- 期刊
Using Dilution and Micropipette Technique to Isolate Oil Producing Algae
利用稀釋／微吸管技術分離淡水油脂藻
摘要
Oleaginous algae has a cellular lipids content over 20% (w/w), making it potential feedstock for biodiesel production during the energy crisis. Taiwan is located in the subtropical hemisphere with long day light for photo-synthesis, which is an optimum environment for algal growth. This study investigates and isolates oleaginous algae from freshwater for biofuel production. Water samples (100ml) collected from fresh water were filtered with membrane filter and enriched with BG11 and 3N-BBM media for 3 weeks. The algal enriched water samples were diluted and the micropipette technique was used to isolate single algal cells. The single algal cells were cultured. Seventeen isolates were successfully isolated from five water samples. From the Nile Red staining results, ten isolates were preliminarily screened for oil producing algae. These isolates were identified according to their morphologies as Botryococcus sp., Chlorella sp., Colastrum sp., Golenkinia sp., Kirchneriella sp., Monoraphidium sp. and Scenedesmus sp. Two out of the 10 isolates having higher oil content than the others were identified as Golenkinia longispicula and Neochloris aquatica by 18S ribosomal RNA sequencing. Golenkinia sp. is the first time reported as an oleaginous alga.
並列摘要
藻細胞油脂含量超過20%（w /w）者可稱為油脂藻，在能源缺乏之際它們可做為生質柴油的料源。台灣處於亞熱帶氣候，長的日照時間非常適於藻株之生長。本研究目的為自淡水水域中分離油脂藻細胞，以便為生質燃料提供更多的料源選擇。採自台中市地區之水樣（100 ml）先以濾膜過濾後，再分別以BG11及3N-BBM培養基積聚。積聚之藻液結合稀釋與微吸管分離法，成功地自五處水樣中分離出17株藻，利用尼羅紅染色法初步篩選出其中10株藻細胞內有油脂分佈。由藻細胞外觀判斷此10株藻之屬名為Bortyococcus sp.、Chlorella sp.、Coelastrum sp.、Golenkinia sp.、Kirchneriella sp、Monoraphidium sp.及Scenedesmus sp.。其中2藻株具備生長快速且油脂含量高的特性，經過18S rRNA鑑定後，此二藻株分別為Neochloris aquatica與Golenkinia longispicula，其中Golenkinia longispicula為首次報導油脂藻。