擬球藻的簡易低溫保存研究   免費試閱

A Simple Cryopreservation Procedure for Nannochloropsis Oculata

擬球藻（Nannochloropsis oculata）培養容易、生長迅速，是水產養殖的重要餌料食物之一。近年來，由於其油脂含量相當高，因而許多研究著重於如何將其轉化為生質柴油，發展綠色能源。在實驗室中，擬球藻也是一個很好的單細胞模式生物。研究者之實驗室最近將從事多項相關研究，包括分析其蛋白質體，運用核磁共振儀分析其代謝體及進行基因轉殖等研究，因此純系的藻種培養及保存，便是研究初期最重要的工作。基於一般低溫保存方法並配合實驗室需求，本研究將開發一套簡易擬球藻低溫保存方法，並探討抗凍劑及複方抗凍劑的保存效果。使用三種常用的滲透性抗凍劑，甲醇（methanol, MeOH）、甘油（glycerol, GLY）及乙二醇（ethylene glycol, EG），以三種不同濃度（20%，40%，60%）處理組合及三種複方組合，結果顯示，以40% 乙二醇、20% 甲醇及複方組合（20% 乙二醇加10% 甘油）有最高的活存率。顯微鏡觀察也檢視到擬球藻在低溫保存前後其外觀上沒有顯著差異。未來將繼續開發修正此一簡易擬球藻低溫保存方法，並著重複方抗凍劑的保存效果研究。

Nannochloropsis oculata, a promising microalga for biodiesel production, was heavily studied in many basic and applied science research fields. In our lab, this microalga was used as a model organism for proteomics and metabolomics. For the short-term conservation of the Nannochloropsis oculata, we developed a simple procedure for cryopreservation using the two-step method protocol with three cryoprotectants. The results showed the highest post-thawed viability was 100% with 40% ethylene glycol (EG) and 20% methanol (MeOH). Among the cryoprotectant combinations, the highest viability achieved was the combination of 20% ethylene glycol and 10% glycerol (GLY). The microscopic observed results showed that the morphology of the microalga was not significantly changed after cryopreservation. Advanced proteomic and metabolomics profiling of this microalga will be further studied.