石油污染土壤富集前后细菌群落组成和共现网络分析

为了探究石油污染土壤中细菌群落在富集过程中的演替规律,试验采用平板划线法、菌落PCR和高通量测序技术,分析了富集前后细菌群落结构、共现网络和核心菌属组成,并对富集后体系中的微生物进行分离鉴定,筛选石油降解菌.研究表明富集体系中可培养微生物隶属于34个属53个种,其中3个为潜在新种微生物,Dietzia maris OS33和Rhodococcus qingshengii OS62-1具有降解石油的能力.高通量测序结果显示,在门分类水平上,富集前后丰度较高的菌门均为Proteobacteria和Actinobacteria,富集后两菌门的丰度可达到97.98%,占据绝对优势;丰度较高菌属由Pseudomonas、Rhodococcus、Bacillus和Xanthomonas转变为Dietzia、Unspecified_Idiomarianceae和Halomonas,标志微生物转变为与石油降解有关的Dietzia.细菌群落共现网络在富集后,网络结构进一步简化且更加稳定,核心微生物转变为与石油降解有关的Pseudomonas、Lysinibacillus、Pseudochrobactrum、Agrobacterium、Lactobacillus,且非石油降解菌P.songnenensis P35可协同石油降解菌D.maris OS33降解石油.     

基于3D响应曲面法与重离子辐射优化海迪茨氏菌降解原油的研究

基于模拟12C6+重离子辐射及生物降解特点,利用中心合成设计及响应曲面法优化了海迪茨氏菌去除原油中有机物过程中互动变量参数因子(辐射剂量、pH值、接种量与温度4个变量参数)及在5个不同水平下所有因素的响应特性,并利用GC-MS进行表征,验证分析降解原油目标函数模型对海迪茨氏菌去除原油样品中有机物的去除率.实验结果表明:二阶回归方程中回归系数β0、β1、β2、β3、β4、β1β2、β1β3、β1β4、β2β3、β2β4、β3β4、β21、β22、β23、β42对海迪茨氏菌降解原油影响效果显著;当辐射剂量与接种量范围分别为15～35Gy和2.5%～7.5%时,对原油的降解率(η)响应值能达到最高值62.6%.降解产物表征结果表明,模拟目标函数中的互动参数,当辐射剂35Gy、温度29℃、pH=6.75、接种量6.75%时,在第7d,原油样品中C16、C19、C25与C26完全被去除;在第14d,原油样品中C21、C22、C24与四甲基完全消失;第36d,原油样品中的C、C、C与3,5-二甲基十二烷的去除率分别达到83.5%、58.9%、65.8%、74.5%.     

Identification of two alkane hydroxylases involved in long-chain n-alkane degradation by Dietzia sp. DQ12-45-1b北大核心CSCD

The aim of this study was to identify genes involved in long-chain alkane degradation in Dietzia sp. DQ12-45-1b. Functional genes were annotated by genome analysis. Induction of alkane hydroxylase genes by C28 n-alkane was analyzed by using quantitative real-time PCR in wild-type Dietzia sp. DQ12-45-1b and its alkW1 gene knockout mutant strain M 5-5. From the genome of Dietzia sp. DQ12-45-1b, two homologues, G1 and G2 genes were annotated, which showed 50% amino acid sequence similarity with AlmA from Acinetobacter sp. DSM17874, and 48% amino acid sequence similarity with LadA from Geobacillus thermodenitrificans NG80-2, respectively. In addition, G1 showed 71% amino acid sequence similarity with G1a, and G2 showed 34% and 87% amino acid sequence similarities with G2a and G2ß, respectively, which were annotated from Dietzia sp. E1 genome. In addition, the alkW1 gene knockout strain M 5-5 could grow with C28 n-alkane as the sole carbon source, indicating the presence of potential long-chain alkane hydroxylase gene(s) other than alkW1 in Diezia sp. DQ12-45-1b. Accordingly, induction of G1 and G2 genes was observed when Dietzia ap. DQ12-45-1b and alkW1 knockout mutant strain M 5-5 grew with C28 n-alkane as sole carbon source. The results indicated that G1 and G2 genes are mostly responsible for the degradation of long-chain alkanes in Dietzia sp. DQ12-45-1b, which has unique multiple alkane hydroxylase systems.