臺灣博碩士論文加值系統

附著性矽藻是九孔育苗時不可或缺的餌料生物，餌藻的質與量，影響苗之生長與活存，如何控制與管理附著板上矽藻膜之技巧，是種苗繁殖上一項關鍵工作。以養殖附著性矽藻生產鮑（九孔）苗其存活成功率高、穩定性佳，但矽藻對鮑（九孔）苗之附著與變態的影響因藻種而異，且同屬不同種也有不同之效益。另一方面，矽藻的形態特徵會隨養殖條件變異，加上種類繁多、同種異名、新種名修正，而使得種名鑑定更是項艱巨的工作。為解決有益於九孔育苗用附著性矽藻種類之鑑定，本研究以附著性矽藻之形態及色素組成來進行探討。由水產試驗所生物技術組取得已分離、保種之附著性矽藻，進行小量培養至對數增殖晚期，經離心濃縮、酸洗後，以光學顯微鏡觀察，再經自然風乾、鍍膜後，以掃描式電子顯微鏡觀察。矽藻分類的形態特徵主要依據其矽質化細胞壁的微細結構，如殼面突出物、殼縫形態及條紋排列等，作為區別。本研究描述已鑑定至種名的8種：卡比曲殼藻（Achnanthes kolbei）、巴拉曲殼藻（Achnanthes parvula）、扁圓卵形藻多孔變種（Cocconeis placentula var. euglypta）、馬氏舟形藻（Navicula margalithii）、細小雙眉藻（Amphora exigua）、麩皮菱形藻（Nitzschia palea）、反轉菱形藻（Nitzschia reversa）、翼繭形藻（Entomoneis alata），僅鑑定到屬的4種：Achnanthes sp.、Seminavis spp. ，以及已知種名的3種：鏡舟形藻（Navicula cf. lenzii）、清晰雙眉藻（Amphora luciae）及左彎菱形藻（Nitzschia laevis）的形態特徵，並討論其分類的依據與區別。同時也利用高效液相層析系統分析，不同藻體內的色素組成，發現利用色素比例無法鑑別矽藻門內種別的差異。

Benthic diatoms are the principal food source for postlarval abalone in hatchery; the survival and growth of postlarvae depend on species and density of diatom. How to manage the diatom film is a key issue for successful postlarval rearing. Postlarval abalones have highly survival rates fed by the cultured benthic diatoms, but their settlement on plates and metamorphosis vary with diatom species. The morphology of diatoms could vary with the culture conditions; in addition, the synonym and new species revision make the identification of diatom species more difficult. To establish the species identification for benthic diatoms beneficial to postlarval abalone, this study aims at the morphology and pigment compositions of 15 benthic diatom species. Algal cells at late exponential growth phase were collected by centrifuge, observed under light microscopic after acid cleaned, and then under scanning electron microscope after air dried on stubs, sputter coated with gold in a apparatus. The morphology of silicate cell wall, such as portulae, raphe system, arrange of striae, are taxonomic characteristics of diatoms. This study discussed the taxonomic characteristics of eight identified species: Achnanthes kolbei, Achnanthes parvula, Cocconeis placentula var. euglypta, Navicula margalithii, Amphora exigua, Nitzschia palea, Nitzschia reversa, Entomoneis alata; four unidentified species: Achnanthes sp. , Seminavis spp. and three known species：Navicula cf. lenzii, Amphora luciae, Nitzschia laevis, The high performance liquid chromatography system was used to analyze the pigments in diatoms. The results showed that the pigment compositions of algal cells can not be used as chemotaxonomic features in species of Bacillariophyta.

摘要........................................................................................................................................I
Abstract...............................................................................................................................III
誌謝........................................................................................................................................V
目錄.......................................................................................................................................VI
圖表目錄.......................................................................................................................................IX
第1章 前言........................................................................................................................................1
第2章 材料與方法........................................................................................................................................5
2.1 藻類來源........................................................................................................................................5
2.2 樣本製配........................................................................................................................................5
2.2.1 活體藻細胞........................................................................................................................................5
2.2.2 矽藻細胞壁處理........................................................................................................................................5
2.2.3 永久玻片製作........................................................................................................................................5
2.3 形態觀察........................................................................................................................................5
2.3.1光學顯微鏡觀察........................................................................................................................................6
2.3.2 掃描式電子顯微鏡（Scanning Electron Microscope）觀察............6
2.4 細胞殼之構造........................................................................................................................................6
2.5 種類之鑑定........................................................................................................................................8
2.5.1 曲殼藻屬（Achnanthes）............................................................................................................................8
2.5.2 卵形藻屬（Cocconeis）.............................................................................................................................8
2.5.3 舟形藻屬（Navicula）..............................................................................................................................9
2.5.4 半舟形藻屬（Seminavis）.............................................................................................................................9
2.5.5 雙眉藻屬（Amphora）...............................................................................................................................9
2.5.6 菱形藻屬（Nitzschia）.............................................................................................................................9
2.5.7 繭形藻屬（Entomoneis）............................................................................................................................9
2.6. 附著性矽藻之色素組成分析..................................................................10
2.6.1 色素之萃取.......................................................................................................................................10
2.6.2 高效液相層析系統（High-Performance Liquid
Chromatography system；HPLC）..............................................................10
2.6.3 色素之定性.......................................................................................................................................10

2.7 統計分析.......................................................................................................................................11
第3章 結果與討論...................................................................................................12
3.1 十五種附著性矽藻之形態........................................................................12
3.1.1 曲殼藻屬（Achnanthes）..............................................................12
3.1.1.1 巴拉曲殼藻（Achnanthes parvula Kützing）............. 12
3.1.1.2 卡比曲殼藻（Achnanthes kolbei Hustedt）................ 15
3.1.1.3 曲殼藻1（Achnanthes sp1 Bory）................................. 17
3.1.2扁圓卵形藻多孔變種(Cocconeis placentula var. euglypta
Ehrenberg) Cleve..............................................................................19
3.1.3 舟形藻屬（Navicula）......................................................................22
3.1.3.1馬氏舟形藻（Navicula margalithii
Lange-Bertalot）..................................................................22
3.1.3.2 鏡舟形藻( Navicula cf. lenzii Hustedt ) .....................24
3.1.4 半舟形藻屬（Seminavis）................................................................26
3.1.4.1 半舟形藻1( Seminavis sp1 Mann )..............................26
3.1.4.2 半舟形藻2( Seminavis sp2 Mann )..............................26
3.1.4.3 半舟形藻3( Seminavis sp3 Mann )..............................26
3.1.5 雙眉藻屬( Amphora）......................................................................30
3.1.5.1 細小雙眉藻( Amphora exigua Gregory）....................30
3.1.5.2 清晰雙眉藻( Amphora luciae Cholnoky）..................32
3.1.6 菱形藻屬（Nitzschia）......................................................................34
3.1.6.1 反轉菱形藻( Nitzschia reversa Smith )......................34
3.1.6.2 麩皮菱形藻( Nitzschia palea Kützing ) Smith..........36
3.1.6.3 左彎菱形藻( Nitzschia laevis Hustedt ) .....................38
3.1.7 翼繭形藻( Entomoneis alata Ehrenberg ).................................39
3.2 十五種附著性矽藻之色素組成...............................................................41
參考文獻........................................................................................................................57
附錄..................................................................................................................................63
附錄1. 台灣九孔養殖產業於1977-2005年之總生產量及總產值............63
附錄2. 台灣九孔苗於2000-2005年之總產量及總產值...............................64

附錄3. F/2培養液配方(Guillard, 1975經水產試驗所生物技術組
修改後使用)...............................................................................................65
附錄4. 矽藻之分類系統........................................................................................66
附錄5. 羽紋型矽藻之構造名詞..........................................................................73
附錄6. 羽紋型矽藻的細胞殼構造.....................................................................75
附錄7. 殼縫在不同殼面上之型態.....................................................................76
附錄8. 殼面之形成.................................................................................................77
附錄9. 殼縫之種類.................................................................................................78
附錄10. 殼面特殊構造.............................................................................................79
附錄11. 羽紋型矽藻條紋之型態..........................................................................80
附錄12. 小孔、孔室及假孔構造............................................................................81
附錄13. 管殼縫之構造.............................................................................................82
附錄14. 矽藻之附著形態........................................................................................83
附錄15. 移動相梯度時間組合表..........................................................................84
作者簡介 ......................................................................................................................85

圖表目錄
圖1. 巴拉曲殼藻（Achnanthes parvula）..........................................................14
圖2. 卡比曲殼藻（Achnanthes kolbei）.............................................................16
圖3. 曲殼藻1（Achnanthes sp1）........................................................................18
圖4. 扁圓卵形藻多孔變種（Cocconeis placentula var. euglypta）.............21
圖5. 馬氏舟形藻（Navicula margalithii）...........................................................23
圖6. 鏡舟形藻（Navicula cf. lenzii）..................................................................25
圖7. 半舟形藻1（Seminavis sp1）.......................................................................27
圖8. 半舟形藻2（Seminavis sp2）......................................................................28
圖9. 半舟形藻3（Seminavis sp3）......................................................................29
圖10. 細小雙眉藻（Amphora exigua）..............................................................31
圖11. 清晰雙眉藻（Amphora luciae）...............................................................33
圖12. 反轉菱形藻（Nitzschia reversa）..............................................................35
圖13. 麩皮菱形藻（Nitzschia palea）................................................................37
圖14. 左彎菱形藻（Nitzschia laevis）...................................................................38
圖15. 翼繭形藻（Entomoneis alata）..................................................................40
圖16. 巴拉曲殼藻（Achnanthes parvula）之層析圖譜................................43
圖17. 卡比曲殼藻（Achnanthes kolbei）之層析圖譜...................................43
圖18. 曲殼藻1（Achnanthes sp1）之層析圖譜..............................................44
圖19. 扁圓卵形藻多孔變種（Cocconeis placentula var. euglypta）
之層析圖譜............................................................................................44
圖20. 馬氏舟形藻（Navicula margalithii）之層析圖譜...................................45
圖21. 鏡舟形藻（Navicula cf. lenzii）之層析圖譜.............................................45
圖22. 半舟形藻1（Seminavis sp1）之層析圖譜.................................................46
圖23. 半舟形藻2（Seminavis sp2）之層析圖譜.................................................46
圖24. 半舟形藻3（Seminavis sp3）之層析圖譜.................................................47
圖25. 細小雙眉藻（Amphora exigua）之層析圖譜...........................................47
圖26. 清晰雙眉藻（Amphora luciae）之層析圖譜...........................................48
圖27. 反轉菱形藻（Nitzschia reversa）之層析圖譜..........................................48
圖28. 麩皮菱形藻（Nitzschia palea）之層析圖譜.............................................49
圖29. 左彎菱形藻（Nitzschia laevis）之層析圖譜.............................................49
圖30. 翼繭形藻（Entomoneis alata）之層析圖譜..............................................50
圖31. chlorophyllide a 之吸收圖譜.....................................................................51
圖32. chlorophyll c1＋c2之吸收圖譜.................................................................51
圖33. fucoxanthin 之吸收圖譜.............................................................................52
圖34. diadinoxanthin 之吸收圖譜.......................................................................52
圖35. diatoxanthin 之吸收圖譜............................................................................53
圖36. chlorophyll a 之吸收圖譜..........................................................................53
圖37. β,β-carotene 之吸收圖譜............................................................................54
表 1. 不同色素之吸收波長....................................................................................55
表 2. 各藻種之不同色素佔層析圖之面積百分比..........................................56

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