The effects of four types of explants removed from 10-cm flower stalks of Doritaenopsis Purple Gem ‘Ching Hua’ (immature apical flower buds, immature lateral flower buds, flower stem nodes, and flower pedicel sections) and combinations of two plant growth regulators [naphthalene acetic acid (NAA) and thidiazuron (TDZ)] on direct in-vitro shoot induction and multiplication were studied. Immature apical flower buds were the only explants that showed induction and multiplication of shoots in vitro. NAA at 5.4 and 10.7 μm combined with either 4.5 or 9.1 μm TDZ provided the fastest and greatest percentages of shoot induction (27% to 40%) and the greatest numbers of shoot multiplication (111–160 shoots per explant). In vitro–induced shoots were rooted on medium containing 5.4 μm NAA and developed into plantlets with normal vegetative and reproductive morphology. Regenerated plantlets were acclimatized, showing 100% survival and establishment in greenhouse. Plantlets were grown to maturity and showed normal flower morphology. No floral off-types were observed. The high rates of shoot multiplication obtained offer a means for mass clonal propagation of this and possibly other related Doritaenopsis hybrids.
Wagner A. Vendrame, Ian Maguire and Virginia S. Carvalho
Wagner A. Vendrame, Virginia S. Carvalho, José M.M. Dias and Ian Maguire
Pollination effectiveness was evaluated for pollen (pollinia) from two Dendrobium hybrids, ‘Sena Red’ and ‘Mini WRL’, submitted to cryopreservation using a vitrification protocol. Parameters evaluated included pollinia exposure to a previtrification solution (PVS2) under ice (0 °C) or room (27 ± 2 °C) temperatures from 1 to 4 hours before cryopreservation (LN). On removal from cryopreservation, pollinia were used to pollinate flowers of the same hybrids to verify viability and germination. All pollinia showed high percentages of germination (greater than 80%) after crosses were performed, except for pollinia from Dendrobium ‘Sena Red’ submitted to 3 hours of precooling (0 °C) in PVS2 followed by LN (60%) and for pollinia submitted to PVS2 for 3 hours at room temperature with no precooling (70%). Capsules were formed for both hybrids and seeds were successfully produced. The seed viability test revealed high viability (90% to 95%) for all treatments for both hybrids. Seeds observed under a microscope contained well-formed embryos and no abnormalities were identified. Seeds from all treatments germinated. Germinating seeds developed into healthy seedlings with well-formed leaves and roots. Cryopreservation of pollinia was successfully accomplished either by direct storage in liquid nitrogen without cryoprotection treatments or by using a PVS2 vitrification protocol.