Abdelnour-Esquivel Ana, Engelmann Florent
Centro de Investigacion en Biotecnologia, Instituto Tecnologico de Costa Rica (ITCR).
Cryo Letters. 2002 Sep-Oct;23(5):299-308.
This paper presents the development of cryopreservation protocols for zygotic embryos and apices of chayote (Sechium edule Jacq. Sw.), a tropical plant species with recalcitrant seeds. Zygotic embryos of two cultivars, Ccocro negro (CN) and Claudio (Cl) could withstand cryopreservation, with survival percentages of 10 and 30 %, after desiccation to 23 and 19 % moisture content (fresh weight basis), respectively. Apices sampled on in vitro plantlets of cultivars Cl, 13 and JM were successfully cryopreserved using a vitrification technique. Optimal conditions included the culture of mother-plants for 22 days on medium containing 0.3 M sucrose, culture of excised apices on the same medium for 1 day, loading of apices for 20 min with 2M glycerol + 0.4M glycerol, treatment with a series of diluted PVS2 solution (60 % PVS2 followed by 80 % PVS2 solution for 15 min (cultivar Cocoro Blanco [CB]) or 30 min (cultivars CN and Cl) at each concentration), rapid freezing and thawing, washing of shoot-tips with a 1.2 M sucrose solution, followed by recovery on media with progressively decreasing sucrose concentrations until the standard concentration of 0.1 M was reached. The highest survival percentages achieved ranged between 17 and 38 %, depending on the cultivar.
本文介绍了佛手瓜(Sechium edule Jacq. Sw.)合子胚和茎尖冷冻保存方案的进展,佛手瓜是一种具有顽拗性种子的热带植物。两个品种Ccocro negro(CN)和Claudio(Cl)的合子胚在分别干燥至含水量为23%和19%(鲜重基础)后能够耐受冷冻保存,存活率分别为10%和30%。使用玻璃化技术成功地对品种Cl、13和JM的离体苗上采集的茎尖进行了冷冻保存。最佳条件包括母株在含0.3M蔗糖的培养基上培养22天,切下的茎尖在同一培养基上培养1天,用2M甘油 + 0.4M甘油加载茎尖20分钟,用一系列稀释的PVS2溶液处理(60% PVS2,然后在每个浓度下用80% PVS2溶液处理15分钟(品种Cocoro Blanco [CB])或30分钟(品种CN和Cl)),快速冷冻和解冻,用1.2M蔗糖溶液洗涤茎尖,然后在蔗糖浓度逐渐降低的培养基上恢复,直至达到0.1M的标准浓度。根据品种不同,实现的最高存活率在17%至38%之间。