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姓名 葉昱宏(Yu-Hung Yeh)  查詢紙本館藏   畢業系所 生命科學系
論文名稱 台灣海邊耐鹽植物濱刺草之內生真菌
(Endophytic fungi from the salt-tolerant coast plant Spinifex littoreus in Taiwan)
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摘要(中) 在過去內生真菌被認為是只存在植物體內而不會對植物造成影響。當植物死去時,內生真菌可以首先腐生死去的植物。在2008年,科學家證明了一種濱海的草本植物需要內生真菌與其共生才能夠抵抗鹽分逆境。此外這些內生真菌有廣泛的宿主範圍包含了單子葉與雙子葉植物。濱刺草是一種濱海植物廣泛生長在台灣海灘沙丘上。這種植物能夠忍受高鹽分、風沙侵襲、低水分與營養缺乏的環境。我們在台灣各地的海灘採集濱刺草,然後將植物的根、莖、葉片、葉鞘用95%乙醇和6%次氯酸鈉進行表面消毒,然後再用無菌水清洗。表面消毒的有效性用imprint technique來驗證。我們全部從3648個植物片段中獲得了1670株菌株。從濱刺草中分離的全部1670株菌株中可被分為59個種類。從植物組織中分離的內生真菌利用菌種型態顯微鏡觀察以及DNA分析來鑑定種類。這些種類對於這種植物來說是新的紀錄,其中還有一種是尚未被發現的新種類Sarocladium sp.。
摘要(英) In the past, endophytic fungi were considered just living in plants without causing impact to the plant. When the plants die, the endophytic fungi can first grow saprobically in dead plants. In 2008, scientists demonstrated that a coastal grass species requires symbiotic fungal endophytes for salt tolerance. In addition, these endophytes have a broad host range encompassing both monocots and eudicots. Spinifex littoreus is a coastal plant in littoral sand dunes common in tropical Asia. The grass is able to tolerate high salinity, sand movement, low water and nutrient availability. We collected Spinifex littoreus at different places in Taiwan. Spinifex littoreus was divided into root, stem, leaf sheath and leaf lamina. Plants were surface-sterilized with 95% ethanol and 6% sodium hypochlorite and then rinsed in sterile water. The effectiveness of surface sterilization was verified by the imprint technique. From 3648 plant segments, altogether 1670 cultures comprising 59 species of fungi were isolated from S. littoreus. Endophytes were isolated from plant tissues and identified based on culture morphology, light microscopy and DNA analysis. The species were all recorded for this plant for the first time worldwide. An undescribed species of Sarocladium was discovered and characterized.
關鍵字(中) ★ 內生真菌
★ 台灣
★ 濱刺草
關鍵字(英) ★ endophytic fungi
★ Taiwan
★ Spinifex littoreus
論文目次 目錄       
中文摘要 i
英文摘要 ii
誌謝 iii
目錄 iv
圖目錄 vi
表目錄 vii
1 Introduction 1
2 Materials and methods 3
2-1 Plants and collection sites 3
2-2 Isolation of fungi 3
2-3 Surface sterilization 4
2-4 Media 4
2-5 DNA isolation 5
2-6 PCR 8
2-7 Agarose gel electrophoresis 10
2-8 Purification of PCR products 10
2-9 DNA sequencing 11
2-10 Morphological and molecular identification 11
2-11 Isolation frequency and colonization rate 12
2-12 Statistical analysis 12
3 Results 13
3-1 Comparison of fungal composition between MEA and SNA 13
3-2 Species richness of endophytic fungi 13
3-3 Comparison of endophytic species composition among the plant organs 13
3-4 Comparison of endophytic species composition among sampling sites 14
3-5 Description of species 15
3-5-1 Alternaria alternata 15
3-5-2 Alternaria tenuissima 16
3-5-3 Aspergillus terreus 17
3-5-4 Fusarium oxysporum 18
3-5-5 Microsphaeropsis arundinis 19
3-5-6 Moniliophthora sp. 20
3-5-7 Phaeosphaeriopsis sp. 21
3-5-8 Rhizoctonia solani 22
3-5-9 Sarocladium sp. nov. 33
4 Discussion 25
5 References 29
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指導教授 羅南德(Roland Kirschner) 審核日期 2013-1-25
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