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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
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dc.contributor.advisor | 劉瑞芬(Ruey-Fen Liou) | |
dc.contributor.author | Fang-Yu Hwu | en |
dc.contributor.author | 胡芳瑜 | zh_TW |
dc.date.accessioned | 2021-06-16T13:01:03Z | - |
dc.date.available | 2018-09-06 | |
dc.date.copyright | 2013-09-06 | |
dc.date.issued | 2013 | |
dc.date.submitted | 2013-08-08 | |
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M., Tham, R., Jamklang, M., Uhrig, J. P. and Gelli, A. 2013. Activity of the calcium channel pore Cch1 is dependent on a modulatory region of the subunit Mid1 in Cryptococcus neoformans. Euk. cell. 12:142-150. 29. Iida, H., Nakamura, H., Ono, T., Okumura, M. S. and Anraku, Y. 1994. MID1, a novel Saccharomyces cerevisiae gene encoding a plasma membrane protein, is required for Ca2+ influx and mating. Mol. Cell. Biol. 14: 8259-8271. 30. Jackson, S. L., and Hardham, A. R. 1996. A transient rise in cytoplasmic free calcium is required to induce cytokinesis in zoosporangia of Phytophthora cinnamomi. Eur. J. Cell Biol. 69: 180-188.. 31. Jackson, S. L., and Heath, I. B. 1993. Roles of calcium ions in hyphal tip growth. Microbio. Rev. 57:367-382. 32. Judelson, H. S., and Roberts, S. 2002. Novel protein kinase induced during sporangial cleavage in the oomycete Phytophthora infestans. Euk. Cell 1: 687-695. 33. Kamoun, S. 2003. Molecular genetics of pathogenic oomycetes. Euk. 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J. Eukaryot. Microbiol. 55: 103–109. 40. Maltese, C. E., Conigliaro, G. and Shaw, D. S. 1995. The Development of Sporangia of Phytophthora infestans. Mycol. Res. 99: 1175-1181. 41. Marks, G. E. 1965. The cytology of Phytophthora infestans. Chromosoma 16: 681-692. 42. Maruoka, T., Nagasoe, Y., Inoue, S., Mori, Y., Goto, J., Ikeda, M. and Iida, H. 2002. Essential hydrophilic carboxyl-terminal regions including cysteine residues of the yeast stretch-activated calcium-permeable channel Mid1. J. Biol. Chem. 277: 11645-11652. 43. Nyochembeng, L. M., Pacumbaba, R. P., and Beyl, C. A. 2002. Calcium Enhanced Zoospore Production of Pythium myriotylum in vitro. J. Phytopathology 150: 396-398 44. Pera, L. M., and Callieri, D. A. 1997. Influence of calcium on fungal growth, hyphal morphology and citric acid production in Aspergillus niger. Folia Microbiol. 42: 551-556. 45. Ribeiro, O. K. 1983. Physiology of asexual sporulation and spore germination in Phytophthora, p. 55–70. In Erwin, D. C., Bartnicki-Garcia, S., and Tsao, P. H. (ed.), Phytophthora, its biology, taxonomy, ecology, and pathology. APS Press, St. Paul, Minn., U. S. A. 46. Rispail, N., Soanes, D. M., Ant, C., Czajkowski, R., Grunler, A., Huguet, R. et al. 2009. Comparative genomics of MAP kinase and calcium-calcineurin signalling components in plant and human pathogenic fungi. Fung. Genet. Biol. 46: 287-298. 47. Robson, G. D., Wiebe, M. G., and Trinci, A. P. J. 1991a. Involvement of Ca2+ in the Regulation of Hyphal Extension and Branching in Fusarium graminearum a-3/5. Exp. Mycol. 15:263-272. 48. Robson, G. D., Wiebe, M. G. and Trinci, A. P. J. 1991b. Low Calcium Concentrations Induce Increased Branching in Fusarium graminearum. Mycol. Res. 95: 561-565. 49. Sackin, H. 1995. Stretch-activated ion channels. Kidney Int int. 48: 1134-1147. 50. Sambrook, J., and Russell, D. W. 2002a. Molecular Cloning: A Laboratory Manual, 3rd ed. volume 1 page 6.4-6.12 Cold Spring Harbor, New York: Cold Spring Harbor Lab. 51. Sambrook, J., and Russell, D. W. 2002b. Molecular Cloning: A Laboratory Manual, 3rd ed. volume 1 page 6.33-6.46 Cold Spring Harbor, New York: Cold Spring Harbor Lab. 52. Sheen, J. 2001. Signal transduction in maize and Arabidopsis mesophyll protoplasts. Plant Physiol. 127: 1466-1475. 53. Shipton, W. A. 1985. Zoospore induction and release in a Pythium causing equine phycomycosis. Trans. Br. mycol. Soc. 84: 147-155 54. Torralba, S., and Heath, I. B. 2001. Cytoskeletal and Ca2+ regulation of hyphal tip growth and initiation. Curr. Top. Dev. Biol. 51:135-187. 55. von Broembsen, S. L., and Deacon, J. W. 1997. Calcium interference with zoospore biology and infectivity of Phytophthora parasitica in nutrient irrigation solutions. Phytopathology 87: 522-528. 56. Walker, C. and van West, P. 2007. Zoospore development in the oomycetes. Fung. Biol. Rev. vol 21, no. 1, pp. 10-18. 57. Wang, S., Cao, J. L., Liu, X., Hu, H. Q., Shi, J., Zhang, S. Z., Keller, N. P. and Lu, L. 2012. 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Biol. 43: 430-438. 62. Zamani, M., Tousley, M.E. and Kane, S.A. Tracking Multiple Objects Using ImageJ with tracking plugins. Online material. | |
dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/61322 | - |
dc.description.abstract | 疫病菌為重要的植物病原卵菌,其寄主範圍廣泛,可於多種重要經濟作物造成危害。病原卵菌外觀形態和真菌相當類似,但進一步的核酸類源關係分析顯示兩者明顯不同,使得分類學將其獨立於真菌界之外。 疫病菌孢囊會進行細胞分裂以產生游走子,是其感染寄主時的初級感染原;在潮濕環境下,游走子會泳動至適合的寄主表面,形成靜止子、發芽並入侵寄主,故孢囊的形成及游走子的釋放在病原傳播過程中扮演了重要的角色,但其中詳細的分子機制目前卻尚未完全明瞭。過去的研究發現,添加鈣離子抑制劑會影響孢囊細胞分裂與游走子釋放,認為鈣離子為此過程的關鍵因子。本研究利用序列比對尋找鈣離子訊息傳遞過程的相關蛋白,獲得一真菌高親和性鈣離子通道次體之同源基因—Ppmid1,並以基因靜默 (gene silencing) 之形式取得Ppmid1基因靜默轉殖株。以即時定量聚合酶連鎖反應偵測Ppmid1,發現其在孢囊中的表現量明顯較高。實驗中觀察發現Ppmid1基因靜默株無法生成正常的孢囊,並且無游走子釋出,所形成的孢囊也常直接發芽,但添加離子至誘導孢囊產生的溶液中,有助於Ppmid1基因靜默株所形成的異常孢囊恢復至正常型態,並且釋放游走子。另外於接種試驗中也發現Ppmid1基因靜默株有致病力下降的情形,在煙草所造成的病斑明顯較野生型P. parasitica (94069) 為小,很可能是因為Ppmid1基因靜默株無法產生正常孢囊所致。這些結果顯示Ppmid1極有可能在疫病菌孢囊形成及游走子釋放過程中扮演重要的角色。 | zh_TW |
dc.description.abstract | Phytophthora parasitica is a notorious oomycete pathogen that causes severe disease in a wide variety of plant species. Plant infection by this pathogen involves mainly its asexual development, whereby papillate sporangia are formed, followed by differentiation and release of biflagellate zoospores. Calcium signaling has been proposed to play a key role in these processes. This study deals with the functional characterization of Ppmid1, which encodes a component of a putative calcium channel. Analysis by quantitative reverse transcriptase-PCR indicated that the expression of Ppmid1 is induced in the stage of sporangia. Moreover, we generated Ppmid1-silenced mutants of P. parasitica, which showed reduced expression of Ppmid1 and exhibited severe defects in its asexual development. Sporangia formed by the mutants lacked papilla and failed to release zoospores in response to cold shock. Instead, they tended to germinate directly. Further analysis revealed that, in response to cold shock for induction of zoospore formation, no sign of cytoplasmic cleavage was detected in the sporangia of Ppmid1-silenced mutants. Interestingly, when the sporangia induction solution was amended with CaCl2, the mutants became able to form papillate sporangia, which released zoospores upon cold shock. Inoculation experiments indicated that Ppmid1-silenced mutants showed reduced virulence toward Nicotiana benthamiana. These results indicated that Ppmid1 plays an important role in the asexual development of P. parasitica. | en |
dc.description.provenance | Made available in DSpace on 2021-06-16T13:01:03Z (GMT). No. of bitstreams: 1 ntu-102-R00633003-1.pdf: 6646744 bytes, checksum: dcda7a27a00ef844b5ff35177032d01a (MD5) Previous issue date: 2013 | en |
dc.description.tableofcontents | 中文摘要 i
ABSTRACT ii CONTENTS iii LIST OF FIGURES vi LIST OF TABLES viii Chapter 1 Introduction 1 Chapter 2 Materials and Methods 5 2.1 Growth of Phytophthora parasitica 5 2.2 Collection of Phytophthora parasitica Each Life Stage 5 2.3 Molecular Cloning and Sequence Analysis of Ppmid1 6 2.4 Isolation of RNA 7 2.5 Quantitative RT-PCR 7 2.6 Southern Blot Hybridization 8 2.7 Generating the Ppmid1-silenced Transfromants 8 2.7.1 Preparation of Silencing Vector 8 2.7.2 Preparation of protoplasts 9 2.7.3 Polyethylene glycerol (PEG)-mediated transformation 9 2.7.4 Selection of transformants 10 2.8 Detached Leaves Inoculation 10 2.9 Microscopy 11 2.9.1 Sporangia Observation 11 2.9.2 Sporangia Cytoplasmic Cleavage Observation 12 2.10 Osmotic Shock Experiments 12 2.11 Oospores Observation 13 2.12 Zoospores Tracking 13 Chapter 3 Results 14 3.1 PpMid1 is a predicted stretch-activated calcium channel subunit. 14 3.2 Expression level of Ppmid1 is highly induced in sporangia 14 3.3 Generation of Ppmid1-silenced transformants. 15 3.4 Profiling the Ppmid1-silenced transformants 16 3.5 Ppmid1-silenced transformant produced highly branched mycelia. 16 3.6 The Ppmid1-silenced transformant produced abnormal sporangia. 17 3.7 The Ppmid1-silenced transformant could not release zoospores and showed defect in cytoplasmic cleavage of sporangia. 18 3.8 The Ppmid1-silenced transformant shows reduced virulence on Nicotiana benthamiana. 19 3.9 The Ppmid1-silenced transformant shows abnormal features in the infected plants. 19 3.10 Addition of CaCl2, MgCl2 or NaCl partially recovered the morphology of sporangia formed by the Ppmid1_7 and their ability of cytoplasmic cleavage 20 3.11 Expression of Ppmid1 is induced by hypoosmotic and hyperionic stress 22 3.12 Silencing of Ppmid1 has no effect on oospore formation. 22 Chapter 4 Discussion 24 4.1 Ppmid1-is a putative stretch-activated calcium channel component. 24 4.2 Polymorphism of Ppmid1-silenced transformants. 24 4.3 Ppmid1 might involve in Phytophthora parasitica mycelia tip growth. 25 4.4 Ppmid1 might be required for normal sporangia formation and sporangia cytoplasmic cleavage of Phytophthora parasitica. 26 4.5 Calcium and other cations supplements relieved the defect of Ppmid1-silenced mutant’s sporangia. 28 4.6 Expression of Ppmid1 were induced in hypoosmotic stimulation 29 4.7 Defect of Ppmid1 impair Phytophthora parasitica virulence for Nicotiana benthamiana 29 4.8 Defect of Ppmid1 in Phytophthora parasitica did not affect the formation of oospores. 30 REFERENCES 51 | |
dc.language.iso | en | |
dc.title | 疫病菌鈣離子通道次單元基因Mid1之功能性分析 | zh_TW |
dc.title | Functional characterization of Mid1 encoding a putative calcium channel subunit in Phytophthora parasitica | en |
dc.type | Thesis | |
dc.date.schoolyear | 101-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 安寶貞(Pao-Jen Ann),李敏惠(Miin-Huey Lee),沈偉強(Wei-Chiang Shen),陳穎練(Ying-Lien Chen) | |
dc.subject.keyword | 鈣離子,孢囊,孢囊細胞質分裂,游走子,Ppmid1,Ppmid1靜默轉植株,stretch-activated calcium channel subunit, | zh_TW |
dc.subject.keyword | calcium,Ppmid1,Ppmid1-silenced transformants,stretch-activated calcium channel subunit,sporangia,sporangia cytoplasmic cleavage,zoospores, | en |
dc.relation.page | 57 | |
dc.rights.note | 有償授權 | |
dc.date.accepted | 2013-08-08 | |
dc.contributor.author-college | 生物資源暨農學院 | zh_TW |
dc.contributor.author-dept | 植物病理與微生物學研究所 | zh_TW |
顯示於系所單位: | 植物病理與微生物學系 |
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