微生物學(xué)報Acta Microbiologica Sinica
48(11)：1479~1485; 4 November 2008
多功能農(nóng)藥降解基因工程菌株m-CDS-1環(huán)境釋放安全評價
蔣建東,，李榮，郭新強,，陳凱,，李順鵬*
(南京農(nóng)業(yè)大學(xué)生命科學(xué)學(xué)院，農(nóng)業(yè)部農(nóng)業(yè)環(huán)境微生物工程重點開放實驗室,，南京 210095)
摘要：【目的】為了評價多功能農(nóng)藥降解基因工程菌株m-CDS-1的環(huán)境釋放中間試驗水平的安全性,。【方法】通過農(nóng)藥檢測,、平板計數(shù),、Most probable number-PCR (MPN-PCR)和Denaturing Gradient Gel Electrophoresis (DGGE)等方法在江蘇大豐進行了工程菌株m-CDS-1田間降解農(nóng)藥效果、定殖動態(tài)和對土壤微生物群落結(jié)構(gòu)影響的研究,?！窘Y(jié)果】投加1.01×107 CFU/g干土的工程菌株m-CDS-1在30 d時均能完全降解10.71 mg/kg的甲基對硫磷和1.29 mg/kg的呋喃丹。平板計數(shù)表明工程菌株m-CDS-1在土壤中快速下降,；MPN-PCR檢測結(jié)果顯示,，在4 d，15 d和30 d時,，0～10 cm混合土壤中該工程菌株的數(shù)目分別為2.15±0.98×106 CFU/g干土,，3.70±4.66×104 CFU/g干土和檢測不出。工程菌株m-CDS-1的投加不會對土壤可培養(yǎng)三大微生物菌群數(shù)量產(chǎn)生顯著影響,；基于細菌16S rDNA V3區(qū)的DGGE分析結(jié)果表明,，施加農(nóng)藥對細菌菌落結(jié)構(gòu)有顯著影響，4 d,，11 d,，30 d時農(nóng)藥施用區(qū)與空白對照區(qū)的圖譜相似性分別為17.16%，49.81%和75.01%,，但到60 d時的相似性為98.62%,。工程菌株m-CDS-1釋放在前期對細菌群落結(jié)構(gòu)有一定影響，4 d,，11 d和30 d工程菌株釋放區(qū)相對于空白的相似性分別為49.57%,，38.30 %和83.30%。在60 d時,，空白,、施藥和施菌小區(qū)的圖譜相似性都在90%以上,。【結(jié)論】工程菌株m-CDS-1不僅可同時高效降解甲基對硫磷和呋喃丹,，仍保持了實驗室內(nèi)的原有特性,，而且不會成為優(yōu)勢菌群長期在土壤環(huán)境中存在，也不會對土壤微生物群落結(jié)構(gòu)造成長期影響,。
關(guān)鍵詞：基因工程菌株m-CDS-1,；環(huán)境釋放；安全評價,；農(nóng)藥降解,；Most probable number-PCR (MPN- PCR)；Denaturing Gradient Gel Electrophoresis (DGGE)
中圖分類號：Q933 文獻標識碼：A 文章編號：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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