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GrainGenes Reference Report: PCT-41-125

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Reference
PCT-41-125
Title
Parameters influencing transient and stable transformation of barley (Hordeum vulgare L.) protoplasts
Journal
Plant Cell, Tissue and Organ Culture
Year
1995
Volume
41
Pages
125-138
Author
Zhang J
[ Show all 8 ]
Abstract
Cat gene expression has been investigated following PEG-mediated plasmid uptake into barley protoplasts. The uptake conditions optimised for transient expression were employed for stable transformation. Transformed protoplast-derived calli of the cvs. Dissa and Igri, were selected on medium containing G418 at 40 micrograms ml-1 or kanamycin sulphate at 250 micrograms ml-1. Absolute transformation frequencies of 28.9 X 10(-5) and 21.3 X 10(-5) were recorded for Dissa with kanamycin sulphate and G418 selection, respectively. The frequency for Igri was 11.5 X 10(-5) with G418 selection. Antibiotic resistant protoplast-derived colonies expressed NPTII activity; Southern hybridisation confirmed integration of the nptII gene into barley genomic DNA.
Keyword
callus
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