微生物学报Acta Microbiologica Sinica
48(11)：1479~1485; 4 November 2008
多功能农药降解基因工程菌株m-CDS-1环境释放安全评价
蒋建东，李荣，郭新强，陈凯，李顺鹏*
(南京农业大学生命科学学院，农业部农业环境微生物工程重点开放实验室，南京 210095)
摘要：【目的】为了评价多功能农药降解基因工程菌株m-CDS-1的环境释放中间试验水平的安全性。【方法】通过农药检测、平板计数、Most probable number-PCR (MPN-PCR)和Denaturing Gradient Gel Electrophoresis (DGGE)等方法在江苏大丰进行了工程菌株m-CDS-1田间降解农药效果、定殖动态和对土壤微生物群落结构影响的研究。【结果】投加1.01×107 CFU/g干土的工程菌株m-CDS-1在30 d时均能完全降解10.71 mg/kg的甲基对硫磷和1.29 mg/kg的呋喃丹。平板计数表明工程菌株m-CDS-1在土壤中快速下降；MPN-PCR检测结果显示，在4 d，15 d和30 d时，0～10 cm混合土壤中该工程菌株的数目分别为2.15±0.98×106 CFU/g干土，3.70±4.66×104 CFU/g干土和检测不出。工程菌株m-CDS-1的投加不会对土壤可培养三大微生物菌群数量产生显著影响；基于细菌16S rDNA V3区的DGGE分析结果表明，施加农药对细菌菌落结构有显著影响，4 d，11 d，30 d时农药施用区与空白对照区的图谱相似性分别为17.16%，49.81%和75.01%，但到60 d时的相似性为98.62%。工程菌株m-CDS-1释放在前期对细菌群落结构有一定影响，4 d，11 d和30 d工程菌株释放区相对于空白的相似性分别为49.57%，38.30 %和83.30%。在60 d时，空白、施药和施菌小区的图谱相似性都在90%以上。【结论】工程菌株m-CDS-1不仅可同时高效降解甲基对硫磷和呋喃丹，仍保持了实验室内的原有特性，而且不会成为优势菌群长期在土壤环境中存在，也不会对土壤微生物群落结构造成长期影响。
关键词：基因工程菌株m-CDS-1；环境释放；安全评价；农药降解；Most probable number-PCR (MPN- PCR)；Denaturing Gradient Gel Electrophoresis (DGGE)
中图分类号：Q933 文献标识码：A 文章编号：0001-6209 (2008) 11-1479-07
Risk Assessment for Environmental Release of Genetically Engineered Mi-croorganism m-CDS-1
Jiandong Jiang, Rong Li, Xinqiang Guo, Kai Chen, Shunpeng Li
(College of Life Sciences, Key Laboratory for Microbiological Engineering of Agricultural Environment of Ministry of Agriculture, Nanjing Agricultural University, Nanjing 210095, China)
Abstract: [Objective] To assess the environmental release risk of genetically engineered microorganism Sphingomonas sp. m-CDS-1. [Methods] Pesticides detection, plates counting, most probable number-PCR (MPN-PCR) and denaturing gradient gel electrophoresis (DGGE) were used to study the pesticides degrading effects, the decline dynamics of m-CDS-1, and the effects of release of m-CDS-1 on community structure of soil microorganisms. [Results] m-CDS-1 (1.01× 107CFU/g dry soil) could degrade both 10.71 mg/kg methyl parathion and 1.29 mg/kg carbofuran in 30 days. m-CDS-1 declined quickly with time, and could not be detected anymore in 30 days. Release of m-CDS-1 would not change the number of the cultivable microorganisms notably. The similarities of PCR-DGGE profiles between the pesticides applied treatments and the controls in 4, 11 and 30 days were 17.16 %, 49.81 % and 75.01 %, and the similarities between the m-CDS-1 released treatments and controls were 49.57 %, 38.3 % and 83.3 %, respectively. In 60 days, DGGE profiles of all treatments and controls had high similarities. [Conclusion] m-CDS-1 could degrade both pesticides remarkably in the field, and would not be the predominant population, and would not affect the community structure of soil microorganisms in the long term.
Keywords: genetically engineered microorganism m-CDS-1; environmental release; risk assessment; pesticides degradation; most probable number-PCR (MPN-PCR); denaturing gradient gel electrophoresis (DGGE)
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