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| DC 欄位 | 值 | 語言 |
|---|---|---|
| dc.contributor.advisor | 張皓巽 | zh_TW |
| dc.contributor.advisor | Hao-Xun Chang | en |
| dc.contributor.author | 陳怡君 | zh_TW |
| dc.contributor.author | Yi-Jyun Chen | en |
| dc.date.accessioned | 2025-02-19T16:10:32Z | - |
| dc.date.available | 2025-02-20 | - |
| dc.date.copyright | 2025-02-19 | - |
| dc.date.issued | 2025 | - |
| dc.date.submitted | 2025-01-23 | - |
| dc.identifier.citation | Ajayi‐Oyetunde, O. O., and Bradley, C. A. 2018. Rhizoctonia solani: Taxonomy, population biology and management of Rhizoctonia seedling disease of soybean. Plant Pathol. 67:3–17.
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| dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/96483 | - |
| dc.description.abstract | 立枯絲核菌 (Rhizoctonia solani) 為一種重要的植物病原菌，能造成多種作物病害以及嚴重的農業經濟損失，而化學防治是當前防治立枯絲核菌病害的主要方法。在諸多化學藥劑中，賓克隆 (pencycuron) 因其對R. solani的專一性防治效果而被廣泛使用；然而，R. solani對賓克隆的抗性機制尚不明確。本研究針對來自臺灣的164株 R. solani AG-7 分離株進行賓克隆敏感性測試，結果顯示菌株對藥劑的反應存在顯著差異。所有分離株均表現出中至高度的抗性，唯獨分離株214對藥劑高度敏感。其中抗性最強的分離株213之EC₅₀值超過2,000 μg/mL，而最敏感的分離株214則顯示EC₅₀值僅為0.47 μg/mL。為理解這二菌株之間的抗藥反應差異，本研究利用比較基因體學和比較轉錄體學，結果顯示這二菌株在基因體上具有顯著差異。菌株213具有34.46 Mb的二倍體 (AB)結構，而菌株214則為94.97 Mb的五倍體 (AAABB)結構且伴隨高雜合度。根據賓克隆處理下的轉錄體分析結果，抗藥性菌株213顯著上調與解毒作用相關的基因，包括細胞色素P450 (cytochrome P450) 和ABC運輸蛋白 (ABC transporter)；相比之下，感藥性菌株214則表現出多元分歧的基因表現反應，其中與DNA複製、脂質生合成及細胞維持相關的途徑顯著下調。後續針對抗藥性菌株213中的細胞色素P450 (RsP450) 和ABC運輸蛋白 (RsABC1) 進行功能性驗證，然因R. solani無法建構穩定基因轉植菌株，故於感藥性的Monilinia fructicola中進行異源表現以驗證R. solani的基因功能。結果顯示，轉殖RsP450及 RsABC1的M. fructicola菌株皆顯著提升對賓克隆的抗藥性，顯示其部分提升了賓克隆的抗藥性。綜合以上結果，本研究針對R. solani AG-7對於殺菌劑賓克隆抗藥性，提供了比較基因體學、比較轉錄體學、及基因功能驗證的嶄新知識。這些結果不僅加深對賓克隆抗藥性機制的理解，亦為立枯絲核菌之病害管理策略及殺菌劑使用提供了重要認識。 | zh_TW |
| dc.description.abstract | Rhizoctonia solani is a destructive plant pathogen, and it causes severe diseases in various crops, leading to substantial economic losses worldwide. Currently, chemical control is the primary strategy for managing Rhizoctonia diseases. Among a diverse range of chemicals, pencycuron is a fungicide widely used due to its specific efficacy against R. solani. Nonetheless, the resistance mechanisms to pencycuron remain poorly understood. In this study, the sensitivity of 164 R. solani AG-7 isolates from Taiwan to pencycuron was investigated, revealing a wide range of sensitivity. All isolates exhibited moderate to high resistance, except for the isolate 214, which was highly sensitive. The most resistant isolate, 213, exhibited an EC₅₀ value exceeding 2,000 μg/mL, whereas the most sensitive isolate, 214, displayed an EC50 value at 0.47 μg/mL. To elucidate the molecular basis of the different pencycuron resistance between these two isolates, comparative genomics and transcriptomics were employed and the results identified that the isolate 213 has a diploid (AB) genome structure of 34.46 Mb, while the isolate 214 exhibits a pentaploid (AAABB) genome structure of 94.97 Mb with high heterozygosity. The resistant isolate 213 significantly upregulated the detoxification-related genes, including cytochrome P450s and ATP-binding cassette (ABC) transporters; in contrast, the sensitive isolate 214 displayed widespread transcriptional responses, such as downregulation of pathways associated with DNA replication, lipid biosynthesis, and cellular maintenance. Subsequently, functional validation of a cytochrome P450 (RsP450) and an ABC transporter (RsABC1) from the isolate 213, was performed using the pencycuron-sensitive Monilinia fructicola as a heterologous host due to the instability of R. solani transformants. The M. fructicola transformants expressing RsP450 or RsABC1 exhibited significantly elevated resistance to pencycuron, indicating their contribution to pencycuron resistance. Collectively, this study provides novel insights into the comparative genomics, transcriptomics, and gene functions of pencycuron resistance in R. solani AG-7. The findings enhance the understanding of pencycuron resistance and offer new prospects to the development of Rhizoctonia disease management and fungicide usage. | en |
| dc.description.provenance | Submitted by admin ntu (admin@lib.ntu.edu.tw) on 2025-02-19T16:10:32Z No. of bitstreams: 0 | en |
| dc.description.provenance | Made available in DSpace on 2025-02-19T16:10:32Z (GMT). No. of bitstreams: 0 | en |
| dc.description.tableofcontents | 摘要 i
Abstract ii Table of Contents iv List of Figures viii List of Tables x Chapter 1 Introduction 1 1.1 Biology of Rhizoctonia solani 1 1.2 Disease symptoms and disease cycle 2 1.3 Management of Rhizoctonia diseases 3 1.4 Fungicide resistance 4 1.5 Fungicide resistance of Rhizoctonia solani 5 1.6 Pencycuron 6 1.7 Research history of discovering the mechanism of action of pencycuron 7 1.8 Transcriptome analysis for studying fungicide resistance mechanisms 9 1.9 Research objectives 11 Chapter 2 Materials and Methods 12 2.1 Chemicals 12 2.2 Fungal materials 12 2.2.1 Rhizoctonia solani isolates 12 2.2.2 Monilinia fructicola isolate 12 2.2.3 Long-term preservation of fungal materials 13 2.3 Screening for pencycuron sensitivity in the R. solani AG-7 population 13 2.4 RNA-Seq analysis 14 2.4.1 RNA extraction, RNA-Seq library construction, and Illumina sequencing 14 2.4.2 RNA-Seq data analysis 16 2.5 Whole-genome sequencing of the R. solani AG-7 isolates 213 and 214 17 2.5.1 DNA extraction, library construction, and PacBio HiFi sequencing 17 2.5.2 Genome survey and ploidy analysis 18 2.5.3 De novo genome assembly and quality assessment 18 2.5.4 Repeat element analysis, gene prediction, and functional annotation 20 2.6 Differentially expressed genes (DEGs) identification 21 2.7 Selection of candidate genes associated with the pencycuron resistance 22 2.8 Vector construction for RsP450 and RsABC1 from genomic DNA of the isolate 213 22 2.9 Functional validation of cytochrome P450 and ABC transporter genes in R. solani and Monilinia fructicola 24 2.9.1 Preparation of the linear DNA fragments for protoplast transformation 24 2.9.2 Protoplast production for R. solani 25 2.9.3 Protoplast production for M. fructicola 26 2.9.4 Polyethyleneglycol (PEG) transformation of protoplasts 26 2.9.5 Pencycuron sensitive assay 28 2.9.6 RT-PCR validation of RsP450 and RsABC1 expression in M. fructicola transformants 28 2.10 Vector construction for RsP450 and RsABC1 from cDNA of the isolate 213 29 2.11 Statistical analyses 31 Chapter 3 Results 32 3.1 Sensitivity of Rhizoctonia solani AG-7 isolates to pencycuron 32 3.2 RNA-Seq analysis 33 3.3 Genome survey of the R. solani AG-7 isolates 213 and 214 33 3.4 De novo assembly of the R. solani AG-7 isolates 213 and 214 34 3.5 Gene prediction and functional annotation 35 3.6 Identification of differentially expressed genes (DEGs) 36 3.7 GO and KEGG pathway enrichment analyses of the pencycuron-responsive DEGs 37 3.8 Candidate genes selection for functional validation in the pencycuron resistance 39 3.9 Functional validation of RsP450 and RsABC1 41 Chapter 4 Discussion 44 4.1 Transcriptomic insights into the pencycuron resistance in the isolate 213 44 4.2 Difficulties in the functional validation of R. solani candidate genes 47 4.3 Functional validation of RsP450 and RsABC1 in Monilinia fructicola 48 4.4 Genomic complexity and transcriptional responses underlying the pencycuron sensitivity in the isolate 214 49 4.5 Conclusion 51 References 52 Tables 69 Figures 87 Supplementary Data 99 1.1 Figures 99 1.2 Tables 105 | - |
| dc.language.iso | en | - |
| dc.subject | 立枯絲核菌 | zh_TW |
| dc.subject | ABC 運輸蛋白 | zh_TW |
| dc.subject | 細胞色素 P450 | zh_TW |
| dc.subject | 外源性物質解毒 | zh_TW |
| dc.subject | 比較轉錄體學 | zh_TW |
| dc.subject | 比較基因體學 | zh_TW |
| dc.subject | 殺菌劑抗藥性 | zh_TW |
| dc.subject | comparative transcriptomics | en |
| dc.subject | Rhizoctonia solani | en |
| dc.subject | ABC transporter | en |
| dc.subject | cytochrome P450 | en |
| dc.subject | xenobiotic detoxification | en |
| dc.subject | comparative genomics | en |
| dc.subject | fungicide resistance | en |
| dc.title | 利用基因體與轉錄體解析立枯絲核菌 AG-7 對殺菌劑賓克隆的反應機制 | zh_TW |
| dc.title | Genomic and Transcriptomic Insights into the Response of Rhizoctonia solani AG-7 to the Fungicide Pencycuron | en |
| dc.type | Thesis | - |
| dc.date.schoolyear | 113-1 | - |
| dc.description.degree | 碩士 | - |
| dc.contributor.oralexamcommittee | 李敏惠;郭志鴻;蔡怡陞 | zh_TW |
| dc.contributor.oralexamcommittee | Miin-Huey Lee;Chih-Horng Kuo;Isheng Jason Tsai | en |
| dc.subject.keyword | 立枯絲核菌,殺菌劑抗藥性,比較基因體學,比較轉錄體學,外源性物質解毒,細胞色素 P450,ABC 運輸蛋白, | zh_TW |
| dc.subject.keyword | Rhizoctonia solani,fungicide resistance,comparative genomics,comparative transcriptomics,xenobiotic detoxification,cytochrome P450,ABC transporter, | en |
| dc.relation.page | 115 | - |
| dc.identifier.doi | 10.6342/NTU202500289 | - |
| dc.rights.note | 同意授權(限校園內公開) | - |
| dc.date.accepted | 2025-01-26 | - |
| 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-lift | 2025-02-20 | - |
| 顯示於系所單位： | 植物病理與微生物學系 | |
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