臺灣博碩士論文加值系統

本研究聚焦於副熱帶黑潮水域中，原核超微植物性浮游生物族群的組成與水層分布的季節性變化，期許這些資料能為黑潮流域的生態背景提出貢獻。進行方式為在臺灣花蓮外海黑潮海域於2012 年 10 月至 2013 年 7 月間進行為期一年四季的採樣觀測。研究顯示在 100 公尺水深內，原核綠藻 (Prochlorococcus) 總存量與硝酸鹽躍層深度呈正相關；聚球藻 (Synechococcus) 總存量與溫度呈負相關。計算得知原核綠藻與聚球藻分別貢獻了全體葉綠素平均濃度的30.2 ± 10.4 % 至 33.0 ± 9.4 %。透過分析表水中的 16S rDNA 序列，發現此水域原核超微浮游生物物種組成在暖季 (夏秋季) 與冷季 (冬春季) 有明顯差異。細看原核超微植物性浮游生物物種組成，可看出夏秋季在表水之中以原核綠藻為主，可達總藍綠菌數量 71 % 以上，聚球藻則介於 2 至 20 %；而在冬春季原核綠藻在不同測站占總藍綠菌數量的22至75 %；反觀聚球藻在冬春季的比例則可達 17 至 54 %。非度量多維度分析 (non-metric multidimemsional scaling, nMDS) 結果也顯示出黑潮表水超微浮游生物四季組成呈現顯著的差異，冬季與夏季其單季節各測站族群結構差異較小但不同季節結構相似度不高，春秋季族群結構為冬季與夏季的過渡。本研究成果首度呈現超微原核植物性浮游生物在副熱帶黑潮水域的群落組成時空分布，以及其對於該水域生地化循環的貢獻程度。

The aim of this study is to reveal the community structure and temporal-seasonal distritribution of prokaryotic picophytoplankton in the surface waters of subtropical Kuroshio Current. The sampling period was from October 2012 to July 2013 in the Kuroshio area off the coast of Hualien, Taiwan. Within the upper 100 m depth, the standing stock of Prochlorococcus positively correlated with the depth of nitracline (p<0.05). The standing stock of Synechococcus negatively correlated with temperature and the depth of the euphotic zone (p<0.05). Overall, Prochlorococcus and Synechococcus respectively occupied 30.2 ± 10.4% to 33.0 ± 9.4% of the chlorophyll concentration. According to the results of 16S rDNA phylogeny, the assemblage composition of prokaryotic picophytoplankton was different in the cold and warm seasons. Prochlorococcus dominated in the surface water in the summer and the autumn. In contrast, Synechococcus was abundant in the cold seasons, and their proportion to total cyanobacteria was 17 to 54%. The results of my study present the community composition of prokaryotic picophytoplankton and their contribution to the biogeochemical cycle of the waters in the subtropical surface Kuroshio Current.

致謝........................................................I
摘要.......................................................II
Abstract.................................................III
目次.......................................................IV
表目次.....................................................VI
圖目次....................................................VII
第一章、研究緒論............................................1
1.1研究背景與文獻回顧........................................1
1.2研究目的.................................................4
第二章、實驗方法與材料.......................................6
2.1研究海域與航次時間........................................6
2.2樣本採集.................................................6
2.2.1營養鹽與葉綠素a樣本.....................................6
2.2.2流式細胞儀樣本.........................................6
2.2.3次世代定序樣本.........................................7
2.3實驗測定.................................................7
2.3.1總溶解性無機氮測定與硝酸鹽躍層深度之計算.................7
2.3.2葉綠素a測定............................................7
2.3.3流式細胞儀樣品測定......................................7
2.3.4次世代定序樣品製備......................................8
2.4次世代定序數據分析........................................9
2.5 統計分析...............................................11
第三章、實驗結果...........................................12
3.1 溫度與鹽度.............................................12
3.2 總無機氮與硝酸鹽躍層深度................................12
3.3 葉綠素 a...............................................13
3.4 流式細胞儀計數結果......................................14
3.4.1 聚球藻...............................................14
3.4.2 原核綠藻.............................................15
3.4.3 超微真核植浮.........................................16
3.4.4 超微植浮的四季變化....................................16
3.4.5 超微原核植浮的葉綠素 a................................16
3.5 次世代定序.............................................17
第四章、討論...............................................19
4.1 水文環境與植浮的關係....................................19
4.1.1 水文環境與聚球藻的關係................................20
4.1.2 水文環境與原核綠藻的關係..............................21
4.1.3 綜合分析.............................................22
4.2 黑潮表水的超微原核生物組成結構分析.......................22
第五章、結論...............................................24
第六章、參考文獻...........................................25

高愷嶸（2014）。蘇拉颱風對亞熱帶西北太平洋貧瘠海域生地化反應之影響。國立臺灣海洋大學海洋環境與生態研究所碩士論文，基隆市。
Behrenfeld MJ, O’Malley RT, Siegel DA, McClain CR, Sarmiento JL, Feldman GC, Milligan AJ, Falkowski PG, Letelier RM, Boss ES (2006) Climate-driven trends in contemporary ocean productivity. Nature 444:752-755
Bouman HA, Ulloa O, Scanlan DJ, Zwirglmaier K, Li WK, Platt T, Stuart V, Barlow R, Leth O, Clementson L (2006) Oceanographic basis of the global surface distribution of Prochlorococcus ecotypes. Science 312:918-921
Callahan BJ, McMurdie PJ, Rosen MJ, Han AW, Johnson AJA, Holmes SP (2016) DADA2: high-resolution sample inference from Illumina amplicon data. Nat Methods 13:581
Chan YF, Chung CC, Gong GC, Hsu CW (2020) Spatial variation of abundant picoeukaryotes in the subtropical Kuroshio Current in winter. Mar Ecol 41:e12579
Chavez FP, Messié M, Pennington JT (2010) Marine primary production in relation to climate variability and change.
Chavez FP, Messie M, Pennington JT (2011) Marine primary production in relation to climate variability and change. Ann Rev Mar Sci 3:227-260
Chen C-C, Hsu S-C, Jan S, Gong G-C (2015) Episodic events imposed on the seasonal nutrient dynamics of an upwelling system off northeastern Taiwan. J Mar Syst 141:128-135
Chen C-C, Jan S, Kuo T-H, Li S-Y (2017) Nutrient flux and transport by the Kuroshio east of Taiwan. J Mar Syst 167:43-54
Chen C-T (1997) The Kuroshio intermediate water is the major source of nutrients on the East China Sea continental shelf. Oceanograph Lit Rev 5:531
Chen Y-LL (2000) Comparisons of primary productivity and phytoplankton size structure in the marginal regions of southern East China Sea. Cont Shelf Res 20:437-458
Chen Y-LL, Chen H-Y, Gong G-C, Lin Y-H, Jan S, Takahashi M (2004) Phytoplankton production during a summer coastal upwelling in the East China Sea. Cont Shelf Res 24:1321-1338
Chen Y-LL, Chen H-Y, Lin Y-H, Yong T-C, Taniuchi Y, Tuo S-h (2014) The relative contributions of unicellular and filamentous diazotrophs to N2 fixation in the South China Sea and the upstream Kuroshio. Deep Sea Res Part I Oceanogr Res Pap 85:56-71
Cheng W-H, Lu H-P, Chen C-C, Jan S, Hsieh C-h (2020) Vertical Beta-Diversity of Bacterial Communities Depending on Water Stratification. Front Microbiol 11:449
Chung C-C, Gong G-C (2019) Attribution of the growth of a distinct population of Synechococcus to the coverage of lateral water on an upwelling. Terr. Atmospheric Ocean Sci 30:575-587
Chung C-C, Gong G-C, Huang C-Y, Lin J-Y, Lin Y-C (2015) Changes in the Synechococcus assemblage composition at the surface of the East China Sea due to flooding of the Changjiang river. Microb Ecol 70:677-688
Chung CC, Chang J, Gong GC, Hsu SC, Chiang KP, Liao CW (2011) Effects of Asian dust storms on synechococcus populations in the subtropical Kuroshio Current. Mar Biotechnol 13:751-763
Clark C (1992) DNA purification from polysaccharide-rich cells. Protocols in protozoology 3
Domingues CM, Church JA, White NJ, Gleckler PJ, Wijffels SE, Barker PM, Dunn JR (2008) Improved estimates of upper-ocean warming and multi-decadal sea-level rise. Nature 453:1090-1093
Donald KM, Scanlan DJ, Carr NG, Mann NH, Joint I (1997) Comparative phosphorus nutrition of the marine cyanobacterium Synechococcus WH7803 and the marine diatom Thalassiosira weissflogii. J Plankton Res 19:1793-1813
Dufresne A, Garczarek L, Partensky F (2005) Accelerated evolution associated with genome reduction in a free-living prokaryote. Genome Biol 6:R14
Fuller NJ, Tarran GA, Yallop M, Orcutt KM, Scanlan DJ (2006) Molecular analysis of picocyanobacterial community structure along an Arabian Sea transect reveals distinct spatial separation of lineages. Limnol Oceanogr 51:2515-2526
Garczarek L, Dufresne A, Rousvoal S, West NJ, Mazard S, Marie D, Claustre H, Raimbault P, Post AF, Scanlan DJ (2007) High vertical and low horizontal diversity of Prochlorococcus ecotypes in the Mediterranean Sea in summer. FEMS Microbiol Ecol 60:189-206
Ghai R, Mizuno CM, Picazo A, Camacho A, Rodriguez-Valera F (2013) Metagenomics uncovers a new group of low GC and ultra-small marine Actinobacteria. Sci Rep 3:1-8
Godhantaraman N, Uye S (2001) Geographical variations in abundance, biomass and trophodynamic role of microzooplankton across an inshore-offshore gradient in the Inland Sea of Japan and adjacent Pacific Ocean. Plankton Biol Ecol 48:19-27.
Gong G-C, Chang J, Wen Y-H (1999) Estimation of annual primary production in the Kuroshio waters northeast of Taiwan using a photosynthesis-irradiance model. Deep Sea Res Part I Oceanogr Res Pap 46:93-108
Gong G-C, Chen Y-LL, Liu K-K (1996) Chemical hydrography and chlorophyll a distribution in the East China Sea in summer: implications in nutrient dynamics. Cont Shelf Res 16:1561-1590
Gong G-C, Shiah F-K, Liu K-K, Chuang W-S, Chang J (1997) Effect of the Kuroshio intrusion on the chlorophyll distribution in the southern East China Sea during spring 1993. Cont Shelf Res 17:79-94
Gong G (1992) Chemical hydrography of the Kuroshio front in the sea northeast of Taiwan. Institute of Oceanography, National Taiwan University, Taipei, Taiwan:204
Heywood J, Zubkov M, Tarran G, Fuchs B, Holligan P (2006) Prokaryoplankton standing stocks in oligotrophic gyre and equatorial provinces of the Atlantic Ocean: evaluation of inter-annual variability. Deep Sea Res Part II Top Stud Oceanogr 53:1530-1547
Hirose M, Katano T, Hayami Y, Kaneda A, Kohama T, Takeoka H, Nakano S-i (2008) Changes in the abundance and composition of picophytoplankton in relation to the occurrence of a Kyucho and a bottom intrusion in the Bungo Channel, Japan. Estuar Coast Shelf Sci 76:293-303
Hoegh-Guldberg O, Bruno JF (2010) The impact of climate change on the world’s marine ecosystems. Science 328:1523-1528
Hung C-C, Gong G-C (2011) Biogeochemical responses in the southern East China Sea after typhoons. Oceanography 24:42-51
Katano T, Kaneda A, Kanzaki N, Obayashi Y, Morimoto A, Onitsuka G, Yasuda H, Mizutani S, Kon Y, Hata K, Takeoka H, Nakano S (2007) Distribution of prokaryotic picophytoplankton from Seto Inland Sea to the Kuroshio region, with special reference to Kyucho events. Aquat Microb Ecol 46:191-201
Lin I, Liu WT, Wu CC, Wong GT, Hu C, Chen Z, Liang WD, Yang Y, Liu KK (2003) New evidence for enhanced ocean primary production triggered by tropical cyclone. Geophys Res Lett 30
Magazzu G, Decembrini F (1995) Primary production, biomass and abundance of phototrophic picoplankton in the Mediterranean Sea: a review. Aquat Microb Ecol 9:97-104
Martin M (2011) Cutadapt removes adapter sequences from high-throughput sequencing reads. EMBnet journal 17:10-12
Moore LR, Rocap G, Chisholm SW (1998) Physiology and molecular phylogeny of coexisting Prochlorococcus ecotypes. Nature 393:464-467
Morel A, Ahn Y-H, Partensky F, Vaulot D, Claustre H (1993) Prochlorococcus and Synechococcus: a comparative study of their optical properties in relation to their size and pigmentation. J Mar Res 51:617-649
Morán XAG, LÓPEZ‐URRUTIA Á, CALVO‐DÍAZ A, Li WK (2010) Increasing importance of small phytoplankton in a warmer ocean. Glob Change Biol 16:1137-1144
Morris RM, Rappé MS, Connon SA, Vergin KL, Siebold WA, Carlson CA, Giovannoni SJ (2002) SAR11 clade dominates ocean surface bacterioplankton communities. Nature 420:806-810
Oksanen J (2015) Vegan: an introduction to ordination. URL http://cran r-project org/web/packages/vegan/vignettes/introvegan pdf 8:19
Omand MM, Mahadevan A (2015) The shape of the oceanic nitracline. Biogeosciences 12
Paerl HW, Willey JD, Go M, Peierls BL, Pinckney JL, Fogel ML (1999) Rainfall stimulation of primary production in western Atlantic Ocean waters: roles of different nitrogen sources and co-limiting nutrients. Mar Ecol Prog Ser 176:205-214
Pai S-C, Tsau Y-J, Yang T-I (2001) pH and buffering capacity problems involved in the determination of ammonia in saline water using the indophenol blue spectrophotometric method. Anal Chim Acta 434:209-216
Pai S-C, Yang C-C, Riley J (1990) Effects of acidity and molybdate concentration on the kinetics of the formation of the phosphoantimonylmolybdenum blue complex. Analytica Chimica Acta 229:115-120
Partensky F, Blanchot J, Vaulot D (1999a) Differential distribution and ecology of Prochlorococcus and Synechococcus in oceanic waters: a review. Bulletin-Institut Oceanographique Monaco-Numero Special- :457-476
Partensky F, Garczarek L (2010) Prochlorococcus: advantages and limits of minimalism. Ann Rev Mar Sci 2:305-331
Partensky F, Hess WR, Vaulot D (1999b) Prochlorococcus, a marine photosynthetic prokaryote of global significance. Microbiol Mol Biol Rev 63:106-127
Qiu B (2001) Kuroshio And Oyashio Currents. Encyclopedia of Ocean Sciences
Qiu B, Lukas R (1996) Seasonal and interannual variability of the North Equatorial Current, the Mindanao Current, and the Kuroshio along the Pacific western boundary. J Geophys Res Oceans 101:12315-12330
Richardson TL, Jackson GA, Ducklow HW, Roman MR (2004) Carbon fluxes through food webs of the eastern equatorial Pacific: an inverse approach. Deep Sea Res Part I Oceanogr Res Pap 51:1245-1274
Rocap G, Distel DL, Waterbury JB, Chisholm SW (2002) Resolution of Prochlorococcus and Synechococcus ecotypes by using 16S-23S ribosomal DNA internal transcribed spacer sequences. Appl Environ Microbiol 68:1180-1191
Rudnick D, Jan S, Centurioni L, Lee C, Lien R-C, Wang J, Lee D-K, Tseng R-S, Kim YY, Chern C-S (2011) Seasonal and Mesoscale Variability of the Kuroshio Near Its Origin. Oceanography 24:52-63
Scanlan DJ, Ostrowski M, Mazard S, Dufresne A, Garczarek L, Hess WR, Post AF, Hagemann M, Paulsen I, Partensky F (2009) Ecological genomics of marine picocyanobacteria. Microbiol Mol Biol Rev 73:249-299
Scanlan DJ, West NJ (2002) Molecular ecology of the marine cyanobacterial genera Prochlorococcus and Synechococcus. FEMS Microbiol Ecol 40:1-12
Schloss PD, Westcott SL, Ryabin T, Hall JR, Hartmann M, Hollister EB, Lesniewski RA, Oakley BB, Parks DH, Robinson CJ (2009) Introducing mothur: open-source, platform-independent, community-supported software for describing and comparing microbial communities. Appl Environ Microbiol 75:7537-7541
Shen M-L, Tseng Y-H, Jan S (2011) The formation and dynamics of the cold-dome off northeastern Taiwan. J Mar Syst 86:10-27
Shimada A, Nishijima M, Maruyama T (1995) Seasonal appearance ofProchlorococcus in Suruga Bay, Japan in 1992–1993. J Oceanogr 51:289-300
Shiozaki T, Chen Y-lL, Lin Y-H, Taniuchi Y, Sheu D-S, Furuya K, Chen H-Y (2014) Seasonal variations of unicellular diazotroph groups A and B, and Trichodesmium in the northern South China Sea and neighboring upstream Kuroshio Current. Cont Shelf Res 80:20-31
Strehl B, Holtzendorff J, Partensky F, Hess WR (1999) A small and compact genome in the marine cyanobacterium Prochlorococcus marinus CCMP 1375: lack of an intron in the gene for tRNA (Leu) UAA and a single copy of the rRNA operon. FEMS Microbiol Lett 181:261-266
Strickland JDH, Parsons TR (1972) A practical handbook of seawater analysis.
Thompson AW, van den Engh G, Ahlgren NA, Kouba K, Ward S, Wilson ST, Karl DM (2018) Dynamics of Prochlorococcus diversity and photoacclimation during short-term shifts in water column stratification at station ALOHA. Front Mar Sci 5:488
Uitz J, Claustre H, Gentili B, Stramski D (2010) Phytoplankton class-specific primary production in the world's oceans: Seasonal and interannual variability from satellite observations. Global Biogeochem Cycles 24
Welschmeyer NA (1994) Fluorometric analysis of chlorophyll a in the presence of chlorophyll b and pheopigments. Limnol Oceanogr 39:1985-1992
Wong GT, Chao S-Y, Li Y-H, Shiah F-K (2000) The Kuroshio edge exchange processes (KEEP) study—an introduction to hypotheses and highlights. Cont Shelf Res 20:335-347
Zwirglmaier K, Heywood JL, Chamberlain K, Woodward EMS, Zubkov MV, Scanlan DJ (2007) Basin‐scale distribution patterns of picocyanobacterial lineages in the Atlantic Ocean. Environ Microbiol 9:1278-1290
Zwirglmaier K, Jardillier L, Ostrowski M, Mazard S, Garczarek L, Vaulot D, Not F, Massana R, Ulloa O, Scanlan DJ (2008) Global phylogeography of marine Synechococcus and Prochlorococcus reveals a distinct partitioning of lineages among oceanic biomes. Environ Microbiol 10:147-161