《咪唑乙烟酸高效降解细菌的筛选及其原生质体制备的研究》-毕业论文（设计）.doc

PAGE I 东北农业大学学士学位论文 　　　　 学号：咪唑乙烟酸高效降解细菌的筛选及其原生质体制备的研究 学生姓名： 指导教师： 教授 所在院系：资源与环境学院 所学专业：生物技术 研究方向：应用微生物 东 北 农 业 大 学 中国·哈尔滨 摘要 除草剂咪唑乙烟酸是咪唑琳酮类性除草剂。咪唑乙烟酸具有杀草谱广、选择性强、活性高等优点，在世界范围内得到广泛应用。自从1993年咪唑乙烟酸国产化，成本下降，用量大幅增加，在我国大豆产区得到大面积的推广和应用。咪唑乙烟酸残效期长，使下茬敏感作物容易产生药害，因此其在土壤中的归趋行为越来越引起重视。 原生质是有组织的生活物质，是细胞生命活动的物质基础。原生质体是由原生质特化而来。原生质体技术的关键是对细胞壁进行消化，以形成大量的原生质体，并使原生质体能高频率的再生细胞壁，回复到正常细胞状态。 本文利用富集驯化技术从长期生产咪唑乙烟酸的农药厂排污口处污泥及污水中驯化培养得到能够降解咪唑乙烟酸的细菌1株, 标记为P310-1，该菌株能够以咪唑乙烟酸为唯一碳源稳定生长。本实验采用化学方法制备筛选所得的降解菌的原生质体，经测定，当稳定剂为pH6.5、再生培养基的pH为7.5时，菌株P310-1原生质体制备及再生的最佳条件为：溶菌酶浓度、温度、酶解时间条件为：2.9 mg/ mL、35℃、3.50 h。并采用药敏片方法测定降解菌的抗药性，并将其作为遗传标记，为对降解菌的后续原生质体融合研究提供试验数据。 关键词：咪唑乙烟酸；降解菌；分离；原生质体制备 Abstract Herbicide imazethapyr is the imidazolinone herbicide. Imazethapyr is a one of broad spectrum herbicides, also has high selectivity and high activity, therefore it had been widely used around the world. Since 1993, the imazethapyr has been localization, and its cost has reduced, application amount has increased, it began to get the promotion and application of large area in Soybean producing areas of our country. The longer residual period of imazethapyr could make drug misadventure for sensitive crops, so the fate of its in the soil has been more and more attention. Archoplasm is the organizational living material, is the the material basis of cell vital movement, all the archoplasm has the similar basic components and characteristics. Protoplast is specializationed from the archoplasm. The key of protoplast technology is to digest the cell wall to form a large number of the protoplasts, and to make protoplast can regeneration cell wall altofrequently, so that to back to the normal cell state. In this paper, the bacteria 1 degrading imazethapyr can be acquired from the sludge and sewage in the long-term producting-imazethapyr factory by enrichment acclimation technique, labeled as P310-1, this bacteria can be stabilizly live with imazethapyr as the sole carbon.In this