CRYOPRESERVATION OF DENDRANTHEMA MORIFOLIUM 'JAPANESE RED' SHOOT TIPS BY DROPLET-VITRIFICATION

Ren-Rui Wang, Xiao-Xia Gao, Long Chen, Qiao-Chun Wang
Chrysanthemum is one of the most popular flower crops worldwide. Development and availability of cryopreservation technique is necessary for long-term conservation of genetic resources. In the present study, we describe a droplet-vitrification procedure for cryopreservation of Dendranthema morifolium ‘Japanese Red’. In vitro stock shoots were maintained on Murashige and Skoog (1962) medium supplemented with 30 g/L sucrose and 7 g/L agar (pH=5.8) and kept at 24±2°C under a 16-h photoperiod with a light intensity of 50 µmol m-2 s-1 provided by fluorescent tubes. Shoot tips (1-2 mm in size with 3-4 leaf primordia) were excised from 4-week-old in vitro shoots and precultured on solidified MS medium containing 0.5 M sucrose for one day. Precultured shoot tips were loaded with a loading solution made of MS containing 2 M glycerol and 0.4 M sucrose for 20 min at room temperature, followed by dehydration with PVS2 for 30 min at 0°C. Each shoot tip was transferred onto a 2.5 µl PVS2 on aluminum foils (2 × 0.8 cm), prior to a direct immersion in liquid nitrogen (LN). After immersed in LN for a few minutes, the foils with shoot tips were transferred into 2 ml cryotubes for cryostorage. Frozen shoot tips were thawed by incubation in a solution composed of MS containing 1.2 M sucrose for 20 min at room temperature, and then post-cultured on MS medium containing 0.05 mg/L GA3 for shoot recovery. With this procedure, 100% of shoot tips survived and 67% of them regenerated shoots following cryopreservation. The droplet-vitrification protocol developed in the present study is being tested for its potential applications to cryopreservation of other four genotypes and cryotherapy for virus eradication of chrysanthemum.
Ren-Rui Wang, , Xiao-Xia Gao, , Long Chen, and Qiao-Chun Wang, (2014). CRYOPRESERVATION OF DENDRANTHEMA MORIFOLIUM 'JAPANESE RED' SHOOT TIPS BY DROPLET-VITRIFICATION. Acta Hortic. 1039, 187-192
DOI: 10.17660/ActaHortic.2014.1039.24
https://doi.org/10.17660/ActaHortic.2014.1039.24
Dendranthema morifolium, cryopreservation, droplet-vitrification, shoot tips
English

Acta Horticulturae