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ARS Home » Pacific West Area » Corvallis, Oregon » National Clonal Germplasm Repository » Research » Publications at this Location » Publication #144999
Title: CRYOPRESERVED STORAGE OF HOPS (HUMULUS L.) GERMPLASM

Author
item Reed, Barbara
item Paynter, Carolyn
item DENOMA, JEANINE - OSU-MONMOUTH
item D Anchino, Jeff
item OKUT, NESE - VOLUNTEER

Submitted to: Society for In Vitro Biology Proceedings
Publication Type: Abstract Only
Publication Acceptance Date: 6/20/2003
Publication Date: 6/20/2003
Citation: REED, B.M., PAYNTER, C.L., DENOMA, J., D ACHINO, J., OKUT, N. CRYOPRESERVED STORAGE OF HOPS (HUMULUS L.) GERMPLASM. SOCIETY FOR IN VITRO BIOLOGY PROCEEDINGS. 2003.

Interpretive Summary: The National Clonal Germplasm Repository (NCGR) stores the US national collection of hops. Hops are usually stored in field collections. These plants are susceptible to loss from environmental stresses and insect or disease attacks. To insure the safety of the collection, a subset of the field collection was established as clean, tissue cultures stored at refrigerator temperatures and also grown in pots in screened houses. Long-term (100 year) storage of this collection as tiny shoot tips frozen in liquid nitrogen (-320 F) will provide additional security. We tested each plant type and stored 100 meristems of each. Shoot tips were grown on a special nutrient gel, and antifreeze solutions were added for additional freeze-protection. Shoot tips were slowly frozen to -40 F, immersed in liquid nitrogen, and stored. Controls were thawed and regrown after 1 hour in liquid nitrogen and recovery ranged from 30 to 80%. The cryopreserved hops will be stored at NCGR Corvallis and at the National Center for Genetic Resources Preservation in Ft. Collins, CO.

Technical Abstract: The USDA-ARS National Clonal Germplasm Repository (NCGR) stores the global diversity of Humulus for the US Plant Germplasm System. The primary collection of hops is stored as trellised plants in a field genebank. To back-up the collection, a subset of the field collection was established as virus-free in vitro cultures stored at 4 C and as potted plants grown in screened houses. In this study we characterized the response of diverse Humulus genotypes to cryopreservation and stored 100 meristems of each in liquid nitrogen. Shoot tips (0.8-1.0 mm) of cold-acclimated in vitro plantlets were dissected and precultured on 5% DMSO medium for 48 h under cold-acclimating conditions. Following cryoprotection, shoot tips were frozen to -40 C at 0.1 C/min in a programmable freezer and immersed in liquid nitrogen. Control vials were thawed in 45 C water for 1 min, then in 23 C water for 2 min. Recovery ranged from 30 to 80%. The cryopreserved meristems will be stored at NCGR Corvallis and at the National Center for Genetic Resources Preservation in Ft. Collins, CO.