台灣海岸植物之內生真菌多樣性研究

以作者查詢圖書館館藏

、以作者查詢臺灣博碩士

、以作者查詢全國書目

、勘誤回報

、線上人數：18

、訪客IP：3.15.219.217

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

台灣海岸植物之內生真菌多樣性研究
(The diversity of endophytic fungi from coastal plants in Taiwan)

相關論文

★ 台灣沉香樹相關的內生菌

檔案

[Endnote RIS 格式]

[Bibtex 格式]

[相關文章]

[文章引用]

[完整記錄]

[館藏目錄]

至系統瀏覽論文 ( 永不開放)

摘要(中)

沙岸植物是一種生存在沙灘這類特殊棲息地，能忍受缺乏營養、高鹽分、強風吹襲和太陽輻射強
的逆境。許多只生長在沙岸的草本植物構成了這一種特殊的植被。這些沙岸植物具有防風、定沙，
保護海岸的功能同時也是扮演沙濱生態系的重要角色。可是這種植被目前卻面臨被破壞的危險。
因為海平面的不斷的上升，加上人為的破壞，可能會使得許多適應這類環境的生物消失。目前已
知內生真菌對於生存在逆境環境的植物有很重要的影響，可是我們對於熱帶/亞熱帶沿海草本植
物的內生菌卻所知甚少。我們的研究目的是檢測和鑑定蔓荊和馬鞍藤這兩種台灣主要海岸植物的
內生真菌並分析真菌是否特異存在於植物物種或是器官和組織。研究兩種植物真菌的組成和多樣
性，預計能提供有關真菌的地理和宿主分佈的新數據，而新發現的菌株和其DNA 數據將被保存
以供未來的研究應用。我們從1782 個蔓荊植物片段中分離了1072 個真菌菌株在這其中有76 個
種類。我們也在2304 個馬鞍藤植物片段中分離了885 個真菌菌株在這其中有140 個種類。這些
種類大多都是其宿主的新的紀錄，在這其中我們也發現一個新物種Alpestrisphaeria sp. nov.，此
物種僅在蔓荊的根和莖中發現。在優勢種中只有Aspergillus terreus 是同時存在於兩種植物體內。
在真菌群落方面，馬鞍藤在自然沿海棲息地和人工植物園之間存在顯著差異。在沿海棲息地，潛
在的致病真菌比腐生真菌更為罕見，這可能表明腐生內生菌在養分循環中的重要性，特別是在植
物被沙子掩埋後腐爛葉片。數據表明內生真菌的種類受到其棲息地、植物物種以及植物器官所影
響。除了內生真菌的研究之外，我們還研究了植物的外生真菌，以解決內生真菌、寄生真菌和腐
生真菌之間是否存在明確分界的問題。在植物園種植的蔓荊中我們發現了Corynespora cassiicola
這株葉片病原體，但在野生外的蔓荊中沒有發現其內生菌紀錄。Dinemasporium spinificis 是台灣
的第一個記錄，為濱刺草上的外生腐生菌，但在先前的濱刺草內生菌研究中沒有其內生真菌的紀
錄。

摘要(英)

Herbaceous vegetation at sand coasts consists of specific plants which can tolerate high salinity,
wind and sand movements, and lack of nutrients, and are, therefore, important for coast protection and
preventing erosion, but are outcompeted in inland habitats. Although this vegetation in Taiwan on the
one hand is endangered by construction, tourism and other human activities, as well as rising sea level,
and on the other hand fungal endophytes are known to have a strong influence on the survival of plants
under such stress factors, few is known about the endophytes of tropical/subtropical coastal herbaceous
plants. The aim of the study was to detect and identify endophytic fungi of Vitex rotundifolia and
Ipomoea pes-caprae, two of the most common plants along the shore of Taiwan, and to analyze
whether fungi were specific to plant species, certain organ or tissues. The study on composition and
diversity of fungi from both plants provided new data about the geographical and host distribution of
fungi, led to the discovery of new species and new fungal records. New isolates and DNA data were
deposited for future systematic and biotechnological studies and applications. We isolated 1072 strains
comprising 76 species from 1782 plant fragments of Vitex rotundifolia and 885 strains comprising 140
species from 2304 fragments of Ipomoea pes-caprae, which were identified with genetic markers as
well as with morphology. These species most are new records for their hosts. Among these species,
Alpestrisphaeria sp. nov. was considered as new and was found only in roots and stems of V.
rotundifolia. Only Aspergillus terreus was a common species in the top three dominant species from
both plants. Fungal communities in I. pes-caprae differed significantly between natural coastal habitats
and artificial plantations in botanical gardens. In coastal habitats, potentially pathogenic fungi were
rarer than saprobic species, which might indicate the importance of saprobic endophytes in nutrient
iii
cycling particularly of decomposing leaves after sand burial. The data indicate that occurrence of
endophytic fungal species is driven by the habitat, plant species, and plant organ. Except for the study
of endophytic fungi, examples of ectophytic fungi were studied for addressing the question whether
there is a clear-cut demarcation between endophytic, parasitic and saprobic fungi. Corynespora
cassiicola was found as leaf pathogen on artificially planted V. rotundifolia, but not as endophyte in
wild populations. Dinemasporium spinificis, a first record from Taiwan, was found only as ectophytic
saprobe on Spinifix littoreus, but was not recorded as endophyte from this plant in previous studies.

關鍵字(中)

★ 沙岸植物
★ 內生菌
★ 生物多樣性
★ 沙灘保護
★ 蔓荊
★ 馬鞍藤

關鍵字(英)

★ sand coast vegetation
★ endophytes
★ biodiversity
★ beach protection
★ beach vitex
★ beach morning glory

論文目次

中文摘要 i
Abstract ii
誌謝 iv
圖目錄 viii
表目錄 x
1. Introduction 1
2. Material and methods 7
2-1 Sample and collection sites 7
2-2 Isolation of endophytic fungi 8
2-3 Isolation of ectophytic fungi 8
2-4 DNA isolation 9
2-4-1 Tissue Dissociation 9
2-4-2 Lysis 9
2-4-3 DNA Binding 9
2-4-4 DNA Wash 10
2-4-5 DNA Elution 10
2-5 PCR 10
2-6 Molecular identification 11
2-7 Phylogenetic analysis 13
2-8 Morphological identification 13
2-9 The identification of Alternaria spp. 14
2-10 Host plant infection experiments 14
2-11 Statistical data analysis 15
3. Results 17
3.1 Endophytic fungal community associated with Vitex rotundifolia 17
3.2 Fungal community comparison between Vitex rotundifolia collected from different sites 19
3.3 Fungal community comparison between organs of Vitex rotundifolia 20
3.4 Endophytic fungal community associated with Ipomoea pes-caprae 23
3-5 Fungal community comparison between Ipomoea pes-caprae collected from different habitats 25
3-6 Fungal community comparison between organs of Ipomoea pes-caprae 27
3-7 Fungal community comparison between Ipomoea pes-caprae and Vitex rotundifolia 30
3-8 Parasitic and saprobic fungi associated with Vitex rotundifolia and Spinifex littoreus 32
3-9 Acrophialophora levis 34
3-10 Alpestrisphaeria sp. nov.. 36
3-11 Aureobasidium melanogenum 40
3-12 Corynespora cassiicola 42
3-13 Dinemasporium spinificis 44
3-14 Hortaea werneckii 48
3-15 Rosellinia sp. 50
4. Discussion 53
5. References 61
6. Appendixes 69
6-1 Appendix I Supplemental Table 69
6-2 Appendix II Conference Presentations 126
6-3 Appendix III List of publications 127

參考文獻

伍淑惠、張元聰、高瑞卿，2010年，台灣海濱植物圖鑑，晨星出版社，台中。

Aoki, T., O’Donnell, K. and Geiser, D. M. J (2014). Systematics of key phytopathogenic Fusarium species: current status and future challenges. Journal of General Plant Pathology 80(3): 189–201 https://doi.org/10.1007/s10327-014-0509-3

Ariyawansa, H. A., Hyde, K. D., Thambugala, K. M., Maharachchikumbura, S. S. N., Al-Sadi, A. M., and Liu, Z. Y. (2016). Additions to Karst Fungi 2: Alpestrisphaeria jonesii from Guizhou Province, China. Phytotaxa 277(3): 255–265.

Beena, K. R., Ananda, K. and Sridhar, K. R. (2000a). Fungal endophytes of three sand dune plant species of west coast of India. Sydowia 52(1): 1–9.

Beena, K. R., Raviraja, N. S. and Sridhar, K. R. (2000b). Seasonal variations of arbuscular mycorrhizal fungal association with lpomoea pes-caprae of coastal sand dunes, Southern India. Journal of Environmental Biology 21(4): 341–347.

Bensch, K., Braun, U., Groenewald, J. Z. and Crous, P. W. (2012). The genus Cladosporium. Studies in Mycology 72: 1–401. DOI: 10.3114/sim0003.

Bills, G. F., Platas, G., Overy, D. P., et al. (2009). Discovery of the parnafungins, antifungal metabolites that inhibit mRNA polyadenylation, from the Fusarium larvarum complex and other hypocrealean fungi. Mycologia 101(4): 449–472. DOI: 10.3852/08-163.

Cabañe, F. J., Bragulat, M. R., and Castellá, G. (2012). Hortaea werneckii isolated from silicone scuba diving equipment in Spain. Medical Mycology 50(8): 852–857.

Cai, L., Hyde, K. D., Taylor, P. W. J., et al. (2009). A polyphasic approach for studying Colletotrichum. Fungal Diversity 39: 183–204.

Carbone, I. and Kohn, L. (1999). A Method for Designing Primer Sets for Speciation Studies in Filamentous Ascomycetes. Mycologia 91(3): 553–556.

Chen, S. l. and Phillips, S. M. (2006). Poaceae. In Wu, Z. G. and Raven, P. H. (Eds), Flora of China 22: 553, Beijing, China, Science Press.

Corkidi, L. and Rincón, E. (1997). Arbuscular mycorrhizae in a tropical sand dune ecosystem on the Gulf of Mexico. Mycorrhiza 7:17–23.

Cousins, M.M., Briggs, J., Gresham, C., Whetstone, J., and Whitwell, T. (2010). Beach Vitex (Vitex rotundifolia): An Invasive Coastal Species. Invasive Plant Science and Management 3: 340–345.

Crous, P. W. and Barber, P.A. (2017). Rosellinia australiensis. Fungal Planet 564: 261.

Crous, P. W., Wingfield, M. J., Guarro, J. et al. (2013). Fungal Planet: 154-213.

Ellis, M. B. (1971). Dematiaceous Hyphomycetes, Commonwealth Mycological Institute Kew, Surrey, England.

Ellis, M. B. (1976). More Dematiaceous Hyphomycetes. CMI, Kew

Fang R. C. and George, S. (1995). CONVOLVULACEAE. In Wu, Z. G. and Raven, P. H. (Eds), Flora of China 16: 308, Beijing, China, Science Press.

Farr, D.F., & Rossman, A.Y. Fungal Databases, U.S. National Fungus Collections, ARS, USDA. Retrieved January 8, 2019, from https://nt.ars-grin.gov/fungaldatabases/

Gams, W. (2016). Are old taxa without living authenticated cultures losing their status? IMA Fungus 7(2): 72–73.

Gardes, M. and Bruns, T. D. (1993). ITS primers with enhanced specificity for basidiomycetes‐application to the identification of mycorrhizae and rusts. Molecular Ecology 2(2): 113–118. https://doi.org/10.1111/j.1365-294X.1993.tb00005.x.

Groenewald JZ, Nakashima C, Nishikawa J, et al. (2012). Species concepts in Cercospora: spotting the weeds among the roses. Studies in Mycology 75(1):115–170.

Guo, S., Mao, W. J., Li, Y. L., Tian, J. H., and Xu, J. (2013). Structural elucidation of the exopolysaccharide produced by fungus Fusarium oxysporum Y24-2. Carbohydrate Research 365: 9–13.

Hawksworth, D.L. (2004). Fungal diversity and its implications for genetic resource collections. Studies in Mycology 50: 9–18.

Hsieh (2016). Diversity of cultivable endophytic fungi of Chinese mesona (Platostoma palustre). Master’s thesis, National Central University, Taoyuan, Taiwan.

Hsieh, C. F., Shen, C. F., and Yang, K. C. (1994). Introduction to the flora of Taiwan, 3: floristics, phytogeography, and vegetation. In Huang, T. C. (Eds.), Flora of Taiwan, 2nd edition, 1: 12. Taipei, Taiwan, the editorial committee of the Flora of Taiwan.

Hu, Y., Hou, T. T., Xin, H. L., Zhang, Q. Y., Zheng, H. C., Rahman, K. and Qin, L. P. (2007). Estrogen-like activity of volatile components from Vitex rotundifolia L. The Indian Journal of Medical Research 126: 68–72.

Huang, Y. L., Zimmerman, N. B. and Arnold, A. E. (2018). Observations on the Early Establishment of Foliar Endophytic Fungi in Leaf Discs and Living Leaves of a Model Woody Angiosperm, Populus trichocarpa (Salicaceae). Journal of Fungi 4(2): 58. DOI: 10.3390/jof4020058.
Hyde, K. D., Cai, L., McKenzie, E.H.C., Yang, Y.L.,

Zhang, J.Z. and Prihastuti, H. (2009). Colletotrichum: a catalogue of confusion. Fungal Diversity 39: 1–17.
Jia, M., Chen, L., Xin, H. L., Zheng, C. J., Rahman, K.,

Han, T., and Qin, L. P. (2016). A Friendly Relationship between Endophytic Fungi and Medicinal Plants: A Systematic Review. Frontiers in Microbiology 7: 906. DOI: 10.3389/fmicb.2016.00906.

Kim, J. W., Ryu, J., and Shim, S. H. (2018). Chemical Investigation on an Endophytic fungus Gibberella moniliformis JS1055 derived from a Halophyte Vitex rotundifolia. Natural Product Sciences 24(3): 189–193 https://doi.org/10.20307/nps.2018.24.3.189.

Kirschner, R. (2018). Fungi on the leaf – a contribution towards a review of phyllosphere microbiology from the mycological perspective. Biochemical Systematics and Ecology 34: 433–448.

Kurtzman, C. & Robnett, C. J. (1997). Identification of clinically important ascomycetous yeasts based on nucleotide divergence in the 5′ End of the Large-Subunit (26S) Ribosomal DNA Gene. Journal of clinical microbiology 35: 1216–1223.

Kurtzman, C.P., Fell, J. W., Boekhout, T. and Robert, V. (2011). The Yeasts, A Taxonomic Study, 5th edn. Amsterdam: Elsevier.

Lin, T. Y., Chen, H. W., and Chen, M. M. (2011). Mapping recent erosional hot spots along the sandy coasts of Taiwan. Journal of Engineering Environment 27: 77–92. http://dx.doi.org/10.6562/JEE.2011.27.5.

Macia-Vicente, J. G., Ferraro, V., Burruano, S. and
Lopez-Llorca, L.V. (2012). Fungal assemblages associated with roots of halophytic and non-halophytic plant species vary differentially along a salinity gradient. Microbial Ecology 64 (3), 668–679.

Manamgoda, D. S., Cai, L., McKenzie, E. H. C. et al. (2012). A phylogenetic and taxonomic re-evaluation of the Bipolaris - Cochliobolus - Curvularia Complex. Fungal Diversity 56: 131. https://doi.org/10.1007/s13225-012-0189-2

Maun, M. A. (2004). Burial of Plants as a Selective Force in Sand Dunes. In Martínez, M.L. & Psuty, N.P. (Eds.) Coastal Dunes, Ecology and Conservation. Ecological Studies 171: 119–135. Springer-Verlag, Berlin/Heidelberg, Germany.

Nair, D. N. and Padmavathy, S. (2014). Impact of Endophytic Microorganisms on Plants, Environment and Humans. The Scientific World Journal Volume 2014, Article ID 250693. http://dx.doi.org/10.1155/2014/250693

Nilsson, R.H., Ryberg, M., Kristiansson, E., Abarenkov, K., Larsson, K. H., and Kõljalg, U. (2006). Taxonomic reliability of DNA sequences in public sequence databases: a fungal perspective. PLOS ONE 1(1): e59. https://doi.org/10.1371/journal.pone.0000059.

Okane, I., and Nakagira, A. (2015). Assemblages of endophytic fungi on Salicornia europaea disjunctively distributed in Japan: towards clarification of the ubiquity of fungal endophytes on halophytes and their ecological roles. Current Science 109: 62–71.

Paliany, A. S., Sivasothy, Y., Awang, K., Rizman-Idid, M. and Alias, S. A. (2014). Marine Derived Fungi of Peninsular Malaysia – a Biochemical Perspective. Chiang Mai Journal of Science 41(4): 894–909.

Park, J. H. and Park, M. J. (2013). First Report of Corynespora Leaf Spot on Beach Vitex Caused by Corynespora cassiicola in Korea. Plant disease 97(11): 1512. https://doi.org/10.1094/PDIS-05-13-0480-PDN

Petrini, L. E. (2013). Rosellinia: a world monograph. Bibliotheca Mycologica 205: 1– 410.

Petrini, O. and Fisher, P. J. (1988). A comparative study of fungal endophytes in xylem and whole stems of Pinus sylvastris and Fagus sylvatica. Transactions of the British Mycological Society 91: 233–238.

Prenafeta-Boldú, F. X., Summerbell, R. C., Boer, W. D., Boschker, H. T. S., and Gams, W. (2014). Biodiversity and ecology of soil fungi in a primary succession of a temperate coastal dune system. Nova Hedwigia 99: 347–372.

Redman, R.S., Sheehan, K. B., Stout, R.G., Rodriguez, R. J. and Henson, J. M. (2002). Thermotolerance conferred to plant host and fungal endophyte during mutualistic symbiosis. Science 298: 1581.

Rodriguez, R. J., J. Henson, E. V. Volkenburgh, M. Hoy, L. Wright, F. Beckwith, Y.-O. Kim and R. S. Redman. (2008). Stress tolerance in plants via habitat-adapted symbiosis. The ISME Journal 2:404–416.

Sandoval-Denis, M., Gené, J., Sutton, D.A., Wiederhold, N.P., and Guarro, J. (2015). Acrophialophora, a poorly known fungus with clinical significance. Journal of Clinical Microbiology 53(5):1549–1555. DOI: 10.1128/JCM.00279-15.

Schoch, C.L., Seifert, K.A., Huhndorf, A., Robert, V., Spouge, J. L., Levesque, C. A., Chen, W., and Fungal Barcoding Consortium. (2012). Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi. PNAS 109: 6241–6246.

Simmons, E.G. (2007). Alternaria: an identification manual. CBS Fungal Biodiversity Centre, The Netherlands.

Sohn, S.-H., Ko, E., Oh, B. G., Kim, S. H., Kim, Y., Shin, M., and Bae, H. (2009). Inhibition effects of Vitex rotundifolia on inflammatory gene expression in A549 human epithelial cells. Annals of Allergy, Asthma and Immunology 103(2):152–159.

Stierle, A., Strobel, G. and Stierle, D. (1993). Taxol and taxane production by Taxomyces andreanae, an endophytic fungus of Pacific yew. Science 260: 214–216.

Tamura K., Stecher G., Peterson D., Filipski A., Kumar S. (2013). MEGA6: Molecular Evolutionary Genetics Analysis version 6.0. Molecular Biology and Evolution 30: 2725−2729.

Tong, X. L. and Lin, T. Y. (2016). Dune restoration experiments during a typhoon season on Taiwan’s Si-Cao coast. Journal of Marine Science and Technology 24(5): 1032–1040. DOI: 10.6119/JMST-016-0526-1.
Tsai, J. N., Ann, P. J., Cheng, H. F., Hsu, Z. H., and

Huang, H. C. (2015). First Report of Corynespora cassiicola Causing Leaf Spot of Papaya in Taiwan. Plant disease 99(11): 1649. https://doi.org/10.1094/PDIS-12-14-1250-PDN

Turhan, G. and Grossmann, F. (1989). Antifungal and Antibacterial Activity of Acrophialophora levis Samson and Tariq Mahmood. Phytopathology124: 200–206.

Waqas, M., Khan, A. L., Hamayun, M., Shahzad, R., Kang, S. M., Kim, J. G., and Lee, I. J. (2015). Endophytic fungi promote plant growth and mitigate the adverse effects of stem rot: an example of Penicillium citrinum and Aspergillus terreus. Journal of Plant Interactions 10(1):280–287. http://dx.doi.org/10.1080/17429145.2015.1079743.

Water Resources Agency, MOEA (2012).The overview of coast in Taiwan. Retrieved February, 2019 from https://web.archive.org/web/20130806042806/http://www.wra.gov.tw/ct.asp?xItem=12592&CtNode=3133.

White, T. J., Bruns, T. D., Lee, S. B., and Taylor, J. W. (1990). Amplification and direct sequencing of fungal ribosomal RNA Genes for phylogenetics. In book: PCR - Protocols and Applications - A Laboratory Manual, pp. 315–322, USA, Academic Press.

Wikee, S., Lombard, L., Nakashima, C., Motohashi, K., Chukeatirote, E., Cheewangkoon, R., McKenzie, E. H. C., Hyde, K. D., and Crous, P. W. (2013). A phylogenetic re-evaluation of Phyllosticta (Botryosphaeriales). Studies in Mycology 76(1): 1–29. DOI: 10.3114/sim0019.

Wilson, D. (1995). Endophyte: The Evolution of a Term, and Clarification of Its Use and Definition. Oikos 73: 274–276.

Woudenberg, J. H., Aveskamp, M. M., de Gruyter, J., Spiers, A. G., and Crous, P. W. (2009). Multiple Didymella teleomorphs are linked to the Phoma clematidina morphotype. Persoonia 22: 56–62.

Yang, J. W. (2016). Study of endophytic and non-endophytic fungi from the coastal grass Spinifex littoreus in Taiwan by combining culture-dependent, culture-independent and opportunistic collection approaches. Master’s thesis, National Central University, Taoyuan, Taiwan.

Yang, J. W., Yeh, Y. H and Kirschner, R. (2016). A new endophytic species of Neostagonospora (Pleosporales) from the coastal grass Spinifex littoreus in Taiwan. Botany 94: 593–598. dx.doi.org/10.1139/cjb-2015-0246.

Yeh, Y. H and Kirschner, R. (2014). Sarocladium spinificis, a new endophytic species from the coastal grass Spinifex littoreus in Taiwan. Botanical Studies 55:25. DOI: 10.1186/1999-3110-55-25.

Yeh, Y. H and Kirschner, R. (2017). First Report of Corynespora Leaf Spot on Vitex rotundifolia Caused by Corynespora cassiicola in Taiwan. Plant Disease 101(8): 1550.

Yeh, Y. H. (2013). Endophytic fungi from the salt-tolerant coast plant Spinifex littoreus in Taiwan. Master’s thesis, National Central University, Taoyuan, Taiwan.

Yin, Y., Ding, Y., Feng, G., et al. (2015). Modification of artificial sea water for the mass production of (+)-terrein by Aspergillus terreus strain PF26 derived from marine sponge Phakellia fusca. Letters in Applied Microbiology 61:580–587.

Zalar, P., Gostinčar, C., de Hoog, G.S., Uršič, V., Sudhadham, M. and Gunde-Cimerman, N. (2008). Redefinition of Aureobasidium pullulans and its varieties. Studies in Mycology 61: 21–38.

Zhang, Y., Liu, F., Wu, W., Cai, L. (2015). A phylogenetic assessment and taxonomic revision of the thermotolerant hyphomycete genera Acrophialophora and Taifanglania. Mycologia 107(4): 768–779.

Zhao, J., Zhou, L., Wang, J., Shan, T., Zhong, L., Liu, X., and Gao, X. (2010). Endophytic fungi for producing bioactive compounds originally from their host plants. Current Research, Technology and Education Topics in Applied Microbiology and Microbial Biotechnology 1: 567–576.

Zhou, Y., Gong, G., Zhang, S., Liu, N., Wang, J., Li, P., Yu, X. (2014). A new species of the genus Trematosphaeria from China. Mycological Progress 13:33–43.

指導教授

羅南德孫維欣(Roland Kirschner Wei-Hsin Sun)

審核日期

2019-4-18

推文