木质素降解酶研究进展与外生菌根真菌

综述了木质素降解酶系统的组成、分子生物学研究和主要影响因子的研究进展,以及国内外在外生菌根真菌产木质素降解酶方面的研究,阐述了其在有机污染土壤修复中的优势。木质素降解酶系统能够降解多环芳烃、氯代芳烃、硝基有机物和染料等多种环境污染物。许多外生菌根真菌都具有木质素降解酶系统。     

萘降解菌的分离及其联合修复作用的研究进展

多环芳烃萘是一类长久存在于环境中的优先污染物,具有致突变性与致癌性。该文综述了不同生态环境中的萘降解菌株的种类,特别是极端环境中分离得到的菌株,不仅具有良好的降解萘的能力,而且具有较高之耐盐,耐压、耐温性,可与植物协同发挥作用,修复受多环芳烃污染的土壤。介绍了萘降解菌株降解机理及其联合修复作用的相关研究进展。降解菌的联合修复作用包括微生物联合化学、物理、生物等方法修复多环芳烃污染的土壤。针对现在萘污染修复的研究趋势,从降解菌种的鉴定保藏、菌株性能优化、联合修复方式的改进等方面的相关生物技术做出了展望。     

氯酚污染土壤的生物强化修复及其微生物种群动态变化的分子生物学监测

在氯酚污染的土壤中接种氯酚降解菌,研究了受污染土壤的生物强化修复.利用传统的微生物计数方法和现代分子生物学手段,研究了生物强化修复过程中微生物种群的动态变化情况.在受氯酚污染的土壤样品中,以氯酚为唯一碳源和能源,分离出了多株对氯酚具有较高降解能力的微生物,利用16S rDNA序列分析方法对部分微生物进行了种属鉴定.土壤中存在的土著的氯酚降解菌可以对低含量的氯酚(100mg·g-1)进行降解;但是当土壤中氯酚含量较高(500 mg·g-1)时,土著微生物的降解能力受到限制,这可能是高浓度的氯酚对土著的氯酚降解菌会产生毒性作用.接种外来微生物后,土壤中可以培养的氯酚降解菌的总数从开始的106CFU·g-1增加到108CFU·g-1,并且,在氯酚含量为100 mg·g-1的土壤样品中,微生物数目的增加比在氯酚含量为500 mg·g-1土壤样品中更快.这表明接种的外来微生物可以在土壤中很好地生长繁殖,有效地促进土壤中氯酚的生物降解.接种外来微生物可以减轻土壤中氯酚对土著微生物初期产生的不利影响.土壤中的氯酚可能会改变微生物种群结构.DGGE分析结果表明,在未受氯酚污染和受氯酚污染的土壤样品中,存在一些共同的DNA谱带,但谱带强度有明显的差异.在受污染的土壤中接种外来微生物进行生物强化,可以促进污染物的生物降解过程,是生物修复过程中的一种重要手段,有着广泛的应用前景,将在我国受污染环境的生物修复中发挥重要作用.     

Fe^0还原土壤中不同结构甲基和氯代硝基苯

采用零价铁（Fe^0）在常温常压下分别还原土壤中邻、间、对三种氯代硝基苯和硝基甲苯。结果表明：相同取代基在苯环上不同取代位置对还原率的影响差别不大。在硝基芳烃化合物浓度约为2．5×10^-6mol／g,Fe^0加入量为50mg／g,25℃条件下,经过5天反应,邻、间、对三种硝基甲苯的还原率均在70％左右,三种氯代硝基苯的还原率则在90％以上。结合计算机模拟所得的六种硝基芳烃的质子亲和力和电子亲和力,对硝还原这两类硝基芳烃的机理进行了初步探讨。     

环糊精对硝基化合物混合体系微生物降解影响

通过对硝基苯和对硝基苯酚混合体系微生物降解过程的研究,探讨了在降解过程中加入-环糊精及其衍生物羧甲基--环糊精(CMCD)对混合体系微生物降解的影响.研究表明,对硝基苯酚不能被单独降解;硝基苯能与对硝基苯酚产生共代谢作用,对硝基苯酚浓度影响混合体系的降解程度.加入环糊精能促进对硝基苯酚的降解率,影响程度与对硝基苯酚的浓度和环糊精的种类及量有关.     

石油污染土壤的生物修复技术及微生物生态效应

利用投菌法和生物刺激法对陕北子长石油污染土壤进行微生物修复研究.通过利用红外分光光度法测定不同处理方法对石油烃的去除效果确定了修复陕北石油污染土壤的最佳方案.修复过程中利用最大可能计数法(MPN)、PCR-琼脂糖电泳法、PCR-DGGE法分别测定了石油烃降解菌数目、催化基因、土壤微生物多样性对土壤微生物生态效应进行研究.结果发现石油污染土壤不同生物处理修复效果为:生物刺激(加入N、P营养物质)生物强化(投加降解菌)其他.土壤中石油烃降解率与可降解石油烃的催化基因含量之间存在正相关关系,修复过程中土壤中的石油烃和烷烃降解菌数量显著多于多环芳烃降解菌数量,投加外源降解菌SZ-1可以显著提高土壤细菌群落的多样性.研究结果有助于深入理解生物修复石油土壤过程中的微生物生态效应变化.     

成晶节杆菌NT16和共生芽孢杆菌NG16联合代谢1-萘酚的特性

1-萘酚是多环芳烃降解过程中极易积累的典型含氧多环芳烃,其难降解、毒性大,直接影响多环芳烃污染环境的修复效果.为探究微生物联合代谢1-萘酚的特性,以前期从含氧多环芳烃污染土壤中分离出的两种共生菌株——成晶节杆菌NT16和芽孢杆菌NG16为受试菌株,结合HPLC、TOC、GC-MS等测定方法进行降解特性的测定.结果表明,NT16菌和NG16菌均能以1-萘酚为唯一碳源和能源生长,两种菌株联合代谢比NT16菌、NG16菌单独降解1-萘酚的生长量分别高2.758×10~9 cfu·mL~(-1)和1.4×10~9 cfu·mL~(-1),对1-萘酚的降解率可提高20%,使体系中TOC值加速降低.NT16菌和NG16菌联合代谢1-萘酚可按照两个途径进行,其一,1-萘酚羟化后开环,进入邻苯二甲酸代谢途径.NT16菌更易进入该途径,而NG16菌则不易通过该途径降解1-萘酚;其二,1-萘酚羟化后开环,进入苯丙酸代谢途径.NG16菌迅速将1-萘酚降解为对羟基苯乙酸,其在降解体系中积累,NT16菌无法高效降解1-萘酚却能够降解对羟基苯乙酸至较短烷基链的小分子化合物.该研究结果将为含氧多环芳烃污染环境实际修复中微生物群落协同作用降解污染物奠定基础.     

碳源及电子受体对厌氧微生物降解对氯硝基苯的影响

为考察氯代硝基苯的共代谢降解过程中,有机碳源类型和电子受体对氯代硝基苯厌氧降解过程的影响,以对氯硝基苯(p-CNB,para-chloronitrobenzene)为目标污染物,通过间歇试验考察了葡萄糖、乙醇、乙酸钠等有机碳源及SO2--4、NO3等竞争性电子受体对厌氧微生物降解对氯硝基苯的影响。结果显示,葡萄糖、乙醇和乙酸钠作为碳源时,菌种获得的还原能力大小依次为:乙醇葡萄糖乙酸钠。SO2-4对菌种降解对氯硝基苯过程不产生竞争性抑制。NO-3对对氯硝基苯降解过程的影响与碳源的量有关,当乙醇-COD为100 mg/L时,硝酸盐产生竞争性抑制,且抑制作用随着NO-3浓度升高而增强;当乙醇-COD为600 mg/L时,硝酸盐对菌种降解对氯硝基苯过程不产生竞争性抑制作用。     

固定化微生物技术修复PAHs污染土壤的研究进展

多环芳烃(PAHs)是环境中普遍存在的一类典型有机污染物,绿色修复PAHs污染土壤已成为国内外环境和土壤界共同关注的热点问题之一.作为一种新型的微生物修复技术,固定化微生物修复有机污染土壤正受到越来越多的关注,国内外相关研究刚刚开始.本文重点评述了近几年国内外有关固定化微生物技术修复PAHs污染土壤的最新进展,总结了固定化微生物技术的修复原理、微生物固定化载体的选择、高效降解菌的筛选、固定化方法及影响因素等方面,并提出今后的发展方向,为我国开展固定化微生物技术修复有机污染土壤的研究提供参考.     

微生物修复油污土壤过程中氮素的变化及菌群生态效应

对陕北地区石油污染土壤进行了微生物修复研究,利用气相色谱-质谱联用技术(gas chromatography-mass spectrometry,GC-MS)测定土壤中烷烃和多环芳烃的含量变化,采用高通量测序技术研究微生物修复对油污土壤微生物群落多样性的影响,对修复过程中土壤不同形态氮含量变化进行了分析测定.结果表明,经过18周的修复,土壤中烷烃含量由初始时的25 987.8mg·kg~(-1)降低为12 788.6 mg·kg~(-1),多环芳烃含量由初始时的5 322.9 mg·kg~(-1)降低为2 917.2mg·kg~(-1).高通量测序结果表明,经过微生物修复处理的土壤微生物群落结构发生了较大变化,一些可降解烃类的菌群如厚壁菌门(Firmicutes)、拟杆菌门(Bacterodetes)、迪茨氏菌属(Dietzia)、不动杆菌属(Acinetobacter)所占丰度增加.经过修复处理的土壤中总氮、氨氮含量呈降低趋势,硝态氮含量在修复前期呈降低趋势,修复后期基本保持不变.研究结果表明,微生物修复处理可去除土壤中的大部分烷烃和多环芳烃,烷烃和多环芳烃的降解效果与土壤中硝氮含量和烃降解菌所占丰度相关.     

Toxic effects of acetochlor, methamidophos and their combination on nifH gene in soil

Toxic effects of two agrochemicals on nifH gene in agricultural black soil were investigated using denaturing gradient gel electrophoresis （DGGE） and sequencing approaches in a microcosm experiment. Changes of soil nifH gene diversity and composition were examined following the application of acetochlor, methamidophos and their combination. Acetochlor reduced the nifH gene diversity （both in gene richness and diversity index values） and caused changes in the nifH gene composition. The effects of acetochlor on nifH gene were strengthened as the concentration of acetochlor increased. Cluster analysis of DGGE banding patterns showed that nifH gene composition which had been affected by low concentration of acetochlor （50 mg/kg） recovered firstly. Methamidophos reduced nifH gene richness that except at 4 weeks. The medium concentration of methamidophos （150 mg/kg） caused the most apparent changes in nifH gene diversity at the first week while the high concentration of methamidophos （250 mg/kg） produced prominent effects on nifH gene diversity in the following weeks. Cluster analysis showed that minimal changes of nifH gene composition were found at 1 week and maximal changes at 4 weeks. Toxic effects of acetochlor and methamidophos combination on nifH gene were also apparent. Different nifH genes （bands） responded differently to the impact of agrochemicals： four individual bands were eliminated by the application of the agrochemicals, five bands became predominant by the stimulation of the agrochemicals, and four bands showed strong resistance to the influence of the agrochemicals. Fifteen prominent bands were partially sequenced, yielding 15 different nifH sequences, which were used for phylogenetic reconstructions. All sequences were affiliated with the alpha- and beta-proteobacteria, showing higher similarity to eight different diazotrophic genera.     

Arsenic uptake by arbuscular mycorrhizal maize （Zea mays L.） grown in an arsenic-contaminated soil with added phosphorus

The effects of arbuscular mycorrhizal (AM) fungus (Glomus mosseae) and phosphorus (P) addition (100 mg/kg soil) on arsenic (As) uptake by maize plants (Zea mays L.) from an As-contaminated soil were examined in a glasshouse experiment.Non-mycorrhizal and zero-P addition controls were included.Plant biomass and concentrations and uptake of As,P,and other nutrients,AM colonization,root lengths,and hyphal length densities were determined.The results indicated that addition of P significantly inhibited root colonization and development of extraradical mycelium.Root length and dry weight both increased markedly with mycorrhizal colonization under the zero-P treatments,but shoot and root biomass of AM plants was depressed by P application.AM fungal inoculation decreased shoot As concentrations when no P was added,and shoot and root As concentrations of AM plants increased 2.6 and 1.4 times with P addition,respectively.Shoot and root uptake of P,Mn,Cu,and Zn increased,but shoot Fe uptake decreased by 44.6%,with inoculation, when P was added.P addition reduced shoot P,Fe,Mn,Cu,and Zn uptake of AM plants,but increased root Fe and Mn uptake of the nonmycorrhizal ones.AM colonization therefore appeared to enhance plant tolerance to As in low P soil,and have some potential for the phytostabilization of As-contaminated soil,however,P application may introduce additional environmental risk by increasing soil As mobility.     

Impact of atrazine and nitrogen fertilizers on the sorption of chlorotoluron in soil and model sorbents

Sorption of chlorotoluron in ammonium sulfate, urea and atrazine multi-solutes system was investigated by batch experiments. The results showed application of nitrogen fertilizers to the soil could affect the behavior of chlorotoluron. At the same concentration of N, sorption of chlorotoluron decreased as the concentration of atrazine increased on the day 0 and 6 in soil, respectively. The sorption of chlorotoluron increased from 0 to 6 d when soils were preincubated with deionized water, ammonium sulfate and urea solution for 6 d. That indicated incubation time was one of the most important factors for the sorption of chlorotoluron in nitrogen fertilizers treatments. The individual sorption isotherms of chlorotoluron in rubbery polymer and silica were strictly linear in single solute system, but there were competition sorption between pesticides or between pesticides and nitrogen fertilizers. That indicated the sorption taken place by concurrent solid-phase dissolution mechanism and sorption on the interface of water-organic matter or water-mineral matter.     

Degradation of metabolites accumulated of benzo[a]pyrene by coupling Penicillium Chrysogenum with KMnO4

Several main metabolites of benzo[a]pyrene （BaP） formed by Penicillium chrysogenum, Benzo[a]pyrene-1,6-quinone （BP 1,6- quinone）, trans-7,8-dihydroxy-7,8-dihydrobenzo[a]pyrene （BP 7,8-diol）, 3-hydroxybenzo[a]pyrene （3-OHBP）, were identified by high-performance liquid chromatography （HPLC）. The three metabolites were liable to be accumulated and were hardly further metabolized because of their toxicity to microorganisms. However, their further degradation was essential for the complete degradation of BaP. To enhance their degradation, two methods, degradation by coupling Penicillium chrysogenum with KMnO4 and degradation only by Penicillium chrysogenum, were compared; Meanwhile, the parameters of degradation in the superior method were optimized. The results showed that （1） the method of coupling Penicillium chrysogenum with KMnO4 was better and was the first method to be used in the degradation of BaP and its metabolites; （2） the metabolite, BP 1,6-quinone was the most liable to be accumulated in pure cultures; （3） the effect of degradation was the best when the concentration of KMnO4 in the cultures was 0.01% （w/v）, concentration of the three compounds was 5 mg/L and pH was 6.2. Based on the experimental results, a novel concept with regard to the bioremediation of BaP-contaminated environment was discussed, considering the influence on environmental toxicity of the accumulated metabolites.     

Potential of plant polyphenol oxidases in the decolorization and removal of textile and non-textile dyes

In this study an effort has been made to use plant polyphenol oxidases; potato （Solanum tuberosum） and brinjal （Solanum melongena）, for the treatment of various important dyes used in textile and other industries. The ammonium sulphate fractionated enzyme preparations were used to treat a number of dyes under various experimental conditions. Majority of the treated dyes were maximally decolorized at pH 3.0. Some of the dyes were quickly decolorized whereas others were marginally decolorized. The initial first hour was sufficient for the maximum decolorization of dyes. The rate of decolorization was quite slow on long treatment of dyes. Enhancement in the dye decolorization was noticed on increasing the concentration of enzymes. The complex mixtures of dyes were treated with both preparations of polyphenol oxidases in the buffers of varying pH values. Potato polyphenol oxidase was significantly more effective in decolorizing the dyes to higher extent as compared to the enzyme obtained from brinjal polyphenol oxidase. Decolorization of dyes and their mixtures, followed by the formation of an insoluble precipitate, which could be easily removed simply by centrifugation.     

Removal of heavy metals from sewage sludge by low costing chemical method and recycling in agriculture

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Isolation and preliminary characterization of a 3-chlorobenzoate degrading bacteria

A study was conducted to compare the diversity of 2-, 3-, and 4-chlorobenzoate degraders in two pristine soils and one contaminated sewage sludge. These samples contained strikingly different populations of mono-chlorobenzoate degraders. Although fewer cultures were isolated in the uncontaminated soils than contaminated one, the ability of microbial populations to mineralize chlorobenzoate was widespread. The 3- and 4-chlorobenzoate degraders were more diverse than the 2-chlorobenzoate degraders. One of the strains isolated from the sewage sludge was obtained. Based on its phenotype, chemotaxonomic properties and 16S rRNA gene, the organism S-7 was classified as Rhodococcus erythropolis. The strain can grow at temperature from 4 to 37℃. It can utilize several （halo）aromatic compounds. Moreover, strain S-7 can grow and use 3-chlorobenzoate as sole carbon source in a temperatures range of 10-30℃ with stoichiometric release of chloride ions. The psychrotolerant ability was significant for bioremediation in low temperature regions. Catechol and chlorocatechol 1,2-dioxygenase activities were present in cell free extracts of the strain, but no （chloro）catechol 2,3- dioxygenase activities was detected. Spectral conversion assays with extracts from R. erythropolis S-7 showed accumulation of a compound with a similar UV spectrum as chloro-cis,cis-muconate from 3-chlorobenzoate. On the basis of these results, we proposed that S-7 degraded 3-chlorobenzoate through the modified ortho-cleave pathway.     

Single and joint effects of pesticides and mercury on soil urease

The influence of two pesticides including chlorimuron-ethyl and furadan and mercury （Hg） on urease activity in 4 soils （meadow burozem and phaeozem） was investigated. The soils were exposed to various concentrations of the two pesticides and Hg individually and simultaneously. Results showed that there was a close relationship between urease activity and organic matter content in soil. Chlorimuron-ethyl and furadan could both activate urease in the 4 soils. The maximum increment of urease activity by chlorimuronethyl was up to 14%-18%. There was almost an equal increase （up to 13%-21%） in the urease activity by furadan. On the contrary, Hg markedly inhibited soil urease activity. A logarithmic equation was used to describe the relationship （P〈0.05） between the concentration of Hg and the activity of soil urease in the 4 tested soils. Semi-effect dose （ED50） values by the stress of Hg based on the inhibition of soil urease in the 4 soils were 88, 5.5, 24 and 20 mg/kg, respectively, according to the calculation of the corresponding equations. The interactive effect of chlorimuron-ethyl or furadan with metal Hg on soil urease was mainly synergic at the highest tested concentrations.     

Organochlorine pesticides in soils under different land usage in the Taihu Lake region, China

A field study was conducted in the Taihu Lake region, China in 2004 to reveal the organochlorine pesticide concentrations in soils after the ban of these substances in the year 1983. Thirteen organochlorine pesticides (OCPs) were analyzed in soils from paddy field, tree land and fallow land. Total organochlorine pesticide residues were higher in agricultural soils than in uncultivated fallow land soils. Among all the pesticides, ΣDDX (DDD, DDE and DDT) had the highest concentration for all the soil samples, ranging from 3.10 ng/g to 166.55 ng/g with a mean value of 57.04 ng/g and followed by ΣHCH, ranging from 0.73 ng/g to 60.97 ng/g with a mean value of 24.06 ng/g. Dieldrin, endrin, HCB and α-endosulfan were also found in soils with less than 15 ng/g. Ratios of p,p'-(DDD DDE)/DDT in soils under three land usages were: paddy field ＞ tree land ＞ fallow land, indicating that land usage inlfuenced the degradation of DDT in soils. Ratios of p,p'-(DDD DDE)/DDT ＞1, showing aged residues of DDTs in soils of the Taihu Lake region. The results were discussed with data from a former study that showed very low actual concentrations of HCH and DDT in soils in the Taihu Lake region, but according to the chemical half-lives and their concentrations in soils in 1980s, the concentration of DDT in soils seemed to be underestimated. In any case our data show that the ban on the use of HCH and DDT resulted in a tremendous reduction of these pesticide residues in soils, but there are still high amounts of pesticide residues in soils, which need more remediation processes.     

Alterations of organ histopathology and metallolhionein mRNA expression in silver barb, Puntius gonionotus during subchronic cadmium exposure

Common silver barb,Puntius gonionotus,exposed to the nominal concentration of 0.06 mg/L Cd for 60 d,were assessed for histopathological alterations(gills,liver and kidney),metal accumulation,and metallothionein(MT)mRNA expression.Fish exhibited pathological symptoms such as hypertrophy and hyperplasia of primary and secondary gill lamellae,vacuolization in hepatocytes,and prominent tubular and glomerular damage in the kidney.In addition,kidney accumulated the highest content of cadmium,more than gills and liver.Expression of MT mRNA was increased in both liver and kidney of treated fish.Hepatic MT levels remained high after fish were removed to Cd-free water.In contrast,MT expression in kidney was peaked after 28 d of treatment and drastically dropped when fish were removed to Cd-free water.The high concentrations of Cd in hepatic tissues indicated an accumulation site or permanent damage on this tissue.