BL10是一個來自台灣北部河口的異營性微藻分離株，具有許多異於其他微細藻類的特性：1. 廣鹽性 (euryhaline)，在2 - 35 ppt的鹽度範圍內有最佳的生長速率。2. 嗜糖性 (saccharophilic)，其生長不受高濃度葡萄糖 (140 gL-1) 的抑制。3. 油脂性 (oleaginous)，含油量可達乾重的80%。4. 容易培養，以0.01gL-1接種，培養72小時後，可達120 gL-1的生物量。5. 有別於標準路徑的長鏈不飽和脂肪酸生合成路徑。6. 特殊的生活史，除了營養細胞 (vegetative cell) 外，尚能產生具有游動能力的游走孢子 (zoospore) 以及爬行能力的阿米巴細胞 (amoeboid cell)。因此，或許能將BL10建立為新型的模式生物，用以研究微藻的滲透壓調節、油脂產生及堆積、長鏈不飽和脂肪酸的生合成路徑、以及細胞遷徙等生命現象。然而欲以其為模式生物，必先了解其屬於哪種藻、能否取得全基因體的資料，以及是否容易進行基因操作。
因此，本研究的目的，在於確認BL10的分類地位、評估與分析基因體大小及染色體核型 (karyotype)，以及研擬全基因體定序及基因操作的策略。首先以18S rDNA的序列與已知Aurantiochytrium以及其他破囊壺菌品系進行親緣關係分析時，發現BL10會與已知的Aurantiochytrium品系形成單系類群 (monophyletic group) 同時和A. limacinum的type species SR21，而非A. mangrovei的type species RCC893有較近似的親緣關係。BL10容易觀察到阿米巴細胞，以及阿米巴細胞在停止移動後不會立即形成游走孢子的形態特徵也和A. limacinum近似，然而BL10不會形成孢子囊，以及單一營養細胞形成孢子囊的特徵與A. limacinum迥異。因此本研究仍暫時將BL10定名為Aurantiochytrium sp.。
此外，我們利用添加0.2% Triton X-100的方法，證明阿米巴細胞為BL10的三種細胞中 (另兩種為游走孢子及營養細胞) 細胞壁最薄 (或唯一缺乏細胞壁) 的一種，同時在特定培養條件 (例如於培養基內添加0.9%硫酸鈉取代海水的作法) 以及特定培養時間 (對數生長期；接種12小時前後) 有較高比例的阿米巴細胞出現 (占總細胞之16%)，可作為利用電穿孔法，將外來基因或雙股RNA送入BL10的勝任細胞來使用。
後續以流式細胞儀，搭配基因體12 Mbp的酵母菌 (Saccharomyces cerevisiae strain W303-1A) 作為內標準，以DNA的相對螢光強度，換算所得之BL10基因體大小為40.5 Mbp，僅為Chlamydomonas (現行微藻模式生物) 的1/3。並以螢光顯微鏡觀察細胞分裂中期細胞的染色體，發現BL10具有3條染色體的核型，並以螢光原位雜交證實為其單套生物體，說明有機會能利用次世代的定序儀，完成BL10全基因組的定序。

BL10 is a marine heterotrophic microalga that isolated from the stream outlet of northern Taiwan, it has many unique characteristics, including (1) euryhaline, it able to grow within the salinity range from 2 to 35 ppt; (2) saccharophilic, its growth not inhibited when the cell cultured in medium containing high glucose concentrations (140 gL-1); (3) oleaginous, its oil accumulated more than 80% of the dry biomass; (4) culture easily, the biomass can reach 120 gL-1 at 72 hour that inoculated with the initial cell concentrations of 0.01gL-1; (5) unique lipid metabolism pathway, and (6) special life cycle in BL10 that contains mobile zoospores and irregular-shaped amoeboid cells (the cell with crawling ability), making BL10 a nice candidate to become a new model organism, which can be used to study osmotic regulation, the production and accumulation of lipid, the long chain polyunsaturated fatty acids synthetic pathway of microalgae, and cell migration. However, the available information is insufficient to establish a model organism, including (1) the taxonomy of BL10 (2) the genome size and karyotype of BL10 (3) gene manipulation of BL10. Therefore, the purpose of this study is to resolve the above issues.
First, in order to identify the species of BL10, the 18S rDNA sequence were used to construct the phylogenetic tree. The results of a molecular phylogenetic analysis showed that BL10 belongs to the genus Aurantiochytrium and closely relates to A. limacinum. From the results of morphological observation, we found that BL10 can be easily observed the formation of amoeboid cells, and the amoeboid cells can divide into numerous zoospores after the cell settlement down. The above results showed that BL10 is similar to A. limacinum, while BL10 is unable to form the sporangium that significant different with A. limacinum.However, we consider that these results can not support the taxonomy of BL10.
On the other hand, we found that amoeboid cell with thin cell wall (or without cell wall), and the percentage of amoeboid cells could be enhanced up to 16% of total cells by regulating the medium composition and acquired at specific time. Those results provide information that it's possible to do gene transformation or dsRNA delivery for genomic functional analysis of BL10.
Then we established the genome size by flow cytometry, and used Saccharomyces cerevisiae W303-1A (12 Mbp) as control to calculate the genome size of BL10 (40.5 Mbp) that based on FL3 intensity. And in the karyotype analysis experiment, we used 8-hydroxyquinoline to arrest the cell cycle at the metaphase, and the chromosomes of BL10 were stained with DAPI and observed by fluorescence microscopy. The experimental results showed that BL10 has three chromosomes. Then we proved that the BL10 is a haploid using fluorescence in situ hybridization with a 45S rDNA probe. According to our results, we believe that the work of whole genome sequencing is possible.

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