以菌株Pseudomonas putida KT2440为宿主构建多功能农药降解菌

发布时间：2018-06-18 14:30

本文选题：同源重组 + 农药微生物降解　；参考：《应用与环境生物学报》2016年06期

【摘要】：通过富集培养法分离到的农药降解菌株存在稳定性差、底物谱窄、安全性不明等问题.Pseudomonas putida KT2440分离自根际土壤,是国际上公认的环境安全型菌株.利用同源重组方法将4种农药降解酶基因pytH、mpd、chd和amy A同时整合到P.putida KT2440的染色体上,获得了一株无抗性基因残留的工程菌P.putida KT-pmca.工程菌KTpmca能够同时降解拟除虫菊酯类杀虫剂、有机磷类杀虫剂、百菌清和氯代酰胺类4种农药,百菌清降解率大于95%,甲氰菊酯、毒死蜱和敌稗的降解率均达到80%以上.此外,传代实验证明4种降解酶基因能够在遗传中稳定存在.本研究表明工程菌KT-pmca可为微生物原位修复消除农药污染提供新的菌株资源.
[Abstract]:Pesticide degradation strains isolated by enrichment culture method have some problems, such as poor stability, narrow substrate spectrum and unclear safety. Pseudomonas putida KT2440 is recognized as an environmentally safe strain from rhizosphere soil. By homologous recombination method, four pesticide degrading enzyme genes, pytHnmpdchd and amy A, were integrated into the chromosome of P.putida KT2440 at the same time. An engineering strain P.putida KT-pmca. without resistance-free gene residues was obtained. KTpmca could simultaneously degrade pyrethroid insecticides, organophosphorus insecticides, chlorothalonil and chloramides. The degradation rate of chlorothalonil was more than 95. The degradation rate of chlorpyrifos, chlorpyrifos and dimethyl reached more than 80%. In addition, four biodegradable enzyme genes were proved to be stable in heredity. This study indicated that the engineering strain KT-pmca could provide a new strain resource for in situ remediation and elimination of pesticide pollution.
【作者单位】：南京农业大学生命科学学院;东南大学医学院附属盐城医院中心实验室;
【基金】：国家高技术发展计划(863计划)项目(2012AA101403) 国家自然科学基金项目(J1210056) 江苏省农业科技创新基金项目(CX(15)1004)资助~~
【分类号】：X172;X592

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