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Routine transformation system for use with CIMMYT wheat varieties

By: Pellegrineschi, A | Centro Internacional de Mejoramiento de Maiz y Trigo (CIMMYT), Mexico DF (Mexico).
Contributor(s): Brito, R.M [coaut.] | Fennell, S [coaut.] | Hernández, R [coaut.] | Hoisington, D.A [coaut.] | Kohli, M.M.|Francis, M [eds.] | McLean, S.D [coaut.] | Olivares, J.J [coaut.] | Salgado, M [coaut.] | Velázquez, L [coaut.].
Material type: materialTypeLabelBookAnalytics: Show analyticsPublisher: Montevideo (Uruguay) CIMMYT : 2000ISBN: 9974-7586-1-0.Subject(s): Chromosome translocation | Gene expression | Gene transfer | Genetic transformation | Research projects | Varieties | CIMMYT | Wheat AGROVOC | Plant breeding AGROVOCSummary: The inability to reach and regenerate the proper cells in the target tissue and problems related to gene expression have been major limiting factors affecting wheat transformations. This paper describes a routine biolistic bombardment-based transformation technology that overcomes these barriers and allows DNA transfer in wheat. The method involves subjecting the plant tissue to microprojectile bombardment with purified DNA and reduced levels of tissue culture manipulation. In vitro culture response of bombarded immature embryos and the production of transgenic plants were investigated in 3 wheat cultivars: Bobwhite, Baviacora, and Attila. In the three genotypes, at least 700 immature embryos were bombarded with a plasmid containing a PPT -resistant gene (Bar) under the control of the ubiquitin promoter that included the reporter gene Gus ({3-glucuronidase). Although the transient expression of the reporter gene {3-glucuronidase fused with the same promoter was similar in the three cultivars tested, the frequency of stable transformations varied with the genotype. The transformation frequency was highest in Bobwhite in which 0.24 to 0.44% of bombarded embryos produced plants. The other two varieties, Baviacora and Attila, also expressed Basta resistance but at a lower frequency than Bobwhite. Preliminary data suggests that the variation in the rate of transformation is more influenced by the differences among the genotypes than by the efficiency of the particle bombardment procedure.||Abbreviations: Phosphoeritromicin (PPT); Dichlorophenoxyacetic Acid (2, 4-D); {3-glucuronidase (Gus),. Phosphinothricin acetyltransferase (Bar); inches cm square (ics).Collection: CIMMYT Staff Publications Collection
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Conference proceedings CIMMYT Knowledge Center: John Woolston Library

Lic. Jose Juan Caballero Flores

 

CIMMYT Staff Publications Collection CIS-2460 (Browse shelf) 1 Available 649048
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The inability to reach and regenerate the proper cells in the target tissue and problems related to gene expression have been major limiting factors affecting wheat transformations. This paper describes a routine biolistic bombardment-based transformation technology that overcomes these barriers and allows DNA transfer in wheat. The method involves subjecting the plant tissue to microprojectile bombardment with purified DNA and reduced levels of tissue culture manipulation. In vitro culture response of bombarded immature embryos and the production of transgenic plants were investigated in 3 wheat cultivars: Bobwhite, Baviacora, and Attila. In the three genotypes, at least 700 immature embryos were bombarded with a plasmid containing a PPT -resistant gene (Bar) under the control of the ubiquitin promoter that included the reporter gene Gus ({3-glucuronidase). Although the transient expression of the reporter gene {3-glucuronidase fused with the same promoter was similar in the three cultivars tested, the frequency of stable transformations varied with the genotype. The transformation frequency was highest in Bobwhite in which 0.24 to 0.44% of bombarded embryos produced plants. The other two varieties, Baviacora and Attila, also expressed Basta resistance but at a lower frequency than Bobwhite. Preliminary data suggests that the variation in the rate of transformation is more influenced by the differences among the genotypes than by the efficiency of the particle bombardment procedure.||Abbreviations: Phosphoeritromicin (PPT); Dichlorophenoxyacetic Acid (2, 4-D); {3-glucuronidase (Gus),. Phosphinothricin acetyltransferase (Bar); inches cm square (ics).

English

R99-00CIMPU|0011|EE|AL-Wheat Program|3

Jose Juan Caballero

CIMMYT Staff Publications Collection

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