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DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 張俊哲(Chun-che Chang) | |
dc.contributor.author | Chun-Wei Lai | en |
dc.contributor.author | 賴峻偉 | zh_TW |
dc.date.accessioned | 2021-06-07T17:41:04Z | - |
dc.date.copyright | 2021-02-20 | |
dc.date.issued | 2021 | |
dc.date.submitted | 2021-02-04 | |
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Wang, Y., Chen, K., Yao, Q., Wang, W., and Zhi, Z. (2007). The basic helix-loop-helix transcription factor family in Bombyx mori. Dev Genes Evol 217, 715-723. Wang, J., and Laurie, G.W. (2004). Organogenesis of the exocrine gland. Dev Biol 273, 1-22. Will, L. (1888). Entwicklungsgeschichte der viviparen Aphiden (Verlag von G. Fischer). Xin, H.H., Zhang, D.P., Chen, R.T., Cai, Z.Z., Lu, Y., Liang, S., and Miao, Y.G. (2015). Transcription factor Bmsage plays a crucial role in silk gland generation in silkworm, Bombyx mori. Archives of insect biochemistry and physiology 90, 59-69. Zhang, Y., Fan, J., Sun, J.-r., and Chen, J.-l. (2015). Cloning and RNA interference analysis of the salivary protein C002 gene in Schizaphis graminum. Journal of Integrative Agriculture 14, 698-705. Zhang, D.B., Wang, Y., Liu, A.K., Wang, X.H., Dang, C.W., Yao, Q., and Chen, K.P. (2013). Phylogenetic analyses of vector mosquito basic helix-loop-helix transcription factors. Insect Molecular Biology 22, 608-621. 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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/15485 | - |
dc.description.abstract | 蚜蟲屬於半翅目中的蚜總科,藉由刺吸式口器取食植物汁液。若取食植物遭受病毒感染,病毒便會感染蚜蟲作為傳播媒介,待下一次的吸食感染新的植物。目前已知蚜蟲會透過具唾液分泌功能的唾腺傳播病毒,然而唾腺在胚胎時期的特化及發育仍是未解之謎。本研究中,我首先針對蚜蟲專屬的唾腺基因 c002 合成反義股探針,探究 c002 mRNA 在豌豆蚜胚胎時期的表現情形。全胚胎整體原位雜合實驗的結果顯示,c002 mRNA 最早被偵測於出現表皮的胚胎時期,表現在外型可辨認的唾腺上。為了調查唾腺是否在更早的發育時期即開始特化,我們在豌豆蚜中選定了黑腹果蠅的唾腺基因 sage 之同源基因 Apsage,並檢視該基因在胚胎時期的表現。全胚胎整體原位雜合實驗的結果顯示 Apsage mRNA 會自肢芽萌發期至表皮發育期間連續表現,並且與 c002 共同表現於胚胎唾腺上。上述結果強烈意味著,第一,於肢芽萌發期,Apsage mRNA 在下唇節中表現的特定位置應為初始唾腺。第二,Apsage 保留著黑腹果蠅 sage 在胚胎唾腺的表現特性。比較黑腹果蠅與豌豆蚜的唾腺發育,兩者的唾腺均在胚帶延長末期開始特化。雖兩昆蟲之初始唾腺均出現於胚胎腹側處,但是豌豆蚜的唾腺卻會在胚胎翻轉後期,進入背部區域。從本研究中可以得出幾個總結:sage 在完全變態或是漸進變態昆蟲中是一個高度保守的唾腺標記分子;蚜蟲專屬的唾腺基因 c002 在晚期胚胎的唾腺即開始表現,顯示了該基因對於若蟲的取食十分重要。最後,唾腺在昆蟲中不同的分佈位置,可能具備特殊的生物意義,例如滿足取食上的需求。 | zh_TW |
dc.description.abstract | Aphids belong to the superfamily Aphidoidea of Hemiptera, feeding on plants with the sucking mouthparts. Meanwhile, aphids can uptake viruses from the plant sap, vectoring them from one plant to another. It has been clear that aphids adopt salivary glands (SGs) to transmit viruses in addition to secreting saliva for digestion. However, when SGs are specified during embryogenesis in aphids remains unknown. In my study, I first synthesized antisense riboprobes of c002, an aphid-specific gene of SGs, investigating how c002 mRNA was expressed during embryogenesis in the pea aphid Acyrthosiphon pisum. Results of the whole mount in situ hybridization (WISH) show that transcripts of c002 were first detected on morphologically identifiable SGs of the embryos with cuticle. To investigate whether SGs were specified in earlier embryos, I monitored the expression of Apsage, a homologue of the SG gene sage in the fly Drosophila melanogaster. WISH results showed that Apsage signals could be continuously monitored from limb-bud formation to cuticle formation, during which signals of c002 and Apsage were colocalized in the embryonic SGs. This strongly suggests that: (1) Apsage transcripts restricted to the labial segments in the embryos with visible limb buds are primordia of SGs; and (2) The feature of sage expression in embryonic SGs of Drosophila is remained in Apsage in the pea aphid. Like D. melanogaster, specification of SGs in A. pisum also started in late germ band extension. However, primordia of SGs in D. melanogaster are located in the ventral side whereas those of SGs in A. pisum were identified in late embryos after katatrepsis. Taken together, we conclude that: (1) sage is a highly conserved SG marker for both holometabolous and hemimetabolous insects; (2) aphid-specific SG gene like c002 is indispensable for feeding of nymphs as c002 is expressed in SGs of late-stage embryos; and (3) locations of SGs may serve special functions to satisfy a particular engulfment need. | en |
dc.description.provenance | Made available in DSpace on 2021-06-07T17:41:04Z (GMT). No. of bitstreams: 1 U0001-0302202113384400.pdf: 80791351 bytes, checksum: 14fb16bba3ddff2a3cb4a41f733500c3 (MD5) Previous issue date: 2021 | en |
dc.description.tableofcontents | 目錄……………………………………………………………………………………i 致謝……………………………………………………………………………………iv 圖目錄…………………………………………………………………………………vi 中英對照表……………………………………………………………………………vii 中文摘要………………………………………………………………………………xii 英文摘要………………………………………………………………………………xiii 第一章 前言…………………………………………………………………………1 1.1. 動物唾腺的外型與功能……………………………………………………1 1.2. 動物的唾腺發育:以老鼠與果蠅為例……………………………………1 1.3. 昆蟲唾腺發育研究:被忽視的唾腺發育及其重要性………………………2 1.4. 研究半翅目蚜蟲之唾腺發育的重要性……………………………………3 1.5. 蚜蟲唾腺基本形態及功能…………………………………………………4 1.6. 蚜蟲唾腺發育的研究進展…………………………………………………4 1.7. 胚胎唾腺之候選標記分子…………………………………………………5 1.8. 唾腺蛋白 C002 及對應基因 c002…………………………………6 1.9. 果蠅的唾腺發育及參與的發育基因……………………………………6 1.10. sage 為合適的唾腺標記候選分子………………………………………8 1.11. sage 在其他昆蟲胚胎發育時期的研究………………………………… 8 1.12. 研究豌豆蚜唾腺發育之策略…………………………………………… 9 第二章 材料與方法……………………………………………………………… 10 2.1. 豌豆蚜飼養…………………………………………………………………10 2.2. 唾腺標記候選基因序列選殖………………………………………………10 2.2.1. 核酸核酸萃取與反轉錄…………………………………………………10 2.2.2. 設計專一性引子…………………………………………………………11 2.2.3. 逆轉錄聚合酶鏈鎖反應和目標核酸序列回收………………………… 11 2.2.4. 接合反應、轉殖和定序…………………………………………………11 2.3. 探針合成……………………………………………………………………12 2.4. 豌豆蚜卵巢的收集和固定…………………………………………………13 2.5. 豌豆蚜唾腺的收集和固定…………………………………………………13 2.6. 原位雜合實驗……………………………………………………………14 2.7. 雙原位雜合實驗……………………………………………………………15 2.8. 唾腺及胚胎封片和影像拍攝、處理……………………………………16 第三章 結果…………………………………………………………………………17 3.1. 豌豆蚜 c002 基因片段選殖及 RNA 探針合成…………………………17 3.2. 豌豆蚜 c002 mRNA於豌豆蚜若蟲唾腺與胚胎時期的表現……………17 3.3. c002 mRNA於胚胎時期 St 18 以前的表現量分析………………………18 3.4. 胚胎中表現 c002 mRNA之組織形態及表現樣式………………………19 3.5. 昆蟲中已知的 sage 直系同源基因序列比較……………………………20 3.6. 豌豆蚜 sage直系同源基因 (Apsage) 片段選殖………………………20 3.7. Apsage 與 c002 於晚期胚胎中的表現分析………………………………21 3.8. Apsage 於胚胎時期中的表現………………………………………………22 第四章 討論…………………………………………………………………………23 4.1. 各齡若蟲唾腺上 c002 的表現建立未來鑑定胚胎唾腺的參考…………23 4.2. c002 的表現標註成熟胚胎的唾腺組織……………………………………24 4.3. c002的表現量不足以標註到早期胚胎的唾腺組織………………………24 4.4. Apsage 表現於原腸化的胚胎前端,也表現於發育中的唾腺……………25 4.5. Apsage無法直接追蹤豌豆蚜唾腺中副腺的發育…………………………26 4.6. 豌豆蚜唾腺發育過程………………………………………………………27 4.7. 果蠅與豌豆蚜於唾腺發育上的異同………………………………………27 4.8. sage 在昆蟲中可能的發育參與角色………………………………………28 參考文獻………………………………………………………………………………29 附錄一 引子對序列資訊……………………………………………………………57 附錄二 藥品配置……………………………………………………………………58 口試提問與建議……………………………………………………………………61 | |
dc.language.iso | zh-TW | |
dc.title | 以 c002 和 Apsage 的表現探究豌豆蚜的唾腺發育 | zh_TW |
dc.title | Formation of salivary glands in the pea aphid Acyrthosiphon pisum: c002 and Apsage expressions | en |
dc.type | Thesis | |
dc.date.schoolyear | 109-1 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 陳俊宏(Chun-hong Chen),李士傑(Shyh-jye Lee),林明德(Ming-der Lin),呂曉鈴(Hsiao-ling Lu) | |
dc.subject.keyword | 豌豆蚜,唾腺,c002,Apsage,孤雌無性胎生, | zh_TW |
dc.subject.keyword | pea aphid,salivary gland,c002,Apsage,asexual viviparous reproduction, | en |
dc.relation.page | 65 | |
dc.identifier.doi | 10.6342/NTU202100450 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2021-02-05 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 基因體與系統生物學學位學程 | zh_TW |
顯示於系所單位: | 基因體與系統生物學學位學程 |
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