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  1. NTU Theses and Dissertations Repository
  2. 生物資源暨農學院
  3. 植物病理與微生物學系
Please use this identifier to cite or link to this item: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/59919
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???org.dspace.app.webui.jsptag.ItemTag.dcfield???ValueLanguage
dc.contributor.advisor陳穎練(Ying-Lien Chen)
dc.contributor.authorLi-Hang Hsuen
dc.contributor.author徐立航zh_TW
dc.date.accessioned2021-06-16T09:45:03Z-
dc.date.available2022-02-16
dc.date.copyright2017-02-16
dc.date.issued2017
dc.date.submitted2017-01-25
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/59919-
dc.description.abstract鐮孢菌屬(Fusarium)包含F. oxysporum、F. solani、F. graminearum與F. verticillioides等,且會在植物及人體造成嚴重的感染。臨床上,鐮孢菌感染發生的頻率在所有侵入性絲狀真菌的感染中排名第三,僅次於麴菌屬(Aspergillus)及毛黴目(Mucorales),而感染的病例中以F. solani及F. oxysporum最多;然而,在植物及人類上,對於鐮孢菌感染的治療仍缺乏有效利器。陽離子態的抗菌胜肽多黏菌素B (polymyxin B) 具有抑制白色念珠菌與新型隱球菌的能力,但針對鐮孢菌的效果仍未知。本研究測試多黏菌素B對12株感染人類及植物(香蕉、番茄、瓜類、豆類、小麥及玉米)之鐮孢菌的抗菌能力。透過稀釋平板法、亞甲藍液染色及XTT還原法測試,此藥劑對所有受測菌株皆具殺真菌能力,其中對大部分菌株的最低殺菌濃度為32或64 μg/mL。多黏菌素B可降低鐮孢菌分生孢子的發芽率,且能造成膜系的缺陷,但厚膜孢子不受藥劑影響。多黏菌素B可與posaconazole產生協同作用(synergistic effect),且具促進藥物fluconazole、voriconazole或amphotericin B對鐮孢菌抑制作用的潛力;多黏菌素B雖在抑制白色念珠菌及隱球菌的效果中與fluconazole有協同作用,然而在鐮孢菌屬中並未發現一樣的情形,顯示酵母菌及絲狀真菌如鐮孢菌在演化中的多樣性。zh_TW
dc.description.abstractThe genus Fusarium comprises many species, including F. oxysporum, F. solani, F. graminearum, and F. verticillioides, and causes severe infections in plants and humans. In clinical settings, Fusarium is the third most frequent mold to cause invasive fungal infections, after Aspergillus and the Mucorales. F. solani and F. oxysporum are the most prevalent Fusarium species to cause clinical disease. However, few effective antifungal drugs are available to treat both human and plant Fusarium infections. The cationic peptide antibiotic polymyxin B (PMB) exhibits antifungal activity against the human fungal pathogens Candida albicans and Cryptococcus neoformans, but its efficacy against Fusarium species is unknown. In this study, we tested the antifungal activity of PMB against 12 Fusarium strains that infect humans and plants (banana, tomato, melon, pea, wheat, and maize). PMB was fungicidal against all 12 Fusarium strains, with minimum fungicidal concentrations of 32 or 64 μg/mL for most strains tested, as evidenced by broth dilution, methylene blue staining, and XTT reduction assays. PMB can reduce the germination rates of conidia, but not chlamydospores, and can cause defects in cell membrane integrity in Fusarium strains. PMB exhibits synergistic activity with posaconazole, and can potentiate the effect of fluconazole, voriconazole, or amphotericin B against Fusarium species. However, PMB does not show synergistic effects with fluconazole against Fusarium species as it does against Candida glabrata and C. neoformans, indicating evolutionary divergence of mechanisms between yeast pathogens and the filamentous fungus Fusarium.en
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Previous issue date: 2017
en
dc.description.tableofcontents致謝 I
摘要 II
Abstract III
Table of Contents IV
1.Introduction 1
2.Materials and Methods
2.1.Strains, growth media, and chemicals 5
2.2.Conidia solution preparation 6
2.3.Determination of conidia germination rate 6
2.4.Determination of minimum inhibitory concentration 6
2.5.Determination of minimum fungicidal concentrations 7
2.6.Determination of fractional inhibitory concentration indices 8
2.7.Propidium iodide staining 9
2.8.RNA extraction and quantitative real-time RT-qPCR analysis 10
2.9.Induction of chlamydospore formation
and determination of germination rate 11
3.Results and Discussion
3.1.Polymyxin B exhibits novel antifungal activity against Fusarium species 12
3.2.Polymyxin B exhibits fungicidal activity against Fusarium species 14
3.3.Conidia, but not chlamydospore, germination of Fusarium strains
is inhibited by polymyxin B 15
3.4.Hypothetical mode of action of polymyxin B 16
3.5.Polymyxin B exhibits synergistic activity with posaconazole,
and potentiates the effect with fluconazole, voriconazole
or amphotericin B against Fusarium strains 17
4.Conclusions 19
5.Acknowledgement 20
 
6.Tables
Table 1.Fusarium strains used in this study 21
Table 2.MICs and MFCs of polymyxin B for 12 Fusarium strains 22
Table 3.Synergistic antifungal activity between polymyxin B (PMB) and
fluconazole (FLC), voriconazole (VRC), posaconazole (PSC) or
amphotericin B (AmB) against 12 Fusarium strains 23
Table 4.Primers used in RT-qPCR 24
7.References 25
8.Figures
Figure 1.Polymyxin B exhibits fungicidal activity against Fusarium strains
infecting humans or plants 30
Figure 2.Polymyxin B inhibits conidia germination of Fusarium strains 32
Figure 3.Polymyxin B damages cell membrane integrity of Fusarium strains 34
Figure 4.The expression of Fusarium CHS1 gene
is slightly affected by polymyxin B 36
Figure 5.Germination rate of chlamydospores produced by
Fusarium oxysporum f. sp. cubense or lycopersici
was not inhibited by polymyxin B 37
Figure S1.Polymyxin B exhibits novel antifungal activity against multiple
Fusarium species infecting humans and plants 39
Figure S2.Polymyxin B demonstrates antibacterial activity against
Pseudomonas aeruginosa that was frequently associated
with the keratitis in humans 40
Figure S3.XTT-based colormetric assays to determine
minimum fungicidal concentration of
polymyxin B against twelve Fusarium species 41
9.Appendix
Appendix A.Roles of PSD2 in Fusarium oxysporum f. sp. lycopersici 4287 43
Appendix B.A clinical case in stingrays infected with Fusarium species 52
dc.language.isoen
dc.subject多黏菌素Bzh_TW
dc.subject鐮孢菌zh_TW
dc.subject抗生素zh_TW
dc.subject抗真菌藥劑zh_TW
dc.subjectAntifungal drugsen
dc.subjectAntibioticsen
dc.subjectFusariumen
dc.subjectPolymyxin Ben
dc.title抗生素多黏菌素B具新穎抗鐮孢菌活性zh_TW
dc.titleThe Antibiotic Polymyxin B Exhibits Novel Antifungal Activity against Fusarium Speciesen
dc.typeThesis
dc.date.schoolyear105-1
dc.description.degree碩士
dc.contributor.oralexamcommittee鍾嘉綾(Chia-Lin Chung),孫培倫(Pei-Lun Sun)
dc.subject.keyword多黏菌素B,鐮孢菌,抗生素,抗真菌藥劑,zh_TW
dc.subject.keywordPolymyxin B,Fusarium,Antibiotics,Antifungal drugs,en
dc.relation.page57
dc.identifier.doi10.6342/NTU201700200
dc.rights.note有償授權
dc.date.accepted2017-01-25
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
Appears in Collections:植物病理與微生物學系

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