Abstract
Since in vitro regeneration and transformation systems in hot pepper (Capsicum annuum L.) have not been available, the application of new genetic manipulations has been limited. Here we report an efficient procedure to regenerate whole pepper plants and to generate transgenic plants expressing a foreign gene was established. High frequency of plant regeneration was observed when hypocotyl and cotyledon explants were cultured on MS/B5 medium supplemented with NAA 0.05 mg·L–1 plus zeatin 2.0 mg·L–1, NAA 0.05 mg·L–1 plus zeatin 2.0 mg·L–1, IBA 10.0 mg·L–1 plus BA 1.0 mg·L–1, IAA 0.02 mg·L–1 plus zeatin 3.0 mg·L–1. An addition of AgNO3 5–10 μm to these media improved the regeneration rate by about 10%. For plant transformation, hypocotyl and cotyledon explants of pepper were preconditioned on kanamycin-free shoot induction medium for 48 hours. Then, co-cultivation with Agrobacterium tumeaacience was done on the co-culture medium for 2 days. The explants were then blotted in sterile filter paper and placed on shoot induction and selection medium containing kanamycin sulfate (100 mg·L–1) and carbenicillin (500 mg·L–1). PCR showed that the introduced ADA gene was integrated and stably expressed in the regenerated plants. ADA enzyme activities were checked by spectrophotometric analysis.
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