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題 名 | 六種水產養殖常用之海洋微細藻類超低溫保存的研究=Study in Cryopreservation of Six Marine Microalgae for Aquaculture |
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作 者 | 張詠政; 李英周; 陳衍昌; 郭金泉; | 書刊名 | 植物種苗 |
卷 期 | 14:2 2012.06[民101.06] |
頁 次 | 頁1-16 |
分類號 | 439.63 |
關鍵詞 | 超低溫保存; 抗凍劑; 海洋微細藻類; 細胞濃度; Cryopreservation; Cryoprotectant; Marine microalgae; Cell concentration; |
語 文 | 中文(Chinese) |
中文摘要 | 本研究探討六種常應用於台灣水產養殖育苗的海洋微細藻類:擬球藻(Nannochloropsis oculata)、扁藻(Tetraselmis elliptica)、簡單雙眉藻(Amphora exigua)、半舟行藻(Seminavis gracilenta)、骨藻(Skeletonema costatum)及等鞭金藻(Isochrysis galbana)的最佳超低溫保存條件。探討5種常用的滲透性抗凍劑,二甲基亞碸(dimethyl sulfoxide;DMSO)、甲醇(methanol;MeOH)、甘油(glycerol;GLY)、乙二醇(ethylene glycol;EG)及丙二醇(propylene glycol;PG)和五種不同濃度(0%;5%;10%;15%;20%),自室溫以每分鐘-1℃降至-40℃,再放入液態氮中保存一天後,經40℃水浴解凍恢復培養之活存率。結果顯示,除了骨藻無法超低溫保存外,擬球藻以20%乙二醇(89%),扁藻以20%丙二醇(57%),簡單雙眉藻以20%二甲基亞碸(90%),半舟行藻以20%二甲基亞碸(94%)及等鞭金藻以15%甘油(72%)做為抗凍劑進行超低溫保存,有最高的活存率,而藻類起始細胞濃度則對藻類活存率沒有顯著影響。 |
英文摘要 | Cryopreservation at ultra-low (-196℃ in LN2) has many advantages over the conventional maintenance of algae strains using serial subculture, it is becoming the method of choice for the long-term conservation of algae nowadays. This study determining the optimal protocol for cryopreservation of six marine microalgae: Nannochloropsis oculata, Tetraselmis elliptica, Seminavis gracilenta, Amphora exigua, Skeletonema costatum and Isochrysis galbana using the two-step method protocol with five cryoprotectants and five concentrations were discussed. The results showed the best cryoprotectant for algae was species-specific. The highest post-thawed viability for N. oculata was 89% with 20% ethylene glycol (EG), for T. elliptica was 57% with 20% propylene glycol (PG), for S. gracilenta was 94% with 20% dimethyl sulfoxide (DMSO), for A. exigua was 90% with 20% dimethyl sulfoxide (DMSO) and for I. galbana was 72% with 15% glycerol (GLY). Two-step method protocol was not suitable for cryo-storage of Skeletonema costatum. The results showed that the cell concentration of the microalgae were not significantly affected in cryopreservation. |
本系統中英文摘要資訊取自各篇刊載內容。