Datura arborea and D. sanguinea hairy roots were produced by cocultivation of leaf fragments with Agrobacterium rhizogenes strain NCPP 1855. Adventitious buds emerged spontaneously-, without exogenous growth regulators, fiom seven hairy root clones of D. arborea and from one hairy, root clone of D. sanguinea. Regenerated plants were successfully acclimatized in the greenhouse. The integration of the bacterial TL-DNA into the genome of the putative transformed plants was confirmed by Southern blot analysis. Transgenie plants displayed increased ability to root in vivo. Morphological traits with relevant ornamental value like plant height, leaf number, size and shape, internode numbe~, and internode length were also affected. Transformation by wild-type Ri TL-DNA provided the chance to study plant growth and differentiation and to select improved genotypes.
Different plant explants of Persian buttercup (Ranunculus asiaticus L.) were screened for callus induction and adventitious shoot regeneration on different media to establish totipotent cultures. Murashige & Skoog (MS) medium was used, supplemented with different concentrations of the following growth regulators: kinetin, benzyladenine (BA), naphthaleneacetic acid (NAA) and indoleacetic acid (IAA). Callus was induced and adventitious buds regenerated only from cotyledonary explants after 4-5 weeks. Subculture of the regenerated buds on the same basal medium in presence of gibberellic acid (GA3) and BA produced well-organized shoots. Rooting was obtained by transferring shoots to growth regulator-free MS medium. A high rate of shoot multiplication has been achieved on medium with high concentration of kinetin and long-day photoperiod. Finally the plants were successfully transferred to soil and grown in a greenhouse.
Buttercup (Ranunculus asiaticus, family Ranunculaceae) hybrids are considered a popular and valuable ornamental crop in Europe and are cultivated both for cut flowers and for pot or border plants. In the Liguria region of northern Italy, Ranunculus hybrid cultivation has dramatically increased over recent years and the related sanitary issues regarding virus infection need to be carefully monitored in order to maintain commercial competitiveness. Presently, ELISA is the most used diagnostic tool for virus diagnosis, but if ‘mother plants’ with high economic value are to be tested, more sensitive molecular diagnostic tools are needed, together with efficient protocols for in vitro culture to eradicate viruses. In this study, new RT‐PCR protocols were tested on virus‐infected Ranunculus hybrid plants, indexed for the 13 most important Ranunculus‐infecting viruses. Rapid, uniform and effective procedures for molecular diagnosis were devised in order to promote an easy technology transfer from scientific institutions to laboratories of small and medium private enterprises. Moreover, an in vitro culture procedure was established for this species, which proved to be extremely effective in virus eradication. Advanced technology and sustainable quality production of high economic value plants are keys for future competitiveness in floriculture.
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