Cryopreservation of Arabidopsis thaliana Seedlings by Vitrification

A procedure has been developed for vitrification of Arabidopsis thaliana (Landsberg erecta) seedlings. The procedure involved:(1) loading of the seedlings with 2M glycerol+0.4M sucrose at 20℃ for 20 min; (2) dehydration of seedlings in vitrification solution (PVS2:30%w/v glycerol +15%w/v ethylene glycol +15%w/v DMSO in MS basal medium containing 0.4M sucrose) at 0℃for 50 min; (3) immersion of seedlings into LN2 directly and thawing at 20℃ for 5~10 min; (4) unloading of the seedlings with M S +1.2M sucrose at 20℃for 30 min. Then reculture vitrified seedlings on MS medium. Seedlings transferred to soil from MS medium and cultured in growth chamber appeared phenotypically identical to untreated control plants and produced normal seeds. No survival were obtained using 3 days old seedlings or older than that with this procedure.