β-CDs recovery strategies is necessary to provide a viable solution to their industrial use. In this work, carboxymethylated and hydroxypropylated β-CDs have been used to form polymers using epichlorohydrin (EPI) as a cross-linking agent. The polymers were coated to Fe3O4 nanoparticles and were jointly used with MJ to elicit V. vinifera suspension-cultured cells. Once elicitation experiments were finished, a magnet easily allowed the recovery of polymers, and t-resveratrol was extracted from them by using ethyl acetate. The results indicated that the production of t-resveratrol in the presence of free carboxymethyl-β-CDs was much lower than that found in the presence of carboxymethyl-β-cyclodextrins-EPI polymer coated magnetic nanoparticles. In addition, the maximal levels of t-resveratrol were found at 168 h of elicitation in the presence of 15 g/L hydroxypropyl-β-CDs polymer coated magnetic nanoparticles and MJ, and non-t-resveratrol was found in the extracellular medium, indicating that all the t-resveratrol produced by the cells and secreted into the culture medium was trapped by the polymer and extracted from it. This work also showed that polymers can be regenerated and reused during three cycles of continuous elicitation since the induction and adsorption capacity of hydroxypropyl-β-CDs polymer-coated magnetic nanoparticles after these cycles of elicitation remained high, allowing high concentrations of t-resveratrol to be obtained.

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Filippi, Antonio, Elisa Petrussa, Francesco Boscutti, Marco Vuerich, Urska Vrhovsek, Zohreh Rabiei, and Enrico Braidot. "Bioactive Polyphenols Modulate Enzymes Involved in Grapevine Pathogenesis and Chitinase Activity at Increasing Complexity Levels." International Journal of Molecular Sciences 20, no. 24 (December 17, 2019): 6357. http://dx.doi.org/10.3390/ijms20246357.

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The reduction of synthetic chemistry use in modern viticulture relies on either the biological control of microorganisms or the induction of pathogenesis-related proteins. In the present study, the effects of hydro-alcoholic plant extracts (PEs) (i.e., by-products of Vitis vinifera L., leaves of Olea europaea L. and Ailanthus altissima (Mill.) Swingle) were tested on purified enzymes activity involved in plant-pathogen interactions. The polyphenolic composition was assayed and analyzed to characterize the extract profiles. In addition, suspension cell cultures of grapevine were treated with PEs to study their modulation of chitinase activity. Application of grape marc’s PE enhanced chitinase activity at 4 g L−1. Additionally, foliar treatment of grape marc’s PE at two doses (4 g L−1 and 800 g L−1) on grapevine cuttings induced a concentration-dependent stimulation of chitinase activity. The obtained results showed that the application of bioactive compounds based on PEs, rich in phenolic compounds, was effective both at in vitro and ex/in vivo level. The overall effects of PEs on plant-pathogen interaction were further discussed by applying a multi-criteria decision analysis, showing that grape marc was the most effective extract.

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Hattori, Tomoki, Yang Chen, Shinichi Enoki, Daisuke Igarashi, and Shunji Suzuki. "Exogenous isoleucine and phenylalanine interact with abscisic acid-mediated anthocyanin accumulation in grape." Folia Horticulturae 31, no. 1 (June 1, 2019): 147–57. http://dx.doi.org/10.2478/fhort-2019-0010.

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AbstractBerry skin colour is a crucial determinant of red/black grape berry quality. We investigated the effects of combination treatments with amino acids and a low concentration of ABA on anthocyanin accumulation in grapes. Among the amino acids tested, isoleucine and phenylalanine resulted in high anthocyanin contents in grape cell cultures. The combination treatments with isoleucine or phenylalanine, and a low concentration of ABA enhanced anthocyanin accumulation in grape cells and detached grape berries. The combination treatment with isoleucine, but not with phenylalanine, and ABA upregulated MybA1 expression. Field-grown grapevines received combination treatments with isoleucine or phenylalanine, and ABA in two growing seasons. In the 2015 growing season, the combination treatments with isoleucine or phenylalanine, and a low concentration of ABA accelerated anthocyanin accumulation in grape berry skins of field-grown grapevines on days 10 and 31 post treatment. The effects on anthocyanin accumulation became negligible at harvest. The effect of the combination treatment with phenylalanine and a low concentration of ABA on anthocyanin accumulation was masked in the 2017 growing season due to the unexpected stimulation of anthocyanin accumulation by the low concentration of ABA, although the combination treatment accelerated anthocyanin accumulation on days 3 and 10 post treatment. Taken together, the results suggested that exogenous isoleucine and phenylalanine interacted with ABA-mediated anthocyanin accumulation in grape berry skins of field-grown grapevines when the activity of ABA used to treat the grapevines was inadequate.

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Repka, V., I. Fischerova, and K. Silharova. "Methyl Jasmonate is a Potent Elicitor of Multiple Defense Responses in Grapevine Leaves and Cell-Suspension Cultures." Biologia plantarum 48, no. 2 (June 1, 2004): 273–83. http://dx.doi.org/10.1023/b:biop.0000033456.27521.e5.

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Lajara, María M., Antonio López-Orenes, María A. Ferrer, and Antonio A. Calderón. "Long-term exposure treatments revert the initial SA-induced alterations of phenolic metabolism in grapevine cell cultures." Plant Cell, Tissue and Organ Culture (PCTOC) 122, no. 3 (June 5, 2015): 665–73. http://dx.doi.org/10.1007/s11240-015-0800-9.

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Fila, Gianni, Jaleh Ghashghaie, Jackson Hoarau, and Gabriel Cornic. "Photosynthesis, leaf conductance and water relations of in vitro cultured grapevine rootstock in relation to acclimatisation." Physiologia Plantarum 102, no. 3 (March 1998): 411–18. http://dx.doi.org/10.1034/j.1399-3054.1998.1020309.x.

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Tran, Daniel, Tingting Zhao, Delphine Arbelet-Bonnin, Takashi Kadono, Patrice Meimoun, Sylvie Cangémi, Camille Noûs, Tomonori Kawano, Rafik Errakhi, and François Bouteau. "Early Cellular Responses Induced by Sedimentary Calcite-Processed Particles in Bright Yellow 2 Tobacco Cultured Cells." International Journal of Molecular Sciences 21, no. 12 (June 16, 2020): 4279. http://dx.doi.org/10.3390/ijms21124279.

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Calcite processed particles (CaPPs, Megagreen®) elaborated from sedimentary limestone rock, and finned by tribomecanic process were found to increase photosynthetic CO2 fixation grapevines and stimulate growth of various cultured plants. Due to their processing, the CaPPs present a jagged shape with some invaginations below the micrometer size. We hypothesised that CaPPs could have a nanoparticle (NP)-like effects on plants. Our data show that CaPPs spontaneously induced reactive oxygen species (ROS) in liquid medium. These ROS could in turn induce well-known cellular events such as increase in cytosolic Ca2+, biotic ROS generation and activation of anion channels indicating that these CaPPs could activate various signalling pathways in a NP-like manner.

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Journal articles on the topic 'Grapevine cell culture'

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