In vitro regeneration and genetic transformation of chrysanthemum (Dendranthema grandiflora var. micromargara)

Abstract

The chrysanthemum stands as one of the main ornamental plants of high commercial demand due to a wide range of flower colors and shapes achieved by genetic breeding. Therefore, it is necessary to establish efficient in vitro culture protocols to achieve genetic transformation of ornamental plants successfully. The objective of this study was to establish an in vitro plant regeneration and genetic transformation system of plants in order to change the color of chrysanthemum var. Micromargara petals. The concentration of 15 g/l was the most suitable for in vitro regeneration of plants, obtaining a greater number of leaves and shoot length compared with 0 and 30 g/l sucrose. The plants obtained in vitro were acclimatized and induced to flower. The flavonoid 3´-hydroxylase gen (f3´h) amplification by RT-PCR in chrysanthemum flower buds presented a 98% similarity to sequences reported in chrysanthemum. Chrysanthemum plants were inoculated with vector pEuMV-YP: ΔAV1: f3'h by biolistics. Plasmid sequencing presented a 99% similarity with Pepper golden mosaic virus- Tamaulipas strain.