Method: low-cost delivery of the cotton leaf crumple virus-induced gene silencing system
<p>Abstract</p> <p>Background</p> <p>We previously developed a virus-induced gene silencing (VIGS) vector for cotton from the bipartite geminivirus<it>Cotton leaf crumple virus</it> (CLCrV). The original CLCrV VIGS vector was designed for biolistic delivery...
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| Format: | Article |
| Language: | English |
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BMC
2012-08-01
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| Series: | Plant Methods |
| Subjects: | |
| Online Access: | http://www.plantmethods.com/content/8/1/27 |
| _version_ | 1818753811114622976 |
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| author | Tuttle John Haigler Candace H Robertson Dominique |
| author_facet | Tuttle John Haigler Candace H Robertson Dominique |
| author_sort | Tuttle John |
| collection | DOAJ |
| description | <p>Abstract</p> <p>Background</p> <p>We previously developed a virus-induced gene silencing (VIGS) vector for cotton from the bipartite geminivirus<it>Cotton leaf crumple virus</it> (CLCrV). The original CLCrV VIGS vector was designed for biolistic delivery by a gene gun. This prerequisite limited the use of the system to labs with access to biolistic equipment. Here we describe the adaptation of this system for delivery by <it>Agrobacterium </it>(<it>Agrobacterium tumefaciens</it>). We also describe the construction of two low-cost particle inflow guns.</p> <p>Results</p> <p>The biolistic CLCrV vector was transferred into two <it>Agrobacterium </it> binary plasmids. Agroinoculation of the binary plasmids into cotton resulted in silencing and GFP expression comparable to the biolistic vector. Two homemade low-cost gene guns were used to successfully inoculate cotton (<it>G. hirsutum) </it> and <it>N. benthamiana</it> with either the CLCrV VIGS vector or the Tomato golden mosaic virus (TGMV) VIGS vector respectively.</p> <p>Conclusions</p> <p>These innovations extend the versatility of CLCrV-based VIGS for analyzing gene function in cotton. The two low-cost gene guns make VIGS experiments affordable for both research and teaching labs by providing a working alternative to expensive commercial gene guns.</p> |
| first_indexed | 2024-12-18T05:13:17Z |
| format | Article |
| id | doaj.art-ea6688c230754f5b9022b1ad3d3c892a |
| institution | Directory Open Access Journal |
| issn | 1746-4811 |
| language | English |
| last_indexed | 2024-12-18T05:13:17Z |
| publishDate | 2012-08-01 |
| publisher | BMC |
| record_format | Article |
| series | Plant Methods |
| spelling | doaj.art-ea6688c230754f5b9022b1ad3d3c892a2022-12-21T21:19:50ZengBMCPlant Methods1746-48112012-08-01812710.1186/1746-4811-8-27Method: low-cost delivery of the cotton leaf crumple virus-induced gene silencing systemTuttle JohnHaigler Candace HRobertson Dominique<p>Abstract</p> <p>Background</p> <p>We previously developed a virus-induced gene silencing (VIGS) vector for cotton from the bipartite geminivirus<it>Cotton leaf crumple virus</it> (CLCrV). The original CLCrV VIGS vector was designed for biolistic delivery by a gene gun. This prerequisite limited the use of the system to labs with access to biolistic equipment. Here we describe the adaptation of this system for delivery by <it>Agrobacterium </it>(<it>Agrobacterium tumefaciens</it>). We also describe the construction of two low-cost particle inflow guns.</p> <p>Results</p> <p>The biolistic CLCrV vector was transferred into two <it>Agrobacterium </it> binary plasmids. Agroinoculation of the binary plasmids into cotton resulted in silencing and GFP expression comparable to the biolistic vector. Two homemade low-cost gene guns were used to successfully inoculate cotton (<it>G. hirsutum) </it> and <it>N. benthamiana</it> with either the CLCrV VIGS vector or the Tomato golden mosaic virus (TGMV) VIGS vector respectively.</p> <p>Conclusions</p> <p>These innovations extend the versatility of CLCrV-based VIGS for analyzing gene function in cotton. The two low-cost gene guns make VIGS experiments affordable for both research and teaching labs by providing a working alternative to expensive commercial gene guns.</p>http://www.plantmethods.com/content/8/1/27CottonVIGSVirus-induced gene silencingCotton leaf crumple virusAgroinoculationParticle inflow gunBombardmentGossypium |
| spellingShingle | Tuttle John Haigler Candace H Robertson Dominique Method: low-cost delivery of the cotton leaf crumple virus-induced gene silencing system Plant Methods Cotton VIGS Virus-induced gene silencing Cotton leaf crumple virus Agroinoculation Particle inflow gun Bombardment Gossypium |
| title | Method: low-cost delivery of the cotton leaf crumple virus-induced gene silencing system |
| title_full | Method: low-cost delivery of the cotton leaf crumple virus-induced gene silencing system |
| title_fullStr | Method: low-cost delivery of the cotton leaf crumple virus-induced gene silencing system |
| title_full_unstemmed | Method: low-cost delivery of the cotton leaf crumple virus-induced gene silencing system |
| title_short | Method: low-cost delivery of the cotton leaf crumple virus-induced gene silencing system |
| title_sort | method low cost delivery of the cotton leaf crumple virus induced gene silencing system |
| topic | Cotton VIGS Virus-induced gene silencing Cotton leaf crumple virus Agroinoculation Particle inflow gun Bombardment Gossypium |
| url | http://www.plantmethods.com/content/8/1/27 |
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