African Journal of Biotechnology Vol. 2 (5), pp. 114-123, May 2003

ISSN 1684-5315  © 2003 Academic Journals

Four gene introduction methods affect the shoot regeneration and localization of transgene expression in greenhouse stem explants and in vitro-grown chrysanthemum stem thin cell layers

J. A. Teixeira da Silva* and S. Fukai

Faculty of Agriculture, Kagawa University, Miki-cho, Kagawa, 761-0795, Japan

*Corresponding Author; E-mail: jaimetex@angelfire.com, Telfax: +81878910747

Accepted 18 April, 2003

Gene introduction method (GIM) affected shoot regeneration capacity (SRC) in standard and spray-type chrysanthemums. SRC was both cultivar and GIM-dependent in both in vitro and greenhouse stem explants, the former significantly higher than the latter. Sonication had an SRC-stimulating effect on in vitro explants. Other GIMs (Agrobacterium, biolistics, Agrolistics) had an SRC-inhibiting effect on greenhouse explants. Genotype-dependence of SRC was observed in both in vitro and greenhouse material. SRC is influenced by the explant and regeneration media, which should be modified if altered by the GIM. Shoots derived from all GIM treatments showed normal growth under in vitro and greenhouse conditions, and flowered normally. In addition, this study further shows that explant origin (in vitro versus greenhouse) and cultivar significantly affect the regeneration process, even when an optimized medium is utilized. The integration of the GUS transgene is also GIM-dependent, but in all cases is shown to occur in the venation.

Keywords: Agroinfection, biolistics, explant survival, regeneration, sonication.