摘要：采集长期施用多菌灵的土壤，从中分离出一株能够有效降解多菌灵的菌株D3。结合菌株形态特征及16S rDNA系统发育分析，将菌株D3鉴定为分枝杆菌属（Mycobacterium sp。）并对菌株D3进行生长和降解特性研究。菌株D3能够利用多菌灵做为唯一碳氮源生长，且在72 h内将50 mg∙L-1的多菌灵几乎全部降解。菌株D3生长的最适pH为6。0-7。0，最适NaCl浓度为0-5g∙L-1,最佳碳源为葡萄糖，最佳有机氮源为蛋白胨，无机氮源为KNO3，具有多粘菌素B和克林霉素抗性。使用响应面法对菌株降解多菌灵的条件进行优化，模拟出最佳降解条件：温度35。07℃，pH值7。17，接种量14。33%。72417

毕业论文关键词：多菌灵；微生物降解；生物学特性；降解特性

ISOLATION AND CHARACTERIZATION OF CARBENDAZIM-DEGRADING STRAINS

Abstract: An efficient carbendazim-degrading bacterial strain D3 was isolated from carbendazim contaminated soil by enrichment culture。 Strain D3 was identified as Mycobacterium sp。 according to physiological and 16S rDNA gene sequence analysis。 Strain D3 could grow with carbendazim as the sole carbon and nitrogen sources, and degraded 50 mg∙L-1 carbendazim within 72 h。 The optimum pH was 6。0-7。0, optimum concentration of NaCl was 0-5g∙L-1, the optimum carbon source was glucose, the optimum organic nitrogen source was peptone, the inorganic nitrogen sources was potassium nitrate and the strain D3 had the ability of resisting Polymyxin B and clindamycin。 Response surface method was used to find the most optimum conditions for carbendazim degradation。 The most optimum conditions for carbendazim degradation by strain D3 was 35。09℃, pH 7。17 and inoculation amount 14。33% 。

Key words: Carbendazim; Biodegradation; Microbial degradation; Identification 

目  录

摘要 1

关键词 1

Abstract 1

Key words 1

引  言 2

1。 材料与方法 2

1。1 培养基与试剂 2

1。2 菌株与质粒 3

1。3 多菌灵降解菌株的分离、筛选及鉴定 3

1。3。1 多菌灵降解菌的分离 3

1。3。2 多菌灵降解菌的筛选 3

1。3。3 菌株基因组总DNA的提取 3

1。3。4 降解菌株16S rDNA基因序列的鉴定 4

1。3。3。1 菌株16S rDNA基因序列扩增 4

1。3。3。2 对PCR产物进行TA克隆 4

1。3。3。3 质粒DNA的小量提取 4

1。3。3。4 降解菌株16S rDNA基因序列测定 5

1。3。3。5 降解菌株系统发育地位的确定 5

1。4 降解菌株生长量的测定 5

1。5 菌株D3对多菌灵降解的研究 5

1。5。1 测量方法 5

1。5。2 多菌灵标准曲线的制作 6

1。6 菌株的培养和接种 6

2。 结果与分析 6

2。1多菌灵降解菌株的分离与筛选 6

2。2 降解菌株D3的分类学鉴定