Organophosphorus pesticides(OPPs) are a set of toxic persistent organic pollutants(POPs) present in the environment. Recently, biochar-mediated bioremediation has exhibited many advantages over conventional methods for the remediation of pesticide-contaminated soil. In the present study, biochar and nitrogen fertilizer(NH_4NO_3)were employed to remediate OPP-contaminated soil and the greenhouse gas(GHG) emission during 90 days of incubation was investigated. After thermal desorption treatment, the content of organophosphorus pesticides reduced from 175.61 μg·kg^(-1) to 62.68 μg·kg^(-1). The addition of NH_4NO_3 in the following bioremediation led to larger reduction(34.35%) of the pesticide concentration than that of biochar(31.90%) for the contaminated soils with thermal desorption treatment, while the simultaneous addition of biochar and NH_4NO_3 led to the largest reduction of pesticide concentration(11.07%) for the soil without thermal desorption treatment. The addition of biochar and NH_4NO_3 only slightly increased the emission rate of GHGs from the soil without thermal treatment,but remarkably increased the emission rate of GHGs from the soil after thermal treatment. In most cases, the addition of NH_4NO_3 is more effective than biochar to promote the degradation of pesticide, but also exhibited higher GHG emission. The microbial community analysis suggests that the enhanced degradation of pesticide is mainly owing to the increased activity of microorganism.
建立了SYBR Green I实时荧光定量聚合酶链式反应(Real Time-qPCR)检测油田污染土壤中烷烃降解基因AlkB和萘降解基因Nah的方法。比对相关降解石油菌株的GenBank序列,设计合成针对烷烃和萘降解基因扩增引物AlkBf/AlkBr和Nahf/Nahr。将纯化的常规PCR胶回收产物与pEASY-T1载体连接,转化到感受态细胞培养。提取并梯度稀释阳性克隆质粒,构建Real Time-qPCR标准测定曲线。25μL扩增体系最佳反应条件:前后引物终浓度为0.2μmol/L,12.5μL 2×TransStart Top Green qPCR SuperMix,AlkB和Nah基因最适退火温度分别为50℃和57℃。Real Time-qPCR技术显示出很高的灵敏性和重复性,比传统PCR技术灵敏度高100倍。对采集于某油田3个功能区的14土壤样品中AlkB定量检测显示,石油污染严重的采油区含有最高的AlkB拷贝数,污染较轻的生活区AlkB拷贝数最少;Nah基因分布均匀。
The present work aimed to develop a novel strategy to bioremediate the petroleum hydrocarbon contaminants in the environment.Salt tolerant bacterium was isolated from Dagang oilfield,China and identified as Corynebacterium variabile HRJ4 based on 16 S r RNA gene sequence analysis.The bacterium had a high salt tolerant capability and biochar was developed as carrier for the bacterium.The bacteria with biochar were most effective in degradation of n-alkanes（C16,C18,C19,C26,C28） and polycyclic aromatic hydrocarbons（NAP,PYR） mixture.The result demonstrated that immobilization of C.variabile HRJ4 with biochar showed higher degradation of total petroleum hydrocarbons（THPs） up to 78.9%after 7-day of incubation as compared to the free leaving bacteria.The approach of this study will be helpful in clean-up of petroleum-contamination in the environments through bioremediation process using eco-friendly and cost effective materials like biochar.
Hairong ZhangJingchun TangLin WangJuncheng LiuRanjit Gajanan GuravKejing Sun
