Micropropagation of poinsettia by organogenesis.

Poinsettia (Euphorbia pulcherrima) is one of the most popular ornamental pot plants. Conventional propagation is by cuttings, generally focused on a period prior to the most intensive time of sales. Rapid multiplication of elite clones, the production of pathogen-free plants and more rapid introduction of novel cultivars (cvs.) with desirable traits, represent important driving forces in the poinsettia industry. In recent years, different strategies have been adopted to micropropagate poinsettia, which could assist breeders to meet consumer demands. The development of reliable in vitro regeneration procedures is likely to play a crucial role in future production systems. Stem nodal explants cultured on semi-solid MS-based medium supplemented with benzylaminopurine (BAP) and naphthalene acetic acid (NAA) develop shoots from adventitious/axillary buds after 7 weeks of culture. Rooting of in vitro regenerated shoots is achieved in semi-solid MS-based medium containing the auxin indole-3-acetic acid (IAA). Four to six weeks after transfer to root-inducing medium, regenerated plants can be transferred to compost and acclimatized in the glasshouse. Direct shoot regeneration from cultured explants is important to minimize somaclonal variation in regenerated plants.

Agrobacterium tumefaciens-mediated transformation of poinsettia, Euphorbia pulcherrima, with virus-derived hairpin RNA constructs confers resistance to Poinsettia mosaic virus.

Agrobacterium-mediated transformation for poinsettia (Euphorbia pulcherrima Willd. Ex Klotzsch) is reported here for the first time. Internode stem explants of poinsettia cv. Millenium were transformed by Agrobacterium tumefaciens, strain LBA 4404, harbouring virus-derived hairpin (hp) RNA gene constructs to induce RNA silencing-mediated resistance to Poinsettia mosaic virus (PnMV). Prior to transformation, an efficient somatic embryogenesis system was developed for poinsettia cv. Millenium in which about 75% of the explants produced somatic embryos. In 5 experiments utilizing 868 explants, 18 independent transgenic lines were generated. An average transformation frequency of 2.1% (range 1.2-3.5%) was revealed. Stable integration of transgenes into the poinsettia nuclear genome was confirmed by PCR and Southern blot analysis. Both single- and multiple-copy transgene integration into the poinsettia genome were found among transformants. Transgenic poinsettia plants showing resistance to mechanical inoculation of PnMV were detected by double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA). Northern blot analysis of low molecular weight RNA revealed that transgene-derived small interfering (si) RNA molecules were detected among the poinsettia transformants prior to inoculation. The Agrobacterium-mediated transformation methodology developed in the current study should facilitate improvement of this ornamental plant with enhanced disease resistance, quality improvement and desirable colour alteration. Because poinsettia is a non-food, non-feed plant and is not propagated through sexual reproduction, this is likely to be more acceptable even in areas where genetically modified crops are currently not cultivated.