Transformation and GUS transient expression on Arabica coffee explants

English. Genetic transformation was conducted on leaf explants and embryogenic calli of arabica coffee Kartika, USDA, S17 clones Beta-glucuronidase (GUS) reporter gene carrying pCAMBIA2301 plasmid were introduced into the young leaf explants using Agrobacterium tumefaciens mediated transformation system in the presence of 100 muM acetosyringone. After 15 minutes inoculation, leaf explants were cocultivated for 2 days. Histochemical GUS assay on leaf explant after 2 days cocultivation showed positive results. Three media were used, namely Lopez-Baez et al. 1 (993) (A), 1/2 MS + 0.5 ppm kinetin (B), and C (Boxtel & Berthoully, 1996) media. Callus induction of the three clones used were observed 14 days after the explants being transfered into the Lopez-Baez et al. (1993) selection medium supplemented with 50 ug/ml kanamysin. In the media B and C the leaf explant became brown, no callus induction but after about 9 months induction of somatic embryogenesis was observed

unknown. Transformasi genetik telah dilakukan pada tanaman kopi arabika klon Kartika, USDA dan S17. Gen report Beta-glucuronidase (GUS) yang terdapat plasmid pCAMBIA2301 diintroduksikan ke dalam eksplan daun muda melalui Agrobacterium tumefaciens dengan penambahan 100 muM asetosiringon. Setelah inokulasi selama 15 menit, eksplan daun dikokultivasi selama 2 hari. Pengujian aktivitas GUS secara histokimia pada eksplan daun yang dilakukan 2 hari setelah kokultivasi menunjukkan hasil positif untuk ketiga klon tersebut. Tiga macam media digunakan, yaitu media A (Lopez-Baez et al., 1993), media B (1/2 MS yang mengandung kinetin 0,5 ppm) dan media C (Boxtel & Berthoully, 1996). Induksi kalus dari ketiga klon yang digunakan terjadi 14 hari setelah pemindahan ke media A yang mengandung 50 ug/ml kanamisin. Pada media B dan C eksplan daun berubah menjadi coklat, tidak terbentuk kalus, tetapi induksi embrio somatik mulai terdeteksi setelah 9 bulan pada media tersebut