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

Citation
T. Hatanaka et al., Transgenic plants of coffee Coffea canephora from embryogenic callus via Agrobacterium tumefaciens-mediated transformation, PL CELL REP, 19(2), 1999, pp. 106-110
Citations number
16
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Plant Sciences","Animal & Plant Sciences
Journal title
PLANT CELL REPORTS
ISSN journal
0721-7714 → ACNP
Volume
19
Issue
2
Year of publication
1999
Pages
106 - 110
Database
ISI
SICI code
0721-7714(199912)19:2<106:TPOCCC>2.0.ZU;2-X
Abstract
Embryogenic calli were induced from leaf explants of coffee (Coffea canepho ra) on McCown's woody plant medium (WPM) supplemented with 5 mu M N-6-(2-is opentenyl)-adenosine (2-iP). These calli were co-cultured with Agrobacteriu m tumefaciens EHA101 harboring pIG121-Hm, containing beta-glucuronidase (GU S), hygromycin phosphotransferase (HPT), and neomycin phosphotransferase II genes. Selection of putative transgenic callus was performed by gradual in crease in hygromycin concentration (5, 50, 100 mg/l). The embryogenic calli surviving on medium containing 100 mg/l hygromycin showed a strong GUS-pos itive reaction with X-Gluc solution. Somatic embryos were formed from these putative transgenic calli and germinated on WPM medium with 5 mu M 2-iP. R egenerated small plantlets with shoots and roots were transferred to medium containing both 100 mg/l hygromycin and 100 mg/l kanamycin for final selec tion of transgenic plants. The selected plantlets exhibited strong GUS acti vity in leaves and roots as indicated by a deep blue color. GUS and HPT gen es were confirmed to be stably integrated into the genome of the coffee pla nts by the polymerase chain reaction.