一株高效MC—RR降解菌的分离鉴定及其降解特性

为了得到一株具有降解微囊藻毒素一RR（MC—RR）特性的产芽孢菌株，采用加热富集芽孢菌的方法，从太湖分离到一株MC．RR降解菌CMl，该菌对MC—RR具有强烈的降解特性。通过形态学特征、生理生化特征及16SrDNA序列分析鉴定该菌株属于耐硼赖氨酸芽孢杆菌（Lysinibacillusb oronitolerans）。通过研究温度和pH值对菌株CMl降解MC—RR能力的影响，发现菌株CMl在60h将MC—RR由12．77μg／mL降解到1．67μg／mL，降解率达86．90％，最适降解温度为37℃，最适pH值为7．0。CMl菌株的胞外物质和胞内物质均能降解MC—RR，但胞内物质具有更强烈的降解特性，12h可以将7．27μg／mL的MC-RR完全降解。为丰富MC-RR降解菌纯菌种研究以及在去除水体中MC—RR应用研究方面提供了理论基础。     

Biodegradation of Chlorimuron-ethyl by the Bacterium Klebsiella jilinsis 2N3

Enrichment culturing of sludge taken from an industrial wastewater treatment pond led to the identification of a bacterium (Klebsiella jilinsis H. Zhang) that degrades chlorimuron-ethyl with high efficiency. Klebsiella jilinsis strain 2N3 grows with chlorimuron-ethyl as the sole nitrogen source at the optimal temperature range of 30–35°C and pH values between 6.0–7.0. In liquid medium, the degradation activity was further induced by chlorimuron-ethyl. Degradation rates followed the pesticide degradation kinetic equation at concentrations between 20 and 200 mg L^?1. Using initial concentrations of 20 and 100 mg L^?1, the degradation rates of chlorimuron-ethyl were 83.5 % and 92.5 % in 12 hours, respectively. At an initial concentration higher than 200 mg L^?1, the degradation rate decreased slightly as the concentration increased. The 2N3 strain also degraded the sulfonylurea herbicides ethametsulfuron, metsulfuron-methyl, nicosulfuron, rimsulfuron, and tribenuron-methyl. This study provides scientific evidence and support for the application of K. jilinsis in bioremediation to reduce environmental pollution.     

稠油降解菌的筛选、鉴定与菌群构建

分别以烷烃、芳烃、胶质沥青质为唯一碳源,从稠油污染过的土壤里分离、筛选可培养的降解菌,将16株组合构建SL-16稠油降解菌群,通过室内摇瓶实验测得该菌群在最佳条件下对陈庄油田稠油降解率可达68%,其适宜的生长及降解温度为35～45℃,pH值为7.0～9.0,含盐量为4 000～14 000 mg/L,接种量为2%,稠油...     

Characterization and catabolic genes detection of three oil-degrading Rhodococcus spp.北大核心CSCD

Oil pollution is one of the major factors causing environmental deterioration. Bioremediation of oil contaminated environments by microorganisms attracts much research attention. This study aimed to screen efficient oil-degrading bacteria from oil contaminated soil and analyze their characteristics and catabolic genes. Oil-degrading bacteria were screened from oil contaminated soil in minimal medium containing crude oil and identified by morphological, physiological and biochemical characteristics and 16S rDNA sequence analysis. Their growth and degradation characteristics were studied with ultraviolet spectroscopy and GC-MS analysis. The surfactant production was studied by adopting culture method. The major oil-degrading related genes were detected by t he PCR a mplification. As a result, t hree oil-degrading bacteria strains named KB1, 2182 and JC3-47 were isolated from the oil contaminated soil samples. The strains could use crude oil as the sole carbon source to degrade oil with a degrading rate of 41.02%, 32.26% and 55.90%, respectively, when cultured in minimal medium containing crude oil for 3 days. The bacteria were identified to belong to genus Rhodococcus. With 100% similarity of 16S rDNA sequences of the three strains with known ones of Rhodococcus, KB1 was preliminarily identified as Rhodococcus erythropolis, 2182 as Rhodococcus equi, and JC3-47 as Rhodococcus qingshengii. They grew well at 10-50 °C, with the initial pH of 3-9 and the NaCl concentration of 0-5%. The optimal temperature for bacterial growth was 35 °C, 35 °C and 30 °C respectively. KB1 and 2182 could grow at pH 2 and 9.0% of NaCl. The bacteria grew well in broth containing different organic substrates as sole carbon source, such as n-dodecane, n-octadecane, benzene, methylbenzene, xylene and naphthaline. KB1 and JC3-47 could grow well in broth containing pyrene. GC-MS analysis revealed that the bacteria could effectively degrade medium- and long-chain alkane components in crude oil. The bacteria produced biosurfactants and decreased the surface tension of the culture broth. They also showed adhesion activities to n-hexadecane. The oil-degrading related genes such as alkane monooxygenase, aromatic-ring-hydroxylating dioxygenase and catechol dioxygenase genes were detected in all the three strains. Besides, biphenyl dioxygenase genes were detected in KB1 and 2182. The isolated Rhodococcus spp. strains could effectively degrade petroleum hydrocarbons with high adaptabilities to extreme environments such as high salt and low temperature. They are supposed to be applied broadly in the bioremediation of oil contaminated soil in such environments.     

Dynamics of bacterial community of Suillus granulatus mushroom shiro in Pine (Pinus tabuliformis) forests北大核心CSCD

To evaluate bacterial community variation in the mushroom shiro of Suillus granulatus during fruiting, we collected soil samples from the mushroom shiro in the pine (Pinus tabuliformis) forest of mountainous area in Beijing from May to November and evaluated the bacterial community using polymerase chain reaction - denaturing gradient gel electrophoresis (PCR-DGGE). Total soil DNA was extracted using a commercial soil DNA isolation kit. PCR amplification and DGGE were performed using bacterial universal primers 338F and 518R. The specific bands were excised from the gel and sequenced. The results revealed that soil bacterial community maintained considerably high level and changed seasonally with the mushroom fruiting. In total, 53 bands of DGGE profiles were sequenced and divided into 5 phyla (Acidobacteria, Actinobacteria, Bacteroidetes, Firmicutes, and Proteobacteria and 22 genera (Acidobacterium, Aminobacter, et al). Species from Proteobacteria and Acidobacteria were the dominant bacterial groups sharing considerably high relative abundance, while class a-Proteobacteria was the most abundant group. The variation of the relative abundance of γ-Proteobacteria species was consistent with the mushroom fruiting season. The relative abundance of Acidobacteria species obviously increased before mushroom flush (in July). The fruiting of S. granulatus and the relative abundance of γ-Proteobacteria were correlated with each other. The present study provided a basis for conservation and domestication of mushroom S. granulatus.     

Isolation,identification and desulfurization mechanism of a sulfur-oxidizing bacterium with salt tolerance北大核心CSCD

Micro-organism with efficient desulfurization performance is a key factor in the biological desulfurization technology. This study aimed to seek such a sulfur-oxidizing strain and understand its desulfurization mechanism. Wastewater in a sewage station of natural gas purification plant was used to screen the sulfide-oxidizing strain, and to identify it based on sequence similarity analysis of 16S rDNA and the morphological characteristics. Thiosulfate was used as substrate for investigating the sulfur oxidation performance and salinity tolerance; the OD600, content change of thiosulfate, sulfate, sulfur, pH and total alkalinity in the cultural system were also investigated. The strain DS-B was found to share the highest sequence similarity with Thioalkalivibrio thiocyanoxidans ARh2, therefore determined as Thioalkalivibrio. At the optimum temperature of 35 °C for growth and degradation, the removal efficiency of thiosulfate could reach 98.7% after 7 days. Strain DS-B had strong resistance to thiosulfate, and the optimal concentration of S2O32- was 2 × 104 mg/L. The analysis for sulfur oxides showed that it could oxidize thiosulfate by the pathway of S2O32-→SO42- / S2O32- → S → SO42-. Therefore the strain DS-B is a sulfur-oxidizing bacterium with great application prospect for its strong salt tolerance and conspicuous removal capability for thiosulfate.     

1株耐冷兼性嗜碱好氧反硝化菌的分离鉴定及反硝化特性

以传统微生物富集分离方法,从垃圾渗滤液活性污泥中筛选到1株高效好氧反硝化菌,通过形态观察、生理生化特征及16S rDNA序列分析,对菌株进行了鉴定,同时对其好氧反硝化特性和异养硝化功能进行了研究.结果表明,筛选到的好氧反硝化菌株为假单胞菌属(Pseudomonas sp.),命名为GL19,GenBank登录号为(KC710974).碳源、C/N、pH及温度对菌株反硝化活性影响较大.在柠檬酸钠为碳源、C/N不低于15、pH 6~10、溶解氧(DO)4.8~7.7 mg·L-1及温度为15~34℃,硝酸盐氮负荷为140 mg·L-1的条件下,硝酸盐去除率均达100%,总氮(TN)平均去除率为96.5%,最终无亚硝酸盐积累;菌株能以亚硝酸盐氮、氨氮为底物进行高效脱氮,20 h内可将140 mg·L-1的亚硝酸盐氮完全去除,28 h内可将280 mg·L-1的氨氮降至3.11 mg·L-1,氨氮去除率达98.9%.显示该菌具有耐冷、高效脱氮特性,可实现同步硝化反硝化,这对南方地区冬季废水处理具有潜在应用价值.     

不同16SrDNA靶序列对DGGE分析活性污泥群落的影响

为探讨不同通用引物扩增16S rDNA靶序列对活性污泥微生物群落分析的影响,更合理的利用变性梯度凝胶电泳(DGGE)技术分析活性污泥样品.从连续流搅拌槽式反应器(CSTR)中获取活性污泥,以3对通用引物341f/534r、968f/1 401r和341f/926r扩增16S rDNA序列,用DGGE分离PCR扩增产物.研究表明采用不同引物对进行DGGE分析时,群落多样性和动态存在显著的差异.341f/534r和968f/1 401r的靶序列分离效果较好,341f/926r的靶序列分离效果较差.引物341f/534r和341f/926r DGGE图谱显示S2和S3相似性高,引物968f/1 401r DGGE图谱显示S1和S2相似性高.由此可见采用不同引物对进行DGGE分析时,群落结构之间的相似性和动态是不一致的.341f/534r的DGGE图谱中条带丰富,多样性最好,968f/1 401r的DGGE图谱次之,341f/926r DGGE图谱条带最少,多样性也较差.因此,在利用DGGE分析活性污泥样品时采用引物341f/534r和968f/1 401r是比较适宜的.     

根瘤菌新类群的全细胞蛋白电泳及16S rDNA全序列分析

应用ＳＤＳ全细胞蛋白电泳技术对根瘤菌３个新类群进行聚类分析，并对３个新类群中心菌株的１６Ｓ ｒＤＮＡ全序列（Ｍｒ≈１．５ｋｂ）进行了测定。将所测序列与ＥＭＢＬ／ＢａｎｋＩＴ／ＤＤＢＪ资料库中根瘤菌及相关菌已知种的１６Ｓ ｒＤＮＡ全序列进行聚类比较，得到系统发育树状图，树状图中，菌株ＳＨ２２６２３和ＳＨ１９３１２与山羊豆根瘤菌、葡萄土壤杆菌、根癌土壤杆菌和悬钩子土壤杆菌在同一个分支上，彼此亲缘关系较     

Respiration induced by substrate and bacteria diversity after application of diuron,hexazinone, and sulfometuron-methyl alone and in mixture

Abstract

After application, herbicides often reach the soil and affect non-target soil microorganisms, decreasing their population, diversity or affecting metabolic activity. Therefore, laboratory studies were performed to evaluate the effects of diuron, hexazinone and sulfometuron-methyl alone and mixed upon carbon transformation by soil microorganisms in clayey and sandy soils and the effect on bacterial diversity and structure. Control treatment without herbicide application was also performed. Sub-samples from the control and herbicide treatments (10?g – in triplicate) were collected before herbicide application and 7, 14, 28 and 42?days after treatment (DAT), then 1?mL of ¹⁴C-glucose solution was applied. The released ¹⁴CO₂ was trapped in 2?M NaOH solution and the radioactivity was analyzed by liquid scintillation counting (LSC), 12?h after glucose application. The effect of herbicides on bacterial diversity was evaluated by T-RFLP. The experiment was conducted in a complete randomized design. Hexazinone did not affect ¹⁴CO₂ evolution. Diuron showed a greater ¹⁴CO₂ evolution in sandy and clayey soil, while sulfometuron-methyl led to an increase in sandy soil, at 42 DAT. A greater evolution of carbon was observed in the treatment with herbicide mixture in sandy soil, compared with the same treatment in clayey soil or control. However, the herbicide mixture application did not affect the soil biological activity measured by the respiration rate induced by substrate. On the other hand, the herbicide mixtures affected the bacterial diversity in both soils, being the strongest effect to diuron and sulfometuron-methyl in clayey soil and hexazinone in sandy soil.