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Development of a gene silencing DNA vector derived from a broad host range geminivirus
BACKGROUND: Gene silencing is proving to be a powerful tool for genetic, developmental, and physiological analyses. The use of viral induced gene silencing (VIGS) offers advantages to transgenic approaches as it can be potentially applied to non-model systems for which transgenic techniques are not...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2009
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2713212/ https://www.ncbi.nlm.nih.gov/pubmed/19573239 http://dx.doi.org/10.1186/1746-4811-5-9 |
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author | Golenberg, Edward M Sather, D Noah Hancock, Leandria C Buckley, Kenneth J Villafranco, Natalie M Bisaro, David M |
author_facet | Golenberg, Edward M Sather, D Noah Hancock, Leandria C Buckley, Kenneth J Villafranco, Natalie M Bisaro, David M |
author_sort | Golenberg, Edward M |
collection | PubMed |
description | BACKGROUND: Gene silencing is proving to be a powerful tool for genetic, developmental, and physiological analyses. The use of viral induced gene silencing (VIGS) offers advantages to transgenic approaches as it can be potentially applied to non-model systems for which transgenic techniques are not readily available. However, many VIGS vectors are derived from Gemini viruses that have limited host ranges. We present a new, unipartite vector that is derived from a curtovirus that has a broad host range and will be amenable to use in many non-model systems. RESULTS: The construction of a gene silencing vector derived from the geminivirus Beet curly top virus (BCTV), named pWSRi, is reported. Two versions of the vector have been developed to allow application by biolistic techniques or by agro-infiltration. We demonstrate its ability to silence nuclear genes including ribulose bisphosphate carboxylase small subunit (rbcS), transketolase, the sulfur allele of magnesium chelatase (ChlI), and two homeotic transcription factors in spinach or tomato by generating gene-specific knock-down phenotypes. Onset of phenotypes occurred 3 to 12 weeks post-inoculation, depending on the target gene, in organs that developed after the application. The vector lacks movement genes and we found no evidence for significant spread from the site of inoculation. However, viral amplification in inoculated tissue was detected and is necessary for systemic silencing, suggesting that signals generated from active viral replicons are efficiently transported within the plant. CONCLUSION: The unique properties of the pWSRi vector, the ability to silence genes in meristem tissue, the separation of virus and silencing phenotypes, and the broad natural host range of BCTV, suggest that it will have wide utility. |
format | Text |
id | pubmed-2713212 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2009 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-27132122009-07-21 Development of a gene silencing DNA vector derived from a broad host range geminivirus Golenberg, Edward M Sather, D Noah Hancock, Leandria C Buckley, Kenneth J Villafranco, Natalie M Bisaro, David M Plant Methods Methodology BACKGROUND: Gene silencing is proving to be a powerful tool for genetic, developmental, and physiological analyses. The use of viral induced gene silencing (VIGS) offers advantages to transgenic approaches as it can be potentially applied to non-model systems for which transgenic techniques are not readily available. However, many VIGS vectors are derived from Gemini viruses that have limited host ranges. We present a new, unipartite vector that is derived from a curtovirus that has a broad host range and will be amenable to use in many non-model systems. RESULTS: The construction of a gene silencing vector derived from the geminivirus Beet curly top virus (BCTV), named pWSRi, is reported. Two versions of the vector have been developed to allow application by biolistic techniques or by agro-infiltration. We demonstrate its ability to silence nuclear genes including ribulose bisphosphate carboxylase small subunit (rbcS), transketolase, the sulfur allele of magnesium chelatase (ChlI), and two homeotic transcription factors in spinach or tomato by generating gene-specific knock-down phenotypes. Onset of phenotypes occurred 3 to 12 weeks post-inoculation, depending on the target gene, in organs that developed after the application. The vector lacks movement genes and we found no evidence for significant spread from the site of inoculation. However, viral amplification in inoculated tissue was detected and is necessary for systemic silencing, suggesting that signals generated from active viral replicons are efficiently transported within the plant. CONCLUSION: The unique properties of the pWSRi vector, the ability to silence genes in meristem tissue, the separation of virus and silencing phenotypes, and the broad natural host range of BCTV, suggest that it will have wide utility. BioMed Central 2009-07-02 /pmc/articles/PMC2713212/ /pubmed/19573239 http://dx.doi.org/10.1186/1746-4811-5-9 Text en Copyright © 2009 Golenberg et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( (http://creativecommons.org/licenses/by/2.0) ), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methodology Golenberg, Edward M Sather, D Noah Hancock, Leandria C Buckley, Kenneth J Villafranco, Natalie M Bisaro, David M Development of a gene silencing DNA vector derived from a broad host range geminivirus |
title | Development of a gene silencing DNA vector derived from a broad host range geminivirus |
title_full | Development of a gene silencing DNA vector derived from a broad host range geminivirus |
title_fullStr | Development of a gene silencing DNA vector derived from a broad host range geminivirus |
title_full_unstemmed | Development of a gene silencing DNA vector derived from a broad host range geminivirus |
title_short | Development of a gene silencing DNA vector derived from a broad host range geminivirus |
title_sort | development of a gene silencing dna vector derived from a broad host range geminivirus |
topic | Methodology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2713212/ https://www.ncbi.nlm.nih.gov/pubmed/19573239 http://dx.doi.org/10.1186/1746-4811-5-9 |
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