Genetic Transformation of Iris germanica Mediated by Agrobacterium tumefaciens

Abstract

<jats:p>A protocol was developed for production of transgenic iris plants (<jats:italic>Iris germanica</jats:italic> L. `Skating Party') from regenerable suspension cultures via <jats:italic>Agrobacterium</jats:italic>-mediated transformation. We tested a series of selection agents, and identified hygromycin and geneticin as the most suitable for selecting transformed iris cells. Suspension cultures of iris were cocultured for 3 days with <jats:italic>A. tumefaciens</jats:italic> LBA 4404(pTOK233) carrying an intron-interrupted uidA (GUS) gene encoding β-glucuronidase, and hpt (hygromycin) and nptII (geneticin) selectable marker genes. Hygromycin- or geneticin-resistant calli having GUS enzyme activity were identified and used to induce plant regeneration. More than 300 morphologically normal transgenic iris plants were obtained in ≈6 months. About 80% of the transformants were GUS-positive and NPTII-positive (paromomycin-resistant). Integration of transgenes into the nuclear genome of iris plants was confirmed by Southern blot analysis. We have, therefore, developed an efficient <jats:italic>A. tumefaciens</jats:italic>-mediated transformation system for <jats:italic>Iris germanica</jats:italic>, which will allow future improvement of this horticulturally important ornamental monocot via genetic engineering.</jats:p>

A review on factors affecting the Agrobacterium-mediated genetic transformation in ornamental monocotyledonous geophytes

“…This promoter was found to be the best for transformation of Iris germanica. This outcome further confirmed that Agrobacteriummediated transformation using CaMV35S can be applied to other important monocotyledonous ornamentals (Jeknic et al, 1999).…”

“…In Iris germanica, LBA4404 gave remarkably higher transformation rates than EHA105 (Jeknic et al, 1999). In Agapanthus praecox, the same LBA4404 was found to be more effective than EHA101 (Suzuki et al, 2001).…”

Micropropagation of iridaceae—a review

“…Hygromycin and geneticin were found to be the best markers for selecting transformed cells. Over 300 morphologically normal transgenic plants carrying the GUS reporter gene were regenerated and stable transgene integration was confirmed by Southern hybridisation (Jeknic et al 1999).…”

Clonal fidelity of Iris sibirica plants regenerated by somatic embryogenesis and organogenesis in leaf-base culture — RAPD and flow cytometer analyses

“…Recently, particle bombardment and Agrobacteriummediated transformation have become the most popular and powerful approaches employed for the creation of horticulturally valuable characteristics such as novel flower colors, leaf shapes, dwarf forms, prolonged cut-flower vase-life, resistance to abiotic or biotic stresses such as viruses/ viroids, pathogens and insects (Teixeira da Silva et al, 2013). Up to date, genetic transformation in genus Iris has been reported only for I. germanica, where stable overexpression of the Lilium lancifolium capsanthin-capsorubin synthase (Llccs) gene in callus tissue resulted in color change from yellow to red-orange (Jeknić et al, 1999(Jeknić et al, , 2012. Genetic transformation of I. sibirica can also be an attractive way to create novel flower colors or to increase capacity for secondary metabolite production or content of essential oils.…”