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  1. NTU Theses and Dissertations Repository
  2. 生命科學院
  3. 生化科技學系
請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/48899
完整後設資料紀錄
DC 欄位值語言
dc.contributor.advisor薛雁冰(Yen-Ping Hsueh)
dc.contributor.authorTsung-Yu Huangen
dc.contributor.author黃孮(每育)zh_TW
dc.date.accessioned2021-06-15T11:11:08Z-
dc.date.available2020-08-21
dc.date.copyright2020-08-21
dc.date.issued2020
dc.date.submitted2020-08-13
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7. Gulshan, K., Rovinsky, S.A., and Moye-Rowley, W.S.J.E.c. (2004). YBP1 and its homologue YBP2/YBH1 influence oxidative-stress tolerance by nonidentical mechanisms in Saccharomyces cerevisiae. Eukaryot Cell 3(2), 318-330.
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9. Hsueh, Y.P., Gronquist, M.R., Schwarz, E.M., Nath, R.D., Lee, C.H., Gharib, S., Schroeder, F.C., and Sternberg, P.W. (2017). Nematophagous fungus Arthrobotrys oligospora mimics olfactory cues of sex and food to lure its nematode prey. eLife 6, e20023
10. Hsueh, Y.P., Mahanti, P., Schroeder, F.C., and Sternberg, P.W. (2013). Nematode-trapping fungi eavesdrop on nematode pheromones. Current Biology 23(1), 83-86.
11. Jasmer, D.P., Goverse, A., and Smant, G.J.A.r.o.p. (2003). Parasitic nematode interactions with mammals and plants. Annu Rev Phytopathol. 41, 245-270.
12. Jeong, P.Y., Jung, M., Yim, Y.H., Kim, H., Park, M., Hong, E., Lee, W., Kim, Y.H., Kim, K., and Paik, Y.K. (2005). Chemical structure and biological activity of the Caenorhabditis elegans dauer-inducing pheromone. Nature 433, 541-545.
13. Li, H., and Durbin, R.J.b. (2009). Fast and accurate short read alignment with Burrows–Wheeler transform. Bioinformatics 25(14), 1754-1760.
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17. Mohd-Yusoff, N.F., Ruperao, P., Tomoyoshi, N.E., Edwards, D., Gresshoff, P.M., Biswas, B., and Batley, J.J.G.G. (2015). Scanning the effects of ethyl methanesulfonate on the whole genome of Lotus japonicus using second-generation sequencing analysis. G3: Genes, Genomes, Genetics 5, 559-567.
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20. Nordbring-Hertz, B.J.P.P. (1973). Peptide‐induced morphogenesis in the nematode‐trapping fungus Arthrobotrys oligospora. Physiologia Plantarum 29, 223-233.
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22. Poplin, R., Ruano-Rubio, V., DePristo, M.A., Fennell, T.J., Carneiro, M.O., Van der Auwera, G.A., Kling, D.E., Gauthier, L.D., Levy-Moonshine, A., and Roazen, D.J.B. (2017). Scaling accurate genetic variant discovery to tens of thousands of samples. bioRxiv, 201178.
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24. Srinivasan, J., Kaplan, F., Ajredini, R., Zachariah, C., Alborn, H.T., Teal, P.E., Malik, R.U., Edison, A.S., Sternberg, P.W., and Schroeder, F.C. (2008). A blend of small molecules regulates both mating and development in Caenorhabditis elegans. Nature 454, 1115-1118.
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29. Vidal-Diez de Ulzurrun, G., Huang, T.Y., Chang, C.W., Lin, H.C., and Hsueh, Y.P. (2019). Fungal feature tracker (FFT): A tool for quantitatively characterizing the morphology and growth of filamentous fungi. PLoS Computational Biology 15, e1007428.
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34. Zhen, Z., Xing, X., Xie, M., Yang, L., Yang, X., Zheng, Y., Chen, Y., Ma, N., Li, Q., Zhang, K.-Q.J.F.G., et al. (2018). MAP kinase Slt2 orthologs play similar roles in conidiation, trap formation, and pathogenicity in two nematode-trapping fungi. Fungal Genetics and Biology 116, 42-50.
dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/48899-
dc.description.abstract線蟲捕捉菌在演化的過程中面對養分缺乏的環境時,選擇了線蟲作為捕食的對象。他們藉由長出複雜的捕蟲構造抓住且殺掉線蟲,並將這些線蟲作為額外的碳與氮的養分來源。我們實驗室以Arthrobotrys oligospora與秀麗隱桿線蟲(Caenorhabditis elegans)為研究對象,分別做為獵食者與被獵食者,探討兩者之間的交互作用關係。為了更進一步地瞭解線蟲捕捉菌如何感知環境中的線蟲與其捕蟲網的生成機制,我們藉由正向遺傳學的篩選(Forward genetic screen)找出無法形成捕捉構造的Arthrobotrys oligospora突變株。利用ethyl methanesulfonate與UV作為致突變劑,我們總共產生了5552個突變株,並在最後挑出16個在捕捉線蟲上具有明顯缺陷的突變株。接著我們對這16個突變株做全基因體定序(whole-genome sequencing) 找到所有突變位置,並以此看哪些突變的基因使他們失去捕捉秀麗隱桿線蟲的能力。經過篩選分析顯示這些突變多與訊息傳遞、轉錄、或膜上運輸有關。我們針對選出的目標基因做基因剃除來檢視這些基因是否參與線蟲捕捉菌捕捉線蟲的過程。目前,我們發現一個具PH domain的基因與線蟲捕捉菌捕捉網的生成有關。預期在這些篩選出的目標基因中,我們除了會找到已知在其它致病性真菌中扮演重要角色的基因,也會發現線蟲捕捉菌自身特有的基因。zh_TW
dc.description.abstractNematode-trapping fungi are a group of nematophagous fungi that have evolved to prey on nematodes by developing complex trap structures under nutrient-poor environments; the traps can capture and kill the nematodes, providing additional nitrogen and carbons sources. We used Arthrobotrys oligospora and Caenorhabditis elegans as models to study the interactions between preys and predators. To identify genes and pathways that are required for sensing the nematodes and trap morphogenesis, we conducted a forward genetic screen in A. oligospora to isolate mutants with defects in trapping C. elegans. We generated 5552 EMS and UV mutants and identified 16 mutants with strong defects in trap morphogenesis. Whole genome sequencing and bioinformatic analyses for the 16 mutants identified the potential mutations that affect trapping ability. Preliminary data demonstrated that mutations in genes that have a role in signaling, transcription or membrane transport might affect trap morphogenesis. Targeted gene deletion mutants were generated to investigate the roles of these candidate genes during nematode-trapping process. So far, we found one unstudied gene predicted to contain a PH domain to influence trap morphogenesis. In addition to genes that conserved in other pathogenic fungi, we have also identified novel genes that are unique in nematode-trapping fungi might play important roles in their predatory behaviors.en
dc.description.provenanceMade available in DSpace on 2021-06-15T11:11:08Z (GMT). No. of bitstreams: 1
U0001-1308202008471800.pdf: 2311380 bytes, checksum: e6f811bc403060127b803007006992e7 (MD5)
Previous issue date: 2020
en
dc.description.tableofcontents口試委員會審定書 ii
致謝 iii
摘要 iv
Abstract v
Chapter 1. Introduction 1
Chapter 2. Materials and Methods 7
2.1 Strains, media, culture conditions 7
2.2 Mutagenesis 8
2.3 Genetic screen on mutants with trapping defects 8
2.4 Trap quantification 9
2.5 Observation on trap morphology 9
2.6 Whole genome sequencing analysis 10
2.7 Transformation 11
2.8 Construction of gene knockout cassettes 12
2.9 Confirmation of gene knockouts in mutants 12
2.10 Rescue assay on EYR41_001410 in TWF1042 mutant 12
Chapter 3. Results 13
3.1 Forward genetic screen identified A. oligospora mutants with trapping defects 13
3.2 Phenotypic characterization of mutants with trapping defects 13
3.3 Whole genome sequencing analysis of 16 mutants in identifying potential candidate genes involved in trapping defects 15
3.4 EYR41_001410 may be the causative gene that leads to trapping defects in TWF1042 17
Chapter 4. Discussion 19
Chapter 5. Future work 24
Chapter 6. Figures 25
References 35
dc.language.isoen
dc.subject線蟲捕捉菌zh_TW
dc.subjectNematode-trapping fungien
dc.title透過正向遺傳篩選選出對線蟲捕食缺陷之線蟲捕捉菌zh_TW
dc.titleForward genetic screen identified mutants with defects in nematode predation in the nematode-trapping fungus Arthrobotrys oligosporaen
dc.typeThesis
dc.date.schoolyear108-2
dc.description.degree碩士
dc.contributor.oralexamcommittee黃慶璨(Ching-Tsan Huang),林晉玄(Ching-Hsuan Lin),吳亘承(Hsuan-Chen Wu)
dc.subject.keyword線蟲捕捉菌,zh_TW
dc.subject.keywordNematode-trapping fungi,en
dc.relation.page37
dc.identifier.doi10.6342/NTU202003198
dc.rights.note有償授權
dc.date.accepted2020-08-13
dc.contributor.author-college生命科學院zh_TW
dc.contributor.author-dept生化科技學系zh_TW
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