Transformation of wheat (Triticum aestivum L.) through electroporation of protoplasts

He D.G., Mouradov A., Yang Y.M., Mouradova E. and Scott K.J. (1994) Transformation of wheat (Triticum aestivum L.) through electroporation of protoplasts. Plant Cell Reports, 14 2-3: 192-196. doi:10.1007/BF00233789


Author He D.G.
Mouradov A.
Yang Y.M.
Mouradova E.
Scott K.J.
Title Transformation of wheat (Triticum aestivum L.) through electroporation of protoplasts
Journal name Plant Cell Reports   Check publisher's open access policy
ISSN 0721-7714
Publication date 1994
Sub-type Article (original research)
DOI 10.1007/BF00233789
Volume 14
Issue 2-3
Start page 192
End page 196
Total pages 5
Publisher Springer-Verlag
Subject 1110 Nursing
Abstract Protoplasts isolated from embryogenic suspension cultures of wheat (Triticum aestivum cv. Hartog) were electroporated in the presence of plasmid pEmuGN and/or pEmuPAT, which contained the reporter gene gus and selectable marker gene bar, respectively. Under optimised electroporation conditions, up to 0.9% of viable protoplasts displayed gus activity two days after electroporation. To select for phosphinothricin (PPT) resistant colonies, electroporated protoplasts were incubated for six weeks in a medium containing 10 μg/ml PPT. The cells surviving the selection were maintained as individual colonies on solid medium or as suspension cultures. More than 60% of these colonies exhibited tolerance to 40 μg/ml PPT when tested 10 months after initial selection. To date, 57 green plants have been regenerated from these colonies and 24 have been transferred to soil. Southern blot analyses of colonies and plants, using the bar gene sequence as the probe, confirmed transformation of the cells. Positive PAT assays of both regenerated colonies and plants indicated the presence of the bar gene product. These results provide a basis for the establishment of routine procedures for transformation of wheat by direct gene transfer into protoplasts.
Keyword gene transfer
selection for phosphinothricin resistance
transgenic plants
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

Document type: Journal Article
Sub-type: Article (original research)
Collection: Scopus Import
 
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