Pollination of Dendrobium hybrids using cryopreserved pollen

Abstract

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.