A Modular Vector for Agrobacterium Mediated Transformation of Wheat

Wheat (cv Chinese Spring) tissues were transformed using Agrobacterium tumefasciens and a new plasmid modular vector, pMVTBP. We constructed pMVTBP with unique restriction sites connecting (1) the CaMV 35S promoter, (2) a Kozak sequence, (3) the FLAG epitope, (4) the (His)^sub 6^ epitope, (5) a codi...

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Bibliographic Details
Published inPlant molecular biology reporter Vol. 17; no. 4; p. 323
Main Authors Peters, Noël R, Ackerman, Steven, Davis, Elizabeth A
Format Journal Article
LanguageEnglish
Published New York Springer Nature B.V 01.01.1999
Subjects
3' Untranslated regions
Bacteria
Callus
Embryos
Epitopes
Explants
Gene expression
Genetic transformation
Geneticin
Modular construction
Plantlets
Reporter gene
Seeds
TATA-binding protein
Tissues
Virulence
Wheat
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Summary:Wheat (cv Chinese Spring) tissues were transformed using Agrobacterium tumefasciens and a new plasmid modular vector, pMVTBP. We constructed pMVTBP with unique restriction sites connecting (1) the CaMV 35S promoter, (2) a Kozak sequence, (3) the FLAG epitope, (4) the (His)^sub 6^ epitope, (5) a coding region (for wheat TATA Binding Protein, wTBP) and (6) the CaMV 35S 3'UTR. This vector thus allows easy exchange of different regulatory or coding sequences. Explants of either germinating mature seeds, or immature embryos, were induced to callus for up to two weeks, treated with virulence-induced bacteria for one hour, then regenerated into plantlets. Transient expression of a GUS reporter gene, assayed at about one week, occurred in 10-12% of calluses. Expression of the FLAG-tagged wTBP was also detected, by immunostaining. Stable expression, by selective growth on geneticin, and by GUS expression at about six weeks, occurred in 1-2% of calluses, quite comparable to that achieved by other methods.
ISSN:0735-9640
1572-9818
DOI:10.1023/A:1007686408369