目录
摘要II
关键词II
Abstract....III
引言
引言1
1材料与方法2
1.1菌株和培养基........2
1.2药品，抗生素和化学试剂2
1.3吡蚜酮的检测方法3
1.4吡蚜酮浓度标准曲线绘制3
1.5吡蚜酮降解菌株的分离与纯化3
1.6降解菌株形态特征鉴定3
1.7降解菌株16S rRNA基因序列测定分析..4
1.8静息细胞的制备5
1.9温度对菌株BYT2降解吡蚜酮的影响研究..5
1.10菌株BYT2对吡蚜酮的生长降解特性研究..............5
1.11菌株BYT2降解吡蚜酮的代谢途径分析5
1.11.1吡蚜酮代谢产物的鉴定....5
1.11.2菌株BYT2以吡蚜酮和NA为唯一碳源的生长特性研究......5
1.12菌株BYT2吡蚜酮水解酶基因的扩增6
2结果与分析6
2.1菌株的分离纯化6
2.2菌株BYT2的菌落形态7
2.3菌株BYT2的16S rRNA基因序列分析.....................7
2.4温度对菌株BYT2降解吡蚜酮的影响9
2.5菌株BYT2对吡蚜酮的生长降解特性9
2.6菌株BYT2对吡蚜酮的代谢途径研究...10
2.7菌株BYT2以吡蚜酮和NA为唯一碳源的生长情况.............11
2.8菌株BYT2吡蚜酮水解酶基因.......12
3讨论 13
致谢14
参考文献15
吡蚜酮降解菌的分离及其降解特性研究
摘 要
吡蚜酮（pymetrozine）是一种高效、低毒、高选择性、无交互抗性、对环境友好的新型吡啶甲亚胺类杀虫剂。但是随着吡蚜酮的使用范围越来越广，在环境和农产品中检测出吡蚜酮的残留，对生态环境和人类健康构成了潜在的威胁。美国环保局（USEPA）已将吡蚜酮定义为可能引 *51今日免费论文网|www.51jrft.com +Q: ^351916072* 
起人类癌症的物质。因此，人们日益关注吡蚜酮降解机制及代谢途径的研究，虽然目前已有人报道过吡蚜酮降解菌及其代谢途径，但有关吡蚜酮降解相关酶和基因尚未有深入研究。本课题通过对从农药厂采集的活性污泥样本进行富集培养，筛选分离获得了一株能以吡蚜酮作为唯一碳源生长的吡蚜酮降解菌株Hydrogenophaga sp. BYT2。该菌株对吡蚜酮的降解效果较好,培养温度为30°C时，在培养24 h后，菌株BYT2对吡蚜酮的降解效率高达90%左右。本课题研究了菌株BYT2对吡蚜酮的生长降解特性，鉴定其代谢产物，解析代谢途径，发现在30°C条件下菌株BYT2能在48 h内完全降解1.40 mM的吡蚜酮，将其降解为4氨基6甲基4,5二氢 [1,2,4]三嗪3酮(AMDT)和烟酸(NA)，最后成功扩增得到了吡蚜酮水解酶基因。
ISOLATION OF PYMETROZINEDEGRADING STRAIN AND RESEARCH OF DEGRADATION CHARACTERISTICS
ABSTRACT
Pymetrozine is a new type of pyridine azomethine insecticide which has highefficiency, lowtoxicity, highselectivity, no cross resistance and environmentfriendliness. With the increasing use of pymetrozine, the residues of pymetrozine can be detected in the environment and agricultural products, and thus poses a potential threat to the ecological environment and human health. The US Environmental Protection Agency (USEPA) has defined pyridoxine as a possible human carcinogen. Therefore, people are paying more and more attention to the research on the degradation mechanism and metabolic pathways of pymetrozine. Although there have been reports of pymetrozinedegrading bacteria and their metabolic pathways, there are no indepth studies on enzymes and genes related to pymetrozine degradation. In this project, an enriched culture of activated sludge samples collected from a pesticide plant was used to obtain a pyrithionedegrading strain Hydrogenophaga sp. BYT2 which can grow with pymetrozine as the sole carbon source. This strain had a good degradation effect on pymetrozine. When the cultivation temperature was 30°C, the degradation efficiency of pymetrozine by strain BYT2 is up to about 90% after 24 hours of cultivation. In this project, the growth and degradation characteristics of pymetrozine by strain BYT2 were studied, the metabolite was identified, and the metabolic pathway was analyzed. It was found that the strain BYT2 could completely degrade 1.40 mM pymetrozine within 48 h at 30°C, degraded to 4amino6methyl4,5dihydro2H[1,2,4] triazin3one (AMDT) and nicotinic acid (NA). Finally the pymetrozine hydrolase gene was successfully amplified.

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