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David J. Robinson - One of the best experts on this subject based on the ideXlab platform.
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Evidence for persistent, symptomless infection of some potato cultivars with Tobacco Rattle Virus
Potato Research, 1998Co-Authors: S. Xenophontos, M. F. B. Dale, David J. Robinson, D. J. F. BrownAbstract:Infection with an M-type (particle producing) isolate of Tobacco Rattle Virus (TRV) was detected in leaves and/or roots of some plants from 11 of 13 potato cultivars grown in soil containing viruliferous trichodorid nematodes. Virus was detected in tubers of 8 of 13 cultivars, although only two (Pentland Dell and Maris Bard) developed spraing symptoms. Six cultivars (Arran Consul, Home Guard, King Edward, Romano, Sante and Wilja) were infected with TRV without developing spraing symptoms. Plants grown from Virus-containing, symptomless tubers became systemically infected with M-type TRV and produced symptomless infected daughter tubers. Virus was maintained through three generations of vegetative propagation, and the plants were sources for acquisition of the Virus by trichodorid nematodes. Distribution of Virus in both spraing-affected and symptomless tubers was erratic. Movement of symptomlessly infected seed tubers may be a means of dissemination of the Virus and of its introduction to previously unaffected sites.
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Susceptibility of transgenic Tobacco plants expressing Tobacco Rattle Virus coat protein to nematode-transmitted and mechanically inoculated Tobacco Rattle Virus
Journal of General Virology, 1993Co-Authors: Antoon T. Ploeg, Derek J. F. Brown, Alexander Mathis, John F. Bol, David J. RobinsonAbstract:Transgenic Samsun NN Tobacco plants expressing the coat protein of Tobacco Rattle Virus were exposed to mechanical leaf inoculation with Tobacco Rattle Virus and to viruliferous trichodorid vector nematodes. Whereas plants were resistant to mechanical inoculation the vector nematodes successfully transmitted Tobacco Rattle Virus to the roots as well as to the leaves of these plants. It is suggested that transgenic resistance is overcome either because vector nematodes inject relatively large numbers of Virus particles into a cell or because they inject destabilized particles. The results indicate that coat protein-mediated resistance is unlikely to be of value for controlling Tobacco Rattle Virus in field crops.
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Nuclear location of the 16K non-structural protein of Tobacco Rattle Virus.
Journal of General Virology, 1991Co-Authors: D. H. Liu, David J. Robinson, G. H. Duncan, Bryan D. HarrisonAbstract:An antiserum, elicited by a synthetic peptide coupled to bovine serum albumin, reacted specifically with the non-structural 16K protein of Tobacco Rattle Virus. The protein was detected in extracts of systemically infected Nicotiana clevelandii leaves, but only in those made with the aid of SDS, urea and 2-mercaptoethanol. Immunogold labelling of ultrathin sections showed that the protein was mainly associated with nuclei, but was also present in the cytoplasm. These observations suggest that the 16K protein binds to macromolecular components of infected cells, especially in nuclei, but do not clarify its function.
Kirankumar S Mysore - One of the best experts on this subject based on the ideXlab platform.
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Tobacco Rattle Virus–based Virus-induced gene silencing in Nicotiana benthamiana
Nature Protocols, 2014Co-Authors: Muthappa Senthil-kumar, Kirankumar S MysoreAbstract:This Protocol describes how to downregulate specific plant genes using Tobacco Rattle Virus Virus-induced gene silencing (TRV-VIGS). The method can be used in a range of plants, but N. benthamiana is used here as an example. Tobacco Rattle Virus (TRV)-based Virus-induced gene silencing (VIGS) is widely used in various plant species to downregulate the expression of a target plant gene. TRV is a bipartite, positive-strand RNA Virus with the TRV1 and TRV2 genomes. To induce post-transcriptional gene silencing (PTGS), the TRV2 genome is genetically modified to carry a fragment of the target gene and delivered into the plant (along with the TRV1 genome) by agroinoculation. TRV1- and TRV2-carrying Agrobacterium strains are then co-inoculated into 3-week-old plant leaves by one of three methods: a needleless syringe, the agrodrench method or by pricking with a toothpick. Target gene silencing occurs in the newly developed noninoculated leaves within 2–3 weeks of TRV inoculation. The TRV-VIGS protocol described here takes only 4 weeks to implement, and it is faster and easier to perform than other gene silencing techniques that are currently available. Although we use Nicotiana benthamiana as an example, the protocol is adaptable to other plant species.
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Tobacco Rattle Virus based Virus induced gene silencing in nicotiana benthamiana
Nature Protocols, 2014Co-Authors: Muthappa Senthilkumar, Kirankumar S MysoreAbstract:Tobacco Rattle Virus (TRV)-based Virus-induced gene silencing (VIGS) is widely used in various plant species to downregulate the expression of a target plant gene. TRV is a bipartite, positive-strand RNA Virus with the TRV1 and TRV2 genomes. To induce post-transcriptional gene silencing (PTGS), the TRV2 genome is genetically modified to carry a fragment of the target gene and delivered into the plant (along with the TRV1 genome) by agroinoculation. TRV1- and TRV2-carrying Agrobacterium strains are then co-inoculated into 3-week-old plant leaves by one of three methods: a needleless syringe, the agrodrench method or by pricking with a toothpick. Target gene silencing occurs in the newly developed noninoculated leaves within 2-3 weeks of TRV inoculation. The TRV-VIGS protocol described here takes only 4 weeks to implement, and it is faster and easier to perform than other gene silencing techniques that are currently available. Although we use Nicotiana benthamiana as an example, the protocol is adaptable to other plant species.
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Tobacco Rattle Virus–based Virus-induced gene silencing in Nicotiana benthamiana
Nature protocols, 2014Co-Authors: Muthappa Senthil-kumar, Kirankumar S MysoreAbstract:Tobacco Rattle Virus (TRV)-based Virus-induced gene silencing (VIGS) is widely used in various plant species to downregulate the expression of a target plant gene. TRV is a bipartite, positive-strand RNA Virus with the TRV1 and TRV2 genomes. To induce post-transcriptional gene silencing (PTGS), the TRV2 genome is genetically modified to carry a fragment of the target gene and delivered into the plant (along with the TRV1 genome) by agroinoculation. TRV1- and TRV2-carrying Agrobacterium strains are then co-inoculated into 3-week-old plant leaves by one of three methods: a needleless syringe, the agrodrench method or by pricking with a toothpick. Target gene silencing occurs in the newly developed noninoculated leaves within 2-3 weeks of TRV inoculation. The TRV-VIGS protocol described here takes only 4 weeks to implement, and it is faster and easier to perform than other gene silencing techniques that are currently available. Although we use Nicotiana benthamiana as an example, the protocol is adaptable to other plant species.
Ying Wang - One of the best experts on this subject based on the ideXlab platform.
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Tobacco Rattle Virus -induced gene silencing in Hevea brasiliensis
2020Co-Authors: Dong Guo, Ying Wang, Jia-hong Zhu, Shi-qing PengAbstract:Abstract BackgroundSince it is very difficult to obtain gene knockouts in rubber tree (Hevea Brasiliensis) due to low genetic transformation efficiency. Virus-induced gene silencing (VIGS) is a powerful gene silencing tool that has been intensively applied in plant. Up to now, the application of VIGS in rubber tree has not yet been reported.ResultsHevea brasiliensis phytoene desaturase (HbPDS) was identified in H. brasiliensis genome. The prediction of small interfering RNAs (siRNAs) from HbPDS and the silencing gene fragment (SGF) were predicted and a length of 409 bp SGF was chosen to be tested. We show that the Tobacco Rattle Virus (TRV) -VIGS is able to induce effective HbPDS silencing in rubber tree. The TRV-VIGS system has the potential for functional gene studies in rubber tree.ConclusionsThis is the first time to report VIGS in rubber tree. The present TRV-VIGS method could be further applied to produce gene silenced rubber tree plants, to advance functional gene of rubber tree. The applied TRV-VIGS method will achieve deeper underground into the natural rubber biosynthesis and regulation in this important rubber-producing plant.
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comprehensive transcriptome analyses reveal tomato plant responses to Tobacco Rattle Virus based gene silencing vectors
Scientific Reports, 2017Co-Authors: Yi Zheng, Ying Wang, Biao Ding, Zhangjun FeiAbstract:In plants, Virus-induced gene silencing (VIGS) is a popular tool for functional genomic studies or rapidly assessing individual gene functions. However, molecular details regarding plant responses to viral vectors remain elusive, which may complicate experimental designs and data interpretation. To this end, we documented whole transcriptome changes of tomato elicited by the application of the most widely used Tobacco Rattle Virus (TRV)-based vectors, using comprehensive genome-wide analyses. Our data illustrated multiple biological processes with functional implications, including (1) the enhanced activity of miR167 in guiding the cleavage of an auxin response factor; (2) reduced accumulation of phased secondary small interfering RNAs from two genomic loci; (3) altered expression of ~500 protein-coding transcripts; and (4) twenty long noncoding RNAs specifically responsive to TRV vectors. Importantly, we unraveled large-scale changes in mRNA alternative splicing patterns. These observations will facilitate future application of VIGS vectors for functional studies benefiting the plant research community and help deepen the understanding of plant-Virus interactions.
B. Dikova - One of the best experts on this subject based on the ideXlab platform.
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IDENTIFICATION OF Tobacco Rattle Virus (TRV) IN SUGAR BEET IN BULGARIA
Biotechnology & Biotechnological Equipment, 2006Co-Authors: B. DikovaAbstract:ABSTRACTBoth indicator and serological methods were used to identify Tobacco Rattle Virus (TRV) in sugar beet in Bulgaria. In mass screening of sugar beet crops, of 440 samples analyzed, 272 (62%) were positive. Beet TRV isolates were identified by the symptoms of 17 types of indicators and hosts of different botanical families. Of the experimental TRV hosts, wheat, barley and potato are the most important for our country. Serologically, the Virus was identified by means of indirect (ACP) and DAS-ELISA with different TRV antisera. Tobacco Rattle Virus, isolated from sugar beet, is immunologically related to the typical Virus strain (TRV type strain, potato isolate K), originating from potato—TRV K.
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ESTABLISHMENT OF Tobacco Rattle Virus (TRV) IN WEEDS AND CUSCUTA
Biotechnology & Biotechnological Equipment, 2006Co-Authors: B. DikovaAbstract:period of 2000 - 2004 the following 12 weed species were proved hosts to Tobacco Rattle Virus (TRV): Amaranthus retroflexus L. - amaranth pigweed, Atriplex patula L. - orache, Chenopodium album L. - lambs-quarters, Cirsium arvense (L.) Scop. - creeping thistle, Convolvulus arvensis L. - bindweed, Datura stramonium L. - thornapple, Poly- gonum lapathifolium L. - pale smartweed, Raphanus raphanistrum L. - wild radish, So- lanum nigrum L. - black dog grape, Sonchus arvensis L. - perennial sow thistle, Sorghum halepensis (L.) Pers. - Johnson-grass and Xanthium strumarium L. - broad cocklebur. The highest percentage of TRV infected plants was found among the species C. album, A. patula, S. halepensis and S. arvensis - 69%, 68%, 67% and 57%, respectively. Positive samples, i.e. TRV carriers, were given by 73 of a total of 149 weed plants, which is 49%. Two production crops in Bulgaria were found infested with the parasite weed dodder, which is another reservoir of TRV infection. Unlike the above mentioned weeds, it possi- bly transmits the Virus to sugar beet without nematode mediators of Trichodoridae family. Based on some morphological data (yellow-orange colored stems and fruit) we assumed this was Cuscuta campestris Yuncker. TRV was identified in self-seeded sugar beet, grown post-harvest on a field of wheat with predecessor sugar beet.
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Tobacco Rattle Virus (TRV) TRANSMISSION BY SUGAR BEET SEEDS
Biotechnology & Biotechnological Equipment, 2005Co-Authors: B. DikovaAbstract:ABSTRACTThe study involves Tobacco Rattle Virus (TRV) detection in racemes of two-year beet plants, seedlings, grown from seeds of these plants and seedlings from commercially available sugar beet seeds. Parts of sugar beet roots with a history of TRV were used for growing of two-year floriferous plants. Since plants had symptoms of yellow mosaic on leaves and raceme stipules, we tested each raceme separately with antiserum to TRV and broad beans wilt Virus (BBWV) by ELISA. BBWV also causes yellow mosaic on leaves and raceme stipules and is usually combined with TRV infection in beet lately. Of 13 flowering plants, 8 were diagnosed with TRV and 2—with BBWV. TRV Virus concentration in racemes was more than 0.3 to more than 0.5 optical units. BBWV showed lower concentration—more than 0.25 optical units in a limited number of plants (only 2), which did not interfere with TRV detection in racemes of flowering beet plants. Artificial vegetative propagation of beet has led to TRV transmission with racemes of tw...
Derek J. F. Brown - One of the best experts on this subject based on the ideXlab platform.
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Molecular diagnostics of some trichodorid nematodes and associated Tobacco Rattle Virus
Plant Pathology, 2004Co-Authors: K. Boutsika, Derek J. F. Brown, Mark S. Phillips, Stuart A. Macfarlane, R. C. Holeva, Vivian C. BlokAbstract:Several Paratrichodorus and Trichodorus (trichodorid) nematode species are natural vectors of Tobacco Rattle Virus (TRV) and cause economically important diseases, especially in potato and ornamental bulbous crops. Identification of trichodorid species based on morphological characters is laborious, time-consuming, and requires the services of highly trained personnel. Molecular diagnostics for trichodorid nematodes, using the ribosomal DNA repeat unit, were successfully developed to distinguish two Paratrichodorus and two Trichodorus species. The complete sequences of the 18S genes and the ITS-1 regions for these species were obtained and species-specific primers successfully designed for them. An RT-PCR assay was developed utilizing isolate-specific primers that amplify serologically distinguishable strains of TRV in individual trichodorid nematodes. The primers were based on the highly conserved RNA-1 segment of the bipartite genome and also on different parts of the RNA-2 segment of the Virus genome.
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Immunogold Localization of Tobacco Rattle Virus Particles within Paratrichodorus anemones.
Journal of nematology, 2000Co-Authors: E. Karanastasi, Stuart A. Macfarlane, N. Vassilakos, I. M. Roberts, Derek J. F. BrownAbstract:Unequivocal evidence of the viral nature of Virus-like particles observed at the specific site of retention of Tobacco Rattle Virus (TRV) in Paratrichodorus and Trichodorus nematodes has not previously been available. A new staining technique using safranin-O, which does not affect viral antigenicity, was used with an antiserum raised against the coat protein of TRV and prepared for use with immunogold labelling. Application of this method enabled the occurrence and localization of particles of TRV to be confirmed in the pharynx of the natural vector of the Virus, Paratrichodorus anemones, and provided unequivocal evidence that the particles observed were TRV particles. The TRV particles were observed attached only to the cuticle lining the posterior tract of the pharyngeal lumen of the vector. Therefore, the specific site of retention of TRV particles in P. anemones is apparently more localized than reported to occur in other vector trichodorid species.
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Susceptibility of transgenic Tobacco plants expressing Tobacco Rattle Virus coat protein to nematode-transmitted and mechanically inoculated Tobacco Rattle Virus
Journal of General Virology, 1993Co-Authors: Antoon T. Ploeg, Derek J. F. Brown, Alexander Mathis, John F. Bol, David J. RobinsonAbstract:Transgenic Samsun NN Tobacco plants expressing the coat protein of Tobacco Rattle Virus were exposed to mechanical leaf inoculation with Tobacco Rattle Virus and to viruliferous trichodorid vector nematodes. Whereas plants were resistant to mechanical inoculation the vector nematodes successfully transmitted Tobacco Rattle Virus to the roots as well as to the leaves of these plants. It is suggested that transgenic resistance is overcome either because vector nematodes inject relatively large numbers of Virus particles into a cell or because they inject destabilized particles. The results indicate that coat protein-mediated resistance is unlikely to be of value for controlling Tobacco Rattle Virus in field crops.
