Department of Agronomy, University of Illinois, Urbana, Illinois 61801.
Plant Physiol. 1985 Jun;78(2):430-2. doi: 10.1104/pp.78.2.430.
Suspension cultures of carrot (Daucus carota, line C1), tobacco (Nicotiana tabacum, line TX1), and Nicotiana plumbaginifolia (line NP) were frozen under controlled conditions with trehalose as the sole cryoprotectant. Maximal post-thaw viability (71-74%), measured by phenosafranin dye exclusion, was obtained with the C1 cells following a 24hour pretreatment with 5 or 10% trehalose and with 40% trehalose as the cryoprotectant during freezing. TX1 cells pretreated for 24 hours with 10% trehalose and cryoprotected with 40% trehalose during freezing showed 47% viability following thawing as determined by phenosafranin dye exclusion. The NP cells required a 3 to 6 day pretreatment with 10% trehalose and 40% trehalose as a cryoprotectant at the time of freezing for the recovery of viable cells. Growing cells were recovered when the C1 and NP cells treated as described were plated on agar-solidified medium following thawing.
用海藻糖作为唯一的冷冻保护剂，在受控条件下对胡萝卜（Daucus carota，C1 系）、烟草（Nicotiana tabacum，TX1 系）和菝葜（Nicotiana plumbaginifolia，NP 系）悬浮培养物进行冷冻。通过酚磺酞染料排除法测定，经过 24 小时用 5%或 10%海藻糖预处理和用 40%海藻糖作为冷冻过程中的冷冻保护剂，C1 细胞获得了最大的解冻后活力（71-74%）。用 10%海藻糖预处理 24 小时并用 40%海藻糖冷冻保护的 TX1 细胞在解冻后用酚磺酞染料排除法测定，活力为 47%。NP 细胞需要用 10%海藻糖预处理 3 至 6 天，并用 40%海藻糖作为冷冻时的冷冻保护剂，才能恢复存活细胞。当按上述方法处理的 C1 和 NP 细胞解冻后在琼脂固化培养基上平板时，可回收生长细胞。
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