Screening of endophytic bacteria against fungal plant pathogens

Bacterial endophytes were found from 6 plant leaves among 35 plant leaves screened. Two of the isolated bacteria showed antagonistic activity against fungal plant pathogens. An isolate named KL1 showed the clear inihibition against plant pathogens, Fusarium oxysporum and Rhizoctonia solani, on PDA as well as TSA plate. Supernatant of the bacterial culture also showed the clear inhibition against the fungal growth on the plate and the antibiotic substance was identified as iturin A by HPLC analysis. KL1 was identified as Bacillus sp. from the 16S rRNA gene analysis. Very thin hyphae of R. solani was miccroscopically observed when the fungus was co-cultivated with KL1.     

城市地表水中肠道病原微生物与粪便污染指示菌的关系研究

本研究选择肠道病毒、伤寒沙门氏菌、志贺氏菌、大肠埃希氏菌作为城市水体中典型的肠道病原微生物,分别建立起相应的实时荧光定量PCR检测方法.对水源水、景观娱乐用水以及城市河流样品进行长期监测,考察典型肠道病原微生物的分布特征和变化规律.肠道病毒在各地表水体中都有检出,浓度大都在103copy·L-1以下.大肠埃希氏菌的含量在保护良好的水体和接纳城市污水的水体中有明显差别,而伤寒沙门氏菌和志贺氏菌通常出现在受粪便污染较重的水体中.统计学的分析结果显示各种水体中的肠道病原微生物检出浓度均符合对数正态分布规律.粪便污染指示菌与肠道病毒之间不存在显著的相关性,但却能在一定程度上反映典型肠道病原菌的存在情况.在大肠菌群或粪大肠菌群浓度为104CFU·L-1以上的样品中,伤寒沙门氏菌、志贺氏菌具有较高的阳性率.     

植物促生菌应用研究进展

植物促生菌即PGPB不仅在农业生产中促进植物生长以及防治植物病害,还逐渐应用于环境保护方面。文章从以下几方面就PGPB的研究进展进行了综述,包括PGPB在农业和环境中的应用、PGPB内生菌、生物防治型PGPB、PGPB的基因工程改良、PGPB剂型等。最后展望了该领域的研究方向。     

水传病原微生物PCR-纳米金显色快速检测方法

水传病原微生物是腹泻等肠胃疾病发病的主要原因,而发展快速、有效的检测方法对于控制和减少该类微生物的传播风险十分重要.因此,本文研究了PCR反应结合不修饰的纳米金颗粒(Gold nano-particles,AuNPs)显色检测方法的最佳盐浓度、探针含量、PCR反应条件等,并从纳米金制备、病原菌种类、检测特异性等方面进行考核,确定了最优检测条件为:10μL PCR扩增产物混合20 pmol探针,95℃反应5 min,55.5℃反应3 min后,40℃杂交10 min;此时,先后加入90μL实验用纳米金和一定浓度的PBS溶液,室温观察颜色变化.结果发现,纳米金溶液由红色变为紫色或近于无色为阳性结果,对应的吸光度值迁移出530 nm,以此可达到快速检测病原微生物的目的.     

Characterization and source analysis of indoor/outdoor culturable airborne bacteria in a municipal wastewater treatment plant

The potential health risks of airborne bacteria emission from a wastewater treatment process have been concerned. However, few studies have investigated the differences in community structure between indoor and outdoor bacteria. In this work, the characterization of airborne bacteria was studied in a municipal wastewater treatment plant in Beijing, China. Two indoor (i.e., fine screen room and sludge dewatering house) and two outdoor (i.e., aeration tank and control site) sampling sites were selected. An Andersen six-stage impactor was used for collecting culturable airborne bacteria in the air, and Illumina MiSeq sequencing was conducted to track the emission source of the culturable airborne bacteria. The results indicate that, compared with the outdoor aeration tank site, the concentrations of culturable airborne bacteria in the indoor fine screen room with poor ventilation were more than ten times higher and the particle size was about twice as large. The community structures of indoor and outdoor culturable airborne bacteria were obviously different. Enterobacteriaceae and opportunistic pathogens were detected in indoor culturable airborne bacteria, with wastewater and sludge dewatering machine identified as the primary sources. Conversely, Enterobacteriaceae and opportunistic pathogens were not detected in outdoor culturable airborne bacteria. Outdoor high wind speed might have resulted in rapid dilution and mixing of culturable airborne bacteria generated from the aeration tank with the ambient air. The results of the present research suggest that covering pollution sources, increasing ventilation rates, and using protective measures for personnel should be implemented to decrease the exposure risk to indoor culturable airborne bacteria.     

生物炭对非常规水源灌溉下土壤-作物病原菌的影响

采用根箱实验,基于定量PCR技术研究非常规水源（再生水和养殖废水）灌溉及生物炭添加后根际土、非根际土及玉米根部病原菌的赋存情况.结果发现,灌溉时间、土壤类型可显著影响土壤中病原菌的整体分布.相较于蒸馏水灌溉下根际土、非根际土的单个病原菌,再生水灌溉下病原菌变化为-0.74~0.17个数量级,养殖废水灌溉下为-0.14~0.60个数量级.生物炭对土壤中病原菌的影响因灌溉水源、灌溉时间而改变,可使再生水及养殖废水灌溉下病原菌检出量最高降低0.35、0.39个数量级.但再生水及养殖废水灌溉可使玉米根部病原菌检出量最高增加1.17、2.20个数量级,生物炭又使非常规水源灌溉下其检出量最高增加0.60、1.08个数量级,带来潜在的健康风险.     

一株DDT降解菌的筛选及其降解特性

以DDT为目标污染物,通过筛选获得了一株效果稳定的DDT降解菌,并对其进行形态学观察,生理生化特性及16S rRNA测序鉴定.经鉴定,该菌株属于甲基菌属（Methylovorus）,命名为Methylovorus sp. XLL03.菌株在pH值为7,温度30℃,外加碳源浓度0.5%,初始DDT浓度20mg/L时生长量最大.在pH值为6,温度30℃下,外加碳源（葡萄糖）浓度0.1%,初始DDT浓度20mg/L时对DDT的降解率最大.在优化条件下,4d后菌株XLL03对DDT最高降解率可达50.4%.利用GC-MS对DDT的降解中间产物进行定性分析,初步推断在菌株XLL03中,DDT最初分别通过脱氯和脱氯化氢生成DDD和DDE,随后DDD和DDE进一步脱氯得到DDMU,最终DDMU开环后又经过一系列反应被彻底矿化.在DDT的代谢过程中,未发现代谢中间产物的积累,表明菌株XLL03在修复受DDT污染的水或土壤中具有一定的应用前景.     

具菲降解特性植物内生细菌的分离筛选及其生物学特性

从长期受石油污染的植物体内分离到内生细菌36株,通过平皿促生和定殖试验筛选出1株能高效降解菲并明显促进小麦生长、能在小麦体内良好定殖的内生细菌7J2菌株.同时,对菌株7J2的生物学特性进行了研究.结果表明,菌株7J2在菲含量为30 mg·L-1的条件下,28℃振荡培养6d,菲的降解率达到99.81%.菌株7J2能够产生吲哚乙酸(IAA)(3.4±0.2)mg·L-1)和铁载体(2,3-二羟基苯甲酸)(4.3±1.6) mg·L-1).振荡培养3d,接活菌的培养基中有效磷浓度比接灭活菌的处理增加了32.5%.另外,菌株7J2对重金属Pb2 、Cr2 、Cu2 、Ni2 、Zn2 、Cd2 均有较强的抗性.通过16SrDNA序列测定,菌株7J2属于肠杆菌属.     

内蒙古自治区环境植物的利用

介绍目前在内蒙古自治区具有一定环境保护价值的环境植物资源,同时发掘其在环境保护之外的其他价值,可以在今后环境保护和园林绿化方面提供一定的借鉴,充分发掘这些植物资源的利用价值。     

O3-BAC-Cl2: A multi-barrier process controlling the regrowth of opportunistic waterborne pathogens in drinking water distribution systems

Simulated drinking water distribution system (DWDS) treated with O₃-BAC-Cl₂ (ozone-biological activated carbon-chlorine) was constructed to study its effects on the regrowth of five typical opportunistic pathogens (OPs). It was found that O₃-BAC-Cl₂ could significantly reduce the regrowth of target OPs in the effluents of DWDS compared with Cl₂ and O₃-Cl₂ with the same residual chlorine levels. However, the effect of O₃-BAC-Cl₂ on the average numbers of target OPs gene markers in the biofilms of DWDS was not apparent, suggesting that OPs in the biofilms of DWDS were tolerant to the upstream disinfection process. The quantification of target OPs in the BAC-filter column demonstrated that OPs decreased with the increase of depth, which was likely due to the organic nutrient gradient and microbial competition inside the BAC-filter. Increase in the ozone dose could further reduce the OPs at the bottom of the BAC-filter. Spearman correlation analysis demonstrated that some significant correlations existed between target microorganisms, suggesting potential microbial ecological relationships. Overall, our results demonstrated that the BAC-filter may act as a “battlefield” suppressing the OPs through microbial competition. O₃-BAC-Cl₂ could be an effective multi-barrier process to suppress the proliferation of OPs in the bulk water of DWDS. However, OPs protected by the biofilms of DWDS should receive further attention because OPs may be detached and released from the biofilms.     

接种内生细菌对龙葵吸收积累镉的影响

通过在LB培养基中加入不同浓度的镉,从镉超富集植物龙葵植株内分离到3株很强的镉抗性内生菌株N1、N2、N4,最高耐受镉浓度达300 mg·L^-1.通过生理生化鉴定和16S rDNA序列分析,初步确定N1为芽孢杆菌属（Bacillus sp.）细菌,N2为肠杆菌属（Enterobacter sp.）细菌,N4为巨大芽孢杆菌（Bacillus megaterium）.将龙葵种植在含有一定镉（50 mg·kg^-1）的土壤中进行盆栽试验,以灌根的方式分别接种分离到的3株细菌及其混菌组合,并对其株高、生物量及镉吸收能力进行分析.结果显示,N4菌株处理的龙葵叶、茎、根部镉含量比不接菌对照分别增加32.94%、14.50%、23.44%,差异显著（p<0.05）;接种N1+N2+N4混菌的处理龙葵叶、茎和根干重分别比不接菌对照高出118.22%、109.83%和113.01%,植株地上和地下镉吸收总量分别增加109.53%和83.01%,差异显著.试验表明供试菌株能显著促进植株生长,强化龙葵吸收土壤中镉的能力.     

一株降低烟草中特有亚硝胺细菌的分离鉴定及特性研究

从白肋烟TRM品种叶片中分离得到一株能降低白肋烟烟草中特有的亚硝胺(TSNA)含量的内生细菌菌株,该菌株能还原硝酸盐和亚硝酸盐.通过形态、生理生化特性分析以及16SrDNA序列进行同源比较,鉴定该菌株为根瘤土壤杆菌属,定名为Agrobaterium tumefaciens.该菌株最佳生长温度为25～30℃,最佳培养基为TSB,最适pH值为7.0～8.0.在采收前一天,对白肋烟品种TN86进行菌株喷洒处理;检测晾制期间TSNA、硝酸盐、亚硝酸盐含量变化.结果表明,在晾制前期,TSNA含量比较低且变化幅度较小;晾制30d时,TSNA含量达到最大;在晾制末期,喷洒菌株处理的烟叶中TSNA含量有明显的降低,比同一时期对照烟叶中TSNA含量降低了81.3%;常规化学成分及烟叶感官质量与对照相似,都达到了优级白肋烟标准.通过分析该菌株对硝酸盐、亚硝酸盐及TSNA的影响,推测该菌株可能通过还原亚硝酸盐,使亚硝酸盐含量减少,从而降低了TSNA的形成.     

高水生植物毒性污染物的初步筛选

针对目前普遍认为水生植物对污染物的敏感性比动物低的观点开展研究. 查阅和筛选了ECOTOX毒性数据库里的污染物水生生物毒性数据,对美国国家水质基准（2009）里的120项优先控制污染物的毒性数据进行了分析. 参考美国的水生生物基准技术指南的数据筛选原则,初步筛选出6种具有高植物毒性的污染物,分别为1,1,1-三氯乙烷、2,4-二氯苯酚、4-硝基苯酚、邻苯二甲酸丁苄酯、邻苯二甲酸二丁酯和N-亚硝基二甲胺. 结果表明,确实存在高植物毒性的污染物,对于这类污染物,水生植物比水生动物更为敏感. 然而,由于水生植物毒性试验远远少于动物毒性试验,并且资料的积累极为欠缺,因此,需要对水生植物毒性效应的研究和应用给予足够的重视.      

The control of red water occurrence and opportunistic pathogens risks in drinking water distribution systems: A review

Many problems in drinking water distribution systems (DWDSs) are caused by microbe, such as biofilm formation, biocorrosion and opportunistic pathogens growth. More iron release from corrosion scales may induce red water. Biofilm played great roles on the corrosion. The iron-oxidizing bacteria (IOB) promoted corrosion. However, when iron-reducing bacteria (IRB) and nitrate-reducing bacteria (NRB) became the main bacteria in biofilm, they could induce iron redox cycling in corrosion process. This process enhanced the precipitation of iron oxides and formation of more Fe₃O₄ in corrosion scales, which inhibited corrosion effectively. Therefore, the IRB and NRB in the biofilm can reduce iron release and red water occurrence. Moreover, there are many opportunistic pathogens in biofilm of DWDSs. The opportunistic pathogens growth in DWDSs related to the bacterial community changes due to the effects of micropollutants. Micropollutants increased the number of bacteria with antibiotic resistance genes (ARGs). Furthermore, extracellular polymeric substances (EPS) production was increased by the antibiotic resistant bacteria, leading to greater bacterial aggregation and adsorption, increasing the chlorine-resistance capability, which was responsible for the enhancement of the particle-associated opportunistic pathogens in DWDSs. Moreover, O₃-biological activated carbon filtration-UV-Cl₂ treatment could be used to control the iron release, red water occurrence and opportunistic pathogens growth in DWDSs.     

Effects of O3/Cl2 disinfection on corrosion and opportunistic pathogens growth in drinking water distribution systems

The effects of O_3/Cl_2 disinfection on corrosion and the growth of opportunistic pathogens in drinking water distribution systems were studied using annular reactors(ARs).The corrosion process and most probable number(MPN) analysis indicated that the higher content of iron-oxidizing bacteria and iron-reducing bacteria in biofilms of the AR treated with O_3/Cl_2 induced higher Fe_3O_4 formation in corrosion scales.These corrosion scales became more stable than the ones that formed in the AR treated with Cl_2 alone.O_3/Cl_2 disinfection inhibited corrosion and iron release efficiently by changing the content of corrosion-related bacteria.Moreover,ozone disinfection inactivated or damaged the opportunistic pathogens due to its strong oxidizing properties.The damaged bacteria resulting from initial ozone treatment were inactivated by the subsequent chlorine disinfection.Compared with the AR treated with Cl_2 alone,the opportunistic pathogens M.avium and L.pneumophila were not detectable in effluents of the AR treated with O_3/Cl_2,and decreased to(4.60 ± 0.14) and(3.09 ± 0.12) log10(gene copies/g corrosion scales) in biofilms,respectively.The amoeba counts were also lower in the AR treated with O_3/Cl_2.Therefore,O_3/C_l2 disinfection can effectively control opportunistic pathogens in effluents and biofilms of an AR used as a model for a drinking water distribution system.     

Characterization of the airborne bacteria community at different distances from the rotating brushes in a wastewater treatment plant by 16S rRNA gene clone libraries

Biological risks of bioaerosols emitted from wastewater treatment processes have attracted wide attention in the recent years. However, the culture-based analysis method has been mostly adopted for detecting the bacterial community in bioaerosols, which may result in the underestimation of total microorganism concentration as not all microorganisms are cultivable. In this study, oligonucleotide fingerprinting of 16S rRNA genes was applied to reveal the composition and structure of the bacterial community in bioaerosols from an Orbal oxidation ditch in a Beijing wastewater treatment plant (WWTP). Bioaerosols were collected at different distances from the aerosol source, rotating brushes, and the sampling height was 1.5 m which is the common respiratory height of a human being. The bacterial communities of bioaerosols were diverse, and the lowest bacterial diversity was found at the sampling site just after the rotating brush rotating brush. A large proportion of bacteria in bioaerosols were affiliated with Proteobacteria and Bacteroidetes. Numerous bacteria present in the bioaerosols also emerged in water, indicating that the bacterial community in the bioaerosols was related to that of the aerosols’ sources. The forced aeration of rotating brushes brought about observably distinct bacterial communities between sampling sites situated before and after the rotating brush. Isolation sources of closest relatives in bioaerosols clone libraries were associated with the aqueous environment in the WWTP. Common potential pathogens in bioaerosols as well as those not reported in previous research were also analyzed in this study. Measures should be adopted to reduce the emission of bioaerosols and prevent their exposure to workers.     

Indigenous bacteria may interfere with the biocontrol of plant diseases

Prodigiosin is a reddish antibiotic pigment that plays an important role in the biocontrol of plant diseases by the bacterium Serratia marcescens. However, its activity is unstable under agricultural conditions; further, it can be degraded by various environmental factors. To examine the effect of epiphytic microbes on the stability of prodigiosin used for biological control processes, we collected a total of 1,280 bacterial isolates from the phylloplane of cyclamen and tomato plants. Approximately 72% of the bacterial strains isolated from the cyclamen plants and 66% of those isolated from the tomato plants grew on minimal agar medium containing 100 μg ml⁻¹ prodigiosin. Certain isolates obtained from both plant species exhibited prodigiosin-degrading activity. We compared the 16S rRNA gene sequences derived from the isolates with sequences in a database. The comparison revealed that the sequences determined for the prodigiosin-degrading isolates were homologous to those of the genera Pseudomonas, Caulobacter, Rhizobium, Sphingomonas, Janthinobacterium, Novosphingobium, and Rathayibacter. These results indicate that indigenous epiphytic microorganisms may interfere with the interaction between plant pathogens and biocontrol agents by degrading the antibiotics produced by the agents.     

Proteomic studies of plant and bacteria interactions during benzene remediation

Phytoremediation is a sustainable remedial approach for removing benzene from environment. Plant associated bacteria could ameliorate the phytotoxic effects of benzene on plant, although the specificity of these interactions is unclear. Here, we used proteomics approach to gain a better understanding of the mechanisms involved in plant-bacteria interactions. Plant associated bacteria was isolated and subsequently inoculated into the sterilized Helianthus annuus, and the uptake rates of benzene b...     

3种耐锑土壤细菌的筛选及对锑的吸附研究

从湖南省冷水江锡矿山尾渣土壤中筛选分离出3株具有较高锑耐性的细菌,通过宏观吸附结合X射线光电子能谱（XPS）,阐明3株细菌对Sb（Ⅲ）和Sb（Ⅴ）的吸附机制.根据生理生化指标和16S rDNA序列同源性对比,鉴定出1株革兰氏阴性菌和2株革兰氏阳性菌,分别为人苍白杆菌（Ochrobactrum anthropi）、芽孢杆菌（Bacillus sp.）及蜡状芽孢杆菌（Bacillus cereus）.3株细菌都呈杆状,且都能在高锑含量营养液中良好生长.3株细菌对锑的等温吸附符合Langmuir模型,对锑的最大吸附量约为11.0~14.3 mg·g^-1,其中,蜡状芽孢杆菌对锑的吸附能力高于其它两株细菌,3株细菌对Sb（Ⅲ）的吸附量略高于Sb（Ⅴ）.XPS结果表明,细菌表面的羧基和氨基在锑吸附过程中发挥主导作用.本研究结果对于理解土壤微生物吸附固定锑机制及其在土壤修复中的应用有重要意义.