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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/53580

Title: 臺灣烈嶼地區廈門老鼠簕真菌群落多樣性與其抗菌和抗發炎活性之探討
Diversity of fungi associated with leaves of Acanthus ilicifolius var. xiamenensis at Liuyu Township, Taiwan and their antimicrobial and anti-inflammatory activities
Authors: Chi, Wei-Chiung
季偉瓊
Contributors: NTOU:Institute of Marine Biology
國立臺灣海洋大學:海洋生物研究所
Keywords: 廈門老鼠簕;抑菌活性;培養體學;內生真菌;高通量定序
Acanthus ilicifolius var. xiamenensis;Endophytes;High throughput sequencing;Anti-microbial activity;Culturomics
Date: 2019
Issue Date: 2020-07-03T08:34:09Z
Abstract: 本論文以金門地區藥用紅樹林植物“廈門老鼠簕"之共生真菌為研究對象,探討其多樣性及其活性物質的可利用性。從傳統培養法中所分離得到的內生真菌約有203株,從中可鑑定出47種不一樣的菌種來:分別有104株來自冬天,可鑑定出的菌種為30種;99株來自夏天,可鑑定出的菌種為26種。子囊菌門(Ascomycota)為主要的門別,而擔子菌門(Basidiomycota)只有出現在夏天的樣本裡。在菌種分類上,分別以Drechslera dematioidea (約佔10.58%)、Colletotrichum sp.3(約佔7.69%) 和Alternaria sp.(約佔7.69%)為冬天主要的種類;在夏天則以Fusarium oxysporum (約佔13.13%)、Diaporthe endophytica (約佔10.10%) 和Colletotrichum sp.1(約佔9.09%)為主要的種類。整體來說,佔有率最高的種類分別為Corynespora cassiicola (約佔6.90%)、Fusarium oxysporum (約佔6.40%) 和Guignardia sp. (約佔6.40%)。在非傳統培養法(高通量定序分析)中,從17片葉子中所得到的菌種分析為有111個操作分類單元,其中有鑑定出屬名的數量有86個;而冬天可得到96個操作分類單元,夏天則有70個操作分類單元。在兩個採樣季節都可得到來自子囊菌門和擔子菌門的基因序列,子囊菌門的基因序列明顯比擔子菌門多。根據序列的豐富度,擁有最高出現頻率的菌種分別為:在冬天是Alternaria sp. (約佔3.46%)、Cladosporium delicatulum (約佔2.56%) 和Pyrenochaetopsis leptospora (約佔1.41%);在夏天則是Aureobasidium sp. (約佔10.72%), Cladosporium sp. (約佔7.90%)、Cladosporium delicatulum (約佔3.45%) 和Hortaea werneckii (約佔3.21%),且此四種菌在整體的佔有率也是最高的。在活性物質篩選方面,針對五種抑制菌株和抗發炎能力來說,則有28株菌株的二次代謝物具有此能力。大部分的菌株之菌液粗萃物都具有抑制格蘭氏陽性菌生長的能力,其中有少部分菌株則會抑制格蘭氏陰性菌中的大腸桿菌:像是Didymella sp. NTOU4354和 NTOU4878、Corynespora cassiicola NTOU4889 和 Xylaria sp. NTOU4900等。在抑制真菌方面,在子囊菌門中的Diaporthales和Glomerellales這兩個目,以及擔子菌門中的Hymenochaetales和 Polyporales這兩個目特別能抑制真菌Candida albicans。Diaporthe endophytica NTOU4920 (Diaporthales)、Phellinus noxius NTOU4917 (Hymenochaetales)和Tinctoporellus epimiltinus NTOU4916 (Polyporales)能抑制真菌Cryptococcus neoformans;菌株Colletotrichum spp.、Phellinus noxius NTOU4917 和Phomopsis sp. 2 NTOU4918則能抑制真菌Candida albicans。在抗發炎方面,有三株菌的二次代謝物的粗萃物具有抑制nitric oxide (NO)低於15%產生的能力和讓細胞存活率高於100%的潛力,分別是Phoma sp. 2 NTOU4338、Nodulisporium sp. NTOU4868 和 Guignardia sp. NTOU4871。根據以上數據結果顯示,藥用紅樹林植物“廈門老鼠簕"之共生真菌不但具有高度的菌種豐富度外還潛藏著發現新穎藥物的潛力。
Diversity and biological activities of the fungi associated with the Kinmen medicinal mangrove plant Acanthus ilicifolius var. xiamenensis were studied. A total of 203 isolates were cultured from surface-sterilized leaves, representing 47 different fungal species: 104 isolates, 30 species in winter and 99 isolates 26 species in summer. Ascomycota was dominant in the leaf samples, with Basidiomycota isolated only in the summer samples. Drechslera dematioidea (10.58%, percentage occurrence), Colletotrichum sp. 3 (7.69%) and Alternaria sp. (7.69%) were dominant in the winter samples; Fusarium oxysporum (13.13%), Diaporthe endophytica (10.10%) and Colletotrichum sp. 1 (9.09%) in the summer samples. Overall, Corynespora cassiicola (6.90%), Fusarium oxysporum (6.40%) and Guignardia sp. (6.40%) had the highest overall percentage occurrence. In the metabarcoding analysis, a total of 111 operational taxonomic units (OTUs) were identified from 17 leaf samples, within which 86 could be given a generic name: 96 OTUs from the winter and 70 OTUs from the summer samples. Sequences belonging to the Ascomycota and the Basidiomycota were detected in both samples but the former phylum was dominant over the latter. Based on sequence abundance, taxa having the highest percentage occurrence included Alternaria sp. (3.46%), Cladosporium delicatulum (2.56%) and Pyrenochaetopsis leptospora (1.41%) in the winter, and Aureobasidium sp. (10.72%), Cladosporium sp. (7.90%), Cladosporium delicatulum (3.45%) and Hortaea werneckii (3.21%) in the summer. These latter four species also had the highest percentage occurrence overall. Among the 168 fungal isolates used for a natural product screening, twenty-eight fungal (16.67%) extracts displayed biological activities to at least one tested bacterial and fungal pathogens and iNOS inhibitory activities. Most extracts were active against the Gram-positive bacteria, but only the extracts of Didymella sp. NTOU4354 and NTOU4878, Corynespora cassiicola NTOU4889 and Xylaria sp. NTOU4900 were active against Escherichia coli. Only the ascomycetous orders Diaporthales, Glomerellales, the basidiomycetous orders Hymenochaetales and Polyporales were able to inhibit growth of the pathogenic fungi, especially Candida albican. Growth of Cryptococcus neoformans were only inhibited by extracts of Diaporthe endophytica NTOU4920 (Diaporthales), Phellinus noxius NTOU4917 (Hymenochaetales) and Tinctoporellus epimiltinus NTOU4916 (Polyporales). For Candida albicans, the extracts of Colletotrichum spp., Phellinus noxius NTOU4917 and Phomopsis sp. 2 NTOU4918 were active against it. Meanwhile, three extracts showed low cytotoxicity (i.e. >100% cell viability) and high iNOS inhibition (<15% of NO production) of cells and they were Phoma sp. 2 NTOU4338, Nodulisporium sp. NTOU4868 and Guignardia sp. NTOU4871. These results aboved indicate that not only the fungal community of A. ilicifolius var. xiamenensis displayed high richness, but also this medicinal plant may be a source for novel drugs.
URI: http://ethesys.lib.ntou.edu.tw/cgi-bin/gs32/gsweb.cgi?o=dstdcdr&s=G0D96340002.id
http://ntour.ntou.edu.tw:8080/ir/handle/987654321/53580
Appears in Collections:[海洋生物研究所] 博碩士論文

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