Cryopreservation of Axillary Buds of Vaccinium in vitro by Droplet Vitrification
The technical parameters were selected during the cryopreservation by droplet vitrification with axillary buds of Vaccinium"Misty"in vitro.The wide spectrum of this method was tested with to-tal 66 genotypes of Vaccinium including 8 spieces.The genetic stability of strains regenerated from 5 cultivars after cryopreservation was tested by ISSR with 10 primers.The results showed that the single axillary bud(length 1-1.5 mm)cut from 30-day-old shoots in vitro was cultured on subculture me-dium(WPM+0.3 mg/L ZT+7 g/L agar+30 g/L sucrose,SCM)and on preculture medium(WPM+7 g/L agar+0.3 mol/L sucrose,PCM)for 1 day,respectively,at(25±2)℃ in the dark.The axillary buds were treated in loading solution(WPM+2.0 mol/L glycerol+1.0 mol/L sucrose,LS)for 30 min(room temperature)and immersed into Plant Vitrification Solution Ⅱ(WPM+300 g/L glycerol+150 g/L ethylene glycol+150 g/L DMSO+0.4 mol/L sucrose,PVS2)for 40 min(0 ℃).The droplets(one bud per droplet)were formed on aluminum foil paper and quickly put into liquid nitrogen(LN).The material taken out from LN was treated immediately in unloading solution(WPM+1.2 mol/L su-crose,ULS)for 20 min(room temperature).The axillary buds were cutltured on SCM medium for regen-eration under the normal culture condition after 1 day in the dark(25±2)℃.The testing results of wide spectrum showed that the average survival rate and regeneration rate were 81.3％and 47％,respec-tively.The molecular marker detection results of 5 bilberry varieties showed no ploymorphic band.
万方数据
维普
