植物體胚細胞、莖頂與花粉冷凍保存之研究

Studies of Plant Cryopreservation with Somatic Embryogenesis Cell, Shoot Tip and Pollen

王昭月(Jau-Yueh Wang) ； 曾夢蛟(Menq-Jiau Tseng)

《植物種苗》 6卷1期 (2004/03) Pp. 1-12

https://doi.org/10.30077/SN.200403.0001

摘要

冷凍保存是一種兼具可安全與集約地將生物材料長期保存於相當低溫(-30℃)或超低溫液態氮中(-196℃)的措施。冷凍保存有助於農業與生物多樣性的維護，尤其對於珍稀物種的保育上特別具有意義。低於-120℃狀態下的冷凍保存，可使遺傳變異機會降到最低，安全地保育其遺傳特質。近二十年間，生物學家業已開發一連串另類的冷凍保護措施，以配合進行植物的細胞、花粉以及微小組織的冷凍保存；包含脫水處理、玻璃質化處理(virtrification)、藻膠膠囊式包埋併脫水處理(encapsulation & dehydration)以及藻膠膠囊式包埋併玻璃質化處理(encapsulation & virtrification)等方法。大致而言，目前係以玻璃質化處理或藻膠膠囊式包埋併玻璃質化處理兩種方式，具有最佳之冷凍保存結果。另外，將植物材料預培養於較高張溶液中(如高濃度之蔗糖溶液)，或在冷凍保存前預經冷健化處理，均可有效地提高冷凍保存後之成活率。而藉由差異溫度分析(DTA：different thermal analysis)、核磁共振(NMR：nuclear magneticresonance)、掃瞄式電子顯微鏡法(SEM：scanning electron microscopy)等技術的應用，亦可精確地監控植物在冷凍逆境下的結冰反應，用以改善冷凍保存實質之成活率。

關鍵字

植物冷凍保存 ； 脫水處理 ； 玻璃質化處理 ； 藻膠膠囊式包理

並列摘要

Cryopreservation is a safe and cost-effective strategy for long term preservation of biological material at very low temperature (below -30 ℃) or directly in ultra-low temperature of liquid nitrogen (-196 ℃). Cryopreserved plant materials could benefit the conservation of agriculture and biodiversity especially for valuable species. Cryogenic conservation (below -120℃) also safeguards for base collections with minimal risk of genetic changes. Recently, alternate cryoprotectant methods have been developed to plant cell, pollen or small tissue that include dehydration, virtrification, encapsulation & dehydration or encapsulation & virtrification. In most cases the virtrification or encapsulation & virtrification showed the most effective than others' procedures did. Furthermore, preculture plant cell in medium with high osmoticum (such as sucrose) or cold hardy (cold acclimation) treatment prior to cryopreservation also revere better regenerating ability. Through the techniques such as different thermal analysis (DTA), nuclear magnetic resonance (NMR), or scanning electron microscopy (SEM) monitoring the response of plants under freeze stress, and improving survival ratio of plant materials after cryopreservation.

並列關鍵字

plant cryopreservation ； dehydration ； virtrification ； encapsulation