Stable Transformation of Wheat by Microprojectile Bombardment
The following protocol describes methods that have been used successfully in the rapid production of multiple independent lines of fertile transgenic wheat (Weeks et al. 1993). Several features of the transformation protocol have been reported and used by other laboratories (Vasil et al. 1993; Becker et al. 1994; Nehra et al. 1994). These features include the biolistics-mediated gene delivery into cell tissue derived from immature embryos and, secondly, the bombardment of calli with microprojectiles coated with DNA containing the selectable marker gene, bar (phosphinothricin acetyl transferase). Selection is carried out using the herbicides bialaphos and Basta.
KeywordsVortex Sucrose Maize Agar Electrophoresis
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- Daines RJ (1990) Dekalb researchers improve maize transformation rates. Biolistic Syst Newsl 1: 1–4Google Scholar
- D’Ovidio R, Tanzarella OA, Porceddu E (1992) Isolation of an alpha-type gliadin gene fromGoogle Scholar
- Triticum durum Desf. and genetic polymorphism at the Gli-2 loci. J Genet Breed 46:41–48Google Scholar
- Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissueGoogle Scholar
- cultures. Physiol Plant 15:473–497Google Scholar
- Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual. Cold Spring Harbor Press, Cold Spring Harbor, New YorkGoogle Scholar
- Vasil V, Srivastava V, Castillo AM, Fromm ME, Vasil IK (1993) Rapid production of transgenic wheat plants by direct bombardment of cultured immature embryos. Bio/Technology 11: 15531558Google Scholar