Статті в журналах: ""grapevine transformation""

1

Baribault, T. J., K. G. M. Skene, and N. Steele Scott. "Genetic transformation of grapevine cells." Plant Cell Reports 8, no. 3 (1989): 137–40. http://dx.doi.org/10.1007/bf00716825.

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2

Li, Z. T., S. Dhekney, M. Dutt, M. Aman, J. Tattersall, K. T. Kelley, and D. J. Gray. "Optimizing Agrobacterium-mediated transformation of grapevine." In Vitro Cellular & Developmental Biology - Plant 42, no. 3 (May 2006): 220–27. http://dx.doi.org/10.1079/ivp2006770.

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3

Dutt, Manjul, Dennis J. Gray, Zhijian T. Li, Sadanand Dhekney, and Marilyn M. Van Aman. "Micropropagation Cultures for Genetic Transformation of Grapevine." HortScience 41, no. 4 (July 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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4

Cutanda, M. C., P. Chatelet, A. Bouquet, G. Lopez, P. Iocco, M. Thomas, O. Botella, F. J. Montero, and L. Torregrosa. "GENETIC TRANSFORMATION OF 'MACABEO' AND 'TEMPRANILLO' GRAPEVINE CULTIVARS." Acta Horticulturae, no. 827 (May 2009): 641–45. http://dx.doi.org/10.17660/actahortic.2009.827.113.

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5

KOVALENKO, P., and A. GALKIN. "Transformation of Grapevine caber net sauvignon by agrobacterium." Cell Biology International Reports 14 (September 1990): 189. http://dx.doi.org/10.1016/0309-1651(90)90855-s.

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6

Kikkert, J. R., J. R. Vidal, and B. I. Reisch. "APPLICATION OF THE BIOLISTIC METHOD FOR GRAPEVINE GENETIC TRANSFORMATION." Acta Horticulturae, no. 689 (August 2005): 459–62. http://dx.doi.org/10.17660/actahortic.2005.689.54.

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7

Guellec, Véronique, Chantal David, Michel Branchard, and Jacques Tempé. "Agrobacterium rhizogenes mediated transformation of grapevine (Vitis vinifera L.)." Plant Cell Tissue and Organ Culture (PCTOC) 20, no. 3 (March 1990): 211–15. http://dx.doi.org/10.1007/bf00041883.

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8

Vidal, Jose R., Julie R. Kikkert, Bruno D. Donzelli, Patricia G. Wallace, and Bruce I. Reisch. "Biolistic transformation of grapevine using minimal gene cassette technology." Plant Cell Reports 25, no. 8 (March 10, 2006): 807–14. http://dx.doi.org/10.1007/s00299-006-0132-7.

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9

Verdugo-Vásquez, Nicolás, Gastón Gutiérrez-Gamboa, Emilio Villalobos-Soublett, and Andrés Zurita-Silva. "Effects of Rootstocks on Blade Nutritional Content of Two Minority Grapevine Varieties Cultivated under Hyper-Arid Conditions in Northern Chile." Agronomy 11, no. 2 (February 12, 2021): 327. http://dx.doi.org/10.3390/agronomy11020327.

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In the 90s, as in other countries, transformation of Chilean viticulture brought about the introduction and spread of European grapevine varieties which has resulted in a massive loss of minor local and autochthonous grapevine varieties traditionally grown in several wine growing regions. Fortunately, in recent years, autochthonous and minority varieties have been revalued due to their high tolerance to pests and diseases and because of their adaptation to thermal and water stress triggered by global warming. In this study, we assessed the nutritional status of two autochthonous grapevines grafted onto four different rootstocks under the hyper-arid climatic conditions of Northern Chile over three consecutive seasons. The results showed that R32 rootstock induced high N, P, Ca, Mg and Mn levels in blades compared to Harmony rootstock. R32 rootstock and to a lesser extent, 1103 Paulsen and 140 Ruggeri rootstocks kept balanced levels of nutrients in blades collected from Moscatel Amarilla and Moscatel Negra grapevine varieties. Additionally, Harmony presented slight nutritional imbalance compared to the rest of studied rootstocks due to its low absorption of Mg, Mn, Ca and P, and its high K absorption, which was exacerbated under warm weather and salinity soil conditions. These results may provide a basis for specific cultivar/rootstock/site combinations, a nutritional guide for the viticulturists of Northern Chile, and options to diversify their production favoring the use of minority and autochthonous varieties that adapt well to hyper-arid conditions of Northern Chile.

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10

Deák, Tamás, Tünde Kupi, Róbert Oláh, Lóránt Lakatos, Lajos Kemény, György Bisztray, and Ernő Szegedi. "Candidate plant gene homologues in grapevine involved in Agrobacterium transformation." Open Life Sciences 8, no. 10 (October 1, 2013): 1001–9. http://dx.doi.org/10.2478/s11535-013-0218-5.

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AbstractThe grapevine (Vitis vinifera) genome was analyzed in silico for homologues of plant genes involved in Agrobacterium transformation in Arabidopsis thaliana and Nicotiana spp. Grapevine homologues of the glucomannan 4-betamannosyltransferase 9 gene CslA-09 involved in bacterial attachment to the cell wall, homologues of reticulon-like proteins BTI1, 2, 3 and RAB8 GTPases, both involved in T-DNA transfer to the host cell, homologues of VirE2 interacting protein VIP1 that contributes to the targeting of T-DNA into the nucleus and to its integration, and homologues of the histone protein H2A, which promotes the expression of T-DNA encoded genes, were selected. Sequences homologous to the arabinogalactan-protein AtAGP17 were not found in the grape genome. Seventeen selected candidates were tested by semiquantitative RT-PCR analysis for changes in their expression levels upon inoculation with Agrobacterium tumefaciens C58. Of the tested homologues, the expression of VvRab8a, VvVip1a and two histone genes (VvHta2 and VvHta10) increased significantly, therefore we hypothesize that these might be involved in Agrobacterium transformation of V. vinifera.

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11

Butiuc-Keul, Anca, and Ana Coste. "Biotechnologies and Strategies for Grapevine Improvement." Horticulturae 9, no. 1 (January 4, 2023): 62. http://dx.doi.org/10.3390/horticulturae9010062.

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Grapevine (Vitis vinifera subsp. Vinifera) is one of the most widespread and economically important perennial fruit crops in the world. Viticulture has changed over the years in response to changing environmental conditions and market demands, triggering the development of new and improved varieties to ensure the crop’s sustainability. The aim of this review is to provide a perspective on the recent developments in biotechnology and molecular biology and to establish the potential of these technologies for the genetic improvement of grapevine. The following aspects are discussed: (i) the importance of molecular marker-based methods for proper cultivar identification and how NGS-based high-throughput technologies have greatly benefited the development of genotyping techniques, trait mapping, and genomic selection; (ii) the recent advances in grapevine regeneration, genetic transformation, and genome editing, such as new breeding technology approaches for enhanced grapevine yield, quality improvement, and the selection of valuable varieties and cultivars. The specific problems and challenges linked to grapevine biotechnology, along with the importance of integrating classical and new technologies, are highlighted.

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12

ZHANG, Xiu-ming, Yi-fei WU, Zhi LI, Chang-bing SONG, and Xi-ping WANG. "Advancements in plant regeneration and genetic transformation of grapevine (Vitis spp.)." Journal of Integrative Agriculture 20, no. 6 (June 2021): 1407–34. http://dx.doi.org/10.1016/s2095-3119(20)63586-9.

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13

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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14

Dutt, Manjul, Zhijian T. Li, Sadanand Dhekney, and Dennis J. Gray. "(285) Characterization of a Composite Promoter from Genomic Sequences of Grapevine." HortScience 41, no. 4 (July 2006): 1053C—1053. http://dx.doi.org/10.21273/hortsci.41.4.1053c.

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Genetic transformation of plants necessitates the use of promoters to control transgene expression. Numerous promoters have been isolated from a wide range of organisms for use in plants. However, many of these natural promoters exhibit relatively low activity and/or have limited use. To provide an alternative, we constructed a composite promoter (EP) using a genomic DNA sequence and a 35 bp TATA-containing fragment from the 2S albumin (VvAlb1) gene core promoter of grapevine. The 0.9-kb genomic sequence was identified after TAIL-PCR, based on the presence of several unique cis-acting elements. The sequence showed no homology to any known plant gene, enhancer, and promoter. Two binary vectors, pEP-EGFP/NPT and pEP-GUS, containing a bifunctional EGFP/NPTII fusion gene and a GUS gene, respectively, were constructed to test transcriptional activity of the composite promoter both qualitatively and quantitatively. Transient GFP expression was observed in somatic embryos (SE) of Vitis vinifera `Thompson Seedless' after Agrobacterium-mediated transformation using pEP-EGFP/NPT. Quantitative GUS assay of stably transformed SE containing pEP-GUS indicated that the EP composite promoter was capable of producing GUS activity as high as 12% of that from a doubly enhanced Cauliflower Mosaic Virus 35S promoter or eight times higher than that from a doubly enhanced Cassava Vein Mosaic Virus promoter. In addition, transformation of Arabidopsis with pEP-GUS yielded comparable GUS activity throughout the plant. These data indicate that the EP composite promoter can be used in transformation studies to provide sustained constitutive gene expression in plants.

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15

Habili, Nuredin, and Forrest W. Nutter. "Temporal and Spatial Analysis of Grapevine Leafroll-Associated Virus 3 in Pinot Noir Grapevines in Australia." Plant Disease 81, no. 6 (June 1997): 625–28. http://dx.doi.org/10.1094/pdis.1997.81.6.625.

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An epidemic of grapevine leafroll disease (GLD), caused by grapevine leafroll-associated virus 3 (GLRaV-3), was monitored over an 11-year period in Nuriootpa, South Australia. Inoculum originated from infected budwood, and initial GLD incidence at the time of transplanting in 1986 was 23.1%. Infected vines were planted in a random spatial pattern. Change in disease incidence was not observed until 8 years after planting, when disease incidence increased to 27.9%. Disease incidence increased to 51.9% by 1996. Disease progress and rate curves (dy/dt versus time) indicated that the logistic (R2 = 96.2) and Gompertz (R2 = 96.3) growth models would best describe disease progress. However, the logistic model, which has a simpler data transformation with fewer model assumptions, was chosen for the purpose of comparing this epidemic (South Australia) with a GLRaV-3 epidemic in Cabernet Sauvignon grapevines in New Zealand. The logistic rate of GLD spread with respect to time was 0.35 logit/year in South Australia and was nearly three times faster (1.19 logits/year) for GLRaV-3 spread in New Zealand. Ordinary runs analyses indicated that the arrangement of infected vines within rows in South Australia was random up to 8 years after transplanting but subsequently became highly aggregated. Thus, GLD-infected plants are contributing to new infections (i.e., there is evidence for plant-to-plant spread), and a biotic vector with a steep dispersal gradient from each point source is likely to be involved.

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16

Li, Zhijian T., S. A. Dhekney, M. Dutt, and D. J. Gray. "An improved protocol for Agrobacterium-mediated transformation of grapevine (Vitis vinifera L.)." Plant Cell, Tissue and Organ Culture 93, no. 3 (April 15, 2008): 311–21. http://dx.doi.org/10.1007/s11240-008-9378-9.

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17

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

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18

Noronha, Henrique, Angélica Silva, Namiki Mitani-Ueno, Carlos Conde, Farzana Sabir, Catarina Prista, Graça Soveral, et al. "The grapevine NIP2;1 aquaporin is a silicon channel." Journal of Experimental Botany 71, no. 21 (June 25, 2020): 6789–98. http://dx.doi.org/10.1093/jxb/eraa294.

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Abstract Silicon (Si) supplementation has been shown to improve plant tolerance to different stresses, and its accumulation in the aerial organs is mediated by NIP2;1 aquaporins (Lsi channels) and Lsi2-type exporters in roots. In the present study, we tested the hypothesis that grapevine expresses a functional NIP2;1 that accounts for root Si uptake and, eventually, Si accumulation in leaves. Own-rooted grapevine cuttings of the cultivar Vinhão accumulated >0.2% Si (DW) in leaves when irrigated with 1.5 mM Si for 1 month, while Si was undetected in control leaves. Real-time PCR showed that VvNIP2;1 was highly expressed in roots and in green berries. The transient transformation of tobacco leaf epidermal cells mediated by Agrobacterium tumefaciens confirmed VvNIP2;1 localization at the plasma membrane. Transport experiments in oocytes showed that VvNIP2;1 mediates Si and arsenite uptake, whereas permeability studies revealed that VvNIP2;1 expressed in yeast is unable to transport water and glycerol. Si supplementation to pigmented grape cultured cells (cv. Gamay Freáux) had no impact on the total phenolic and anthocyanin content, or on the growth rate and VvNIP2;1 expression. Long-term experiments should help determine the extent of Si uptake over time and whether grapevine can benefit from Si fertilization.

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19

Nakano, M., Y. Hoshino, and 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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20

Sabbadini, S., L. Capriotti, C. Limera, O. Navacchi, G. Tempesta, and B. Mezzetti. "A plant regeneration platform to apply new breeding techniques for improving disease resistance in grapevine rootstocks and cultivars." BIO Web of Conferences 12 (2019): 01019. http://dx.doi.org/10.1051/bioconf/20191201019.

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Worldwide grapevine cultivation is based on the use of elite cultivars, in many cases strictly linked to local important wine brands. Most of Vitis viniferacultivars have high susceptibility to fungal and viral diseases therefore, new breeding techniques (e.g. Cisgenesis, RNAi and gene editing) offer the possibility to introduce new clones of the main cultivars with increased diseases resistance, in order to reduce environmental impact and improve quality in the intensive wine grape industry. This study is finalized to develop efficient in vitro regeneration and transformation protocols to extend the application of these technologies in wine grape cultivars and rootstocks. With this aim, in vitro regeneration protocols based on the production of meristematic bulks (Mezzetti et al., 2002) were optimized for different grapevine cultivars (Glera, Vermentino, Sangiovese, Thompson Seedless) and rootstocks (1103 Paulsen, and 110 Richter). The meristematic bulks were then used as explants for Agrobacteriummediated genetic transformation protocols, by comparing the use of NPTII and e-GFP as marker genes. Results confirmed the efficiency of meristematic bulks as the regenerating tissue to produce new modified plants in almost all the above genotypes. The highest regeneration efficiency in some genotypes allowed the selection of stable modified lines/calli with only the use of e-GFP marker gene. This protocol can be applied in the use of MYB marker gene for the production of cisgenic lines. Genotypes having the highest regeneration and transformation efficiency were also used for transformation experiments using a hairpin gene construct designed to silence the RNA-dependent RNA polymerase (RpRd) of the GFLV and GLRaV3, which would induce multiple virus resistances, and the Dicer-like protein 1 (Bc-DCL1) and Bc-DCL2 to control B. cinerea infection.

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21

Le Gall, O., L. Torregrosa, Y. Danglot, T. Candresse, and 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 (January 1994): 161–70. http://dx.doi.org/10.1016/0168-9452(94)90034-5.

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22

Jardak, Rahma, Ahmed Mliki, Abdelwahed Ghorbel, and Götz M. Reustle. "Transient expression of uidA gene in grapevine protoplasts after PEG-mediated transformation." OENO One 36, no. 2 (June 30, 2002): 93. http://dx.doi.org/10.20870/oeno-one.2002.36.2.975.

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Leaf protoplasts, isolated from Vitis vinifera «Sakasly» and «Muscat d’Alexandrie» and protoplasts from embryogenic tissue of Vitis sp. «Seyval blanc» were incubated in the presence of PEG in a transformation solution containing the plasmid pBI426 which carries the β-glucuronidase (gus) and the neomycin phosphotransferase II (nptII) genes. The treated protoplasts were cultivated in CPW13 medium without kanamycin selection. 48h after the PEG transformation, transient expression of gus gene was detected histochemically and fluorimetrically in the protoplast cultures.

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23

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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24

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

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25

Jardak-Jamoussi, Rahma, Patrick Winterhagen, Badra Bouamama, Cornelia Dubois, Ahmed Mliki, Thierry Wetzel, Abdelwahed Ghorbel, and Goetz M. Reustle. "Development and evaluation of a GFLV inverted repeat construct for genetic transformation of grapevine." Plant Cell, Tissue and Organ Culture (PCTOC) 97, no. 2 (February 24, 2009): 187–96. http://dx.doi.org/10.1007/s11240-009-9514-1.

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26

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

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27

Shin, Hye Young, Gi Hoon Kim, Sang Jae Kang, Jeung-Sul Han, and Cheol Choi. "Optimization of Agrobacterium-mediated transformation procedure for grapevine ‘Kyoho’ with carrot antifreeze protein gene." Journal of Plant Biotechnology 44, no. 4 (December 31, 2017): 388–93. http://dx.doi.org/10.5010/jpb.2017.44.4.388.

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28

Müller, Carsten, Kristina Ullmann, and Pablo Steinberg. "The Grapevine-Shoot Extract Vineatrol30 Inhibits the Chemically Induced Malignant Transformation of BALB/c-3T3 Cells." Journal of Medicinal Food 14, no. 1-2 (January 2011): 34–39. http://dx.doi.org/10.1089/jmf.2010.0022.

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29

Sanjurjo, Laura, José Ramón Vidal, Antonio Segura, and Francisco de la Torre. "Genetic transformation of grapevine cells using the minimal cassette technology: The need of 3′-end protection." Journal of Biotechnology 163, no. 4 (February 2013): 386–90. http://dx.doi.org/10.1016/j.jbiotec.2012.11.014.

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30

Creasap, J. E., C. L. Reid, M. C. Goffinet, R. Aloni, C. Ullrich, and T. J. Burr. "Effect of Wound Position, Auxin, and Agrobacterium vitis Strain F2/5 on Wound Healing and Crown Gall in Grapevine." Phytopathology® 95, no. 4 (April 2005): 362–67. http://dx.doi.org/10.1094/phyto-95-0362.

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Agrobacterium vitis is the causal agent of crown gall disease in grapevine, which can be severe in many regions worldwide. Vitis vinifera cultivars are highly susceptible to freeze injury, providing the wounds necessary for infection by A. vitis. Wound position in relation to the uppermost bud of cuttings was determined to be important in tumor development. Inoculated wounds below buds developed tumors, whereas wounds opposite the bud did not, implying that indole-3-aectic acid flow contributes to tumor formation. If auxin was applied to wounds prior to inoculation with a tumorigenic A. vitis strain, all sites of inoculation developed tumors, accompanied by an increased amount of callus in the cambium. Wounds inoculated with an A. vitis biological control strain F2/5 prior to application of the pathogen did not develop galls. A closer examination of these wounds determined that callus cells formed in the cambium during wound healing are susceptible to transformation by the pathogen. Although the mechanism by which F2/5 prevents transformation is unknown, our observations suggest that F2/5 inhibits normal wound healing by inducing necrosis in the cambium.

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31

Gribaudo, Ivana, Valentina Scariot, Giorgio Gambino, Andrea Schubert, Richard Göller, and Margit Laimer. "TRANSFORMATION OF VITIS VINIFERA L. CV NEBBIOLO WITH THE COAT PROTEIN GENE OF GRAPEVINE FANLEAF VIRUS (GFLV)." Acta Horticulturae, no. 603 (April 2003): 309–14. http://dx.doi.org/10.17660/actahortic.2003.603.40.

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32

Martinelli, L., D. Costa, V. Poletti, S. Festi, N. Buzkan, A. Minafra, P. Saldarelli, G. P. Martelli, and A. Perl. "GENETIC TRANSFORMATION OF TOBACCO AND GRAPEVINE FOR RESISTANCE TO VIRUSES RELATED TO THE RUGOSE WOOD DISEASE COMPLEX." Acta Horticulturae, no. 528 (May 2000): 323–30. http://dx.doi.org/10.17660/actahortic.2000.528.44.

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33

Khan, Nadeem, Fizza Fatima, Muhammad Salman Haider, Hamna Shazadee, Zhongjie Liu, Ting Zheng, and Jinggui Fang. "Genome-Wide Identification and Expression Profiling of the Polygalacturonase (PG) and Pectin Methylesterase (PME) Genes in Grapevine (Vitis vinifera L.)." International Journal of Molecular Sciences 20, no. 13 (June 28, 2019): 3180. http://dx.doi.org/10.3390/ijms20133180.

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In pectin regulation, polygalacturonases (PGs) and pectin methylesterases (PMEs) are critical components in the transformation, disassembly network, and remodeling of plant primary cell walls. In the current study, we identified 36 PG and 47 PME genes using the available genomic resources of grapevine. Herein, we provide a comprehensive overview of PGs and PMEs, including phylogenetic and collinearity relationships, motif and gene structure compositions, gene duplications, principal component analysis, and expression profiling during developmental stages. Phylogenetic analysis of PGs and PMEs revealed similar domain composition patterns with Arabidopsis. The collinearity analysis showed high conservation and gene duplications with purifying selection. The type of duplications also varied in terms of gene numbers in PGs (10 dispersed, 1 proximal, 12 tandem, and 13 segmental, respectively) and PMEs (23 dispersed, 1 proximal, 16 tandem, and 7 segmental, respectively). The tissue-specific response of PG and PME genes based on the reported transcriptomic data exhibited diverged expression patterns in various organs during different developmental stages. Among PGs, VvPG8, VvPG10, VvPG13, VvPG17, VvPG18, VvPG19, VvPG20, VvPG22, and VvPG23 showed tissue- or organ-specific expression in majority of the tissues during development. Similarly, in PMEs, VvPME3, VvPME4, VvPME5, VvPME6, VvPME19, VvPME21, VvPME23, VvPME29, VvPME31, and VvPME32 suggested high tissue-specific response. The gene ontology (GO), Kyoto Encyclopedia of Genes and Genomics (KEGG) enrichment, and cis-elements prediction analysis also suggested the putative functions of PGs and PMEs in plant development, such as pectin and carbohydrate metabolism, and stress activities. Moreover, qRT-PCR validation of 32 PG and PME genes revealed their role in various organs of grapevines (i.e., root, stem, tendril, inflorescence, flesh, skins, and leaves). Therefore, these findings will lead to novel insights and encourage cutting-edge research on functional characterization of PGs and PMEs in fruit crop species.

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Landi, Lucia, Sergio Murolo, and Gianfranco Romanazzi. "Colonization of Vitis spp. Wood by sGFP-Transformed Phaeomoniella chlamydospora, a Tracheomycotic Fungus Involved in Esca Disease." Phytopathology® 102, no. 3 (March 2012): 290–97. http://dx.doi.org/10.1094/phyto-06-11-0165.

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To evaluate wood colonization and interactions with Vitis spp. of Phaeomoniella chlamydospora, a fungal agent involved in Esca disease, isolate CBS 229.95 was transformed using a pCT74 construct which contained the genetic markers for synthetic green fluorescent protein (sGFP) and hygromycin B phosphotransferase. Nine stable P. chlamydospora fungal transformants (Pch-sGFP lines) were obtained using polyethylene-glycol-mediated transformation of protoplasts. These were characterized for sgfp and hygromycin B phosphotransferase (hph) genome insertions and for sGFP fluorescence emission, using quantitative polymerase chain reaction and fluorimetric systems, respectively. No correlation was observed between sgfp copy number genome insertion and sGFP fluorescence expression. Cuttings of Vitis vinifera ‘Montepulciano’, ‘Verdicchio’, ‘Sangiovese’, ‘Biancame’, and ‘Cabernet Sauvignon’; and the grapevine rootstocks ‘Kober 5BB’, ‘SO4’, ‘420A’, ‘1103P’, and V. rupestris were inoculated by immersion in a conidial suspension of the selected fungal Pch-sGFP71 line and incubated at 4 ± 1 and 25 ± 1°C. Wood colonization was estimated through epifluorescence microscopy and was affected by incubation temperature. After 6 months at 4 ± 1°C, the fungal growth was completely inhibited. At 25 ± 1°C, the highest extent of wood colonization was recorded in Montepulciano and Verdicchio, with the lowest in the rootstocks SO4 and V. rupestris. The expression of the Pch-sGFP71 transformed line was localized in the xylem area, primarily around the vessels. The use of sGFP-transformed P. chlamydospora helped to clarify different aspects associated with the location of this pathogen in grapevine tissue, before disease symptom expression.

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Felber, Aretusa Cristina, Julio Cesar Polonio, Ravely Casarotti Orlandelli, Alessandra Tenório Costa, Eliane Papa Ambrosio-Albuquerque, Renata Assis Castro, Maria Carolina Quecine-Verdi, João Lúcio de Azevedo, João Alencar Pamphile, and Káthia Socorro Mathias Mourão. "Agrobacterium-Mediated Transformation of Diaporthe schini Endophytes Associated with Vitis labrusca L. and Its Antagonistic Activity Against Grapevine Phytopathogens." Indian Journal of Microbiology 59, no. 2 (March 14, 2019): 217–24. http://dx.doi.org/10.1007/s12088-019-00787-0.

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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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López-Pérez, A. J., J. Carreño, and M. Dabauza. "TRANSFORMATION OF EMBRYOGENIC CALLUS AND TRANSGENIC PLANT REGENERATION IN TABLE GRAPEVINE 'SUGRAONE' (VITIS VINIFERA L.): EFFECT OF AGROBACTERIUM TUMEFACIENS STRAIN." Acta Horticulturae, no. 827 (May 2009): 415–20. http://dx.doi.org/10.17660/actahortic.2009.827.71.

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38

Fan, Chaohong, Ni Pu, Xiping Wang, Yuejin Wang, Li Fang, Weirong Xu, and Jianxia Zhang. "Agrobacterium-mediated genetic transformation of grapevine (Vitis vinifera L.) with a novel stilbene synthase gene from Chinese wild Vitis pseudoreticulata." Plant Cell, Tissue and Organ Culture 92, no. 2 (December 8, 2007): 197–206. http://dx.doi.org/10.1007/s11240-007-9324-2.

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39

Bouamama, Badra, Asma Ben Salem-Fnayou, Hatem Ben Jouira, Abdelwahed Ghorbel, and Ahmed Mliki. "Influence of the flower stage and culture medium on the induction of somatic embryogenesis from anther culture in Tunisian grapevine cultivars." OENO One 41, no. 4 (December 31, 2007): 185. http://dx.doi.org/10.20870/oeno-one.2007.41.4.835.

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Aims: The aim of this research is to identify the optimal flower developmental stage, in order to establish an efficient regeneration protocol via somatic embryogenesis in several local grapevine (Vitis vinifera L.) cultivars, using anther culture.Methods and results: Immature anthers sampled on fruity cuttings, at three flower developmental stages were cultured on three media [MS (1962), NN (1969) and CP (1987)] to which several phytohormonal combinations were added. Anthers, sampled at the « separated floral buds » stage, corresponding to the tetrad stage of microsporogenesis revealed to be the most appropriate for somatic embryogenesis compared to anthers sampled at the « separated clusters » and « early bloom » stages. Therefore, a unique medium, CP (1987) supplemented with 9 μM of 2,4-D and 11.35 μM of thidiazuron was established for the induction as well as the development of somatic embryos.Conclusion: The identification of an appropriate flower developmental stage and the optimization of a unique medium allowed establishing a reliable and repetitive regeneration system via somatic embryogenesis, in several local grapevine cultivars.Significance and impact of study: These embryogenic lines would be particularly appropriate as a target for the genetic transformation, directed towards the induction of tolerance to several biotic and abiotic stresses as well as for bioprospecting.

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40

Monteiro, Patrı́cia B., Diva C. Teixeira, Renê R. Palma, Monique Garnier, Joseph-Marie Bové, and Joël Renaudin. "Stable Transformation of the Xylella fastidiosa Citrus Variegated Chlorosis Strain withoriC Plasmids." Applied and Environmental Microbiology 67, no. 5 (May 1, 2001): 2263–69. http://dx.doi.org/10.1128/aem.67.5.2263-2269.2001.

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ABSTRACT Xylella fastidiosa is a gram-negative, xylem-limited bacterium affecting economically important crops (e.g., grapevine, citrus, and coffee). The citrus variegated chlorosis (CVC) strain ofX. fastidiosa is the causal agent of this severe disease of citrus in Brazil and represents the first plant-pathogenic bacterium for which the genome sequence was determined. Plasmids for the CVC strain of X. fastidiosa were constructed by combining the chromosomal replication origin (oriC) of X. fastidiosa with a gene which confers resistance to kanamycin (Kanr). In plasmid p16KdAori, the oriCfragment comprised the dnaA gene as well as the two flanking intergenic regions, whereas in plasmid p16Kori theoriC fragment was restricted to thednaA-dnaN intergenic region, which contains dnaA-box like sequences and AT-rich clusters. In plasmid p16K, no oriC sequence was present. In the three constructs, the promoter region of one of the two X. fastidiosa rRNA operons was used to drive the transcription of the Kanr gene to optimize the expression of kanamycin resistance in X. fastidiosa. Five CVC X. fastidiosa strains, including strain 9a5c, the genome sequence of which was determined, and two strains isolated from coffee, were electroporated with plasmid p16KdAori or p16Kori. Two CVC isolates, strains J1a12 and B111, yielded kanamycin-resistant transformants when electroporated with plasmid p16KdAori or p16Kori but not when electroporated with p16K. Southern blot analyses of total DNA extracted from the transformants revealed that, in all clones tested, the plasmid had integrated into the host chromosome at the promoter region of the rRNA operon by homologous recombination. To our knowledge, this is the first report of stable transformation in X. fastidiosa. Integration of oriC plasmids into the X. fastidiosa chromosome by homologous recombination holds considerable promise for functional genomics by specific gene inactivation.

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Martínez-Márquez, Ascensión, Jaime Morante-Carriel, Karla Ramírez-Estrada, Rosa Cusido, Susana Sellés-Marchart, Javier Palazon, Maria Angeles Pedreño, and Roque Bru-Martínez. "A reliable protocol for the stable transformation of non-embryogenic cells cultures of grapevine (Vitis vinifera L.) and Taxus x media." Journal of Biological Methods 2, no. 2 (June 24, 2015): 21. http://dx.doi.org/10.14440/jbm.2015.51.

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42

Lizamore, Darrell, and Chris Winefield. "The addition of an organosilicone surfactant to Agrobacterium suspensions enables efficient transient transformation of in vitro grapevine leaf tissue at ambient pressure." Plant Cell, Tissue and Organ Culture (PCTOC) 120, no. 2 (September 30, 2014): 607–15. http://dx.doi.org/10.1007/s11240-014-0627-9.

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Esterio, M., J. Auger, C. Ramos, and H. García. "First Report of Fenhexamid Resistant Isolates of Botrytis cinerea on Grapevine in Chile." Plant Disease 91, no. 6 (June 2007): 768. http://dx.doi.org/10.1094/pdis-91-6-0768c.

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Botrytis cinerea Pers. (teleomorph Botryotinia fuckeliana (de Bary) Whetzel) is a haploid, filamentous ascomycete that causes gray mold on many economically important crops in temperate regions, especially grapevine. The management of gray mold on table grape in Chile involves cultural and chemical methods. Currently, protection programs are based on several fungicide families (dicarboximides, anilinopyrimidines, mixture of anilinopyrimidines and phenylpyrroles, and hydroxyanilides [fenhexamid]). During the last 25 years, B. cinerea developed resistance to virtually all specific fungicides used to control gray mold. Field resistance to benzimidazoles, phenylcarbamates, and dicarboximides was detected soon after their introduction. Recent studies using PCR-duplex and specific primers for the detection of transposable elements on Chilean B. cinerea isolates recovered from different table grape cultivars corroborated the presence of two sibling cryptic populations, transposa and vacuma (3). Some vacuma isolates have shown natural resistance to fenhexamide (HydR1) and it has been separated into two groups on a molecular basis using a marker gene (Bc-hch): Group I, fenhexamid-resistant vacuma isolates; Group II, vacuma and transposa isolates sensitive to this fungicide (HydS) (2). Group I and II isolates can not interbred (1,2). Other B. cinerea resistant phenotypes, HydR2 and HydR3, have been reported as belonging to Group II (1,4). Single-spore isolates of B. cinerea (472) were collected from different table grape cultivars from 13 locations in the Chilean Central Valley. The isolation was done during harvest time from rotting berries. Fenhexamid (Teldor; Bayer CropScience, Monheim, Germany) was diluted to 10 μg a.i./ml and added to the solid medium (10 g of glucose, 1.5 g of K2HPO4, 2 g of KH2PO4, 1 g of (NH4)2SO4, 0.5 g of MgSO4·H2O, 2 g of yeast extract, and 12.5 g of agar in 1 liter) to reach concentrations of 0, 0.025, 0.05, and 0.1 μg a.i./ml. A 5-mm mycelial plug from each isolate of B. cinerea was cut from the edge of 4-day-old colonies placed in the center of petri dishes with the described fungicide-amended medium and incubated at 20°C for 5 days. Two measurements, octogonal diameters, were taken from each of three replicates per treatment. Means were calculated and the diameter of the inoculated plug was subtracted from each mean. For each isolate, a linear regression of the percent inhibition of mycelial growth versus the Log10 transformation for each of the four concentrations of fenhexamid was obtained. The 50% effective concentration of fenhexamid (EC50) was calculated with the regression equation for each isolate. So, 95.3% of B. cinerea isolates were sensitive (EC50 under 0.083 μg/ml), 1.9% were less sensitive (EC50 between 0.084 and 0.1 μg/ml), and 2.8% (13 isolates) were resistant EC50 values ranging from 0.1 to 8.4 μg/ml. Through PCR-restriction fragment length polymorphism, according to the Bc-hch gene restriction pattern, all resistant isolates analyzed belong to Group II of B. cinerea (Bc-hch2) (2). To our knowledge, this is the first report of fenhexamid resistant isolates of B. cinerea on grapevine in Chile and South America. It would be necessary to study the population dynamics of these isolates, although failure of botrytis control in the field with this compound has not been reported. References: (1) C. Albertini et al. Mycol. Res. 106:1171, 2002. (2) E. Fournier et al. Mycologia 97:1251, 2005. (3) T. Giraud et al. Mol. Biol. Evol. 14:1177, 1997. (4) P. Leroux et al. Phytoma 599:31, 2006.

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Szankowski, I., K. Briviba, J. Fleschhut, J. Sch�nherr, H.-J. Jacobsen, and H. Kiesecker. "Transformation of apple ( Malus domestica Borkh.) with the stilbene synthase gene from grapevine ( Vitis vinifera L.) and a PGIP gene from kiwi ( Actinidia deliciosa )." Plant Cell Reports 22, no. 2 (September 1, 2003): 141–49. http://dx.doi.org/10.1007/s00299-003-0668-8.

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45

Chen, Qiuju, Bohan Deng, Jie Gao, Zhongyang Zhao, Zili Chen, Shiren Song, Lei Wang, et al. "Comparative Analysis of miRNA Abundance Revealed the Function of Vvi-miR828 in Fruit Coloring in Root Restriction Cultivation Grapevine (Vitis vinifera L.)." International Journal of Molecular Sciences 20, no. 16 (August 20, 2019): 4058. http://dx.doi.org/10.3390/ijms20164058.

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Root restriction cultivation leads to early maturation and quality improvement, especially in the anthocyanin content in grapevine. However, the molecular mechanisms that underlie these changes have not been thoroughly elucidated. In this study, four small RNA libraries were constructed, which included the green soft stage (GS) and ripe stage (RS) of ‘Muscat’ (Vitis vinifera L.) grape berries that were grown under root restriction (RR) and in traditional cultivation (no root restriction, CK). A total of 162 known miRNAs and 14 putative novel miRNAs were detected from the four small RNA libraries by high-throughput sequencing. An analysis of differentially expressed miRNAs (DEMs) revealed that 13 miRNAs exhibited significant differences in expression between RR and CK at the GS and RS stages, respectively. For different developmental stages of fruit, 23 and 34 miRNAs showed expression differences between the GS and RS stages in RR and CK, respectively. The expression patterns of the eight DEMs and their targets were verified by qRT-PCR, and the expression profiles of target genes were confirmed to be complementary to the corresponding miRNAs in RR and CK. The function of Vvi-miR828, which showed the down regulated expression in the RS stage under root restriction, was identified by gene transformation in Arabidopsis. The anthocyanin content significantly decreased in transgenic lines, which indicates the regulatory capacity of Vvi-miR828 in fruit coloration. The miRNA expression pattern comparison between RR and CK might provide a means of unraveling the miRNA-mediated molecular process regulating grape berry development under root restricted cultivation.

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Zhang, Xiaowei, Fangxiu Xue, Zepeng Wang, Jian Wen, Cheng Guan, Feng Wang, Ling Han, and Na Ying. "A Novel Method of Hyperbola Recognition in Ground Penetrating Radar (GPR) B-Scan Image for Tree Roots Detection." Forests 12, no. 8 (July 30, 2021): 1019. http://dx.doi.org/10.3390/f12081019.

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Ground penetrating radar (GPR), as a newly nondestructive testing technology (NDT), has been adopted to explore the spatial position and the structure of the tree roots. Due to the complexity of soil distribution and the randomness of the root position in the natural environment, it is difficult to locate the root in the GPR B-Scan image. In this study, a novel method for root detection in the B-scan image by considering both multidirectional features and symmetry of hyperbola was proposed. Firstly, a mixed dataset B-Scan images were employed to train Faster RCNN (Regions with CNN features) to obtain the potential hyperbola region. Then, the peak area and its connected region were filtered from the four directional gradient graphs in the proposed region. The symmetry test was applied to segment the intersecting hyperbolas. Finally, two rounds of coordinate transformation and line detection based on Hough transform were employed for the hyperbola recognition and root radius and position estimation. To validate the effectiveness of this approach for tree root detection, a mixed dataset was made, including synthetic data from gprMax as well as field data collected from 35 ancient tree roots and fresh grapevine controlled experiments. From the results of hyperbola recognition as well as the estimation for the radius and position of the root, our method shows a significant effect in root detection.

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Sedlo, Jiří, and Pavel Tomšík. "Strategic development of varietal vineyards in the Czech Republic." Acta Universitatis Agriculturae et Silviculturae Mendelianae Brunensis 60, no. 2 (2012): 325–34. http://dx.doi.org/10.11118/actaun201260020325.

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The paper describes strategic changes in the structure of grapevine (Vitis vinifera L.) varieties grown in the Czech Republic. In 2004–2005, (i.e. after the admission of the Czech Republic into the EU) expenditures associated with restructuralisation and transformation of vineyards amounted for CZK 25,423 thous. The authors examine the development taking place in this domain within the last 50 years (i.e. from 1960 to 2010) and pay detailed attention to the period of 1989 to 2010. The paper analyses reasons of these changes and tries to describe the future development expected after 2010. The current production potential of the Czech Republic are 19,633.45 hectares of vineyards. For the time being, there are in average 1.07 wine growers per hectare of vineyards. As compared with 1960, the acreage of vineyards has doubled up and the number of the most frequent varieties has also increased. Within the period of 1989–1990, four varieties (i.e. Müller Thurgau, Green Veltliner, Italian Riesling and Sankt Laurent) occupied more than 60 % of the total vineyards area in the Czech Republic, whereas at present there are altogether 8 varieties (Müller Thurgau, Green Veltliner, Italian Riesling, Rhein Riesling, Sauvignon, Sankt Laurent, Blaufrankish, and Zweigeltrebe) at the nearly the same acreage.As far as the percentages of Müller Thurgau, Green Veltliner, Italian Riesling and Sankt Laurent varieties is concerned, it is anticipated that their acreages will further decrease, whereas those of Rhein Riesling, Sauvignon, Blaufrankish and Zweigeltrebe are expected to grow. The industry is under pressure of all Porter’s five forces of competition from external sources.

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Biasi, Rita, Roberta Farina, and Elena Brunori. "Family Farming Plays an Essential Role in Preserving Soil Functionality: A Study on Active Managed and Abandoned Traditional Tree Crop-Based Systems." Sustainability 13, no. 7 (April 2, 2021): 3967. http://dx.doi.org/10.3390/su13073967.

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In traditional agricultural areas, where traditional crops (TCs) are cultivated, small farms are still highly represented. Located prevalently in marginal and sensitive areas, agricultural areas have undergone deep transformation. Smallholders have maintained the traditional asset of cultivation (extensive and low input requirement management) only to some extent. In some cases they have adapted traditional orchards into more intensive planting systems. Frequently, they have abandoned agriculture. The land use and management influence soil functions, i.e., the capability of a specific soil to provide key functions in terrestrial ecosystems. In order to assess whether small farms are environmentally sustainable, we used a set of soil quality indicators in three traditional tree crops in the Latium region (central Italy), like hazelnut, grapevine, and Citrus. The soil parameters, chemical, biological, and biochemical, were quantified under three different management models: extensive cultivation, intensive cultivation, and abandonment. The selected set of indicators proved to be able to discriminate adequately between the management models and to be suitable for the soil health assessment. Results proved that hazelnut orchards stored more organic C, independently from farming management, while vineyard showed the lower total organic carbon (TOC). The microbial carbon vs. organic carbon ratio (Cmic-to Corg ratio) was higher for vineyards and Citrus groves, denoting a more active degradation of soil organic matter. Soil enzymes (ESs) involved in C cycle were variable along the different treatments and mainly influenced by the C inputs to soil and soil cover, whereas those involved in N, P, and S cycles were higher in abandoned and extensive TCs. Overall, extensive cultivation performed better in terms of soil quality than intensive or abandonment. This study suggests that a transition to an agriculture based on agroecological principles and toward extensification would provide significant soil-based environmental benefits in marginal sensitive areas.

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Bai, Yunhe, Xiaowen Zhang, Xuxian Xuan, Ehsan Sadeghnezhad, Fei Liu, Tianyu Dong, Dan Pei, Jinggui Fang, and Chen Wang. "miR3633a-GA3ox2 Module Conducts Grape Seed-Embryo Abortion in Response to Gibberellin." International Journal of Molecular Sciences 23, no. 15 (August 7, 2022): 8767. http://dx.doi.org/10.3390/ijms23158767.

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Seedlessness is one of the important quality and economic traits favored by grapevine consumers, which are mainly affected by phytohormones, especially gibberellin (GA). GA is widely utilized in seedless berry production and could effectively induce grape seed embryo abortion. However, the molecular mechanism underlying this process, like the role of RNA silencing in the biosynthesis pathway of GA remains elusive. Here, Gibberellin 3-β dioxygenase2 (GA3ox2) as the last key enzyme in GA biosynthesis was predicated as a potential target gene for miR3633a, and two of them were identified as a GA response in grape berries. We also analyzed the promoter regions of genes encoding GA biosynthesis and found the hormone-responsive elements to regulate grape growth and development. The cleavage interaction between VvmiR3633a and VvGA3ox2 was validated by RLM-RACE and the transient co-transformation technique in tobacco in vivo. Interestingly, during GA-induced grape seed embryo abortion, exogenous GA promoted the expression of VvmiR3633a, thereby mainly repressing the level of VvGA3ox2 in seed embryos. We also observed a negative correlation between down-regulated VvGA20ox2/VvGA3ox2 and up-regulated VvGA2ox3/VvGA2ox1, of which GA inactivation was greater than GA synthesis, inhibited active GA content, accompanied by the reduction of VvSOD and VvCAT expression levels and enzymatic activities. These series of changes might be the main causes of grape seed embryo abortion. In conclusion, we have preliminarily drawn a schematic mode of GA-mediated VvmiR3633a and related genes regulatory network during grape seed abortion induced by exogenous GA. Our findings provide novel insights into the GA-responsive roles of the VvmiR3633a-VvGA3ox2 module in the modulation of grape seed-embryo abortion, which has implications for the molecular breeding of high-quality seedless grape berries.

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Scorza, R., J. M. Cordts, D. J. Gray, D. W. Ramming, and R. L. Emershad. "Transformation of Grape (Vitis vinifera L.)." HortScience 30, no. 4 (July 1995): 876F—876. http://dx.doi.org/10.21273/hortsci.30.4.876f.

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Transgenic grapevines were regenerated from somatic embryos produced from immature zygotic embryos of two seedless grape selections and from leaves of in vitro-grown plants of `Thompson Seedless'. Somatic embryos were bombarded with gold microparticles using the Biolistic PDS-1000/He device (Bio-Rad Labs) and then exposed to engineered A. tumefaciens EHA101 (E. Hood, WSU). Alternately, somatic embryos were exposed to A. tumefaciens without bombardment. Following cocultivation, secondary embryos multiplied on Emershad and Ramming proliferation medium under kan selection. Transgenic embryos were identified after 3 to 5 months and developed into rooted plants on woody plant medium with 1 mM N6-benzyladenine, 1.5% sucrose, and 0.3% activated charcoal. Seedless selections were transformed with plasmids pGA482GG (J. Slightom, Upjohn) and pCGN7314 (Calgene), which carry GUS and NPTII genes. `Thompson Seedless' was transformed with pGA482GG and pGA482GG/TomRSVcp-15 (D. Gonsalves, Cornell Univ.) containing the tomato ringspot virus coat protein gene. Integration of foreign genes into grapevines was verified by growth on kan, GUS, and PCR assays, and Southern analyses.

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