产生抗真菌活性物质的植物内生真菌的分离纯化及鉴定
毕业论文关键词：内生真菌、分离、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.
Key words: Endophytic fungi, Separation, Antifungal activity
目录
摘要 I
目录 1
引言 3
1 材料与方法 3
1.1 实验材料 3
1.1.1 样品采集 3
1.1.2 药品 3
1.1.3 培养基 3
1.1.4 菌种 4
1.1.5主要的仪器设备 4
1.2 实验方法 5
1.2.1 样品采集 5
1.2.2 清洗 5
1.2.3 取样 5
1.2.4 表面灭菌 5
1.2.5 平板接种 5
1.2.6 室温培养 6
1.2.7 菌种纯化 6
1.2.8 菌种保藏 7
1.2.9 菌种发酵 7
1.2.10过滤发酵液 7
1.2.11抗真菌试验 8
1.2.12菌丝形态鉴定 9
1.2.13菌种DNA的提取、扩增、鉴定 9
2 结果 11
2.1 11号菌-拟茎点霉Phomopsis sp. 11
2.1.1 11号菌青霉抑制梯度实验结果 11
2.1.2 11号菌菌丝及产孢结构拍照 12
2.1.3 11号菌菌种鉴定 13
2.2 164号菌-曲霉属Aspergillus 13
2.2.1 164号菌青霉抑制梯度实验结果 13
2.2.2 164号菌菌丝及产孢结构拍照 13
2.2.3 164号菌菌种鉴定 15
2.3 268号菌-水葫芦黑斑病菌Alternaria alternata 15