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產生抗真菌活性物質的植物內生真菌的分離純化及鑒定

時間:2019-04-19 21:21來源:畢業論文
通過對采集的三十種不同的植物進行內生真菌的分離純化,再通過真菌抑制實驗篩選出具有抗青霉活性的內生真菌,然后提取目標內生真菌的DNA,PCR擴增其ITS并測序,并結合菌絲形態和產

摘要:植物內生真菌普遍認為是從表面已經滅菌的健康植物中分離純化得到的,存在于健康植物的組織器官中,對植物本身不產生明顯侵染的一類真菌[1]。隨著研究的深入,人們發現植物內生真菌的次生代謝產物中含有各種不同的生物活性物質。目前己有多種植物內生真菌被研究出具有抑菌活性。本實驗通過對采集的三十種不同的植物進行內生真菌的分離純化,再通過真菌抑制實驗篩選出具有抗青霉活性的內生真菌,然后提取目標內生真菌的DNA,PCR擴增其ITS并測序,并結合菌絲形態和產孢結構鑒定目標內生真菌的分類地位。通過該實驗分離出了三種具有抑制真菌活性的內生真菌,分別是從桂花中分離得到的擬莖點霉Phomopsis sp.和曲霉屬Aspergillus,從黃楊中分離到的水葫蘆黑斑病菌Alternaria alternata。34671
畢業論文關鍵詞:內生真菌、分離、抑菌活性
Separation Purification and Identification of Antifungal Endophytic fungi in Plant
Abstract:Endophytic fungus is generally considered to be isolated and purified from healthy plants which the surface have been sterilized. They exist in healthy tissues and organs of plants, and they do not produce a significant infestation. As studying deep mycorrhizal fungi in plant, we found secondary metabolites in endophytic fungi contain various biologically active substances. Currently, antibacterial activity substances had been found in a variety of endophytic fungi. In this study, endophytic fungi were isolated and purified from thirty different plants. Antifungal activity fungi were screened through fungi inhibition experiments. Then, DNA of these endophytic fungi was purified. ITS was amplified and sequenced. Their sequence was blasted in NCBI database. At last, the classification status of these endophytic fungi wereidentified by the result of sequence and mycelium morphology. Three antifungal activity endophytic fungi were purified and identified in this experiment. They are Phomopsis sp. and Aspergillus from Osmanthus and Alternaria alternata from Boxwood.
源`自*六)維[論*文'網www.mmeqir.tw

Key words:  Endophytic fungi, Separation, Antifungal activity
目錄
摘要    I
目錄    1
引言    3
1 材料與方法    3
1.1  實驗材料    3
1.1.1 樣品采集    3
1.1.2 藥品    3
1.1.3 培養基    3
1.1.4 菌種    4
1.1.5主要的儀器設備    4
1.2 實驗方法    5
1.2.1 樣品采集    5
1.2.2 清洗    5
1.2.3 取樣    5
1.2.4 表面滅菌    5
1.2.5 平板接種    5
1.2.6 室溫培養    6
1.2.7 菌種純化    6
1.2.8 菌種保藏    7
1.2.9 菌種發酵    7
1.2.10過濾發酵液    7
1.2.11抗真菌試驗    8
1.2.12菌絲形態鑒定    9
1.2.13菌種DNA的提取、擴增、鑒定    9
2 結果    11
    2.1  11號菌-擬莖點霉Phomopsis sp.    11
2.1.1  11號菌青霉抑制梯度實驗結果    11
        2.1.2  11號菌菌絲及產孢結構拍照    12
        2.1.3  11號菌菌種鑒定    13
2.2  164號菌-曲霉屬Aspergillus    13
2.2.1  164號菌青霉抑制梯度實驗結果    13
2.2.2  164號菌菌絲及產孢結構拍照    13
2.2.3  164號菌菌種鑒定    15
2.3  268號菌-水葫蘆黑斑病菌Alternaria alternata    15 產生抗真菌活性物質的植物內生真菌的分離純化及鑒定:http://www.mmeqir.tw/shengwu/20190419/32260.html
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