Propagation of Rosa hybrida L. cv. Coolwater Under Tissue Culture and Transformation of the RhAA Gene via Agrobacterium tumefaciens

Abstract

Rose is the most favorite cut flowers all over the world. Production of high-quality flowers, prevention, and delay of flower senescence, is a major goal in floriculture. Now a day, biotechnological approaches have been used to improve ornamental attributes. Tissue culture and genetic transformation appear to offer valuable advancements for operating floral characteristics. In this study, after optimizing sterilization for the first step of tissue culture, lateral buds of Rosa hybrida cv. Coolwater were cultured on MS medium supplemented with different concentrations and combinations of BA (Benzyladenine). The results indicated that the highest growth rate and establishment were 80% on 1.5 mg l-1 BA. In the second stage, explants were transferred to MS medium containing various concentrations of BA, NAA (Naphthaleneacetic acid) and IAA (Indole acetic acid) hormones. The maximum number of shoots per each explant (8.00 ± 0.18) belonged to 3 mg l-1 BA with 0.5 mg l-1 NAA. Explants were also transferred to the rooting medium induction with different concentration of IBA (Indole butyric acid) and Phl (Phloroglucinol). The best rooting induction was selected in MS/2 with 3 mg l-1 IBA (62.22%). In order to the production of transgenic plants, the lateral buds and Agrobacterium tumefaciens LBA 4404 were used for transformation. Two parameters affecting Agrobacterium infection efficiency were investigated, including inoculation, media. The optimum time for infection was 10 minutes. The effects demonstrated that the best medium in inoculation was sucrose 3% and the percentage of transgenic was 10%. Transgenic plants were confirmed by PCR (Polymerase chain reaction). The appearance of the 470bp band revealed that plants were transgenic with RhAA. According to studies, it seems that the gene in the roses affects the vase life of the flower.