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1

Gölles, R., R. Moser, H. Pühringer, H. Katinger, M. Laimer da Câmara Machado, A. Minafra, V. Savino, P. Saldarelli et G. P. M. da Câmara Machado. « TRANSGENIC GRAPEVINES EXPRESSING COAT PROTEIN GENE SEQUENCES OF GRAPEVINE FANLEAF VIRUS, ARABIS MOSAIC VIRUS, GRAPEVINE VIRUS A AND GRAPEVINE VIRUS B ». Acta Horticulturae, no 528 (mai 2000) : 307–14. http://dx.doi.org/10.17660/actahortic.2000.528.42.

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Yu, Yanyan, Yong Ni, Tian Qiao, Xiaomin Ji, Jinghao Xu, Bo Li et Qinghua Sun. « Overexpression of VvASMT1 from grapevine enhanced salt and osmotic stress tolerance in Nicotiana benthamiana ». PLOS ONE 17, no 6 (16 juin 2022) : e0269028. http://dx.doi.org/10.1371/journal.pone.0269028.

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Salt and drought stresses are major environmental conditions that severely limit grape growth and productivity, while exogenous melatonin can alleviate the drought and salt damage to grapevines. N-acetylserotonin methyltransferase (ASMT) is the key enzyme in melatonin synthesis, which plays a critical role in regulating stress responses. However, the roles of ASMTs from grapevine under drought and salt stresses responses remain largely unclear. In this study, the VvASMT1 gene was isolated from grapevine, and its physiological functions in salt and mimic drought stress tolerance were investigated. Expression pattern analysis revealed that VvASMT1 was significantly induced by different salt and osmotic stresses. Ectopic expression of VvASMT1 in Nicotiana benthamiana significantly enhanced melatonin production in transgenic plants. Compared with wild-type plants, the transgenic lines exhibited a higher germination ratio, longer root length, lower degree of leaf wilting and relative water content (RWC) under salt and osmotic stresses. In addition, under salt and osmotic stresses, overexpression of VvASMT1 improved proline and malondialdehyde (MDA) contents, increased the activity of antioxidant enzymes and decreased the accumulation of reactive oxygen species (ROS). Taken together, our results demonstrate the explicit role of VvASMT1 in salt and osmotic stress responses, which provides a theoretical foundation for the genetic engineering of grapevine.
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Gribaudo, I., G. Gambino, S. Leopold et M. Laimer. « MOLECULAR CHARACTERIZATION OF TRANSGENIC GRAPEVINE PLANTS ». Acta Horticulturae, no 689 (août 2005) : 485–92. http://dx.doi.org/10.17660/actahortic.2005.689.59.

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Levenko, B. A., et M. A. Rubtsova. « HERBICIDE RESISTANT TRANSGENIC PLANTS OF GRAPEVINE ». Acta Horticulturae, no 528 (mai 2000) : 339–42. http://dx.doi.org/10.17660/actahortic.2000.528.46.

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Gray, D. J., Z. T. Li, D. L. Hopkins, M. Dutt, S. A. Dhekney, M. M. Van Aman, J. Tattersall et K. T. Kelley. « Transgenic Grapevines Resistant to Pierce's Disease ». HortScience 40, no 4 (juillet 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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Krastanova, S., K. S. Ling, H. Y. Zhu, B. Xue, T. J. Burr et D. Gonsalves. « DEVELOPMENT OF TRANSGENIC GRAPEVINE ROOTSTOCKS WITH GENES FROM GRAPEVINE FANLEAF VIRUS AND GRAPEVINE LEAFROLL ASSOCIATED CLOSTEROVIRUSES 2 AND 3 ». Acta Horticulturae, no 528 (mai 2000) : 367–72. http://dx.doi.org/10.17660/actahortic.2000.528.52.

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Dutt, Manjul, Dennis J. Gray, Zhijian T. Li, Sadanand Dhekney et Marilyn M. Van Aman. « Micropropagation Cultures for Genetic Transformation of Grapevine ». HortScience 41, no 4 (juillet 2006) : 972C—972. http://dx.doi.org/10.21273/hortsci.41.4.972c.

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A major drawback to the use of embryogenic cultures for transformation of grapevine is that their ability to undergo genetic transformation is cultivar-dependent. Also, depending on cultivar, embryogenic cultures are difficult to impossible to maintain over time, reducing their utility for use in genetic transformation. An alternative to the use of embryogenic cultures for transformation of grapevine is the use of micropropagation cultures, which are easier to initiate from a wide range of grapevine cultivars and can be maintained over time without loss of function. Vitis vinifera `Thompson Seedless' was used as a model for genetic transformation using micropropagation cultures. In vitro cultures were initiated from apical meristems of actively growing vines and maintained in C2D medium containing 4 μM of 6-benzylaminopurine (C2D4B). Shoot tips and nodes were collected from proliferating in vitro cultures for transformation studies. A variety of wounding techniques, including nicking, sonication, and fragmenting of meristematic tissues was employed in order to enable Agrobacterium infection. We used a construct containing a bidirectional 35S promoter complex with a marker gene composed of a bifunctional fusion between an enhanced green fluorescent protein (EGFP) gene and a neomycin phosphotransferase (NPTII) gene in one direction and a hybrid lytic peptide gene in the other. Transgenic shoots growing in C2D4B medium containing 200 mg·L-1 each of carbenicillin and cefotaxime and 20 mg·L-1 of kanamycin were selected based on GFP fluorescence. Transgenic shoots were rooted and transferred to a greenhouse. To date, 18 transgenic lines have been generated. Details on the transformation procedure will be discussed.
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Aleynova, Olga A., Konstantin V. Kiselev, Zlata V. Ogneva et 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 (26 octobre 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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Li, Wei, Changxi Dang, Yuxiu Ye, Zunxin Wang, Laibao Hu, Fan Zhang, Yang Zhang et al. « Overexpression of Grapevine VvIAA18 Gene Enhanced Salt Tolerance in Tobacco ». International Journal of Molecular Sciences 21, no 4 (15 février 2020) : 1323. http://dx.doi.org/10.3390/ijms21041323.

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In plants, auxin/indoleacetic acid (Aux/IAA) proteins are transcriptional regulators that regulate developmental process and responses to phytohormones and stress treatments. However, the regulatory functions of the Vitis vinifera L. (grapevine) Aux/IAA transcription factor gene VvIAA18 have not been reported. In this study, the VvIAA18 gene was successfully cloned from grapevine. Subcellular localization analysis in onion epidermal cells indicated that VvIAA18 was localized to the nucleus. Expression analysis in yeast showed that the full length of VvIAA18 exhibited transcriptional activation. Salt tolerance in transgenic tobacco plants and Escherichia. coli was significantly enhanced by VvIAA18 overexpression. Real-time quantitative PCR analysis showed that overexpression of VvIAA18 up-regulated the salt stress-responsive genes, including pyrroline-5-carboxylate synthase (NtP5CS), late embryogenesis abundant protein (NtLEA5), superoxide dismutase (NtSOD), and peroxidase (NtPOD) genes, under salt stress. Enzymatic analyses found that the transgenic plants had higher SOD and POD activities under salt stress. Meanwhile, component analysis showed that the content of proline in transgenic plants increased significantly, while the content of hydrogen peroxide (H2O2) and malondialdehyde (MDA) decreased significantly. Based on the above results, the VvIAA18 gene is related to improving the salt tolerance of transgenic tobacco plants. The VvIAA18 gene has the potential to be applied to enhance plant tolerance to abiotic stress.
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Rubtsova, M. A., et B. A. Levenko. « PHOSPHINOTHRICIN- AND CROWN GALL-RESISTANT TRANSGENIC PLANTS OF GRAPEVINE ». Acta Horticulturae, no 625 (septembre 2003) : 465–72. http://dx.doi.org/10.17660/actahortic.2003.625.55.

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Zok, A., I. Forgács, A. Pedryc, R. Oláh et E. Szegedi. « Agrobacterium tumefaciens virE1Inhibits crown gall development in transgenic grapevine ». Acta Alimentaria 41, Supplement 1 (septembre 2012) : 214–18. http://dx.doi.org/10.1556/aalim.41.2012.suppl.21.

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Voegel, Tanja M., Jeremy G. Warren, Ayumi Matsumoto, Michele M. Igo et Bruce C. Kirkpatrick. « Localization and characterization of Xylella fastidiosa haemagglutinin adhesins ». Microbiology 156, no 7 (1 juillet 2010) : 2172–79. http://dx.doi.org/10.1099/mic.0.037564-0.

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Xylella fastidiosa is a Gram-negative, xylem-inhabiting, plant-pathogenic bacterium responsible for several important diseases including Pierce's disease (PD) of grapevines. The bacteria form biofilms in grapevine xylem that contribute to the occlusion of the xylem vessels. X. fastidiosa haemagglutinin (HA) proteins are large afimbrial adhesins that have been shown to be crucial for biofilm formation. Little is known about the mechanism of X. fastidiosa HA-mediated cell–cell aggregation or the localization of the adhesins on the cell. We generated anti-HA antibodies and show that X. fastidiosa HAs are present in the outer membrane and secreted both as soluble proteins and in membrane vesicles. Furthermore, the HA pre-proteins are processed from the predicted molecular mass of 360 kDa to a mature 220 kDa protein. Based on this information, we are evaluating a novel form of potential resistance against PD by generating HA-expressing transgenic grapevines.
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Ju, Yan-lun, Zhuo Min, Xiao-feng Yue, Yu-lin Zhang, Jun-xiang Zhang, Zong-qin Zhang et Yu-lin Fang. « Overexpression of grapevine VvNAC08 enhances drought tolerance in transgenic Arabidopsis ». Plant Physiology and Biochemistry 151 (juin 2020) : 214–22. http://dx.doi.org/10.1016/j.plaphy.2020.03.028.

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Boss, Paul K., Lekha Sreekantan et Mark R. Thomas. « A grapevine TFL1 homologue can delay flowering and alter floral development when overexpressed in heterologous species ». Functional Plant Biology 33, no 1 (2006) : 31. http://dx.doi.org/10.1071/fp05191.

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Grapevines (Vitis vinifera L.) have unusual plant architecture in that the shoot apical meristem produces both vegetative structures and primordia that are capable of forming inflorescences at regular intervals. These primordia are termed ‘uncommitted’ and differentiate into inflorescences or tendrils depending on the environment in which they are produced. To investigate the molecular relationship between tendrils and inflorescences and vine architecture, we cloned a TFL1 homologue from grapevine (VvTFL1). VvTFL1 is expressed in shoot apices early in latent bud development and in buds soon after bud burst. The grapevine homologue of LEAFY, VFL, is expressed at the same stages as VvTFL1 as well as in the later stages of inflorescence development. Neither VvTFL1 nor VFL were detected in tendrils. VvTFL1 was overexpressed in tobacco and Arabidopsis to confirm that it was functionally similar to TFL1 and not the close homologue FT. Flowering was delayed significantly in tobacco and Arabidopsis transformants overexpressing VvTFL1. However, an unexpected phenotype was observed in some of the transgenic Arabidopsis lines where the floral meristem became indeterminate and a new inflorescence would emerge from within the developing silique. Our findings suggest that VvTFL1 is a repressor of floral development. The nucleotide sequence reported in this paper has been submitted to GenBank under the accession number AF378127 (VvTFL1).
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Vigne, Emmanuelle, Véronique Komar et Marc Fuchs. « Field Safety Assessment of Recombination in Transgenic Grapevines Expressing the Coat Protein Gene of Grapevine fanleaf virus ». Transgenic Research 13, no 2 (avril 2004) : 165–79. http://dx.doi.org/10.1023/b:trag.0000026075.79097.c9.

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Vandelle, Elodie, Pietro Ariani, Alice Regaiolo, Davide Danzi, Arianna Lovato, Claudia Zadra, Nicola Vitulo, Giorgio Gambino et Annalisa Polverari. « The Grapevine E3 Ubiquitin Ligase VriATL156 Confers Resistance against the Downy Mildew Pathogen Plasmopara viticola ». International Journal of Molecular Sciences 22, no 2 (19 janvier 2021) : 940. http://dx.doi.org/10.3390/ijms22020940.

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Downy mildew, caused by Plasmopara viticola, is one of the most severe diseases of grapevine (Vitis vinifera L.). Genetic resistance is an effective and sustainable control strategy, but major resistance genes (encoding receptors for specific pathogen effectors) introgressed from wild Vitis species, although effective, may be non-durable because the pathogen can evolve to avoid specific recognition. Previous transcriptomic studies in the resistant species Vitis riparia highlighted the activation of signal transduction components during infection. The transfer of such components to V. vinifera might confer less specific and therefore more durable resistance. Here, we describe the generation of transgenic V. vinifera lines constitutively expressing the V. riparia E3 ubiquitin ligase gene VriATL156. Phenotypic and molecular analysis revealed that the transgenic plants were less susceptible to P. viticola than vector-only controls, confirming the role of this E3 ubiquitin ligase in the innate immune response. Two independent transgenic lines were selected for detailed analysis of the resistance phenotype by RNA-Seq and microscopy, revealing the profound reprogramming of transcription to achieve resistance that operates from the earliest stages of pathogen infection. The introduction of VriATL156 into elite grapevine cultivars could therefore provide an effective and sustainable control measure against downy mildew.
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Dalla Costa, Lorenza, Daniela Vinciguerra, Lisa Giacomelli, Umberto Salvagnin, Stefano Piazza, Katia Spinella, Mickael Malnoy, Claudio Moser et Ugo Marchesi. « Integrated approach for the molecular characterization of edited plants obtained via Agrobacterium tumefaciens-mediated gene transfer ». European Food Research and Technology 248, no 1 (2 novembre 2021) : 289–99. http://dx.doi.org/10.1007/s00217-021-03881-0.

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AbstractAgrobacterium tumefaciens-mediated gene transfer—actually the most used method to engineer plants—may lead to integration of multiple copies of T-DNA in the plant genome, as well as to chimeric tissues composed of modified cells and wild type cells. A molecular characterization of the transformed lines is thus a good practice to select the best ones for further investigation. Nowadays, several quantitative and semi-quantitative techniques are available to estimate the copy number (CN) of the T-DNA in genetically modified plants. In this study, we compared three methods based on (1) real-time polymerase chain reaction (qPCR), (2) droplet digital PCR (ddPCR), and (3) next generation sequencing (NGS), to carry out a molecular characterization of grapevine edited lines. These lines contain a knock-out mutation, obtained via CRISPR/Cas9 technology, in genes involved in plant susceptibility to two important mildew diseases of grapevine. According to our results, qPCR and ddPCR outputs are largely in agreement in terms of accuracy, especially for low CN values, while ddPCR resulted more precise than qPCR. With regard to the NGS analysis, the CNs detected with this method were often not consistent with those calculated by qPCR and ddPCR, and NGS was not able to discriminate the integration points in three out of ten lines. Nevertheless, the NGS method can positively identify T-DNA truncations or the presence of tandem/inverted repeats, providing distinct and relevant information about the transgene integration asset. Moreover, the expression analysis of Cas9 and single guide RNA (sgRNA), and the sequencing of the target site added new information to be related to CN data. This work, by reporting a practical case-study on grapevine edited lines, explores pros and cons of the most advanced diagnostic techniques available for the precocious selection of the proper transgenic material. The results may be of interest both to scientists developing new transgenic lines, and to laboratories in charge of GMO control.
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Aguero, C. B., A. M. Dandekar et C. P. Meredith. « TRANSGENIC GRAPEVINE PLANTS EXPRESSING GREEN FLUORESCENT PROTEINS TARGETED TO THE APOPLAST ». Acta Horticulturae, no 689 (août 2005) : 475–780. http://dx.doi.org/10.17660/actahortic.2005.689.57.

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Li, Peiying, Dongdong Yu, Bao Gu, Hongjuan Zhang, Qiying Liu et Jianxia Zhang. « Overexpression of the VaERD15 gene increases cold tolerance in transgenic grapevine ». Scientia Horticulturae 293 (février 2022) : 110728. http://dx.doi.org/10.1016/j.scienta.2021.110728.

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Burger, Anita L., Leonora Watts et Frederik C. Botha. « Grapevine promoter directs gene expression in the nectaries of transgenic tobacco ». Physiologia Plantarum 126, no 3 (mars 2006) : 418–34. http://dx.doi.org/10.1111/j.1399-3054.2006.00598.x.

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Harst, Margit, Beatrix-Axinja Cobanov, Ludger Hausmann, Rudolf Eibach et Reinhard Töpfer. « Evaluation of pollen dispersal and cross pollination using transgenic grapevine plants ». Environmental Biosafety Research 8, no 2 (avril 2009) : 87–99. http://dx.doi.org/10.1051/ebr/2009012.

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Li, Hui, Zhen Gao, Qiuju Chen, Qin Li, Meng Luo, Jiyuan Wang, Liping Hu et al. « Grapevine ABA receptor VvPYL1 regulates root hair development in Transgenic Arabidopsis ». Plant Physiology and Biochemistry 149 (avril 2020) : 190–200. http://dx.doi.org/10.1016/j.plaphy.2020.02.008.

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Jiu, SongTao, Chen Wang, Ting Zheng, Zhongjie Liu, XiangPeng Leng, Tariq Pervaiz, Abolfazl Lotfi, JingGui Fang et XiaoMin Wang. « Characterization of VvPAL-like promoter from grapevine using transgenic tobacco plants ». Functional & ; Integrative Genomics 16, no 6 (25 août 2016) : 595–617. http://dx.doi.org/10.1007/s10142-016-0516-x.

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Vigne, Emmanuelle, Marc Bergdoll, Sébastien Guyader et Marc Fuchs. « Population structure and genetic variability within isolates of Grapevine fanleaf virus from a naturally infected vineyard in France : evidence for mixed infection and recombination ». Journal of General Virology 85, no 8 (1 août 2004) : 2435–45. http://dx.doi.org/10.1099/vir.0.79904-0.

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The nematode-borne Grapevine fanleaf virus, from the genus Nepovirus in the family Comoviridae, causes severe degeneration of grapevines in most vineyards worldwide. We characterized 347 isolates from transgenic and conventional grapevines from two vineyard sites in the Champagne region of France for their molecular variant composition. The population structure and genetic diversity were examined in the coat protein gene by IC-RT-PCR-RFLP analysis with EcoRI and StyI, and nucleotide sequencing, respectively. RFLP data suggested that 55 % (191 of 347) of the isolates had a population structure consisting of one predominant variant. Sequencing data of 51 isolates representing the different restrictotypes confirmed the existence of mixed infection with a frequency of 33 % (17 of 51) and showed two major predominant haplotypes representing 71 % (60 of 85) of the sequence variants. Comparative nucleotide diversity among population subsets implied a lack of genetic differentiation according to host (transgenic vs conventional) or field site for most restrictotypes (17 of 18 and 13 of 18) and for haplotypes in most phylogenetic groups (seven of eight and six of eight), respectively. Interestingly, five of the 85 haplotypes sequenced had an intermediate divergence (0·036–0·066) between the lower (0·005–0·028) and upper range (0·083–0·138) of nucleotide variability, suggesting the occurrence of homologous RNA recombination. Sequence alignments clearly indicated a mosaic structure for four of these five variants, for which recombination sites were identified and parental lineages proposed. This is the first in-depth characterization of the population structure and genetic diversity in a nepovirus.
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Li, Min, Si-qi Shen, Yi-bin Xing, Wan-wan Jiao, Yong-rui Zhan, Ya-dan Sun, Da-long Guo et Yi-He Yu. « Vitis vinifera VvPUB17 functions as a E3 ubiquitin ligase and enhances powdery mildew resistance via the salicylic acid signaling pathway ». Journal of Berry Research 11, no 3 (27 août 2021) : 419–30. http://dx.doi.org/10.3233/jbr-210709.

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BACKGROUND: Powdery mildew affects grapevine growth and development and reduces grapevine fruit yield and quality. Plant U-box (PUB) E3 ubiquitin ligases play important roles in ubiquitin/proteasome-mediated protein degradation during plant development and in the plant defense response. OBJECTIVE: We cloned the VvPUB17 gene from Vitis vinifera and analyzed that VvPUB17 enhanced the resistance of grapevine to powdery mildew through the SA signal pathway. METHODS: Pathogen inoculation of Arabidopsis thaliana and grapevine plants was carried out by the tableting method. Gene expression was analyzed by quantitative real-time PCR. Sequence analysis and in vitro ubiquitination experiments show the structure and characteristics of VvPUB17. Exogenous salicylic acid, methyl jasmonate, ethylene and powdery mildew induced the expression of VvPUB17 in Arabidopsis leaves to verify the resistance of VvPUB17 to powdery mildew. RESULTS: Sequence analysis and in vitro ubiquitination experiments show that VvPUB17 contains U-box and Armadillo repeats (ARM repeat) and has E3 ubiquitin ligase activity dependent on the conserved U-box motif. Transgenic plants showed elevated levels of key genes related to the SA defense response pathway and high concentrations of salicylic acid. CONCLUSIONS: VvPUB17 functions as an E3 ubiquitin ligase that enhances the resistance of grapes to powdery mildew through the SA signal pathway.
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Arrey-Salas, Oscar, José Carlos Caris-Maldonado, Bairon Hernández-Rojas et Enrique Gonzalez. « Comprehensive Genome-Wide Exploration of C2H2 Zinc Finger Family in Grapevine (Vitis vinifera L.) : Insights into the Roles in the Pollen Development Regulation ». Genes 12, no 2 (20 février 2021) : 302. http://dx.doi.org/10.3390/genes12020302.

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Some C2H2 zinc-finger proteins (ZFP) transcription factors are involved in the development of pollen in plants. In grapevine (Vitis vinifera L.), it has been suggested that abnormalities in pollen development lead to the phenomenon called parthenocarpy that occurs in some varieties of this cultivar. At present, a network involving several transcription factors types has been revealed and key roles have been assigned to members of the C2H2 zinc-finger proteins (ZFP) family in model plants. However, particularities of the regulatory mechanisms controlling pollen formation in grapevine remain unknown. In order to gain insight into the participation of ZFPs in grapevine gametophyte development, we performed a genome-wide identification and characterization of genes encoding ZFP (VviZFP family). A total of 98 genes were identified and renamed based on the gene distribution into grapevine genome. The analysis performed indicate significant changes throughout VviZFP genes evolution explained by high heterogeneity in sequence, length, number of ZF and presence of another conserved domains. Moreover, segmental duplication participated in the gene family expansion in grapevine. The VviZFPs were classified based on domain and phylogenetic analysis into three sets and different groups. Heat-map demonstrated differential and tissue-specific expression patterns of these genes and k-means clustering allowed to identify a group of putative orthologs to some ZFPs related to pollen development. In transgenic plants carrying the promVviZFP13::GUS and promVviZFP68::GUS constructs, GUS signals were detectable in the anther and mature pollen grains. Expression profiling of selected VviZFP genes showed differential expression pattern during flower development and provides a basis for deepening in the understanding of VviZFPs role on grapevine reproductive development.
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Monier, C., P. Barbier et B. Walter. « PROTECTION AGAINST GRAPEVINE FANLEAF VIRUS IN TRANSGENIC TOBACCO CONTAINING NON-TRANSLATABLE SEQUENCES ». Acta Horticulturae, no 528 (mai 2000) : 379–84. http://dx.doi.org/10.17660/actahortic.2000.528.54.

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Zok, A., R. Oláh, É. Hideg, V. G. Horváth, P. B. Kós, P. Majer, Gy Váradi et E. Szegedi. « Effect of Medicago sativa ferritin gene on stress tolerance in transgenic grapevine ». Plant Cell, Tissue and Organ Culture (PCTOC) 100, no 3 (4 décembre 2009) : 339–44. http://dx.doi.org/10.1007/s11240-009-9641-8.

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Martinelli, L., et G. Mandolino. « Genetic transformation and regeneration of transgenic plants in grapevine (Vitis rupestris S.) ». Theoretical and Applied Genetics 88, no 6-7 (août 1994) : 621–28. http://dx.doi.org/10.1007/bf01253963.

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Hanif, Muhammad, Mati Rahman, Min Gao, Jinhua Yang, Bilal Ahmad, Xiaoxiao Yan et Xiping Wang. « Heterologous Expression of the Grapevine JAZ7 Gene in Arabidopsis Confers Enhanced Resistance to Powdery Mildew but Not to Botrytis cinerea ». International Journal of Molecular Sciences 19, no 12 (5 décembre 2018) : 3889. http://dx.doi.org/10.3390/ijms19123889.

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Jasmonate ZIM-domain (JAZ) family proteins comprise a class of transcriptional repressors that silence jasmonate-inducible genes. Although a considerable amount of research has been carried out on this gene family, there is still very little information available on the role of specific JAZ gene members in multiple pathogen resistance, especially in non-model species. In this study, we investigated the potential resistance function of the VqJAZ7 gene from a disease-resistant wild grapevine, Vitis quinquangularis cv. “Shang-24”, through heterologous expression in Arabidopsis thaliana. VqJAZ7-expressing transgenic Arabidopsis were challenged with three pathogens: the biotrophic fungus Golovinomyces cichoracearum, necrotrophic fungus Botrytis cinerea, and semi-biotrophic bacteria Pseudomonas syringae pv. tomato DC3000. We found that plants expressing VqJAZ7 showed greatly reduced disease symptoms for G. cichoracearum, but not for B. cinerea or P. syringae. In response to G cichoracearum infection, VqJAZ7-expressing transgenic lines exhibited markedly higher levels of cell death, superoxide anions (O2¯, and H2O2 accumulation, relative to nontransgenic control plants. Moreover, we also tested the relative expression of defense-related genes to comprehend the possible induced pathways. Taken together, our results suggest that VqJAZ7 in grapevine participates in molecular pathways of resistance to G. cichoracearum, but not to B. cinerea or P. syringae.
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Laquitaine, Laurent, Eric Gomès, Julie François, Chloé Marchive, Stéphanie Pascal, Saïd Hamdi, Rossitza Atanassova, Serge Delrot et Pierre Coutos-Thévenot. « Molecular Basis of Ergosterol-Induced Protection of Grape Against Botrytis cinerea : Induction of Type I LTP Promoter Activity, WRKY, and Stilbene Synthase Gene Expression ». Molecular Plant-Microbe Interactions® 19, no 10 (octobre 2006) : 1103–12. http://dx.doi.org/10.1094/mpmi-19-1103.

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Type I lipid transfer proteins (LTPs) are basic, 9-kDa cystein-rich proteins believed to be involved in plant defense mechanisms. A 2,100-bp fragment containing the coding region of Vitis vinifera lipid transfer protein 1 (VvLTP1) and 1,420-bp of its promoter region was isolated by screening a grape genomic library. In silico analysis revealed several putative, defense-related, cis-regulatory elements such as W- and MYB-boxes, involved in the binding of WRKY and MYB transcription factors, respectively. The 5′-truncated versions of the VvLTP1 promoter were generated, cloned in front of the β-glucuronidase (GUS) reporter gene, and introduced in tobacco plants and grapevine cell suspensions using Agrobacterium spp. Single MYB- and the W-boxes identified on the 0.250-kbp fragment were sufficient to induce GUS activity in transgenic tobacco plants after transient expression of MYB and WRKY. Ergosterol, a nonspecific fungal elicitor, induced GUS activity in transgenic grapevine cell suspensions transformed with the 1,420- and 750-bp promoter containing a palindromic arrangement of two Wboxes but not the 650- or 250-bp fragment, where only one W-box was present. Moreover, ergosterol triggered WRKY, VvLTP1, and stilbene synthase gene expression in grape plantlets and enhanced protection against Botrytis cinerea. The molecular basis of ergosterol-induced protection is discussed.
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Zhu, Ziguo, Guirong Li, Chaohui Yan, Li Liu, Qingtian Zhang, Zhen Han et Bo Li. « DRL1, Encoding A NAC Transcription Factor, Is Involved in Leaf Senescence in Grapevine ». International Journal of Molecular Sciences 20, no 11 (31 mai 2019) : 2678. http://dx.doi.org/10.3390/ijms20112678.

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The NAC (for NAM, ATAF1,2, and CUC2) proteins family are plant-specific transcription factors, which play important roles in leaf development and response to environmental stresses. In this study, an NAC gene, DRL1, isolated from grapevine Vitis vinifera L. “Yatomi Rose”, was shown to be involved in leaf senescence. The quantity of DRL1 transcripts decreased with advancing leaf senescence in grapevine. Overexpressing the DRL1 gene in tobacco plants significantly delayed leaf senescence with respect to chlorophyll concentration, potential quantum efficiency of photosystem II (Fv/Fm), and ion leakage. Moreover, exogenous abscisic acid (ABA) markedly reduced the expression of DRL1, and the ABA and salicylic acid (SA) concentration was lower in the DRL1-overexpressing transgenic plants than in the wild-type plants. The DRL1 transgenic plants exhibited reduced sensitivity to ABA-induced senescence but no significant change in the sensitivity to jasmonic acid-, SA- or ethylene-induced senescence. Transcriptomic analysis and RNA expression studies also indicated that the transcript abundance of genes associated with ABA biosynthesis and regulation, including 9-cis-epoxycarotenoid dioxygenase (NCED1), NCED5, zeaxanthin epoxidase1 (ZEP1), ABA DEFICIENT2 (ABA2), ABA4, and ABA INSENSITIVE 2 (ABI2), was markedly reduced in the DRL1-overexpressing plants. These results suggested that DRL1 plays a role as a negative regulator of leaf senescence by regulating ABA synthesis.
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Nakano, M., Y. Hoshino et M. Mii. « Regeneration of transgenic plants of grapevine (Vitis viniferaL.) viaAgrobacteriumrhizogenesmediated transformation of embryogenic calli ». Journal of Experimental Botany 45, no 5 (1994) : 649–56. http://dx.doi.org/10.1093/jxb/45.5.649.

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Yamamoto, T., H. Iketani, H. Ieki, Y. Nishizawa, K. Notsuka, T. Hibi, T. Hayashi et N. Matsuta. « Transgenic grapevine plants expressing a rice chitinase with enhanced resistance to fungal pathogens ». Plant Cell Reports 19, no 7 (13 juin 2000) : 639–46. http://dx.doi.org/10.1007/s002999900174.

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Galambos, A., A. Zok, A. Kuczmog, R. Oláh, P. Putnoky, W. Ream et E. Szegedi. « Silencing Agrobacterium oncogenes in transgenic grapevine results in strain-specific crown gall resistance ». Plant Cell Reports 32, no 11 (1 août 2013) : 1751–57. http://dx.doi.org/10.1007/s00299-013-1488-0.

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Zhu, Ziguo, Guirong Li, Li Liu, Qingtian Zhang, Zhen Han, Xuesen Chen et Bo Li. « A R2R3-MYB Transcription Factor, VvMYBC2L2, Functions as a Transcriptional Repressor of Anthocyanin Biosynthesis in Grapevine (Vitis vinifera L.) ». Molecules 24, no 1 (27 décembre 2018) : 92. http://dx.doi.org/10.3390/molecules24010092.

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In grapevine, the MYB transcription factors play an important role in the flavonoid pathway. Here, a R2R3-MYB transcription factor, VvMYBC2L2, isolated from Vitis vinifera cultivar Yatomi Rose, may be involved in anthocyanin biosynthesis as a transcriptional repressor. VvMYBC2L2 was shown to be a nuclear protein. The gene was shown to be strongly expressed in root, flower and seed tissue, but weakly expressed during the fruit development in grapevine. Overexpressing the VvMYBC2L2 gene in tobacco resulted in a very marked decrease in petal anthocyanin concentration. Expression analysis of flavonoid biosynthesis structural genes revealed that chalcone synthase (CHS), dihydroflavonol 4-reductase (DFR), leucoanthocyanidin reductase (LAR) and UDP glucose flavonoid 3-O-glucosyl transferase (UFGT) were strongly down-regulated in the VvMYBC2L2-overexpressed tobacco. In addition, transcription of the regulatory genes AN1a and AN1b was completely suppressed in transgenic plants. These results suggested that VvMYBC2L2 plays a role as a negative regulator of anthocyanin biosynthesis.
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Takuhara, Yuki, Masayuki Kobayashi et Shunji Suzuki. « Low-temperature-induced transcription factors in grapevine enhance cold tolerance in transgenic Arabidopsis plants ». Journal of Plant Physiology 168, no 9 (juin 2011) : 967–75. http://dx.doi.org/10.1016/j.jplph.2010.11.008.

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Krastanova, Stoyanka V., Vasudevan Balaji, Michele R. Holden, Mary Sekiya, Baodi Xue, Esengul A. Momol et Thomas J. Burr. « Resistance to crown gall disease in transgenic grapevine rootstocks containing truncated virE2 of Agrobacterium ». Transgenic Research 19, no 6 (25 février 2010) : 949–58. http://dx.doi.org/10.1007/s11248-010-9373-x.

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39

Thomzik, J. E., K. Stenzel, R. Stöcker, P. H. Schreier, R. Hain et D. J. Stahl. « Synthesis of a grapevine phytoalexin in transgenic tomatoes (Lycopersicon esculentumMill.) conditions resistance againstPhytophthora infestans ». Physiological and Molecular Plant Pathology 51, no 4 (octobre 1997) : 265–78. http://dx.doi.org/10.1006/pmpp.1997.0123.

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Kikkert, Julie R., Dominique Hébert-Soulé, Patricia G. Wallace, Michael J. Striem et Bruce I. Reisch. « Transgenic plantlets of ‘Chancellor’ grapevine (Vitis sp.) from biolistic transformation of embryogenic cell suspensions ». Plant Cell Reports 15, no 5 (janvier 1996) : 311–16. http://dx.doi.org/10.1007/bf00232362.

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41

Grimmig, Bernhard, Roland Schubert, Regina Fischer, Rüdiger Hain, Peter H. Schreier, Christian Betz, Christian Langebartels, Dieter Ernst et Heinrich Sandermann. « Ozone- and ethylene-induced regulation of a grapevine resveratrol synthase promoter in transgenic tobacco ». Acta Physiologiae Plantarum 19, no 4 (décembre 1997) : 467–74. http://dx.doi.org/10.1007/s11738-997-0043-4.

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42

Kikkert, Julie R., Dominique H�bert-Soul�, Patricia G. Wallace, Michael J. Striem et Bruce I. Reisch. « Transgenic plantlets of 'Chancellor' grapevine ( Vitis sp.) from biolistic transformation of embryogenic cell suspensions ». Plant Cell Reports 15, no 5 (1 janvier 1996) : 311–16. http://dx.doi.org/10.1007/s002990050023.

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43

Cheng, Jing, Keji Yu, Mingyue Zhang, Ying Shi, Changqing Duan et Jun Wang. « The Effect of Light Intensity on the Expression of Leucoanthocyanidin Reductase in Grapevine Calluses and Analysis of Its Promoter Activity ». Genes 11, no 10 (30 septembre 2020) : 1156. http://dx.doi.org/10.3390/genes11101156.

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To investigate the effect of light intensity on flavonoid biosynthesis, grapevine calluses were subjected to high light (HL, 250 μmol m−2 s−1) and dark (0 μmol m−2 s−1) in comparison to 125 μmol m−2 s−1 under controlled conditions (NL). The alteration of flavonoid profiles was determined and was integrated with RNA sequencing (RNA-seq)-based transcriptional changes of the flavonoid pathway genes. Results revealed that dark conditions inhibited flavonoid biosynthesis. Increasing light intensity affected flavonoids differently—the concentrations of flavonols and anthocyanins as well as the expressions of corresponding genes were less affected, whereas flavan-3-ol concentrations were predominantly increased, which caused enhanced trans-flavan-3-ol concentrations. Moreover, genes encoding leucoanthocyanidin reductase (LAR) exhibited different response patterns to light intensity changes—VviLAR1 expression increased with an increased light intensity, whereas VviLAR2 expression was insensitive. We further confirmed that the known transcription factors (TFs) involved in regulating flavan-3-ol biosynthesis utilized VviLAR1 as a target gene in grapevine calluses. In addition, VviLAR1 promoter activity was more sensitive to light intensity changes than that of VviLAR2 as determined using a transgenic Arabidopsis leaf system. These results suggested that light intensity had the most prominent effect on trans-flavan-3-ols in grapevine calluses and demonstrated that the two LAR genes had different response patterns to light intensity changes.
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44

Le Gall, O., L. Torregrosa, Y. Danglot, T. Candresse et A. Bouquet. « Agrobacterium-mediated genetic transformation of grapevine somatic embryos and regeneration of transgenic plants expressing the coat protein of grapevine chrome mosaic nepovirus (GCMV) ». Plant Science 102, no 2 (janvier 1994) : 161–70. http://dx.doi.org/10.1016/0168-9452(94)90034-5.

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45

Yoshikawa, N., S. Gotoh, M. Umezawa, N. Satoh, H. Satoh, T. Takahashi, T. Ito et K. Yoshida. « Transgenic Nicotiana occidentalis Plants Expressing the 50-kDa Protein of Apple chlorotic leaf spot virus Display Increased Susceptibility to Homologous Virus, but Strong Resistance to Grapevine berry inner necrosis virus ». Phytopathology® 90, no 3 (mars 2000) : 311–16. http://dx.doi.org/10.1094/phyto.2000.90.3.311.

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The 50-kDa protein (P50) encoded by the open reading frame 2 of Apple chlorotic leaf spot virus (ACLSV), a putative movement protein, was expressed in transgenic Nicotiana occidentalis plants. P50 in transgenic plants was mainly detected in a modified form in the cell wall fraction, similar to that in infected leaves. The P50-expressing plants (P50 plants) complemented the systemic spread of the P50-defective mutants of an infectious cDNA clone of ACLSV (pCLSF), indicating that P50 in transgenic plants was functional. Severity of symptoms was greatly enhanced and accumulation of virus in upper leaves was increased in P50 plants inoculated with pCLSF or ACLSV compared with that in nontransgenic control plants (NT plants). Conversely, transgenic plants expressing the coat protein of ACLSV (CP plants) showed a significant delay in symptom development and a reduction of virus accumulation. However, most P50 plants inoculated with Grapevine berry inner necrosis virus (GINV), another species of the genus Trichovirus, neither developed obvious symptoms nor supported virus accumulation in inoculated or upper leaves. In contrast, systemic symptoms developed and virus accumulated equally in NT and CP plants inoculated with GINV. After inoculation with Apple stem grooving virus or Apple stem pitting virus, there was no difference in symptom development and virus accumulation among P50, CP, and NT plants. Our results indicate that transgenic plants expressing a functional P50 were more susceptible to homologous virus and, on the contrary, showed strong resistance to the heterologous virus GINV.
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Zhang, Zhan, Luming Zou, Chong Ren, Fengrui Ren, Yi Wang, Peige Fan, Shaohua Li et Zhenchang Liang. « VvSWEET10 Mediates Sugar Accumulation in Grapes ». Genes 10, no 4 (28 mars 2019) : 255. http://dx.doi.org/10.3390/genes10040255.

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Sugar accumulation is a critical event during grape berry ripening that determines the grape market values. Berry cells are highly dependent on sugar transporters to mediate cross-membrane transport. However, the role of sugar transporters in improving sugar accumulation in berries is not well established in grapes. Herein we report that a Sugars Will Eventually be Exported Transporter (SWEET), that is, VvSWEET10, was strongly expressed at the onset of ripening (véraison) and can improve grape sugar content. VvSWEET10 encodes a plasma membrane-localized transporter, and the heterologous expression of VvSWEET10 indicates that VvSWEET10 is a hexose-affinity transporter and has a broad spectrum of sugar transport functions. VvSWEET10 overexpression in grapevine calli and tomatoes increased the glucose, fructose, and total sugar levels significantly. The RNA sequencing results of grapevine transgenic calli showed that many sugar transporter genes and invertase genes were upregulated and suggest that VvSWEET10 may mediate sugar accumulation. These findings elucidated the role of VvSWEET10 in sugar accumulation and will be beneficial for the improvement of grape berry quality in the future.
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Hily, Jean-Michel, Sandrine Demanèche, Nils Poulicard, Mélanie Tannières, Samia Djennane, Monique Beuve, Emmanuelle Vigne et al. « Metagenomic-based impact study of transgenic grapevine rootstock on its associated virome and soil bacteriome ». Plant Biotechnology Journal 16, no 1 (9 août 2017) : 208–20. http://dx.doi.org/10.1111/pbi.12761.

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48

Yoshikawa, N., Y. Saitou, A. Kitajima, T. Chida, N. Sasaki et M. Isogai. « Interference of Long-Distance Movement of Grapevine berry inner necrosis virus in Transgenic Plants Expressing a Defective Movement Protein of Apple chlorotic leaf spot virus ». Phytopathology® 96, no 4 (avril 2006) : 378–85. http://dx.doi.org/10.1094/phyto-96-0378.

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Transgenic Nicotiana occidentalis plants expressing a movement protein (P50) and partially functional deletion mutants (ΔA and ΔC) of the Apple chlorotic leaf spot virus (ACLSV) showed resistance to Grapevine berry inner necrosis virus (GINV). The resistance is highly effective and GINV was below the level of detection in both inoculated and uninoculated upper leaves. In contrast, GINV accumulated in inoculated and uninoculated leaves of nontransgenic (NT) plants and transgenic plants expressing a dysfunctional mutant (ΔG). On the other hand, in some plants of a transgenic plant line expressing a deletion mutant (ΔA', deletion of the C-terminal 42 amino acids), GINV could spread in inoculated leaves, but not move into uninoculated leaves. In a tissue blot hybridization analysis of ΔA'-plants inoculated with GINV, virus could be detected in leaf blade, midribs, and petiole of inoculated leaves, but neither in stems immediately above inoculated leaves nor in any tissues of uninoculated leaves. Immunohistochemical analysis of GINV-inoculated leaves of ΔA'-plants showed that GINV could invade into phloem parenchyma cells through bundle sheath of minor veins, suggesting that the long-distance transport of GINV might be inhibited between the phloem cells and sieve element (and/or within sieve element) rather than bundle sheath-phloem interfaces. Immunogold electron microscopy using an anti-P50 antiserum showed that P50 accumulated on the parietal layer of sieve elements and on sieve plates. The results suggested that resistance in P50-transgenic plants to GINV is due to the interference of both long-distance and cell-to-cell movement of the virus.
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Ritzenthaler, C., C. Laporte, F. Gaire, P. Dunoyer, C. Schmitt, S. Duval, A. Piéquet et al. « Grapevine Fanleaf Virus Replication Occurs on Endoplasmic Reticulum-Derived Membranes ». Journal of Virology 76, no 17 (1 septembre 2002) : 8808–19. http://dx.doi.org/10.1128/jvi.76.17.8808-8819.2002.

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ABSTRACT Infection by Grapevine fanleaf nepovirus (GFLV), a bipartite RNA virus of positive polarity belonging to the Comoviridae family, causes extensive cytopathic modifications of the host endomembrane system that eventually culminate in the formation of a perinuclear “viral compartment.” We identified by immunoconfocal microscopy this compartment as the site of virus replication since it contained the RNA1-encoded proteins necessary for replication, newly synthesized viral RNA, and double-stranded replicative forms. In addition, by using transgenic T-BY2 protoplasts expressing green fluorescent protein in the endoplasmic reticulum (ER) or in the Golgi apparatus (GA), we could directly show that GFLV replication induced a depletion of the cortical ER, together with a condensation and redistribution of ER-derived membranes, to generate the viral compartment. Brefeldin A, a drug known to inhibit vesicle trafficking between the GA and the ER, was found to inhibit GFLV replication. Cerulenin, a drug inhibiting de novo synthesis of phospholipids, also inhibited GFLV replication. These observations imply that GFLV replication depends both on ER-derived membrane recruitment and on de novo lipid synthesis. In contrast to proteins involved in viral replication, the 2B movement protein and, to a lesser extent, the 2C coat protein were not confined to the viral compartment but were transported toward the cell periphery, a finding consistent with their role in cell-to-cell movement of virus particles.
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Valat, Laure, Marc Fuchs et Monique Burrus. « Transgenic grapevine rootstock clones expressing the coat protein or movement protein genes of Grapevine fanleaf virus : Characterization and reaction to virus infection upon protoplast electroporation ». Plant Science 170, no 4 (avril 2006) : 739–47. http://dx.doi.org/10.1016/j.plantsci.2005.11.005.

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