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研究生: 孫培峰
Sun, Pei-Feng
論文名稱: 微生物在臺灣小毛氈苔的分布及其影響
The role of microbes inside Drosera spatulata’s mucilage
指導教授: 蔡怡陞
Tsai, Isheng Jason
口試委員: 蔡怡陞
Tsai, Isheng Jason
羅南德
Roland Kirschner
陳盈嵐
Chen, Ying-Lan
林盈仲
Lin, Ying-Chung Jimmy
陳可萱
Chen, Ko-Hsuan
口試日期: 2023/04/11
學位類別: 博士
Doctor
系所名稱: 生物多樣性國際研究生博士學位學程
Taiwan International Graduate Program on Biodiversity
論文出版年: 2024
畢業學年度: 112
語文別: 英文
論文頁數: 192
英文關鍵詞: Acrodontium crateriforme, Drosera spatulata, interaction, mucilage, traps
研究方法: 調查研究比較研究田野調查法
DOI URL: http://doi.org/10.6345/NTNU202400545
論文種類: 學術論文
相關次數: 點閱:81下載:1
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  • Carnivorous plants secrete digestive fluid for nutrient acquisition. Although the fluid provides extreme conditions with low pH value and hydrolytic enzymes, several studies have found that the microbial community inside the mucilage plays an important role for prey digestion. Drosera spatulata is a carnivorous plant that secretes mucilage to stick to insects. Its leaves are covered with “tentacles” ending in glandular heads. These heads include glandular cells which produce sticky mucilage. Unlike pitcher plants containing digestive fluid inside modified foliar structures, the mucilage of D. spatulata is exposed to the environment. External influences are especially important in determining the distribution and abundance of microorganisms. In this study, we characterised the microbial communities of D. spatulata mucilage from northern Taiwan by using amplicon sequencing. To identify the relationship between D. spatulata and microorganisms, we inoculated microbes on D. spatulata and analyzed their gene expression. As the result, we found that the fungus Acrodontium crateriforme is the ecologically dominant species in D. spatulata mucilage. Based on the transcriptomes when encountering prey insects, we revealed a high degree of genetic co-option in each species during fungus-plant coexistence and digestion. Expression patterns of the holobiont during digestion further revealed synergistic effects in several gene families including fungal aspartic and sedolisin peptidases, which facilitate the digestion of sundew’s prey, as well as transporters and dose-dependent responses in plant genes involved in the jasmonate signalling pathway. This study shows that botanical carnivory is defined by multidimensional adaptations that correlate with interspecific interactions.

    Chapter1: Introduction 1 1.1 Microbe-phyllosphere interaction 1 1.2 Carnivorous plants 1 1.3 Drosera 2 1.4 Objectives 3 Chapter2: Persistence of dominant fungus Acrodontium crateriforme in mucilage of sundew Drosera spatulata 4 2.1 Introduction 4 2.2 Methods and Materials 11 2.2.1 Collection of sundew mucilage and surrounding plants 11 2.2.2 Genomic DNA extraction and metabarcoding of environmental samples 11 2.2.3 Amplicon sequencing data processing 12 2.2.4 Morphology observation of D. spatulata stalk gland 13 2.3 Results 14 2.3.1 Fungal communities in Drosera mucilage were distinct from those in surrounding plants 14 2.3.2 Ecological dominance of A. crateriforme in Drosera plant 17 2.4 Discussion 24 Chapter3: The genomic basis behind sundew Drosera spatulata’s response to A. crateriforme and its prey 26 3.1 Introduction 26 3.2 Methods and Materials 27 3.2.1 Isolation and identification of fungal species from sundew mucilage 27 3.2.2 Preparation of sterilised Drosera spatulata 28 3.2.3 Preparation of different substrates for feeding experiment 28 3.2.4 A. crateriforme growth conditions in different condition 28 3.2.5 Genome sequencing, assembly and annotation of A. crateriforme 29 3.2.6 Greenhouse experiment for plant-microbe interaction 29 3.2.7 RNA extraction 29 3.2.8 Gene prediction of A. crateriforme 30 3.2.9 Inoculation experiment 31 3.2.10 Western blot 31 3.2.11 Feeding experiment of D. spatulata 31 3.2.12 Comparative genomics and phylogenomics 32 3.2.13 Transcriptome analysis 32 3.2.14 Phytohormone analysis 33 3.3 Results 38 3.3.1 A. crateriforme enhanced prey digestion in D. spatulata 38 3.3.2 Genome of A. crateriforme as an extremophilic fungus 41 3.3.3 Digestion genes were co-opted and retained ancestral expression trends from plant-microbial coexistence 52 3.3.4 Transcriptome dynamic of holobiont digestion in nature 61 3.3.5 Synergistic expression in fungal peptidases and transporters 77 3.3.6 Dosage dependent response of genes involved in Jasmonate (JA) signalling pathway 81 3.4 Discussion 84 Chapter4: Conclusion 86 References 180

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