Development of Efficient Plant Regeneration and Transformation System for Impatiens Using Agrobacterium tumefaciens and Multiple Bud Cultures as Explants

Simple item page
dc.contributor.authorDan, Yinghuien
dc.contributor.authorBaxter, Aaronen
dc.contributor.authorZhang, Songen
dc.contributor.authorPantazis, Christopher J.en
dc.contributor.authorVeilleux, Richard E.en
dc.contributor.departmentSchool of Plant and Environmental Sciencesen
dc.date.accessioned2019-11-13T13:49:26Zen
dc.date.available2019-11-13T13:49:26Zen
dc.date.issued2010-08-09en
dc.description.abstractBackground: Impatiens (Impatiens walleriana) is a top selling floriculture crop. The potential for genetic transformation of Impatiens to introduce novel flower colors or virus resistance has been limited by its general recalcitrance to tissue culture and transformation manipulations. We have established a regeneration and transformation system for Impatiens that provides new alternatives to genetic improvement of this crop. Results: In a first step towards the development of transgenic INSV-resistant Impatiens, we developed an efficient plant regeneration system using hypocotyl segments containing cotyledonary nodes as explants. With this regeneration system, 80% of explants produced an average of 32.3 elongated shoots per initial explant plated, with up to 167 elongated shoots produced per explant. Rooting efficiency was high, and 100% of shoots produced roots within 12 days under optimal conditions, allowing plant regeneration within approximately 8 weeks. Using this regeneration system, we developed an efficient Agrobacterium-mediated Impatiens transformation method using in vitro multiple bud cultures as explants and a binary plasmid (pHB2892) bearing gfp and nptII genes. Transgenic Impatiens plants, with a frequency up to 58.9%, were obtained within 12 to 16 weeks from inoculation to transfer of transgenic plants to soil. Transgenic plants were confirmed by Southern blot, phenotypic assays and T1 segregation analysis. Transgene expression was observed in leaves, stems, roots, flowers, and fruit. The transgenic plants were fertile and phenotypically normal. Conclusion: We report the development of a simple and efficient Agrobacterium-mediated transformation system for Impatiens. To the best of our knowledge, there have been no reports of Agrobacterium-mediated transformation of Impatiens with experimental evidence of stable integration of T-DNA and of Agrobacterium-mediated transformation method for plants using in vitro maintained multiple bud cultures as explants. This transformation system has the advantages of 1) efficient, simple and rapid regeneration and transformation (with no need for sterilization or a greenhouse to grow stock plants), 2) flexibility (available all the time) for in vitro manipulation, 3) uniform and desirable green tissue explants for both nuclear and plastid transformation using Agrobacteriummediated and biolistics methods, 4) no somaclonal variation and 5) resolution of necrosis of Agrobacteriuminoculated tissues.en
dc.description.sponsorshipThis work was supported through funds from the “High Value Horticulture and Forestry-Virginia” Special Grants (Project No. 2003-38891- 02112) from the United States Department of Agriculture CSREES, the Department of Horticulture, the Virginia Tobacco Indemnification and Community Revitalization Commission and operating funds from the Commonwealth of Virginia to the Institute for Advanced Learning and Research.en
dc.format.extent12 pagesen
dc.format.mimetypeapplication/pdfen
dc.identifier.citationDan et al.: Development of Efficient Plant Regeneration and Transformation System for Impatiens Using Agrobacterium tumefaciens and Multiple Bud Cultures as Explants. BMC Plant Biology 2010 10:165.en
dc.identifier.doihttps://doi.org/10.1186/1471-2229-10-165en
dc.identifier.urihttp://hdl.handle.net/10919/95527en
dc.identifier.volume10en
dc.language.isoenen
dc.publisherBMCen
dc.rightsCreative Commons Attribution 2.0 Genericen
dc.rights.urihttp://creativecommons.org/licenses/by/2.0/en
dc.titleDevelopment of Efficient Plant Regeneration and Transformation System for Impatiens Using Agrobacterium tumefaciens and Multiple Bud Cultures as Explantsen
dc.title.serialBMC Plant Biologyen
dc.typeArticle - Refereeden
dc.type.dcmitypeTexten

Files

Original bundle
Now showing 1 - 1 of 1
Thumbnail Image
Name:
1471-2229-10-165.pdf
Size:
5.17 MB
Format:
Adobe Portable Document Format
Description:
Download

Collections

Scholarly Works, School of Plant and Environmental Sciences