Transgenic plants of coffee Coffea canephora from embryogenic callus via Agrobacterium tumefaciens-mediated transformation

T. Hatanaka, Y. E. Choi, T. Kusano, H. Sano

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    42 Citations (Scopus)

    Abstract

    Embryogenic calli were induced from leaf explants of coffee (Coffea canephora) on McCown's woody plant medium (WPM) supplemented with 5 μM N6-(2-isopentenyl)-adenosine (2-iP). These calli were co-cultured with Agrobacterium tumefaciens EHA101 harboring pIG121-Hm, containing β-glucuronidase (GUS), hygromycin phosphotransferase (HPT), and neomycin phosphotransferase II genes. Selection of putative transgenic callus was performed by gradual increase in hygromycin concentration (5, 50, 100 mg/l). The embryogenic calli surviving on medium containing 100 mg/l hygromycin showed a strong GUS-positive reaction with X-Gluc solution. Somatic embryos were formed from these putative transgenic calli and germinated on WPM medium with 5 μM 2-iP. Regenerated small plantlets with shoots and roots were transferred to medium containing both 100 mg/l hygromycin and 100 mg/l kanamycin for final selection of transgenic plants. The selected plantlets exhibited strong GUS activity in leaves and roots as indicated by a deep blue color. GUS and HPT genes were confirmed to be stably integrated into the genome of the coffee plants by the polymerase chain reaction.

    Original languageEnglish
    Pages (from-to)106-110
    Number of pages5
    JournalPlant Cell Reports
    Volume19
    Issue number2
    DOIs
    Publication statusPublished - 1999 Dec

    Keywords

    • Agrobacterium tumefaciens
    • Coffea canephora
    • Transformation

    ASJC Scopus subject areas

    • Agronomy and Crop Science
    • Plant Science

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