Selected fungicolous Hypocreales in northern Taiwan

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魏亦晴(I-Chin Wei) 查詢紙本館藏

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生命科學系

論文名稱

(Selected fungicolous Hypocreales in northern Taiwan)

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摘要(中)

菌生真菌為一種真菌分別以寄生、片利共生、腐生的形式生長在另一種真菌上，目前仍無法明確得知其菌生菌與宿主間的相互作用。子囊菌門當中的肉座菌目(Hypocreales)，有很大一部分為菌生真菌的種類。本研究以隨機採集的方式，鑑定及分析北台灣菌生真菌種類，同時記錄台灣新的真菌種類。利用真菌型態學及DNA分子技術鑑定出真菌種類；比較真菌培養於玉米瓊酯培養基及自然基質下，真菌型態的不同，同時利用Internal transcribed spacer (ITS)，Large subunit (LSU) rDNA或 small subunit (SSU) rDNA 鑑定真菌種類。此研究採集及鑑定的菌生真菌有14種，分別是: Acremonium sp., Acremonium citrinum, Cladobotryum semicirculare, Hypomyces chrysospermus, Hypomyces albidus, Hypomyces corticiicola, Lecancillium psallioate, Nectriopsis rexiana‏, Ramularia coleosporii, Ramularia sp.?, Simplicillium lanosoniveum, Sphaerostilbella sp., Trichoderma harzianum, 及 Trichoderma cf. asperellum。菌生真菌的宿主多樣性中：有子囊菌門，擔子菌門及黏菌門。而宿主種類中含有可食用的菇類，分解木材的多孔菌類，叢枝菌根共生菌，腐生及植物寄生菌及黏菌。此篇研究中的菌生菌，有些是台灣新紀錄：Acremonium citrinum 於荷葉上的子囊菌分離, 此菌種在台灣目前沒有紀錄,而 Nectriopsis rexiana發現生長於黏菌上，在台灣還未發現此種類；Trichoderma 種類，於馬勃上分離出來，在先前的台灣紀錄由植物或土壤分離，馬勃作為宿主為台灣新的記錄。於葡萄科葉片上的銹病(Phakopsoraceae)分離出的似Ramularia sp.菌株，利用光學顯微鏡，掃描式及穿透式電子顯微鏡，同時透過分析LSU和SSU rDNA 序列進行比較。型態及電顯結果鑑定為Ramularia sp.，但是DNA序列結果顯示菌株較接近黑粉菌亞門（Ustilaginomycotina）的種類。

摘要(英)

The fungicolous fungi refer to species of fungi that consistently are associated with other fungi, including fungi growing on other fungi as parasites (mycoparasites), commensals or saprobes. Hypocreales (Ascomycota) is an order of fungi which has a large proportion of mycoparasitic or mycosaprotrophic species, but we still not know the details of interactions between the fungi. We randomly collected the samples from Yilan, Taipei, Taoyuan, Hsinchu, Natou, Kinmen and Guangxi (China). Molecular analyses were used to determine the fungicolous species. The morphology of the species on natural substrata was compared with that in culture on Corn Meal Agar (CMA). In the DNA analysis, Basic Local Alignment Search Tool (BLAST) for ± 97% identities of Internal transcribed spacer (ITS), large subunit (LSU) rDNA and small subunit (SSU) rDNA sequences were used for species identification as well as A phylogenetic analysis 14 fungicolous fungi were collected, including Acremonium sp., Acremonium citrinum, Cladobotryum semicirculare, Hypomyces chrysospermus, Hypomyces albidus, Hypomyces corticiicola, Lecancillium psallioate, Nectriopsis rexiana‏, Ramularia coleosporii, Ramularia sp.?,Simplicillium lanosoniveum, Sphaerostilbella sp., Trichoderma harzianum, and Trichoderma cf. asperellum. 3 of 14 fungicolous fungi species could only be identified to the genus. Host fungi were diverse, including Basidiomycota, Myxomycota, and Ascomycota with diverse genera. The host species include species of edible mushroom (Auricularia, Suillus), wood-decomposing polypores (Fomitella, Microporus), mycorrhiza fungi (Scleroderma, Suillus), saprobic and plant parasitic microfungi (Coleosporium, unidentified Phakopsoraceae, Gonatophragmium, Hemileia, Puccinia, Talaromyces) and slime mold (Arcyria). Some species were records for Taiwan; Acremonium citrinum which collected from a sooty mold and Nectriopsis rexiana on the mycomycete Arcyria denudata are new records, and Scleroderma is a new host genus for Trichoderma harzianum and Trichoderma cf. asperellum in Taiwan. One strain we collected from a rust fungus (Phakopsoraceae) on leaves of Ampelopsis brevipedunculata is a Ramularia-like species. Material of this fungus was studied on the natural substrate and in culture with light, scanning and transmission electron microscopy, and analyses of LSU and SSU rDNA sequences. The morphology and ultrastructure suggest identification as Ramularia species, but the DNA sequences indicate a relationship to Ustilaginomycotina.

關鍵字(中)

★ 菌生真菌

關鍵字(英)

★ fungicolous fungi

論文目次

中文摘要 V
英文摘要 VI
致謝 VIII
目錄 IX
圖目錄 XI
表目錄 XIII
List of abbreviations XIV
1 Introduction 1
1-1 What are fungicolous fungi 1
1-2 Types of fungicolous associations 1
1-3 Fungicolous Hypocreales 3
1-4 Studies of fungicolous fungi in Taiwan 4
1-5 Fungicolous fungi used in applied mycology 5
1-6 Aim of this study 6
2 Material and Methods 7
2-1 Specimen preservation 7
2-2 Light- microscopes 7
2-3 Isolation of fungi 8
2-4 Medium 8
2-5 Strains and growth condition 8
2-6 DNA isolation 9
2-6-1 Lysis 9
2-6-2 DNA binding 10
2-6-3 Wash 11
2-6-4 DNA Elution 11
2-7 PCR 11
2-7-1 Agarose gel electrophoresis 13
2-7-2 DNA sequencing 13
2-8 Morphological identification 14
2-9 Preparation of material for scanning electron microscopy 14
2-9-1 Fixation 14
2-9-2 Dehydration 15
2-9-3 Critical point drying 15
2-9-4 Specimen mounting and gold coating 16
2-10 Preparation of material for transmission electron microscopy 16
2-10-1 Fixation 16
2-10-2 Dehydration 17
2-10-3 Resin infiltration and embedment 17
2-10-4 Sectioning 18
2-10-5 Staining 18
3 Results 20
3-1 Acremonium sp. 21
3-2 Acremonium citrinum 23
3-3 Hypomyces semicirculare 25
3-4 Hypomyces chrysospermus 27
3-5 Hypomyces albidus 30
3-6 Hypomyces corticiicola 32
3-7 Lecanicillium psalliotae 35
3-8 Nectriopsis rexiana 37
3-9 Ramularia coleosporii 39
3-10 Ramularia sp. 41
3-11 Simplicillium lanosoniveum 45
3-12 Sphaerostilbella sp. 47
3-13 Trichoderma harzianum 49
3-14 Trichoderma cf. asperellum 51
4 Discussion 53
5 References 59
圖目錄

Fig. 1 Acremonium sp. grown in nature and culture 71
Fig. 1-1 Acremonium sp. grown on different substrates 72
Fig. 2 Acremonium citrinum grown in nature 73
Fig. 2-1 Acremonium citrinum grown in culture 74
Fig. 2-2 Acremonium citrinum grown on different substrates 75
Fig. 3 Cladobotryum semicirculare in nature 76
Fig. 3-1 Cladobotryum semicirculare grown on different substrata 77
Fig. 4 Hypomyces chrysospermus grown in nature 78
Fig. 4-1 Hypomyces chrysospermus grown in culture 79
Fig. 4-2 Hypomyces chrysospermus grown on different substrata 80
Fig. 5 Hypomyces albidus grown in nature 81
Fig. 5-1 Hypomyces albidus grown in culture 82
Fig. 5-2 Hypomyces albidus grown in culture 83
Fig. 6 Hypomyces corticiicola in culture 84
Fig. 6-1 Hypomyces corticiicola on natural substrate 85
Fig 6-2 Phylogenetic tree of related species of Hypomyces cf corticiicola……....86
Fig. 7 Lecancillium psallioate grown in nature 87
Fig. 7-1 Lecancillium psallioate grown in different substrata 88
Fig. 8 Nectriopsis rexiana grown in nature 89
Fig. 8-1 Nectriopsis rexiana grown in culture 90
Fig. 9 Ramularia coleosporii grown in nature 91
Fig. 9-1 Ramularia coleosporii grown in culture 92
Fig 10 Ramularia sp. ? grown in nature 93
Fig 10-1 Ramularia sp. ? grown in culture 94
Fig 10-2 Ramularia sp. ? on rust fungi (Phakopsoraceae) on
Ampelopsi brevipedunculata seen with scanning electron microscopy (SEM) 95
Fig 10-3 Ramularia sp. ? seen with Transmission Electron Microscopy (TEM) 96
Fig. 11 Simplicillium lanosoniveum grown in nature 97
Fig. 11-1 Simplicillium lanosoniveum grown in culture 98
Fig. 11-2 Phylogenetic tree of related species of Simplicillium lanosonivem 99
Fig. 12 Sphaerostilbella sp. grown in nature 100
Fig. 12-1 Sphaerostilbella sp. grown in culture 101
Fig. 13 Trichoderma harzianum in culture 102
Fig. 14 Trichoderma cf. asperellum in culture 103
Fig. 14-1 Trichoderma cf. asperellum grown in different substrata 104
表目錄

Table 1 Comparison of morphological among the Acremonium species 105
Table 2 Comparison of the morphology among the species of Hypomyces (anamorph: Sepedonium) in culture and nature 106
Table 2-1 Comparison of the morphology among the species of Hypomyces (anamorph: Sepedonium) in culture 107
Table 3 Morphological comparison of Cladobotryum anamorphs of Hypomyces corticiicola, H. polyporinus, and H. stereicola based on Carey and Rogerson (1981); Gray and Jones (1980); Põldmaa and Samuels (1999)
108
Table 4 Comparison of the morphology among the species of Sphaerostilbella 109

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指導教授

羅南德(Roland Kirschner)

審核日期

2016-12-1

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