A study of gene introduction methods on various chrysanthemum cultivars

Abstract


Okada Tsuyoshi*, Kishi Keigo and Takashi Naito

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. Genotypedependence 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 GIMdependent, but in all cases is shown to occur in the venation.

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