Twigs of Salix species are candidates for cryopreservation procedures because they become tolerant of freezing temperatures during mid-winter. We examined several variables in developing a two-step cryopreservation procedure for sections from these twigs. Samples of Salix triandra cooled to-30°C or-35°C and then transferred to the vapor phase over liquid nitrogen gave the greatest percent shoot formation. Cooling rate to-35°C had a major influence on shoot formation. Samples cooled at rates greater than 10°C/hr showed no shoot formation. The highest percent of shoot formation was achieved by cooling at 0.21°C/hr. Cooling rate from-35°C to liquid nitrogen did not influence shoot formation. Warming procedures affected shoot formation. Transferring samples from-160°C either to a +2°C cold room or to-3°C methanol gave similar levels of shoot formation. No shoot formation occurred either with warming in +40°C water or very slowly in a Styrofoam box. Eight of eleven Salix taxa tested using the established protocol had significant levels of shoot formation after cryogenic treatment.
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Document Type: Regular Paper
January 1, 2004
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CryoLetters is a bimonthly international journal for low temperature sciences, including cryobiology, cryopreservation or vitrification of cells and tissues, chemical and physical aspects of freezing and drying, and studies involving ecology of cold environments, and cold adaptation
The journal publishes original research reports, authoritative reviews, technical developments and commissioned book reviews of studies of the effects produced by low temperatures on a wide variety of scientific and technical processes, or those involving low temperature techniques in the investigation of physical, chemical, biological and ecological problems.