Location: National Clonal Germplasm RepositoryTitle: Alternative storage for germplasm of native hawaiian berries Author
Submitted to: Acta Horticulturae
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 4/1/2011
Publication Date: 2/3/2012
Citation: Reed, B.M., Uchendu, E., Wada, S., Zee, F.T. 2012. Alternative storage for germplasm of native hawaiian berries. Acta Horticulturae. 918:113-120. Interpretive Summary: Ohelo is a small, native Hawaiian shrub commonly found in disturbed, open sites at 640 to 3700 meter elevation on the islands of Hawaii and Maui. Plants are very hardy and have a compact form and bright waxy red new growth. Three Ohelo berry cultivars: Kilauea, Red Button, and Nene were selected for ornamental use due to their good growth habit and color. This study tested storage of these plants in liquid nitrogen (cryopreservation) and cold storage of tissue cultured shoots for medium and long-term storage. Shoot cultures in tissue-culture bags were maintained in good condition at refrigerator temperatures for more than a year. For cryopreservation shoot cultures were tested with three protocols that are also used for other types of berries. All three protocols had regrowth of >40%. This was high enough for long-term storage of the germplasm. Regrowth ranged from 42% up to 75% depending on the protocol used. These studies show that Ohelo plants can be successfully cold-stored or cryopreserved with any of these techniques.
Technical Abstract: Ohelo (Vaccinium reticulatum Smith) is a small, native shrub commonly found in disturbed, open sites at 640 to 3700 meter elevation on the islands of Hawaii and Maui. The shrub is very hardy with compact form and bright waxy red new growth. Three Ohelo berry cultivars: Kilauea, Red Button, and Nene were selected for ornamental use. This study tested cryopreservation and in vitro cold storage of these cultivars for germplasm preservation. Shoot cultures in StarPac tissue-culture bags were maintained in good condition in 4 ºC cold storage for more than a year. For cryopreservation, shoot cultures of the three cultivars were cold acclimated with alternating temperatures, then the shoot tips were cryopreserved with controlled rate cooling, encapsulation dehydration and PVS2 vitrification. Regrowth following controlled rate cooling was 42 to 51%; encapsulation dehydration 59 to 75% and PVS2 vitrification 51 to 61%. Bead encapsulated shoot tips were moderately desiccation sensitive and could only be dried to 31% moisture content before losing most viability. These results indicate that germplasm from each of the three cultivars retained viability and regrowth capacity under these storage conditions. Ohelo plant germplasm could be efficiently preserved using these techniques.