Abstract
In vitro plantlets were used for axillary shoot tip isolation to avoid microbial contamination which often occurred from use of greenhouse-grown plants. Periodic subculturing gave a supply of uniform plantlets necessary for obtaining shoot tips for cryogenic experiments. Previous results have shown that all cells within a shoot tip did not survive cryogenic exposure and the regrowth percentage depended upon the composition of the culture medium. A medium containing 0.5 mg/liter zeatin, 0.2 mg/liter gibberellic acid, and 0.5 mg/liter indoleacetic acid gave regeneration of a multipleshoot mass from control and some frozen shoot tips. The two-step cooling procedure (0.2-0.3 °C/min to −35 °C followed by immersion in liquid nitrogen) gave high percentages of regrowth in the cultivars Norland and Red Pontiac. Shoots were obtained from treated materials in both cultivars.
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