Cryopreservation of rose in vitro shoot tips rose by vitrification.

The cryopreservation technique of rose shoot tips by vitrification was studied to provide theoretical and practical basis for the cryopreservation of rose shoot tips.Four rose cultivars, Gold Medal, Pink Maya,Han Hong and Bo were selected for cryopreservation by the simple vitrification and vitrification.The simple vitrification method made significant difference in vitro shoot tips of different rose cultivars.The survival rate of Golden Medal's shoot tips was the highest, 55.3% . The effects of preculture, loading and plant vitrification solution 2 treatment time on survival rate were tested on Pink Maya's shoot tips. A method of the vitrification cryopreservation procedure of rose shoot tips was constructed. Shoot tips about 1cm in length were precultured for 5-6d on MS medium supplemented with 0.3mol·L-1 sucrose. Dissected shoot apices about 1.5mm in length were loaded with solution A(MS+2mol·L-1glycerol+0.4mol·L-1sucrose) for 30min or solution B[60%PVS2+40%(MS+0.15 mol·L-1sucrose)] for 40min at room temperature.These excised shoot tips were sufficiently dehydrated in a highly concentrated plant vitrification solution (PVS2:300g·L-1glycerol+150g·L-1ethylene glycol+150g·L-1dimethyl sulfoxide+0.4mol·L-1sucrose+MS) for 40min at 0℃ and then plunged into liquid nitrogen and conserved for 24h. After rapid thawed in water at 40℃ for 90s, the tips were rinsed with 1.2mol·L-1 sucrose solution and then plated on MS medium supplemented with 1.0mol·L-1 6-BA and 0.1mol·L-1 IBA. The survival rate of shoot tips reached 53.3%.