PMID: 11902001Mar 21, 2002Paper

Growth and differentiation of transgenic callus regulated by phytohormones and antibiotics in transformation of loblolly pine

Yi chuan xue bao = Acta genetica Sinica
Wei TangVanessa Samuels

Abstract

Mature zygotic embryos of loblolly pine (Pinus taeda L.) were transformed by Agrobacterium tumefaciens strain LBA 4404 harbouring the plasmid pBI121 which carried the selectable marker gene, neomycin phosphotransferase II (npt II) controlled by the promoter of the nopaline synthase gene, and the uidA reporter gene, encoding beta-glucuronidase (GUS) driven by the cauliflower mosaic virus 35S promoter. Organogenic transgenic calli and transgenic regenerated plantlets were produced on selection medium containing 15 mg/L kanamycin, and confirmed by GUS histochemical staining, polymerase chain reaction (PCR), and southern blot analysis. Influences of phytohormone (BA/IBA) and antibiotics on growth and differentiation of organogenic transgenic calli were investigated. Of the phytohormone (BA/IBA) and antibiotics administered, 500 mg/L carbenicillin combined with 2 mg/L BA and 0.5 mg/L IBA (BA/IBA = 4) resulted in a 54.2% higher increase in the growth of transgenic calli as well as in the differentiation of transgenic calli, which was 45.7% more than that of control on the 6th week of culture. Claforan at 500 mg/L combined with 2 mg/L BA and 0.5 mg/L IBA resulted in a 40.8% increase in the growth of transgenic calli, and 38.7% increas...Continue Reading

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