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PLANT GENETIC RESOURCES

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

^* Corresponding author (corbr{at}ars-grin.gov).

Genetic conservation of vegetatively-propagated grasses requires intensive care of pot cultures or carefully separated field plots. Even with intensive care, there is high risk of mechanical contamination and loss of plants to biotic and abiotic stresses. Medium- and long-term storage of this germplasm would be more cost effective and provide a backup for field or greenhouse germplasm collections. Cynodon spp. (bermudagrass and stargrass) germplasm is typically maintained as growing plants in breeders' collections. The development of a long-term storage protocol for Cynodon in liquid nitrogen (LN) could provide a secure backup of these collections. A diverse group of Cynodon taxa was evaluated for long-term storage in LN at –196°C. The encapsulation and dehydration (ED) cryopreservation protocol was most effective when combined with a 1- to 4-wk cold-acclimation period and dehydration to 19 to 23% moisture before exposure to LN. Nineteen of the 25 accessions (76%) had >40% regrowth. Thirty shoot tips of each of 25 Cynodon accessions are now stored at the National Clonal Germplasm Repository (NCGR), Corvallis, and 50 shoot tips are held at the National Center for Germplasm Resources Preservation (NCGRP) in Fort Collins, CO, as a long-term backup in LN.

Abbreviations: CA, cold acclimation or cold acclimated • ED, encapsulation and dehydration • KT, kinetin • LN, liquid nitrogen • MS, Murashige and Skoog • NCGR, National Clonal Germplasm Repository • NCGRP, National Center for Germplasm Resources Preservation