Pennycooke J C, Towill L E
Department of Horticulture and Landscape Architecture, Colorado State University, Fort Collins, CO 80523, USA, , , , , , US.
USDA/ARS National Seed Storage Laboratory, 1111 S. Mason St., Fort Collins, CO 80521, USA e-mail:
Plant Cell Rep. 2000 Jun;19(7):733-737. doi: 10.1007/s002999900171.
Routine cryopreservation of shoot tips from sweet potato [Ipomoea batatas (L.) Lam] has been hampered by their survival variability after cryogenic exposure. We examined the effects of light conditions on stock plants, sucrose preculture and cryoprotectant loading on survival after vitrification using PVS2 solution. The survival of vitrified sweet potato shoot tips cooled to approximately -208 °C was increased by preculturing with 0.3 M sucrose for 24 h at 22 °C. Survival was also enhanced by excising shoot tips immediately after the 8-h dark photoperiod. The best survival after cryogenic exposure was obtained using 2 M glycerol +0.4 M sucrose for 1 h at 22 °C followed by dehydration with PVS2 for 16 min at 22 °C. Rapid cooling was used and achieved by the immersion of foil strips into partially solidified nitrogen. Successfully vitrified and warmed shoot tips directly developed shoots on a medium containing 1 μM NAA, 0.5 μM BA and 0.1 μM kinetin with only minimum callus formation. Shoot formation occurred in all surviving shoot tips. This procedure shows promise for cryopreserving sweet potato shoot tips.
甘薯[Ipomoea batatas (L.) Lam]茎尖的常规冷冻保存一直受到低温处理后存活变异性的阻碍。我们研究了光照条件对母株的影响、蔗糖预培养以及使用PVS2溶液玻璃化处理后冷冻保护剂加载对存活的影响。在22°C下用0.3 M蔗糖预培养24小时,可提高冷却至约-208°C的玻璃化甘薯茎尖的存活率。在8小时黑暗光周期后立即切除茎尖也可提高存活率。在22°C下使用2 M甘油+0.4 M蔗糖处理1小时,然后在22°C下用PVS2脱水16分钟,可获得低温处理后最佳的存活率。采用快速冷却,即将铝箔条浸入部分凝固的液氮中实现。成功玻璃化并解冻的茎尖在含有1 μM萘乙酸、0.5 μM苄氨基腺嘌呤和0.1 μM激动素的培养基上直接发育成芽,仅形成少量愈伤组织。所有存活的茎尖都形成了芽。该方法有望用于甘薯茎尖的冷冻保存。