一株拮抗烟草黑胫病菌放线菌的分子鉴定
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一株拮抗烟草黑胫病菌放线菌的分子鉴定(7000字)
摘 要:目前,对于烟草黑胫病的防治主要以化学防治为主,但化学农药的使用易对环境和生态造成污染,为了寻找能对烟草黑胫病菌有拮抗作用的放线菌,于湖南农业大学烟草基地试验田的土样中分离得到一株F菌株对烟草黑胫病菌有明显抑菌活性,为明确该放线菌的种类,利用原核生物16SrRNA基因通用引物对该放线菌进行PCR扩增和测序,利用MAGE构建系统发育树,再结合形态学、培养特征等对其进行鉴定,初步判断其为暗黑链霉菌(Streptomyces atratus)。
关键词: 放线菌;拮抗作用;16SrRNA
Molecular Identification of a Actinomycetes Strain Antagonistic Tobacco Black Tibial Germs
Abstract:Nowadays, the main means to resist Tobacco black shank is by means of chemical. However, using chemical pesticides will pollute the environment and ecology. In order to find the antagonistic actinomycetes with antagonism to Tobacco black shank, we have a strain numbering F selected from the experimental field of Tobacco black shank based in Hunan Agriculture University which obviously has antibacterial activity to Tobacco black shank germ. By using the universal primers of prokaryote gene 16SrRNA to amplify and sequence the actinomycete, and then use the MAGE to construct phylogenetic tree; and finally, according to morphology, culture characteristic, we preliminary identify it is belong to Streptomyces atratus.
Key words:Actinomycetes; antagonism; 16SrRNA
目 录
摘 要 4
关键词 4
1 前 言 5
2 材料与方法 6
2.1 主要试剂 6
2.2 主要仪器设备 6
2.3 基因组DNA的提取 6
2.4 PCR扩增 7
2.5 琼脂糖凝胶电泳 8
2.6 系统发育分析 8
3 结果与分析 8
3.1 DNA的提取 8
3.2 琼脂糖凝胶电泳 9
3.3 16SrRNA序列分析 10
3.4 系统发育树的构建 11
3.5 F菌株的形态 11
4 结论与讨论 12
4.1 DNA的提取 12
4.2 聚合酶链式反应 13
4.3 结果 14
参考文献 14
致 谢 16
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