Genetic transformation ofPelargonium X hortorum
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Summary
TransgenicPelargonium X hortorum have been producedvia Agrobacterium tumefaciens-mediated transformation. The regeneration protocol used provided a regeneration frequency approximately to 95 percent. Clumps of regenerants, from cotyledons and hypocotyls ofPelargonium X hortorum seedlings, were inoculated with the disarmed strain EHA101 ofAgrobacterium tumefaciens. This strain contains a binary vector carrying neomycin phosphotransferase II, hygromycin B phosphotransferase and ß-glucuronidase genes. Selection on the regeneration medium supplemented with hygromycin allowed production of transgenic plants in up to 20% of the inoculated explants. The insertion of foreign DNA was demonstrated by Southern and polymerase chain reaction analysis: these experiments indicated that the inserted T-DNA is not full length for most of the plants. All RO transgenic plants exhibited a normal phenotype and are fertile.
Keywords
Polymerase Chain Reaction Transgenic Plant Full Length Regeneration Frequency HygromycinAbbreviations
- GUS
ß-glucuronidase coding sequence
- PCR
polymerase chain reaction
- CaMV
cauliflower mosaic virus
- NPTII
neomycin phosphotransferase coding sequence
- NOS
nopaline synthase gene promoter and terminator
- HPH
hygromycin B phosphotransferase coding sequence
- SDS
sodium dodecyl sulphate
- EDTA
(ethylenedinitro trilo)tetra-acetic acid disodium salt
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