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  1. NTU Theses and Dissertations Repository
  2. 生物資源暨農學院
  3. 植物醫學碩士學位學程
請用此 Handle URI 來引用此文件: http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/101678
完整後設資料紀錄
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dc.contributor.advisor林乃君zh_TW
dc.contributor.advisorNai-Chun Linen
dc.contributor.author白宇傑zh_TW
dc.contributor.authorYu-Jie Baien
dc.date.accessioned2026-02-26T16:38:36Z-
dc.date.available2026-02-27-
dc.date.copyright2026-02-26-
dc.date.issued2026-
dc.date.submitted2026-02-23-
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dc.identifier.urihttp://tdr.lib.ntu.edu.tw/jspui/handle/123456789/101678-
dc.description.abstract青蔥 (Allium fistulosum L.) 為臺灣重要之香辛蔬菜,近年受 Colletotrichum siamense 所引起的新興病害青蔥捲葉型炭疽病 (anthracnose-twister disease of green onion) 危害,造成嚴重經濟損失。由於該病原具潛伏感染特性且現行化學防治效果有限,需發展替代性防治策略。本研究首先建立一穩定的病原性試驗系統,並分別篩選具促進植物生長與病原拮抗潛力之有益根部內生細菌 (beneficial root endophytes, BREs),評估其合成菌群 (synthetic microbial community, SynCom) 於青蔥捲葉型炭疽病管理上之應用潛力。研究結果顯示,C. siamense Col-413 具穩定且較強之致病力,因此選定為標準病原菌株。透過比較不同接種濃度、接種方式及植株週齡,成功確立可穩定誘發中度病徵之接種條件。另外,不同青蔥品種 (系) 對青蔥捲葉型炭疽病之感病性不同,其中「北蔥」於實生苗與分蘗株苗階段皆展現較佳抗病性,而「吉品蔥」與「乙品小蔥」等四季蔥品種則表現更為感病。本研究成功分離出四株具顯著促進植物生長效果且非人體伺機病原之菌株,分別為Fictibacillus enclensis (S3R3)、Pseudomonas atacamensis (V3R2)、P. nitroreducens (V3K2) 及 Comamonas terrigena (O3N3)。於盆栽試驗中,上述菌株所組成之 SynComs (如 All 與 All-S3R3 組合) 於多數生長指標上皆顯著優於單一菌株與未處理對照組,顯示其具備促進青蔥生長之潛力;然而這些 SynComs 組合於試驗條件下未能顯著抑制病害發展。病原拮抗潛力與內生性菌株篩選中鑑定出 Bacillus velezensis 與 B. subtilis,其中 B. velezensis (C-KBM-453) 於實生苗防治試驗中可顯著降低罹病度,但在分蘗苗試驗中,所有處理組 (包含SynComs) 皆無顯著防治效果。綜言之,本研究確立穩定的感病系統與評估基礎,並證實 SynComs 在促進青蔥生長方面具應用潛力。未來透過優化內生菌接種濃度與施用策略,應可提升有益根部內生菌於青蔥整合性管理中之生物防治潛力。zh_TW
dc.description.abstractGreen onion (Allium fistulosum L.) is an important culinary herb in Taiwan. Recently, its production has been severely threatened by an emerging disease, anthracnose-twister disease caused by Colletotrichum siamense, resulting in substantial economic losses. Because this pathogen frequently establishes latent infections and current chemical control measures are often ineffective, the development of alternative management strategies is urgently needed. In this study, a stable pathogenicity assay system was first established, and beneficial root endophytes (BREs) with plant growth-promoting or antagonistic activities were subsequently screened to evaluate their application potential as synthetic microbial communities (SynComs) for managing anthracnose-twister disease of green onion. The results showed that C. siamense Col-413 exhibited stable and relatively high virulence and was therefore selected as the standard pathogen strain. By comparing various inoculum concentrations, inoculation methods, and plant ages, conditions that consistently resulted in moderate disease severity were successfully established. In addition, green onion cultivars (land variety) differed in their susceptibility to anthracnose-twister disease: both the seed-propagated seedlings and tiller-propagated plants of the “Beicong” cultivar exhibited comparatively higher resistance, whereas several “Sijicong” cultivars, including “Jipincong” and “Yipincong” were more susceptible. Four plant growth-promoting bacteria, showing no potential as human pathogens, were successfully isolated and identified as Fictibacillus enclensis (S3R3), Pseudomonas atacamensis (V3R2), P. nitroreducens (V3K2), and Comamonas terrigena (O3N3). In pot experiments, SynComs composed of these strains (e.g., the All and All-S3R3) performed significantly better than single-strain treatments and the untreated control in most growth-related parameters, indicating strong growth-promoting potential in green onion. However, under the experimental conditions tested, these SynComs did not significantly suppress disease development. During the screening of antagonistic and endophytic strains, Bacillus velezensis and B. subtilis were identified. Among them, B. velezensis (C-KBM-453) significantly reduced disease severity in seed-propagated seedlings, whereas no significant disease control effects were observed in tiller-propagated plants across all treatment, including the SynComs. Overall, this study establishes a stable and reproducible pathogenicity assay system and demonstrates the potential of the SynComs for promoting green onion growth. Further optimization of endophyte inoculum concentrations and application strategies may enhance the biocontrol potential of the beneficial root endophytes within integrated disease management programs for green onion.en
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dc.description.tableofcontents口試委員會審定書 I
誌謝 II
摘要 III
Abstract V
目次 VII
表次 XIII
圖次 XIV
附表次 XVI
附圖次 XVII
壹、 前人研究 1
一、青蔥介紹 1
1. 青蔥簡史、營養價值與產業重要性 1
2. 青蔥栽培品種與特性 1
(a) 北蔥 1
(b) 四季蔥 2
(c) 大蔥 3
二、青蔥栽培管理概要 3
三、青蔥炭疽病概述 6
四、植物內生細菌與根部內生細菌之定義 9
五、有益根部內生細菌之功能與應用潛力 10
1. 促進植物生長之根部內生菌 (Plant growth-promoting root endophytic bacteria) 10
(a) 提升植物可利用養分 11
(b) 合成植物生長調節物質 11
2. 抑制病害之根部內生菌 (Disease-suppressive endophytic bacteria) 11
(a) 抗生素與次級代謝物的產生 11
(b) 細胞壁水解酵素的分泌 12
(c) 空間與營養競爭 13
(d) 誘導性系統抗性 (Induced systemic resistance, ISR) 13
3. 減緩逆境壓力之根部內生菌 (Stress-mitigating endophytic bacteria) 14
六、合成菌群 (synthetic microbial community, SynCom) 15
1. SynCom 的出現與定義 15
2. 菌相分析與 SynCom組合技術的成熟 16
3. SynCom 組合應用策略的演進 18
4. 根部內生SynCom 於植物生長促進與病害管理的應用現況 19
貳、研究動機與目的 21
參、材料與方法 23
一、供試植物與栽培條件 23
1. 青蔥品種來源 23
2. 栽培介質組成與滅菌方式 23
3. 實生苗之栽培流程 23
4. 分蘗苗之栽培與肥培管理 24
二、病原菌株來源、培養及接種源之製備 24
1. 病原菌株來源與保存 24
2. 產孢培養優化—scallion powder agar (SPA) 產孢培養基製備 25
3. 病原菌孢子懸浮液之製備與濃度調整 25
三、病原菌系統與病原性測試流程之建立 25
1. 病原菌系統建立策略與參考依據 25
2. 病原菌菌株篩選試驗 26
(a) 實生苗病原菌菌株篩選試驗 26
(b) 分蘗苗病原菌菌株篩選試驗 26
3. 接種濃度篩選試驗 26
4. 接種方式比較試驗 27
5. 不同植株週齡對感病性之影響 27
6. 不同青蔥品種之感病性評估 28
7. 病徵評估標準與罹病指數計算 28
四、促進植物生長潛力之內生菌篩選 29
1. 土壤樣本採集與保存 29
2. 促進植物生長菌株篩選流程 29
(a) 土壤萃取液製備、青蔥種子表面消毒與發芽處理 30
(b) 土壤萃取液處理青蔥種苗 30
(c) 根部組織液製備 30
(d) 促進植物生長潛力之內生菌分離、保存與培養 30
(e) 促進植物生長潛力之內生菌二次篩選 31
3. 評估單一菌株之促進植物生長能力 31
五、病原拮抗潛力菌株之篩選與內生能力確認 32
1. 候選菌株對病原菌之拮抗效果初步篩選 32
2. 候選菌株對病原菌拮抗能力之定量測試 32
3. 病原拮抗潛力候選菌株於植物根部內生能力確認 33
六、菌種鑑定 34
1. Genomic DNA 萃取 34
2. 聚合酶連鎖反應 (Polymerase chain reaction, PCR) 35
3. 電泳、純化與定序分析 35
七、促進植物生長候選菌株與病原菌之對峙培養試驗 36
八、候選菌株間之相容性與交互作用評估 36
1. 相容性測定 (Cross-streak test) 36
2. 吸引測試 (Strain attraction test) 37
3. 交叉餵養測試 (Cross-feeding) 37
(a) 條件培養液 (Conditioned medium, CM) 之製備 38
(b) 交叉餵養試驗設計與生長評估 38
九、促進植物生長潛力與病原拮抗潛力內生菌株及其 SynComs 之病害防治效果評估 38
1. 促進植物生長潛力內生菌株與其 SynComs 組合設計 39
2. 促進植物生長潛力內生菌株與其 SynComs 之接種與病害防治試驗 39
3. 病原拮抗潛力內生菌株與其 SynComs 組合設計 40
4. 病原拮抗潛力內生菌株與其 SynComs 之接種與病害防治試驗 40
(a) 實生苗防治試驗 40
(b) 分蘗苗防治試驗 40
十、統計分析 41
肆、結果 42
一、病原菌系統建立與病原菌致病力評估 42
1. 產孢培養條件之優化 (SPA 製備) 42
2. 不同 C. siamense 菌株之致病力比較 42
3. 接種濃度篩選與標準條件建立 43
4. 不同接種方式之比較 44
5. 不同植株週齡對感病性之影響 45
6. 不同青蔥品種對 C. siamense Col-413 之感病性 45
二、促進植物生長潛力內生菌之篩選與植株生長影響 46
1. 不同篩選批次之植株生長影響 46
2. 促進植物生長潛力之內生菌分離 47
3. 促進植物生長潛力之內生菌二次篩選 47
4. 評估單一菌株之促進植物生長能力 48
三、促進植物生長之候選菌株鑑定 49
四、促進植物生長候選菌株與病原菌之對峙培養 50
五、促進植物生長候選菌株間相容性與交互作用評估 50
1. 相容性測定 50
2. 吸引測試 50
3. 交叉餵養測試 51
(a) 純條件培養液 (CM) 下之菌株生長表現 51
(b) 條件培養液與新鮮培養液等體積混合 (CM:KBM = 1:1) 下之菌株生長表現 51
六、促進植物生長內生菌株及其 SynComs 對青蔥生長之影響 52
七、促進植物生長內生菌株及其 SynComs 之病害防治效果評估 53
八、病原拮抗潛力菌株之篩選與內生能力確認 54
1. 候選菌株對病原菌之拮抗效果初步篩選 54
2. 候選菌株對病原菌之拮抗能力定量測試 54
3. 病原拮抗候選菌株之根部內生能力確認 54
九、病原拮抗潛力之候選菌株鑑定 55
十、病原拮抗候選菌株間相容性與交互作用評估 55
1. 相容性測定 55
2. 吸引測試 55
3. 交叉餵養測試 56
(a) 純條件培養液下之菌株生長表現 56
(b) 條件培養液與新鮮培養基等體積混合 (CM:KBM = 1:1) 下之菌株生長表現 56
十一、病原拮抗潛力內生菌株及其 SynComs 之病害防治效果評估 57
1. 實生苗防治試驗 57
2. 分蘗苗防治試驗 57
伍、討論 58
陸、結論 70
柒、參考文獻 72
捌、表 102
玖、圖 109
拾、附表 151
拾壹、附圖 155
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dc.language.isozh_TW-
dc.subject青蔥捲葉型炭疽病-
dc.subjectColletotrichum siamense-
dc.subject生物防治-
dc.subject植物內生菌-
dc.subject合成菌群-
dc.subjectAnthracnose-twister disease of green onion-
dc.subjectColletotrichum siamense-
dc.subjectBiocontrol-
dc.subjectEndophytic bacteria-
dc.subjectSynthetic microbial community (SynCom)-
dc.title根部內生細菌對青蔥捲葉型炭疽病生物防治潛力之評估zh_TW
dc.titleEvaluation of the biocontrol potential of root endophytic bacteria against anthracnose-twister disease in green onionen
dc.typeThesis-
dc.date.schoolyear114-1-
dc.description.degree碩士-
dc.contributor.oralexamcommittee鍾嘉綾;黃晋興zh_TW
dc.contributor.oralexamcommitteeChia-Lin Chung;Jin-Hsing Huangen
dc.subject.keyword青蔥捲葉型炭疽病,Colletotrichum siamense生物防治植物內生菌合成菌群zh_TW
dc.subject.keywordAnthracnose-twister disease of green onion,Colletotrichum siamenseBiocontrolEndophytic bacteriaSynthetic microbial community (SynCom)en
dc.relation.page159-
dc.identifier.doi10.6342/NTU202600764-
dc.rights.note同意授權(限校園內公開)-
dc.date.accepted2026-02-23-
dc.contributor.author-college生物資源暨農學院-
dc.contributor.author-dept植物醫學碩士學位學程-
dc.date.embargo-lift2031-01-28-
顯示於系所單位:植物醫學碩士學位學程

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