Development of a stable transformation procedure for protoplasts of Coffea arabica C.V. Colombia
Abstract
The purpose of the present study was to develop a stable transformation procedure for Coffea arabica c.v. Colombia. Protoplasts isolated from embryogenic suspension cultures were used to study transient expression of a reporter gene, the $\beta$-glucuronidase (gusA). Conditions were optimized for PEG-mediated-DNA uptake and for transient expression of gusA. Once transient assay conditions were optimized, stable transformation was pursued using an antibiotic resistance gene neomycin phosphotransferase (neo). Kanamycin and Geneticin (G418) resistant calli were obtained. Based on NPTII enzymatic activity, PCR, and Southern analysis, it could be demonstrated that these calli were stably transformed.
Degree
Ph.D.
Advisors
Hodges, Purdue University.
Subject Area
Botany|Molecular biology
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