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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
請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/97594
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dc.contributor.advisor歐海仁zh_TW
dc.contributor.advisorHiran A. Ariyawansaen
dc.contributor.author卓義敦zh_TW
dc.contributor.authorYi-Tun Choen
dc.date.accessioned2025-07-03T16:09:59Z-
dc.date.available2025-07-04-
dc.date.copyright2025-07-03-
dc.date.issued2025-
dc.date.submitted2025-06-18-
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/97594-
dc.description.abstract青蔥是台灣重要作物,廣泛應用於蔬菜與香料。近年來,新興青蔥葉部病原菌如 Stemphylium vesicarium, Colletotrichum spaethianum 與 C. circinans 在台灣大量發生並造成顯著損失。在我們先前的研究中,從健康青蔥葉部中分離出一株Bacillus velezensis GFB08 菌株,具有在實驗室與溫室條件下降低青蔥葉部病害的能力。本篇研究的主要目標為透過基因體學與代謝組體學評估 B. velezensis GFB08 在青蔥上的抑病機制。透過結合illumina與Nanopore定序進行基因體組裝,B. velezensis GFB08具有單一3.89 mb的染色體,其中包含3,735編碼序列,藉由比較基因體分析,GFB08具有完整且高度保守的fengycin及bacillomycin D二次代謝物基因簇。代謝物純化與質譜分析進一步證實GFB08產生的fengycin對三種青蔥葉部病原菌皆具有廣效的抗真菌活性,而bacillomycin D 則表現出物種專一性的抗真菌活性,特別是fengycin與bacillomycin D 在抑制C. spaethianum上呈現出加成的抗菌效果。此外,GFB08產生的揮發性有機化合物(VOCs),如乙酸、乙偶姻、1-丁醇與3-甲基丁醇,均能顯著抑制C. spaethianum的菌絲生長。為了評估GFB08的實際應用潛力,本研究同時進行了植物生長促進試驗、發酵條件最佳化與田間防治試驗,結果顯示GFB08 僅在種子時期能夠顯著促進青蔥生長。5 L發酵試驗成果顯示,GFB08的最適產孢條件為1% 糖蜜, 0.5% 大豆粉, 28°C, pH 7, 與200 rpm,內生孢子濃度可達2×109 CFU/mL。田間防治試驗中,B. velezensis GFB08 稀釋液100倍可抑制青蔥黑腐病的罹病嚴重度。整體而言,本研究為生物防治菌株B. velezensis GFB08 提供了功能上的參考依據,並揭示其在永續農業上的應用潛力。zh_TW
dc.description.abstractThe Welsh onion is an essential crop in Taiwan, widely consumed as a vegetable or spice. In recent years, foliar fungal pathogens Stemphylium vesicarium, Colletotrichum spaethianum, and C. circinans have been identified as emerging threats, causing significant damage to the Welsh onion in Sangxing, Taiwan. In our recent study, Bacillus velezensis GFB08 isolated from healthy Welsh onion leaves demonstrated the ability to reduce foliar diseases under laboratory and greenhouse conditions. The main objective of this study was to elucidate the disease suppression mechanisms of B. velezensis GFB08 through genomic and metabolomic approaches. Using Illumina and Nanopore hybrid assembly, the present study found that B. velezensis GFB08 genome harbored a 3.89 mb chromosome with 3,735 coding sequences. Comparative genomic analysis revealed complete and highly conserved biosynthetic gene clusters (BGCs) for fengycin and bacillomycin D. Metabolite profiling confirmed that fengycin produced by GFB08 exhibited broad-spectrum antifungal activity against all three foliar pathogens, while bacillomycin D displayed species-specific effects. Notably, fengycin and bacillomycin D displayed additive antifungal activity against C. spaethianum. Additionally, volatile organic compounds (VOCs) such as acetic acid, 2-pentanone, acetoin, 1-butanol and 3-methyl- were identified as major antifungal VOCs from GFB08, significantly inhibiting C. spaethianum growth under in vitro. To facilitate the practical application of GFB08, plant growth promotion assays, fermentation optimization, and field evaluations were also conducted. GFB08 enhanced Welsh onion growth only during seed stage. Based on 5 L fermentation optimization, B. velezensis GFB08 achieved highest endospore production under the following conditions: 1% molasses, 0.5% soybean flour, 28°C, pH 7, and 200 rpm, reaching a concentration of 2×109 CFU/mL. Field trials demonstrated that a 100-fold dilution of B. velezensis GFB08 effectively reduced the disease severity of Welsh onion leave blight. Overall, this study provides a comprehensive functional characterization of the biocontrol agent B. velezensis GFB08 and highlight its potential for sustainable agriculture.en
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dc.description.tableofcontents口試委員會審定書 i
致謝 ii
摘要 iii
Abstract iv
Table of Contents vi
List of Figures ix
List of Tables xi
List of Supplementary Figures xii
List of Supplementary Tables xiii
Chapter 1. Introduction 1
Welsh onion and foliar diseases 1
Foliar disease management of Welsh onion 1
Bacillus velenzsis, a well-known beneficial microbe 3
Aims of this study 5
Chapter 2. Materials and methods 6
Bacillus velenzsis GFB08 cultivation and genomic DNA extraction 6
Whole genome sequencing, assembly, and annotation 7
Comparative genomics 8
Isolation, identification, and bioactivity assay of lipopeptides 8
Collection, identification, and bioactivity assay of VOCs 13
Plant growth promotion (PGP) assay 14
Optimization of fermentation conditions 16
Field trials 18
Statistical analysis 19
Chapter 3. Results 20
Genome features of Bacillus velenzsis GFB08 20
Comparative genomics 21
Isolation, identification, and bioactivity assay of lipopeptides 23
Collection, identification, and bioactivity assay of VOCs 27
Plant growth promotion assay 29
Optimization of fermentation condition 29
Field trials 31
Chapter 4. Discussion 33
General Conclusions 38
References 40
Figures 54
Tables 66
Supplementary materials 70
-
dc.language.isoen-
dc.subject生物活性二次代謝物zh_TW
dc.subject發酵條件最佳化zh_TW
dc.subject生物防治zh_TW
dc.subject比較基因體分析zh_TW
dc.subject植物生長促進zh_TW
dc.subjectPlant growth promotionen
dc.subjectFermentation optimizationen
dc.subjectBioactive secondary metabolitesen
dc.subjectBiocontrol agenten
dc.subjectComparative genomic analysisen
dc.title透過基因體學與代謝組體學評估 Bacillus velezensis GFB08 在青蔥上的抑病機制zh_TW
dc.titleElucidating Disease Suppression Mechanisms of Bacillus velezensis GFB08 in Welsh Onion through Genomic and Metabolomic Approachesen
dc.typeThesis-
dc.date.schoolyear113-2-
dc.description.degree碩士-
dc.contributor.oralexamcommittee楊玉良;陳賢明zh_TW
dc.contributor.oralexamcommitteeYu-Liang Yang;Hieng-Ming Tingen
dc.subject.keyword生物防治,生物活性二次代謝物,比較基因體分析,植物生長促進,發酵條件最佳化,zh_TW
dc.subject.keywordBiocontrol agent,Bioactive secondary metabolites,Comparative genomic analysis,Plant growth promotion,Fermentation optimization,en
dc.relation.page98-
dc.identifier.doi10.6342/NTU202501190-
dc.rights.note同意授權(全球公開)-
dc.date.accepted2025-06-19-
dc.contributor.author-college生物資源暨農學院-
dc.contributor.author-dept植物病理與微生物學系-
dc.date.embargo-lift2027-07-01-
顯示於系所單位:植物病理與微生物學系

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