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Cryopreservation of Bermudagrass Germplasm by Encapsulation Dehydration

^a USDA-ARS National Clonal Germplasm Repository, 33447 Peoria Road, Corvallis, OR 97333
^b USDA-ARS National Forage Seed Production Research Center, 3450 SW Campus Way, Corvallis, OR 97331

View larger version (17K): [in a new window]   Fig. 1. (A) Regrowth of 4-wk CA Cynodon transvaalensis ‘Uganda’ shoot tips following encapsulation in alginate beads, air drying for 0 to 7 h, and 10 min exposure in liquid N followed by rehydration and plating on recovery medium. Data were adjusted so that control shoot tip regrowth equaled 100%. (B) Bead moisture content at each time period.

View larger version (14K): [in a new window]   Fig. 2. Response of three Cynodon accessions to 0 to 4 wk of cold acclimation followed by 6-h air drying and cryopreservation with the encapsulation and dehydration protocol. Data were adjusted so that control shoot tip regrowth equaled 100%.

View larger version (34K): [in a new window]   Fig. 3. Mean regrowth of liquid nitrogen (LN) exposed and control shoot tips of (A) Cynodon dactylon species and cultivars and (B) Cynodon species other than C. dactylon, cold acclimation for 4 wk, encapsulated in alginate beads, and dried for 6 h in the laminar flow hood. Shoot tips in beads were exposed to LN for 10 min, rewarmed at room temperature for 20 min, rehydrated for 5 min, and plated on recovery medium. Data presented are the mean recovery of vials from a single storage experiment. Two vials were rewarmed on site at the National Clonal Germplasm Repository (NCGR) while the other was sent by overnight mail to the National Center for Germplasm Resources Preservation (NCGRP) in Ft. Collins, CO, transferred to long-term LN storage tanks, and later rewarmed. Data were taken 4 wk after plating on recovery medium (three replicates of 10, two reps at NCGR, and one rep at NCGRP). Liquid N means without error bars are averages of two vials. Controls were dried and rehydrated but not exposed to LN (one replicate of 10).