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... Isolation and Characterization of Methanofollis Species from Potential Cold Seep Area-Four Way Closure Ridge Offshore SW Taiwan. 指導教授: 賴美津. 資料載入處理中... 跳到主要內容 臺灣博碩士論文加值系統 ::: 網站導覽| 首頁| 關於本站| 聯絡我們| 國圖首頁| 常見問題| 操作說明 English |FB專頁 |Mobile 免費會員 登入| 註冊 功能切換導覽列 (167.99.71.17)您好！臺灣時間：2021/12/2715:07 字體大小：       ::: 詳目顯示 recordfocus 第1筆/ 共1筆  /1頁 論文基本資料 摘要 外文摘要 目次 參考文獻 電子全文 QRCode 本論文永久網址: 複製永久網址Twitter研究生:鄧乃瑄研究生(外文):Nai-HsuanTeng論文名稱:台灣西南海域四方圈合海脊冷泉區甲烷泡菌屬新種之純化與特性分析論文名稱(外文):IsolationandCharacterizationofMethanofollisSpeciesfromPotentialColdSeepArea-FourWayClosureRidgeOffshoreSWTaiwan指導教授:賴美津口試委員:湯森林、黃啟裕口試日期:2018-07-09學位類別:碩士校院名稱:國立中興大學系所名稱:生命科學系所學門:生命科學學門學類:生物學類論文種類:學術論文論文出版年:2018畢業學年度:106語文別:中文論文頁數:86中文關鍵詞:古菌、甲烷古菌、甲烷古菌、甲烷泡菌屬、四方圈合海脊、冷泉、天然氣水合物外文關鍵詞:Archaea、Methanogen、Methanofollis、FourWayClosureRidge、ColdSeep、GasHydrate相關次數: 被引用:2點閱:71評分:下載:0書目收藏:0 四方圈合海脊為一個背斜海脊，是一個收集甲烷的良好地形。

應用高解析的測深儀(bathymetry)、深拖側掃聲納圖像(deep-towedsidescansonarimagery)、高解析震波剖面探測(high-resolutionseismicprofiling)以及海底拖式攝影機(towedvideoobservations)分析均確認四方圈合海脊是天然氣水合物累積的潛力區。

四方圈合海脊亦具有強甲烷滲漏點，造就了以甲烷碳源為基礎的典型冷泉(coldseep)化學自養菌群落(chemosyntheticcommunity)和高密度具甲烷氧化菌共生的貽貝與蝦蟹。

2015年海洋研究船三號OR3-1900航次自台灣四方圈合海脊C5-6測站使用V-Corer(Video-guidedmulticorer)獲得的岩心底泥樣品，經增殖培養後純化出甲烷古菌FWC-SCC2。

菌株FWC-SCC2與MethanofollistationisChile9T的16SrDNA序列達98.2%相似性，與其它甲烷泡菌屬已發表菌株的16SrDNA相似性達96-98%。

但菌株FWC-SCC2與甲烷泡菌屬的物種分析平均核苷酸一致性(Averagenucleotideidentity,ANI)與基因組對基因組距離分析(Genometogenomedistanceanalysis,GGDA)，結果分別為78.2-79.8%以及17.8-23.8%，支持菌株FWC-SCC2為甲烷泡菌屬的新種。

菌株FWC-SCC2細胞為直徑0.8-1.2μm的不規則球菌，其表層蛋白分子量為116.4kDa；可以利用甲酸和H2/CO2作為碳源進行甲烷化作用。

菌株FWC-SCC2最適生長條件範圍為37C、0.17M氯化鈉及pH6.7-7.0。

在基因體的分析上發現菌株FWC-SCC2與其他甲烷泡菌屬菌株相比具38個獨特的蛋白質直系同源群(ClustersofOrthologousGroupsofproteins,COGs)，在38個COGs中包含有鐵離子的ABC運輸蛋白(ABC-typeirontransportsystemFetAB)、對苯二酚單氧化酶YgiN(QuinolmonooxygenaseYgiN)兩種和耐氧性相關的COGs以及維持細胞膜完整性的噬菌體休克蛋白A(PhageshockproteinA)。

FourWayClosureRidgeisananticlinalridgethatrepresentsanidealtrapformethanehydrateaccumulationsthroughtheanalysesofhigh-resolutionbathymetry,deep-towedsidescansonarimagery,high-resolutionseismicprofilingandtowedvideoobservationsoftheseafloor.FourWayClosureRidgeisoneofthesiteswithintensivemethaneseepagestobringupatypicalcoldseepusedmethaneassubstratechemosyntheticcommunityandwithhighlydensemusselbedwithshrimps,crabsandclamswasfoundatthissite.Amesophilic,hydrogenotrophicmethanogen,strainFWC-SCC2,wasisolatedfromdeep-seasedimentobtainedattheC5-6stationatFourWayClosureRidgeregionbytheR/VOceanResearcherIIIduringORIII-1900cruisein2015.PhylogeneticanalysisrevealedthatstrainFWC-SCC2wasmostcloselyrelatedtoMethanofollistationisChile9T(98.2%16SrRNAgenesequencesimilarity)andshare96-98%16SrRNAgenesequencesimilaritywithothervalidMethanofollisspecies.GenomerelatednessbetweenstrainFWC-SCC2andMethanofollisgenuswascomputedusingbothaveragenucleotideidentity(ANI)andgenome-to-genomedistanceanalysis(GGDA)withvaluesof78.2-79.8%and17.8-23.8%,respectively.Basedonthemorphological,phenotypicandphylogeneticcharacteristicspresentedhere,itissuggestedthatstrainFWC-SCC2representsanewspeciesofgenusMethanofollis.Thecellswereirregularcocci,non-motilewith0.8-1.2μmindiameter.TheMrofsurface-layerproteinwas116.4kDa.StrainFWC-SCC2usedH2/CO2orformateascatabolicsubstrates.Thegrowthconditionswereat20-40C,0-0.85MNaClandpH5.9-8.2.ComparativegenomicanalysisbetweenstrainFWC-SCC2andotherMethanofollisrevealsstrainFWC-SCC2have38uniqueCOGs,includedABC-typeirontransportsystemFetAB,quinolmonooxygenaseYgiN,twotypeaerotolerantCOGsandphageshockproteinAthatcanmaintaincellmembraneintegrity. 中文摘要iAbstractii目錄iii表目錄vi圖目錄vii壹、前言1貳、前人研究3一、台灣西南外海四方圈合海脊的地質特色及天然氣水合物蘊藏區3二、四方圈合海脊冷泉區生物3三、四方圈合海脊冷泉區微生物多樣性5四、深海底泥中以甲烷為主的生態系6五、甲烷古菌(Methanogen,Methanoarchaea)8(一)甲烷古菌分類91.甲烷囊菌屬(Methanoculleus)102.甲烷泡菌屬(Methanofollis)12六、研究目的與重要結果14參、材料與方法15一、採樣時間與地點15二、除氧操作系統、藥劑與培養基15(一)除氧操作系統(Hungatestation)15(二)MB/W及MM/Wmedium配製15(三)還原劑與碳源製備17(四)抗生素制備17(五)TGC(thioglycollate)培養基配製18三、甲烷古菌增殖培養與厭氧轉殖接種18(一)接菌18(二)氣象層析儀偵測樣品甲烷氣體產量19(三)抗生素添加20(四)連續稀釋法20(五)厭氧操作箱(Coychamber)20(六)Roll-tube方法20四、微生物形態觀察21(一)位相差顯微鏡觀察細胞形態21(二)穿透式電子顯微鏡經負染方式觀察細胞形態21(三)場發射掃描電子顯微鏡觀察細胞22五、生長與生理生化特性分析23(一)甲烷古菌使用碳源的配製與測試23(二)抗生素抗性測試23(三)生長曲線的測定24(四)溫度生長範圍測試24(五)鹽濃度生長範圍測試24(六)酸鹼度生長範圍測試25(七)SDS感受性測試25(八)表面蛋白(Surfacelayer)之製備26(九)蛋白質電泳分析(SDS-PAGE)26六、核酸萃取與分析27(一)甲烷古菌增殖樣品DNA萃取27(二)甲烷古菌染色體DNA萃取28(三)核酸純度與定量分析29(四)聚合酶連鎖反應29(五)核酸膠體電泳29(六)PCR增幅樣品回收30(七)DNA黏合反應31(八)勝任細胞製備31(九)質體轉形作用32(十)質體DNA萃取與純化32七、核酸定序與親緣關係分析33(一)16SrRNA基因定序與序列分析33(二)染色體DNA定序與序列比較34肆、結果與討論36一、四方圈合海脊底泥甲烷古菌之增殖培養與純化36二、自四方圈合海脊底泥純化之甲烷古菌菌株FWC-SCC236(一)細胞形態37(二)16SrRNA基因序列與系統演化分析38(三)生理特性分析38(四)基因體分析411.菌株FWC-SCC2與甲烷泡菌屬菌株基因體資訊412.菌株FWC-SCC2與甲烷泡菌屬菌株間蛋白質直系同源群比較423.菌株FWC-SCC2鞭毛基因434.甲烷代謝途徑44三、部分純化甲烷古菌45(一)PR1-G0-0-10_C2_A45(二)PR1-G2-0-10_C2_M45伍、結論46陸、表與圖47柒、參考文獻67捌、附錄78 鄭婉言、林曉武、陳麗雯、楊燦堯和戚務正。

2015。

台灣鄰近海底冷泉與泥火山：能源與生物寶藏。

自然科學簡訊。

27:30-35.蔣孟庭。

2016。

底棲性有孔蟲群集組成與冷泉生地化環境之關連―以臺灣西南海域四方圈合海脊為例。

國立臺灣大學地質科學研究所碩士學位論文。

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