骆驼刺根际细菌的植物促生能力

采用纯培养方法使用5种培养基分析沙漠干旱植物骆驼刺的根际可培养细菌群落,并用盆钵试验验证这些菌株的植物促生能力.共从骆驼刺根际土中分离纯化了120株细菌,根据16S rRNA基因序列划分成32个16S r RNA基因型,分布在Actinobacteria、Firmicutes、Proteobacteria和Bacteroidetes等4个门的17个属内.其中,Actinobacteria占全部分离菌株的77.3%,是骆驼刺根际的优势微生物;该门的Streptomyces和Arthrobacter两个属是分离菌株最多的属.从16个型(55株)的细菌中检测到了固氮酶nif H基因,占全部分离菌株的45.8%、全部型的50%.盆钵试验中,菌株Microbacterium sp.WLJ053、Streptomyces sp.WLJ079、Paenibacillus sp.WLJ097、Sphingomonas sp.WLJ118和Chryseobacterium sp.WLJ119能显著提高玉米的株高、鲜重和干重,具有植株促生能力.使用营养丰富的LB和WS培养基获得的微生物种类和特有微生物数量都更多,含nif H基因和具有植物促生能力的菌株比例更高.本研究说明沙漠植物根际蕴含了大量微生物种质资源,具有较好的开发前景.     

无苯酚培养基分离到的降酚菌多样性的分子分析

用3种不含苯酚的培养基(YPG、10%YPG和LB)从上海焦化厂废水处理系统(A2/O法)好氧池的悬浮污泥分离到24株降酚菌株.通过16SrDNAPCR扩增产物的限制性酶切分析(amplifiedribosomalDNArestrictionanalysis,ARDRA)、ERICPCR指纹图谱分析、多组分苯酚羟化酶大亚基(thelargestsubunitofthemulticomponentphenolhydroxylase,LmPH)基因的PCR扩增及16SrRNA基因测序的方法对这些降酚菌株进行表征.通过ARDRA分型,将这24株降酚菌株分为8个类型;利用ERICPCR可以将这24个菌株分为17种类型,说明同一ARDRA类型内菌株具有多样性.对17个ERIC类型代表菌株的16SrDNA扩增产物进行克隆并测序,测序结果在GenBank和RDP中进行比对.结果表明,与这17个代表菌株同源性最高的菌中,有6株是未见报道具有降酚功能的菌株.在这24株降酚菌中,有19株在苯酚含量为200mg/L的MP培养基中培养5d,苯酚降解率为20%左右,其余5株苯酚降解率达到100%,且均属于Rhodococcus属.本研究在降酚菌生物多样性上作了有益的探索.图2表1参20     

海洋苯酚降解菌Candida sp.P5的分离鉴定及其降解特性

从海洋沉积物中分离、筛选到一株能以苯酚作为唯一碳源和能源的酵母菌P5．根据菌落特征、菌体形态、生理生化特性和18SrDNA序列分析，确定菌株P5为假丝酵母菌属（Candicla　sp．）．该菌株最适宜生长和降解苯酚的条件为：温度25℃，pH6．0～7．0，摇床转速100r／min，需氧；菌株P5能在较高浓度的苯酚条件下生长，在72h内可以降解95％以上的苯酚．对苯酚代谢途径和相关酶的研究发现，菌株P5主要在邻苯二酚1，2-双加氧酶作用下通过邻位途径进行苯酚代谢．图7表2参24     

活性炭负载TiO2催化臭氧氧化去除水中酚的研究

本文采用活性炭负载TiO2作催化剂对臭氧氧化去除含酚废水进行了研究．研究了不同的酚初始浓度、pH值及臭氧浓度对苯酚去除率的影响，获得了反应动力学常数．实验结果表明：苯酚初始浓度越小，去除率越高．在本实验条件下，向150mL苯酚溶液中通入浓度为3．48mg/L、流量为0．05m^3／h的臭氧化空气反应30min去除率即可达到99％以上。     

石油降解菌株G5 16S rDNA全序列的系统学分析

用PCR方法扩增了一株石油降解菌株G5的16S rRNA基因全序列，并对其进行了克隆和测序．对该序列在GenBank中的BLAST结果表明，所有与该序列高度同源的序列都是假单胞菌的16S rRNA基因．其中假单胞菌的代表菌株Pseudomonas aeruginosa，P．fluoroscens，P .putida，P.syringae的16S rRNA基因序列与G5的16S rRNA基因序列同源性分别为93．4％，98．4％，96．3％，97．5％．对G5和其他39株假单胞菌的16S rRNA基因序列进行聚类分析，获得的系统发育树与RDP(Ribosomal Database Project)报道的系统发育树基本一致，其中菌株G5与5株P．chlororaphis聚类在一起．图2参7     

海洋苯酚降解菌Candida sp.P5的分离鉴定及其降解特性

从海洋沉积物中分离、筛选到一株能以苯酚作为唯一碳源和能源的酵母菌P5.根据菌落特征、菌体形态、生理生化特性和18S rDNA序列分析,确定菌株P5为假丝酵母菌属(Candida sp.).该菌株最适宜生长和降解苯酚的条件为:温度25℃,pH6.0～7.0,摇床转速100r/min,需氧;菌株P5能在较高浓度的苯酚条件下生长,在72h内可以降解95%以上的苯酚.对苯酚代谢途径和相关酶的研究发现,菌株P5主要在邻苯二酚1,2-双加氧酶作用下通过邻位途径进行苯酚代谢.图7表2参24     

苏云金芽胞杆菌菌株WB9编码活性因子的基因分析

采用PCR—RFLP及PCR技术对苏云金芽胞杆菌新分离菌株WB9的ICP、VIP、几丁质酶和肠毒素4种活性因子的编码基因进行了分析．结果表明，该菌株含有编码ICP的cry1Aa、cry1Ab、cry1Cb、cry1Fa和cry1GaS个cry1基因型，编码Vip3A蛋白的vip3A基因以及编码3种肠毒素的hblA、bceT和entS基因；不含编码几丁质酶的基因．将vip3A、hblA、bceT和entS基因PCR产物回收纯化后直接测序，经在线的BLAST软件进行同源性分析，前两个基因片段与Bt已公布基因序列的相应片段同源性分别为99％和96％～98％，bceT基因片段与蜡质芽孢杆菌bceT相应片段同源性为97％．     

2株硝化细菌的分离鉴定与脱氮性能研究

本研究以沙洋太平河生活排污口黑臭水体为研究对象,验证其在乳糖、淀粉、甲酸、葡萄糖及NaHCO₃等不同碳源下的硝化效率及产物类型,以获得其优势碳源,并进一步分离纯化具有高效硝化功能的单菌。结果表明,不同碳源下氮素去除效果不尽相同,作用第4 d的氨氮去除率依次为乳糖>NaHCO₃>葡萄糖>甲酸>淀粉,亚硝态氮去除率依次为乳糖≈葡萄糖>淀粉>NaHCO₃≈甲酸。添加不同碳源的硝化产物具有较大差异,以葡萄糖为碳源时硝化活性最大,氨氮脱除率最高,而以分子量较小、化学结构简单的甲酸为碳源的培养基氨氮脱除率不如以葡萄糖为碳源的培养基高。经过多轮传代富集培养筛选出2株硝化细菌,克隆其16S rRNA基因以获得其分类地位,观察其形态验证其功能发现,16S rRNA基因测序结果表明2株纯菌分别为植物固氮菌属(Phytobacter sp.)和肠杆菌属(Gibbsiella sp.),故分别命名为Phytobacter sp.R3和Gibbsiella sp.R4,以葡萄糖、乳糖、NaHCO₃     

麦草畏降解菌株Ochrobactrum sp.3-3的分离鉴定及其降解特性

麦草畏是理想的抗除草剂转基因工程的靶标除草剂;发掘新的麦草畏高效降解菌株和基因具有非常重要的理论和应用价值.从南京土壤样品中分离到一株麦草畏高效降解菌株,命名为3-3.根据生理生化特征和16S r DNA序列相似性分析,将其初步鉴定为苍白杆菌属(Ochrobactrum sp.).菌株3-3在48 h内完全降解100 mg/L的麦草畏.该菌株降解麦草畏的最适温度为30℃,最适p H为7.0.代谢产物高效液相和质谱鉴定结果表明该菌株降解麦草畏的起始步骤是脱甲基,形成没有除草活性的3,6-二氯水杨酸(DCSA).菌株粗酶液只在NADH存在时才有麦草畏脱甲基酶活性.PCR扩增和该菌基因组生物信息学分析表明该菌株没有已报道的麦草畏脱甲基酶基因DMO、Mtv及Dmt或其同源序列.总之,本研究首次分离筛选到苍白杆菌属的麦草畏降解菌,且该菌可能存在一个新的氧化酶类麦草畏脱甲基酶基因.     

硫杆菌的分离鉴定及其对煤矿废弃物的氧化脱硫特性

从宁夏大武口高硫煤矸石山的酸性废水中分离得到一株自养硫氧化细菌CMTF-32．该菌极端耐酸，能氧化低价态硫化物，氧化单质硫的能力强，在d11时单质硫的氧化率达到65．6％，日产硫酸能力最高时能达3．0g／L．培养基pH值在培养过程中明显下降．16S rDNA序列分析表明，该菌株与Acidithiobacillus thiooxidans相似性为99％，结合形态、生理生化实验结果，确定菌株CMTF-32为嗜酸氧化硫硫杆菌．煤矸石的脱硫实验表明，在煤矸石粒径小于2mm，初始pH为1．5，30℃培养条件下脱硫效果最佳，d14时脱硫量为2．72g／L，脱硫率可达到84．5％，使煤矸石的硫污染得到了有效的控制．图5表1参13     

Quantification of polyphenols during retting and characterization of bacteria from the Kadinamkulam Backwaters, Kerala

The retting environment which provides a competitive niche for specialized microbes is speculated to harbour a variety of microbes with high biodegradation potential. In this context, an effort has been made to isolate and identify bacterial species having high tolerance to phenol In vitro. Maximum polyphenol (1.897 mg l(-1)) as observed during the initial period of retting, which decreased as retting proceeded. Based on biochemical characterization, the isolated bacterial strains were identified as Micrococcus sp., Moraxella sp. strain MP1, Moraxella sp. strain MP2 and Moraxella sp. strain MP3, Pseudomonas sp. strain PP1 and Pseudomonas sp. strain PP2, Amphibacillus sp., Brucella sp. strain BP1 and Brucella sp. strain BP2, Aquaspirillum sp., Escherichia coli strain EP1 and Escherichia coli strain EP2, Campylobacter sp., Aeromonas sp., Neisseria sp., Vibrio sp., Erwinia sp. and Mesophilobacter sp. These strains were found to tolerate maximum concentration of phenol viz. 200 to 1000 mg l(-1). Plasmid analysis of phenol resistant bacterial isolates showed that almost all the cultures had at least one plasmid of size > 1Kb. Studies on the protein profile of isolated bacterial cultures showed the presence of proteins with molecular sizes ranging from 10 to 85 KDa with exception of Mesophilobacter and Neisseria having still high molecular weight protein (95 KDa). Bacterial strains isolated from coir-ret-liquor showed tolerance to high phenol concentration.     

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.     

ERIC-PCR:一种快速鉴别环境细菌菌株的方法

以 Ｅ Ｒ Ｉ Ｃ（ 肠杆菌基因间共有重多序列） 序列设计的特异引物对３ 株大肠杆菌、３ 株软腐欧氏杆菌、２ 株草生欧氏杆菌、１ 株梨火疫欧氏杆菌、１ 株催娩克氏菌、１ 株阴沟肠杆菌和９ 株具有降酚能力的细菌等２０ 种细菌的基因组 Ｄ Ｎ Ａ 进行 Ｐ Ｃ Ｒ 分析．每种菌株均存在数目不等的各自独特的带型,能够区分同一种类中极为相近的菌株．经多次重复,各特异性扩增的主要带型能重复稳定出现．聚类分析的结果表明,供试菌株多数按照分类地位归到相应的组中．９ 种具有降酚能力的细菌中,２ 种分离自处理焦化废水池活性污泥,７ 种来自土壤样品． Ｅ Ｒ Ｉ Ｃ Ｐ Ｃ Ｒ 指纹图显示,前２ 种属于一类,都有一条大小约１ ．１ ｋｂ 的主带,其它７ 种土壤中分离出来的细菌除其中１ 种有１ 条约１ ．２ ｋｂ 大小的主带外,其余６ 种都有１ 条大小约１ ．４ ｋｂ 的带．聚类分析将从土壤中分离出来的细菌归为３ 种不同的类型． Ｅ Ｒ Ｉ Ｃ Ｐ Ｃ Ｒ 可以从分子水平对细菌基因组进行快速指纹图分析,在对环境细菌分离物进行快速鉴定和归类等方面具有实用价值     

Assessment of phenol-degrading ability of Acinetobacter sp. B9 for its application in bioremediation of phenol-contaminated industrial effluents

Successful bioremediation of a phenol-contaminated environment requires application of those microbial strains that have acquired phenol tolerance and phenol-degrading abilities. A newly isolated strain B9 of Acinetobacter sp. was adapted to a high phenol concentration by growing sequentially from low- to high-strength phenol. The acclimatised strain was able to grow and completely degrade up to 14?mM of phenol in 136?h. The degradation rates were found to increase with an increase in the phenol concentration from 2.0 to 7.5?mM. The strain preferred neutral to alkaline pH range for growth and phenol degradation, with the optimum being pH 8.0. The optimum temperature for phenol degradation was found to be in the range of 30–35°C. Transmission electron micrographs showed a disorganised and convoluted cell membrane in the case of phenol-stressed cells, showing a major effect of phenol on the membrane. Enzymatic and gas chromatography-mass spectrometry studies show the presence of an ortho-cleavage pathway for phenol degradation. Efficient phenol degradation was observed even in the presence of pyridine and heavy metals as co-toxicants showing the potential of strain in bioremediation of industrial wastes. Application of strain B9 to real tannery wastewater showed 100% removal of initial 0.5?mM phenol within 48?h of treatment.     

苯酚降解菌UW7的鉴定及对苯酚的降解作用

从焦化厂污水处理曝气池泥样中分离出具有降解苯酚能力的UW7菌株,根据形态、生理生化性状初步鉴定为不动杆菌属(Acinetobacter).该菌16S rRNA基因序列(在GenBank中的登录号为GU083586)与多株鲁氏不动杆菌(A.lowffii)的相似性在99%以上.结合形态、生理生化特性,鉴定UW7菌株为鲁氏不动杆菌(A.lowffii UW7),该种细菌具有降解苯酚的特性尚未见报道.该菌株降解苯酚的最适温度为30℃,最适生长pH 7.0,对2.5 g/L浓度的苯酚能够有效降解,对3.5～4.0 g/L浓度的苯酚有较强的耐受能力,是处理高浓度苯酚废水的良好菌种资源.     

葡枝根霉(Rhizopus stolonifer)YF6脂肪酶基因的克隆及其序列分析

筛选并鉴定了1株高产脂肪酶菌株葡枝根霉YF6,并采用简并PCR等分子技术从中克隆到1个新的脂肪酶基因lipRs及其对应的全长cDNA,序列分析表明,该基因编码区全长1173bp,不含内含子序列,编码1段由26个氨基酸残基组成的信号肽和1个由365个氨基酸残基组成的成熟蛋白,该成熟蛋白计算分子量(Mr)为39268.27,等电点为pH7.66,有5个可能的N-糖基化位点,含有脂肪酶特征序列GHSLGGA,与来自米根霉(Rhizopus oryzae)的脂肪酶(AAF32408)同源性最高,一致性为83%,相似性为89%.该基因序列已提交GenBank,登录号为DQ139862.     

Sequential anaerobic and aerobic treatment of pulp and paper mill effluent in pilot scale bioreactor

In the present study sequential anaerobic and aerobic treatment in two step bioreactor was performed for removal of colour in the pulp and paper mill effluent. In anaerobic treatment, colour 50%, lignin 62%, COD 29%, absordable organic halides (AOX) 25% and phenol 29% were reduced in eight days. The anaerobically treated effluent was separately applied in bioreactor in presence of fungal strain, Paecilomyces sp., and bacterial strain, Microbrevis luteum. Data of study indicated reduction in colour 80%, AOX 74%, lignin 81%, COD 93% and phenol 76 per cent by Paecilomyces sp. where as Microbrevis luteum showed removal in colour 59%, lignin 71%, COD 86%, AOX 84% and phenol 88% by day third when 7 days anaerobically treated effluent was further treated by aerobic microorganisms. Change in pH of the effluent and increase in biomass of microorganism's substantiated results of the study, which was concomitant to the treatment method.     

一株菲降解菌的特性及相关降解基因的克隆

从石油污染土壤中分离到一株菲降解菌2F5-2.根据该菌株生理生化特征和16S rDNA序列相似性分析,将其初步鉴定为鞘氨醇杆菌属(Sphingobium sp.).该菌株在10 h内对100 mg/L的菲的降解率为100%.降解菲的最适温度为30℃,最适pH为7.对降解途径的初步研究显示,该菌株通过水杨酸途径降解菲.克隆了编码芳香烃双加氧酶α亚基的基因phdA,它与菌株Sphingomonas sp.P2、Sphingobium yanoikuyae B1、Sphingomonas sp.ZP1中phdA的同源性分别为97.9%、98%和100%,表明该基因具有保守性.图6参16     

丁草胺高效降解细菌的分离

通过瓶培养法富集培养,从肥东县一块单晚稻田土中分离出一株丁草胺高效降解细菌ＷＹ３０６,经鉴定,该菌为节杆菌属菌Ａｒｔｈｒｏｂａｃｔｅｒ ｓｐ．ＷＹ３０６．Ａｒｔｈｒｏｂａｃｔｅｒ ｓｐ．ＷＹ３０６降解丁草胺的影响因素研究表明：丁草胺降解半衰期与初始菌量近似成反比;当丁草胺添加浓度为５、９ｍｇ／Ｌ时,其降解半衰期分别为０．９７ｈ和１．８６ｈ,随浓度的增大而增大,而当丁草胺添加浓度为０．８ｍｇ／Ｌ或其