由於全球暖化與人類活動導致海平面上升，沙岸棲息地逐漸受到威脅。此現象對於生物多樣性和生物資源的影響尚不清楚。沿海植物能夠適應極端的氾濫和乾旱、土壤養分貧乏、高鹽度、強風和太陽輻射，對保護和穩定台灣西部沙岸遭受侵蝕威脅的土地至關重要。當這些棲息地喪失時，一些沿海植物仍然在植物園(非原生環境)中保存。於為期三年的研究中，我們將研究遷地保育對於與沙灘沿海植物密切相關的微生物生物多樣性的影響。其中，我們選擇馬鞍藤(Ipomoea pes-caprae)作為代表性的沿海植物，其廣泛分佈於熱帶沙岸，並以四個天然海濱和三個植物園作為採集地點，且將該植物體內之內生真菌作為代表性微生物。在依賴培養法中，內生真菌將在無菌條件下分離，並依據某些屬和種之特定分類學標準以ITS序列以及其他DNA鑑定條碼(Barcode)（TEF, TUB, RPB2, CAL and other genes）進行物種鑑定。本研究除了會產生超過一千種新菌株之外，也將以ITS序列資料作為基礎的次世代定序進行小規模的測試，進而了解不依賴培養法是否有其他物種(例如：依賴培養法未發現之物種)及物種數量。此外，於本計畫的平行合作中，於依賴培養法所得之菌株將進行降解污染水源中合成染料(synthetic dyes)的測試以篩選其應用潛力。最後，本研究將比較自然與人工採集地之間的物種組成和物種多樣性的差異，以提供關於非原生環境植物保護對相關微生物生物資源影響的數據，作為自然保護和生物保護的決策基礎。 ;The sand coast habitats are endangered by rising sea levels due to global warming and human activities. The effects on biodiversity and biological resources are widely unknown. Coastal plants are adapted to extremes of inundation and drought, poor soil nutrients, high salinity, strong wind and solar irradiation and are of utmost importance for protecting and stabilizing the threatened land along the sand shore of western Taiwan. When these habitats are lost, some coastal plants are still preserved ex-situ in botanical gardens. In our study designed for three years, we investigate the effect of such ex-situ conservation of on the biodiversity of microbes intimately associated with such plants at sand beaches. We choose Ipomoea pes-caprae as representative coastal plant which is widespread at tropical sand coasts, four natural coastal populations and two botanical gardens as collection sites, and the endophytic fungi associated with this plant as representative microbes. In a culture-dependent approach, endophytic fungi are isolated under axenic conditions and identified to species with the universal ITS marker as well as additional barcodes (TEF, TUB, RPB2, CAL and other genes) according to the specific taxonomic requirements in certain genera and species groups of fungi. In addition to this main topic yielding over one thousand new strains, next generation sequencing based on ITS markers is applied for testing at small scale whether this culture-independent method will reveal other species or species numbers. Also in collaboration parallel to this project, strains from the culture-dependent approach are to be screened for their application potential by their ability to degrade synthetic dyes which cause wastewater pollution. The species compositions and diversities between the natural and artificial collection sites are compared in order to provide data about the effects of ex-situ plant conservation on the associated microbial bioresources as foundation for decision-making in nature protection and bioconservation.
