A cryopreservation method comprising sorbitol pre-growth treatment, DMSO cryoprotection and two-step controlled rate cooling has been optimized for differentiated embryogenic suspensor masses (ESM) of Picea sitchensis. The protocol was applied to clonal cultures from five different half-sibling families each represented by five different genotypes and their responses to cryopreservation assessed over 3 years. Nineteen of the 25 clonal lines tested survived LN and were capable of regrowth and producing stage 2-4 somatic embryos. Following the second subculture cycle of ESM after they had been retrieved from cryogenic storage, post-cryopreservation regrowth was comparable with that of controls. A General Linear Model, multifactorial Main Effects Plot revealed no significant differences between genotypes (P>0.05) in response to cryopreservation, whereas significant differences between families (P<0.05) were detected.
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Document Type: Research Article
Publication date: July 1, 2007
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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.