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Agrobacterium -mediated large-scale transformation of wheat ( Triticum aestivum L.) using glyphosate selection

Agrobacterium -mediated large-scale transformation of wheat ( Triticum aestivum L.) using... An Agrobacterium -mediated transformation system with glyphosate selection has been developed for the large-scale production of transgenic plants. The system uses 4-day precultured immature embryos as explants. A total of 30 vectors containing the 5-enol-pyruvylshikimate-3-phosphate synthase gene from Agrobacterium strain CP4 ( aroA :CP4), which confers resistance to glyphosate, were introduced into wheat using this system. The aroA :CP4 gene served two roles in this study—selectable marker and gene of interest. More than 3,000 transgenic events were produced with an average transformation efficiency of 4.4%. The entire process from isolation of immature embryos to production of transgenic plantlets was 50–80 days. Transgenic events were evaluated over several generations based on genetic, agronomic and molecular criteria. Forty-six percent of the transgenic events fit a 3:1 segregation ratio. Molecular analysis confirmed that four of six lead transgenic events selected from Agrobacterium transformation contained a single insert and a single copy of the transgene. Stable expression of the aroA: CP4 gene was confirmed by ELISA through nine generations. A comparison of Agrobacterium -mediated transformation to a particle bombardment system demonstrated that the Agrobacterium system is reproducible, has a higher transformation efficiency with glyphosate selection and produces higher quality transgenic events in wheat. One of the lead events from this study, no. 33391, has been identified as a Roundup Ready wheat commercial candidate. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Plant Cell Reports Springer Journals

Agrobacterium -mediated large-scale transformation of wheat ( Triticum aestivum L.) using glyphosate selection

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References (42)

Publisher
Springer Journals
Copyright
Copyright © 2003 by Springer-Verlag
Subject
LifeSciences
ISSN
0721-7714
eISSN
1432-203X
DOI
10.1007/s00299-003-0617-6
pmid
12835912
Publisher site
See Article on Publisher Site

Abstract

An Agrobacterium -mediated transformation system with glyphosate selection has been developed for the large-scale production of transgenic plants. The system uses 4-day precultured immature embryos as explants. A total of 30 vectors containing the 5-enol-pyruvylshikimate-3-phosphate synthase gene from Agrobacterium strain CP4 ( aroA :CP4), which confers resistance to glyphosate, were introduced into wheat using this system. The aroA :CP4 gene served two roles in this study—selectable marker and gene of interest. More than 3,000 transgenic events were produced with an average transformation efficiency of 4.4%. The entire process from isolation of immature embryos to production of transgenic plantlets was 50–80 days. Transgenic events were evaluated over several generations based on genetic, agronomic and molecular criteria. Forty-six percent of the transgenic events fit a 3:1 segregation ratio. Molecular analysis confirmed that four of six lead transgenic events selected from Agrobacterium transformation contained a single insert and a single copy of the transgene. Stable expression of the aroA: CP4 gene was confirmed by ELISA through nine generations. A comparison of Agrobacterium -mediated transformation to a particle bombardment system demonstrated that the Agrobacterium system is reproducible, has a higher transformation efficiency with glyphosate selection and produces higher quality transgenic events in wheat. One of the lead events from this study, no. 33391, has been identified as a Roundup Ready wheat commercial candidate.

Journal

Plant Cell ReportsSpringer Journals

Published: Jun 1, 2003

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