The Experts below are selected from a list of 231 Experts worldwide ranked by ideXlab platform
Luis Rubio - One of the best experts on this subject based on the ideXlab platform.
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Broad bean wilt virus 1 encoded VP47 and SCP are suppressors of plant post-transcriptional gene silencing
European Journal of Plant Pathology, 2020Co-Authors: Caterina Carpino, Inmaculada Ferriol, Luis Rubio, Salvatore Davino, Laura Elvira-gonzález, Ezio Peri, Luis GalipiensoAbstract:Broad bean wilt virus 1 (BBWV-1, genus Fabavirus , family Secoviridae ) is a bipartite positive single-stranded RNA (+ssRNA) virus infecting important horticultural and ornamental crops worldwide. RNA1 encodes proteins involved in virus replication, whereas RNA2 encodes the large and small coat proteins (LCP, and SCP, respectively) and two putative movement proteins with overlapping C-terminal but different sizes: 47.2 kDa (VP47) and 37 kDa (VP37). Post-transcriptional gene silencing (PTGS) is a mechanism of gene regulation and defense against pathogens such as viruses. However, most plant viruses encode proteins called viral suppressors of RNA silencing (VSRs) which able to inhibit PTGS. Previously, BBWV-1 VP37 was identified as VSR and here, we have demonstrated that VP47 and SCP, but not LCP, are also VSRs by two approaches: (i) transient expression in Nicotiana benthamiana 16c plants expressing the green fluorescent protein (GFP) constitutively, and (ii) movement complementation assays using a viral vector based on Turnip crinkle virus sequence containing GFP (pTCV-GFP). In contrast LCP but not SCP of Broad been wilt virus 2 (BBWV-2, genus Fabavirus , family Secoviridae ) is a VSR, which illustrates that viruses of the same genus can interact differently with the host.
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RNA2-encoded VP37 protein of Broad bean wilt virus 1 is a determinant of pathogenicity, host susceptibility, and a suppressor of post-transcriptional gene silencing.
Molecular plant pathology, 2020Co-Authors: Caterina Carpino, Luis Rubio, Salvatore Davino, Laura Elvira-gonzález, Ezio Peri, Inmaculada Ferriol Safont, Vicente Medina, Luis Galipienso TorregrosaAbstract:Broad bean wilt virus 1 (BBWV-1, genus Fabavirus, family Secoviridae) is a bipartite, single-stranded positive-sense RNA virus infecting many horticultural and ornamental crops worldwide. RNA1 encodes proteins involved in viral replication whereas RNA2 encodes two coat proteins (the large and small coat proteins) and two putative movement proteins (MPs) of different sizes with overlapping C-terminal regions. In this work, we determined the role played by the small putative BBWV-1 MP (VP37) on virus pathogenicity, host specificity, and suppression of post-transcriptional gene silencing (PTGS). We engineered a BBWV-1 35S-driven full-length cDNA infectious clone corresponding to BBWV-1 RNA1 and RNA2 (pBBWV1-Wt) and generated a mutant knocking out VP37 (pBBWV1-G492C). Agroinfiltration assays showed that pBBWV1-Wt, as the original BBWV-1 isolate, infected broad bean, tomato, pepper, and Nicotiana benthamiana, whereas pBBWV1-G492C did not infect pepper and tomato systemically. Also, pBBWV1-G492C induced milder symptoms in broad bean and N. benthamiana than pBBWV1-Wt. Differential retrotranscription and amplification of the (+) and (-) strands showed that pBBWV1-G492C replicated in the agroinfiltrated leaves of pepper but not in tomato. All this suggests that VP37 is a determinant of pathogenicity and host specificity. Transient expression of VP37 through a potato virus X (PVX) vector enhanced PVX symptoms and induced systemic necrosis associated with programmed cell death in N. benthamiana plants. Finally, VP37 was identified as a viral suppressor of RNA silencing by transient expression in N. benthamiana 16c plants and movement complementation of a viral construct based on turnip crinkle virus (pTCV-GFP).
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A comparative study of viral infectivity, accumulation and symptoms induced by broad bean wilt virus 1 isolates
Journal of Plant Pathology, 2019Co-Authors: Caterina Carpino, Luis Rubio, Salvatore Davino, Laura Elvira-gonzález, Ezio Peri, Luis GalipiensoAbstract:Broad bean wilt virus 1 (BBWV-1, genus Fabavirus , family Secoviridae ) is a bipartite positive-sense single-stranded RNA virus distributed worldwide infecting many herbaceous species. Until now, scarce information regarding biological properties of BBWV-1 isolates is available. This work shows a comparative study on virus infectivity (proportion of infected plants over inoculated plants), virus accumulation and symptoms induced by four genetically different BBWV-1 isolates (Ben, B41/99, NSRV and PV0548) which were mechanically inoculated on several herbaceous hosts. The four BBWV-1 isolates infected broad bean, tomato, pepper and Nicotiana benthamiana plants, whereas none of them infected cucumber, common bean and melon. Infectivity ranged from 40 to 60% in tomato and from 75% to 95% in pepper, whereas it was 100% in broad bean and N. benthamiana for the four BBWV-1 isolates. Symptoms showed differences depending on the host plant, the viral isolate and the infection time. Virus accumulation was determined in broad bean and pepper plants and showed differences among host species and BBWV-1 isolates. No association between plant symptom manifestation and viral titre was observed.
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Molecular and biological characterization of highly infectious transcripts from full-length cDNA clones of broad bean wilt virus 1.
Virus research, 2016Co-Authors: Inmaculada Ferriol, Silvia Ambrós, Dorivaldo M. Da Silva, Bryce W. Falk, Luis RubioAbstract:Broad bean wilt virus 1 (BBWV-1), genus Fabavirus, has a genome composed of two single-stranded positive-sense RNAs of ∼5.8 (RNA1) and 3.4kb (RNA2). Full-length cDNA clones of both genomic RNAs (pBenR1 and pBenR2) from BBWV-1 isolate Ben were constructed under the control of the T7 promoter. In vitro derived capped transcripts were infectious in Nicotiana benthamiana, Chenopodium quinoa and Vicia faba plants. The biological activity of viral transcripts was not affected by extra bases at the 5'-terminus introduced during in vitro transcription. Virions derived from the infectious cDNA clones displayed similar viral infectivity and accumulation, as well as symptom induction as the wild-type BBWV-1 isolate.
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Rapid detection and discrimination of Fabaviruses by flow-through hybridisation with genus- and species-specific riboprobes
Annals of Applied Biology, 2015Co-Authors: Inmaculada Ferriol, Stefano Panno, Ezequiel A. Rangel, Antonio Olmos, Chenggui Han, Salvatore Davino, Luis RubioAbstract:Viruses cause significant damage in agricultural crops worldwide. Disease management requires sensitive and specific tools for virus detection and identification. Also, detection techniques need to be rapid to keep pace with the continuous emergence of new viral diseases. The genus Fabavirus is composed of five viruses infecting many economically important crops worldwide. This research describes the development of a procedure based on flow-through hybridisation (FTH), which is faster than and as sensitive as conventional hybridisation for virus detection in tissue-prints from infected plants. Six digoxigenin-labelled RNA probes were synthesised with two levels of specificity: (a) five specific for each viral species within this genus, and (b) a genus-specific probe that hybridises with a nucleotide sequence signature only found in the 5′-untranslated region of the genus Fabavirus, which is the first of this type reported for plant viruses. The new procedure developed is useful for rapid detection and discrimination of the five Fabaviruses identified so far and opens the possibility of discovering new species of this genus.
Inmaculada Ferriol - One of the best experts on this subject based on the ideXlab platform.
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Broad bean wilt virus 1 encoded VP47 and SCP are suppressors of plant post-transcriptional gene silencing
European Journal of Plant Pathology, 2020Co-Authors: Caterina Carpino, Inmaculada Ferriol, Luis Rubio, Salvatore Davino, Laura Elvira-gonzález, Ezio Peri, Luis GalipiensoAbstract:Broad bean wilt virus 1 (BBWV-1, genus Fabavirus , family Secoviridae ) is a bipartite positive single-stranded RNA (+ssRNA) virus infecting important horticultural and ornamental crops worldwide. RNA1 encodes proteins involved in virus replication, whereas RNA2 encodes the large and small coat proteins (LCP, and SCP, respectively) and two putative movement proteins with overlapping C-terminal but different sizes: 47.2 kDa (VP47) and 37 kDa (VP37). Post-transcriptional gene silencing (PTGS) is a mechanism of gene regulation and defense against pathogens such as viruses. However, most plant viruses encode proteins called viral suppressors of RNA silencing (VSRs) which able to inhibit PTGS. Previously, BBWV-1 VP37 was identified as VSR and here, we have demonstrated that VP47 and SCP, but not LCP, are also VSRs by two approaches: (i) transient expression in Nicotiana benthamiana 16c plants expressing the green fluorescent protein (GFP) constitutively, and (ii) movement complementation assays using a viral vector based on Turnip crinkle virus sequence containing GFP (pTCV-GFP). In contrast LCP but not SCP of Broad been wilt virus 2 (BBWV-2, genus Fabavirus , family Secoviridae ) is a VSR, which illustrates that viruses of the same genus can interact differently with the host.
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Molecular and biological characterization of highly infectious transcripts from full-length cDNA clones of broad bean wilt virus 1.
Virus research, 2016Co-Authors: Inmaculada Ferriol, Silvia Ambrós, Dorivaldo M. Da Silva, Bryce W. Falk, Luis RubioAbstract:Broad bean wilt virus 1 (BBWV-1), genus Fabavirus, has a genome composed of two single-stranded positive-sense RNAs of ∼5.8 (RNA1) and 3.4kb (RNA2). Full-length cDNA clones of both genomic RNAs (pBenR1 and pBenR2) from BBWV-1 isolate Ben were constructed under the control of the T7 promoter. In vitro derived capped transcripts were infectious in Nicotiana benthamiana, Chenopodium quinoa and Vicia faba plants. The biological activity of viral transcripts was not affected by extra bases at the 5'-terminus introduced during in vitro transcription. Virions derived from the infectious cDNA clones displayed similar viral infectivity and accumulation, as well as symptom induction as the wild-type BBWV-1 isolate.
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Rapid detection and discrimination of Fabaviruses by flow-through hybridisation with genus- and species-specific riboprobes
Annals of Applied Biology, 2015Co-Authors: Inmaculada Ferriol, Stefano Panno, Ezequiel A. Rangel, Antonio Olmos, Chenggui Han, Salvatore Davino, Luis RubioAbstract:Viruses cause significant damage in agricultural crops worldwide. Disease management requires sensitive and specific tools for virus detection and identification. Also, detection techniques need to be rapid to keep pace with the continuous emergence of new viral diseases. The genus Fabavirus is composed of five viruses infecting many economically important crops worldwide. This research describes the development of a procedure based on flow-through hybridisation (FTH), which is faster than and as sensitive as conventional hybridisation for virus detection in tissue-prints from infected plants. Six digoxigenin-labelled RNA probes were synthesised with two levels of specificity: (a) five specific for each viral species within this genus, and (b) a genus-specific probe that hybridises with a nucleotide sequence signature only found in the 5′-untranslated region of the genus Fabavirus, which is the first of this type reported for plant viruses. The new procedure developed is useful for rapid detection and discrimination of the five Fabaviruses identified so far and opens the possibility of discovering new species of this genus.
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Detection and identification of Fabavirus species by one-step RT-PCR and multiplex RT-PCR.
Journal of virological methods, 2013Co-Authors: Stefano Panno, Inmaculada Ferriol, Ezequiel A. Rangel, Antonio Olmos, Chenggui Han, Federico Martinelli, Luis Rubio, Salvatore DavinoAbstract:The genus Fabavirus of the family Secoviridae comprises a group of poorly characterized viruses. To date, only five species have been described: Broad bean wilt virus 1 (BBWV-1), Broad bean wilt virus 2 (BBWV-2), Lamium mild mosaic virus (LMMV), Gentian mosaic virus (GeMV) and Cucurbit mild mosaic virus (CuMMV). The development is described of two RT-PCR procedures for the detection and identification of Fabavirus species: a one-step RT-PCR using a single pair of conserved primers for the detection of all Fabaviruses, and a one-step multiplex RT-PCR using species-specific primers for the simultaneous detection and identification of the above-mentioned species of the genus Fabavirus. These methods were applied successfully to field samples and the results were compared with those obtained by molecular hybridization and ELISA. The combination of the two techniques enables rapid, sensitive and reliable identification of the five known Fabavirus species, as well as the possibility of discovering new species of this genus.
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The complete genome sequence of Lamium mild mosaic virus, a member of the genus Fabavirus
Archives of Virology, 2013Co-Authors: Ezequiel A. Rangel, Stefano Panno, Inmaculada Ferriol, Antonio Olmos, Salvatore Davino, Luis RubioAbstract:Lamium mild mosaic virus (LMMV) is the only one of the five members of the genus Fabavirus for which there are no nucleotide sequence data. In this study, the complete genome sequence of LMMV was determined and compared with the available complete genome sequences of other members of the genus Fabavirus . The genome was the largest of the genus but maintained the typical organization, with RNA 1 of 6080 nucleotides (nt), RNA 2 of 4065 nt, and an unusually long 3′ untranslated region in RNA 2 of 603 nt. Phylogenetic analysis of the amino acid sequences of the protease-polymerase (Pro-Pol) region and the two coat proteins confirmed that LMMV belongs to a distinct species within the genus Fabavirus .
Salvatore Davino - One of the best experts on this subject based on the ideXlab platform.
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Broad bean wilt virus 1 encoded VP47 and SCP are suppressors of plant post-transcriptional gene silencing
European Journal of Plant Pathology, 2020Co-Authors: Caterina Carpino, Inmaculada Ferriol, Luis Rubio, Salvatore Davino, Laura Elvira-gonzález, Ezio Peri, Luis GalipiensoAbstract:Broad bean wilt virus 1 (BBWV-1, genus Fabavirus , family Secoviridae ) is a bipartite positive single-stranded RNA (+ssRNA) virus infecting important horticultural and ornamental crops worldwide. RNA1 encodes proteins involved in virus replication, whereas RNA2 encodes the large and small coat proteins (LCP, and SCP, respectively) and two putative movement proteins with overlapping C-terminal but different sizes: 47.2 kDa (VP47) and 37 kDa (VP37). Post-transcriptional gene silencing (PTGS) is a mechanism of gene regulation and defense against pathogens such as viruses. However, most plant viruses encode proteins called viral suppressors of RNA silencing (VSRs) which able to inhibit PTGS. Previously, BBWV-1 VP37 was identified as VSR and here, we have demonstrated that VP47 and SCP, but not LCP, are also VSRs by two approaches: (i) transient expression in Nicotiana benthamiana 16c plants expressing the green fluorescent protein (GFP) constitutively, and (ii) movement complementation assays using a viral vector based on Turnip crinkle virus sequence containing GFP (pTCV-GFP). In contrast LCP but not SCP of Broad been wilt virus 2 (BBWV-2, genus Fabavirus , family Secoviridae ) is a VSR, which illustrates that viruses of the same genus can interact differently with the host.
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RNA2-encoded VP37 protein of Broad bean wilt virus 1 is a determinant of pathogenicity, host susceptibility, and a suppressor of post-transcriptional gene silencing.
Molecular plant pathology, 2020Co-Authors: Caterina Carpino, Luis Rubio, Salvatore Davino, Laura Elvira-gonzález, Ezio Peri, Inmaculada Ferriol Safont, Vicente Medina, Luis Galipienso TorregrosaAbstract:Broad bean wilt virus 1 (BBWV-1, genus Fabavirus, family Secoviridae) is a bipartite, single-stranded positive-sense RNA virus infecting many horticultural and ornamental crops worldwide. RNA1 encodes proteins involved in viral replication whereas RNA2 encodes two coat proteins (the large and small coat proteins) and two putative movement proteins (MPs) of different sizes with overlapping C-terminal regions. In this work, we determined the role played by the small putative BBWV-1 MP (VP37) on virus pathogenicity, host specificity, and suppression of post-transcriptional gene silencing (PTGS). We engineered a BBWV-1 35S-driven full-length cDNA infectious clone corresponding to BBWV-1 RNA1 and RNA2 (pBBWV1-Wt) and generated a mutant knocking out VP37 (pBBWV1-G492C). Agroinfiltration assays showed that pBBWV1-Wt, as the original BBWV-1 isolate, infected broad bean, tomato, pepper, and Nicotiana benthamiana, whereas pBBWV1-G492C did not infect pepper and tomato systemically. Also, pBBWV1-G492C induced milder symptoms in broad bean and N. benthamiana than pBBWV1-Wt. Differential retrotranscription and amplification of the (+) and (-) strands showed that pBBWV1-G492C replicated in the agroinfiltrated leaves of pepper but not in tomato. All this suggests that VP37 is a determinant of pathogenicity and host specificity. Transient expression of VP37 through a potato virus X (PVX) vector enhanced PVX symptoms and induced systemic necrosis associated with programmed cell death in N. benthamiana plants. Finally, VP37 was identified as a viral suppressor of RNA silencing by transient expression in N. benthamiana 16c plants and movement complementation of a viral construct based on turnip crinkle virus (pTCV-GFP).
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A comparative study of viral infectivity, accumulation and symptoms induced by broad bean wilt virus 1 isolates
Journal of Plant Pathology, 2019Co-Authors: Caterina Carpino, Luis Rubio, Salvatore Davino, Laura Elvira-gonzález, Ezio Peri, Luis GalipiensoAbstract:Broad bean wilt virus 1 (BBWV-1, genus Fabavirus , family Secoviridae ) is a bipartite positive-sense single-stranded RNA virus distributed worldwide infecting many herbaceous species. Until now, scarce information regarding biological properties of BBWV-1 isolates is available. This work shows a comparative study on virus infectivity (proportion of infected plants over inoculated plants), virus accumulation and symptoms induced by four genetically different BBWV-1 isolates (Ben, B41/99, NSRV and PV0548) which were mechanically inoculated on several herbaceous hosts. The four BBWV-1 isolates infected broad bean, tomato, pepper and Nicotiana benthamiana plants, whereas none of them infected cucumber, common bean and melon. Infectivity ranged from 40 to 60% in tomato and from 75% to 95% in pepper, whereas it was 100% in broad bean and N. benthamiana for the four BBWV-1 isolates. Symptoms showed differences depending on the host plant, the viral isolate and the infection time. Virus accumulation was determined in broad bean and pepper plants and showed differences among host species and BBWV-1 isolates. No association between plant symptom manifestation and viral titre was observed.
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Rapid detection and discrimination of Fabaviruses by flow-through hybridisation with genus- and species-specific riboprobes
Annals of Applied Biology, 2015Co-Authors: Inmaculada Ferriol, Stefano Panno, Ezequiel A. Rangel, Antonio Olmos, Chenggui Han, Salvatore Davino, Luis RubioAbstract:Viruses cause significant damage in agricultural crops worldwide. Disease management requires sensitive and specific tools for virus detection and identification. Also, detection techniques need to be rapid to keep pace with the continuous emergence of new viral diseases. The genus Fabavirus is composed of five viruses infecting many economically important crops worldwide. This research describes the development of a procedure based on flow-through hybridisation (FTH), which is faster than and as sensitive as conventional hybridisation for virus detection in tissue-prints from infected plants. Six digoxigenin-labelled RNA probes were synthesised with two levels of specificity: (a) five specific for each viral species within this genus, and (b) a genus-specific probe that hybridises with a nucleotide sequence signature only found in the 5′-untranslated region of the genus Fabavirus, which is the first of this type reported for plant viruses. The new procedure developed is useful for rapid detection and discrimination of the five Fabaviruses identified so far and opens the possibility of discovering new species of this genus.
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Detection and identification of Fabavirus species by one-step RT-PCR and multiplex RT-PCR.
Journal of virological methods, 2013Co-Authors: Stefano Panno, Inmaculada Ferriol, Ezequiel A. Rangel, Antonio Olmos, Chenggui Han, Federico Martinelli, Luis Rubio, Salvatore DavinoAbstract:The genus Fabavirus of the family Secoviridae comprises a group of poorly characterized viruses. To date, only five species have been described: Broad bean wilt virus 1 (BBWV-1), Broad bean wilt virus 2 (BBWV-2), Lamium mild mosaic virus (LMMV), Gentian mosaic virus (GeMV) and Cucurbit mild mosaic virus (CuMMV). The development is described of two RT-PCR procedures for the detection and identification of Fabavirus species: a one-step RT-PCR using a single pair of conserved primers for the detection of all Fabaviruses, and a one-step multiplex RT-PCR using species-specific primers for the simultaneous detection and identification of the above-mentioned species of the genus Fabavirus. These methods were applied successfully to field samples and the results were compared with those obtained by molecular hybridization and ELISA. The combination of the two techniques enables rapid, sensitive and reliable identification of the five known Fabavirus species, as well as the possibility of discovering new species of this genus.
Xueping Zhou - One of the best experts on this subject based on the ideXlab platform.
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Broad bean wilt virus 2 encoded VP53, VP37 and large capsid protein orchestrate suppression of RNA silencing in plant
Virus research, 2014Co-Authors: Lingfang Kong, Yaqin Wang, Xiuling Yang, Garry Sunter, Xueping ZhouAbstract:Viruses encode RNA silencing suppressors to counteract host RNA silencing-mediated defense responses. In this study, we demonstrate that VP53, VP37 and LCP encoded by RNA2 of broad bean wilt virus 2 (BBWV-2), a member of the genus Fabavirus, are strong suppressors of RNA silencing triggered by single-stranded sense RNA. They, however, had no effect on suppression of RNA silencing induced by double-stranded RNA. We provide evidence that these three suppressors can significantly limit the accumulation of small interfering RNAs (siRNAs) in tissues where the GFP gene has been silenced, and prevent the long distance spread of the induced silencing signal. Gel mobility shift assays showed that all three suppressors could bind ssRNA in a size-specific manner. Interestingly, VP37 and LCP, but not VP53, could reverse the silencing of a GFP gene in leaf tissue. Furthermore, these three proteins are capable of enhancing pathogenicity of potato virus X. Collectively, our findings indicate that viruses employ a more sophisticated strategy to overcome the host defense response mediated through suppression of RNA silencing during virus infection. As far as we are aware, this is the first report of RNA silencing suppressors encoded by a virus in the genus Fabavirus.
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Cell-to-cell trafficking, subcellular distribution, and binding to coat protein of Broad bean wilt virus 2 VP37 protein.
Virus research, 2009Co-Authors: Chengke Liu, Chunmei Meng, Li Xie, Jian Hong, Xueping ZhouAbstract:Broad bean wilt virus 2 (BBWV 2) is a member of the genus Fabavirus of the family Comoviridae. To date, a movement protein (MP) of BBWV 2 has not been described. Here we demonstrate that the green fluorescent protein (GFP)-VP37 fusion protein can move from initial bombarded cells to neighboring cells in Nicotiana benthamiana epidermal leaves. In addition, the GFP-VP37 fusion protein localizes as a halo around the nucleus and as punctate spots on the cell periphery in N. benthamiana epidermal leaf cells and BY-2 suspension cells. Fluorescence near the nucleus also was co-localized with the endoplasmic reticulum in BY-2 cells. Fibrous networks were found in GFP-VP37 agro-infiltrated N. benthamiana epidermal leaf cells. Deletion analyses indicated that the C-terminal region of the VP37 protein is essential for localization at the cell periphery. Using a blot overlay assay and bimolecular fluorescence complementation assay, the purified 6×His-tagged VP37 protein was shown to bind specifically to the small coat protein of BBWV 2. The above results indicate that VP37 is a movement protein.
Antonio Olmos - One of the best experts on this subject based on the ideXlab platform.
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Recent Advances on Detection and Characterization of Fruit Tree Viruses Using High-Throughput Sequencing Technologies
Viruses, 2018Co-Authors: Varvara I. Maliogka, Angelantonio Minafra, Pasquale Saldarelli, Ana B. Ruiz-garcía, Miroslav Glasa, Nikolaos I. Katis, Antonio OlmosAbstract:Perennial crops, such as fruit trees, are infected by many viruses, which are transmitted through vegetative propagation and grafting of infected plant material. Some of these pathogens cause severe crop losses and often reduce the productive life of the orchards. Detection and characterization of these agents in fruit trees is challenging, however, during the last years, the wide application of high-throughput sequencing (HTS) technologies has significantly facilitated this task. In this review, we present recent advances in the discovery, detection, and characterization of fruit tree viruses and virus-like agents accomplished by HTS approaches. A high number of new viruses have been described in the last 5 years, some of them exhibiting novel genomic features that have led to the proposal of the creation of new genera, and the revision of the current virus taxonomy status. Interestingly, several of the newly identified viruses belong to virus genera previously unknown to infect fruit tree species (e.g., Fabavirus, Luteovirus) a fact that challenges our perspective of plant viruses in general. Finally, applied methodologies, including the use of different molecules as templates, as well as advantages and disadvantages and future directions of HTS in fruit tree virology are discussed.
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Rapid detection and discrimination of Fabaviruses by flow-through hybridisation with genus- and species-specific riboprobes
Annals of Applied Biology, 2015Co-Authors: Inmaculada Ferriol, Stefano Panno, Ezequiel A. Rangel, Antonio Olmos, Chenggui Han, Salvatore Davino, Luis RubioAbstract:Viruses cause significant damage in agricultural crops worldwide. Disease management requires sensitive and specific tools for virus detection and identification. Also, detection techniques need to be rapid to keep pace with the continuous emergence of new viral diseases. The genus Fabavirus is composed of five viruses infecting many economically important crops worldwide. This research describes the development of a procedure based on flow-through hybridisation (FTH), which is faster than and as sensitive as conventional hybridisation for virus detection in tissue-prints from infected plants. Six digoxigenin-labelled RNA probes were synthesised with two levels of specificity: (a) five specific for each viral species within this genus, and (b) a genus-specific probe that hybridises with a nucleotide sequence signature only found in the 5′-untranslated region of the genus Fabavirus, which is the first of this type reported for plant viruses. The new procedure developed is useful for rapid detection and discrimination of the five Fabaviruses identified so far and opens the possibility of discovering new species of this genus.
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Detection and identification of Fabavirus species by one-step RT-PCR and multiplex RT-PCR.
Journal of virological methods, 2013Co-Authors: Stefano Panno, Inmaculada Ferriol, Ezequiel A. Rangel, Antonio Olmos, Chenggui Han, Federico Martinelli, Luis Rubio, Salvatore DavinoAbstract:The genus Fabavirus of the family Secoviridae comprises a group of poorly characterized viruses. To date, only five species have been described: Broad bean wilt virus 1 (BBWV-1), Broad bean wilt virus 2 (BBWV-2), Lamium mild mosaic virus (LMMV), Gentian mosaic virus (GeMV) and Cucurbit mild mosaic virus (CuMMV). The development is described of two RT-PCR procedures for the detection and identification of Fabavirus species: a one-step RT-PCR using a single pair of conserved primers for the detection of all Fabaviruses, and a one-step multiplex RT-PCR using species-specific primers for the simultaneous detection and identification of the above-mentioned species of the genus Fabavirus. These methods were applied successfully to field samples and the results were compared with those obtained by molecular hybridization and ELISA. The combination of the two techniques enables rapid, sensitive and reliable identification of the five known Fabavirus species, as well as the possibility of discovering new species of this genus.
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The complete genome sequence of Lamium mild mosaic virus, a member of the genus Fabavirus
Archives of Virology, 2013Co-Authors: Ezequiel A. Rangel, Stefano Panno, Inmaculada Ferriol, Antonio Olmos, Salvatore Davino, Luis RubioAbstract:Lamium mild mosaic virus (LMMV) is the only one of the five members of the genus Fabavirus for which there are no nucleotide sequence data. In this study, the complete genome sequence of LMMV was determined and compared with the available complete genome sequences of other members of the genus Fabavirus . The genome was the largest of the genus but maintained the typical organization, with RNA 1 of 6080 nucleotides (nt), RNA 2 of 4065 nt, and an unusually long 3′ untranslated region in RNA 2 of 603 nt. Phylogenetic analysis of the amino acid sequences of the protease-polymerase (Pro-Pol) region and the two coat proteins confirmed that LMMV belongs to a distinct species within the genus Fabavirus .
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The complete genome sequence of Lamium mild mosaic virus, a member of the genus Fabavirus.
Archives of virology, 2013Co-Authors: Ezequiel A. Rangel, Stefano Panno, Inmaculada Ferriol, Antonio Olmos, Salvatore Davino, Luis RubioAbstract:Springer-Verlag Wien 2013 Abstract Lamium mild mosaic virus (LMMV) is the only one of the five members of the genus Fabavirus for which there are no nucleotide sequence data. In this study, the complete genome sequence of LMMV was determined and compared with the available complete genome sequences of other members of the genus Fabavirus. The genome was the largest of the genus but maintained the typical orga- nization, with RNA 1 of 6080 nucleotides (nt), RNA 2 of 4065 nt, and an unusually long 3 0 untranslated region in RNA 2 of 603 nt. Phylogenetic analysis of the amino acid sequences of the protease-polymerase (Pro-Pol) region and the two coat proteins confirmed that LMMV belongs to a distinct species within the genus Fabavirus. The genus Fabavirus, subfamily Comovirinae, family Secoviridae (21) consists of five viral species: Broad bean wilt virus 1 (BBWV-1), Broad bean wilt virus 2 (BBWV- 2), Gentian mosaic virus (GeMV), Cucurbit mild mosaic virus (CuMMV) and Lamium mild mosaic virus (LMMV). Fabaviruses infect a wide range of hosts, including dicot- yledonous plants and some families of monocotyledonous
