Cryopreservation of dormant buds from diverse Fraxinus species

Ash (Fraxinus) is an economically important tree genus in the landscape industry, as well as a key component of North American forests, especially in the North Central United States and adjacent regions in Canada. In recent years, the Emerald Ash Borer beetle (Agrilus planipennis) has significantly threatened the survival of native North American Fraxinus species. A dormant-bud cryopreservation technique has been developed as a method to conserve specific clones of ash. Dormant buds of three ash species were successfully cryopreserved when desiccated on their stem sections to 30 percent moisture content (w/v) and then cooled at rates of either -1 degree C per h or -5 degree C per day to either -30 or -35 degree C before immersion in liquid nitrogen vapor (LNV). Stem sections were removed from LNV, warmed, and rehydrated, and their buds grafted onto rootstocks to evaluate survival. Recovery percentages ranged from 34 to 100 percent after LNV exposure and were dependent upon accession and cooling rate. The cryopreservation methods proposed herein can complement seed-collection efforts aimed at conserving diversity, supplementing ex situ genebank and botanic-garden collections.