实时荧光定量PCR法检测环境假单胞菌属细菌丰度

应用实时荧光定量PCR法,建立了总细菌及假单胞菌属的标准曲线,并以环境样本中假单胞菌属与总细菌的比值反映其细菌丰度.应用该方法评价人工除藻反应器中组合填料、无纺布、弹性填料等介质对太湖水中溶藻细菌之一的假单胞菌属的富集情况.结果显示:该方法对总细菌的检测范围为103~108个基因拷贝/μL(R2=0.997);假单胞菌属的检测范围为1~105个基因拷贝/μL(R2=0.994).对除藻反应器溶藻细菌富集效果的评价显示,所建立的方法能有效检测反应器中填料对该类溶藻细菌的富集程度.初步证明所建立的方法可有效定量假单胞菌属在环境样本中的丰度.     

太湖溶藻细菌的分离及评价

从放置于太湖梅梁湾流域的除藻中试装置的人工介质上分离到一株溶藻细菌,经生理、生化和分子生物学鉴定,该株细菌属于芽孢杆菌属.经测定该菌具有较强的溶解铜绿微囊藻和降解微囊藻毒素LR(MC-LR)的功能:该菌(105个/mL)对铜绿微囊藻液(4.5×107个/mL)作用第16,32,48h的溶藻率分别为48%、63%和96%;对MC-LR(2.649μg/L)作用第18,36,54和72h藻毒素降解率分别为15%、29%、73%和100%.应用新建立的实时荧光定量PCR法(RTQ-PCR)对人工介质上的该溶藻芽孢杆菌丰度进行定量检测,结果显示,人工介质上该细菌的丰度约为1%,在7,8,9三个月中呈现递增趋势,其分布具有一定的时空变化规律.     

氧化铁纳米颗粒固定化溶藻菌的除藻作用

为了探讨氧化铁纳米颗粒固定化溶藻菌后对铜绿微囊藻的溶藻作用和降解微囊藻毒素LR(MC-LR)的作用,从太湖水体中分离获得一株属于芽孢杆菌属的溶藻菌,将其与氧化铁纳米颗粒固定化后,以普通Fe2O3作为实验对照,测定其溶藻作用和降解MC-LR作用.以激光粒度仪测定氧化铁纳米颗粒与溶藻菌两者单独和同时存在时的粒径大小及粒径分布,以获得氧化铁纳米颗粒固定化溶藻菌的证据.分析结果表明:在太湖水体中分离获得的一株芽孢杆菌具有较强的溶藻和降解MC-LR的作用,作用24 h后的溶藻率和MC-LR降解率分别达到44.3%和20.5%.同时加入氧化铁纳米颗粒后,溶藻率达到53.0%,MC-LR的降解率达到33.0%.激光粒度仪检测发现,氧化铁纳米颗粒与细菌混合后的颗粒物的平均直径增加到氧化铁纳米颗粒的12.3倍,同时氧化铁纳米颗粒和细菌单独存在时的粒径分布峰消失.氧化铁纳米颗粒是一种具有良好性能的生物固定化材料,可能通过其特有的途径有效地固定化溶藻菌,并增强溶藻菌的溶藻和降解藻毒素作用.     

膜生物反应器净化污水的硝化反硝化性能

比较了膜生物反应器(MBR)和传统活性污泥工艺(CAS)在相同运行条件下处理生活污水的硝化和反硝化性能.结果表明,MBR对NH4 -N和TN的去除率分别比CAS高54.8%和37.3%.2种工艺的亚硝化、反硝化作用均呈零级反应,对应降解速率常数MBR分别约为CAS的2.2倍和2.5倍;CAS中硝化作用为零级反应,而MBR中硝化作用随时间推移趋于平缓.MBR中的细菌总数、硝酸菌、亚硝酸菌和反硝化菌数量分别比CAS工艺中相应菌种高1~2个数量级.通过控制曝气强度或减小回流通道断面限制缺氧区溶解氧质量浓度,可提高MBR中的反硝化效果.     

Biogenesis of β-barrel membrane proteins in bacteria and eukaryotes: evolutionary conservation and divergence

Membrane-embedded β-barrel proteins span the membrane via multiple amphipathic β-strands arranged in a cylindrical shape. These proteins are found in the outer membranes of Gram-negative bacteria, mitochondria and chloroplasts. This situation is thought to reflect the evolutionary origin of mitochondria and chloroplasts from Gram-negative bacterial endosymbionts. β-barrel proteins fulfil a variety of functions; among them are pore-forming proteins that allow the flux of metabolites across the membrane by passive diffusion, active transporters of siderophores, enzymes, structural proteins, and proteins that mediate protein translocation across or insertion into membranes. The biogenesis process of these proteins combines evolutionary conservation of the central elements with some noticeable differences in signals and machineries. This review summarizes our current knowledge of the functions and biogenesis of this special family of proteins.     

N-acetylmuramic acid 6-phosphate lyases (MurNAc etherases): role in cell wall metabolism, distribution, structure, and mechanism

MurNAc etherases cleave the uniqued-lactyl ether bond of the bacterial cell wall sugar N-acetylmuramic acid (MurNAc). Members of this newly discovered family of enzymes are widely distributed among bacteria and are required to utilize peptidoglycan fragments obtained either from the environment or from the endogenous cell wall (i.e., recycling). MurNAc etherases are strictly dependent on the substrate MurNAc possessing a free reducing end and a phosphoryl group at C6. They carry a single conserved sugar phosphate isomerase/sugar phosphate- binding (SIS) domain to which MurNAc 6-phosphate is bound. Two subunits form an enzymatically active homodimer that structurally resembles the isomerase module of the double-SIS domain protein GlmS, the glucosamine 6-phosphate synthase. Structural comparison provides insights into the two-step lyase-type reaction mechanism of MurNAc etherases: β-elimination of the D-lactic acid substituent proceeds through a 2,3-unsaturated sugar intermediate to which water is subsequently added. Received 31 August 2007; received after revision 12 October 2007; accepted 1 November 2007     

石榴枯萎病的生防木霉菌株的筛选及鉴定

目的 石榴枯萎病是由甘薯长喙壳菌引起,对石榴有严重危害的土传病害。利用拮抗微生物抑制病原菌的繁殖是石榴枯萎病绿色防控的有效手段。通过筛选具有对石榴枯萎病原菌产生拮抗效果的微生物,研究其抑制菌作用,以期为石榴枯萎病生物防控提供菌种资源。方法 从石榴的根际土壤中分离出真菌,根据产孢量、生长速率、拮抗系数及平板对峙结果进行筛选,对防效较好的真菌根据形态学和分子系统学进行鉴定,通过盆栽实验评估其对石榴枯萎病的防治效果。结果 能有效抑制石榴枯萎病菌菌丝的生长的菌株T9经鉴定为非洲哈茨木霉菌（Trichoderma afroharzianum）,盆栽防治效果最高可达71.35%。结论 该菌株展现出了对石榴枯萎病的生物防治效果,具有应用价值。     

7种化学杀菌剂和拮抗菌代谢物对苹果斑点落叶病菌的抑制活性及增效组合筛选

目的 为苹果斑点落叶病的防控提供新颖的杀菌剂及其增效组合。方法 采用菌丝生长速率法测定了7种化学杀菌剂、拮抗菌发酵代谢物及其复配剂对苹果斑点落叶病菌的抑制活性。结果 7种化学杀菌剂均对病菌有较强的抑制活性,其中戊唑醇、苯醚甲环唑和丙环唑3种药剂对病菌抑制效果最佳,EC₅₀分别达0.21、0.30和0.81 μg/mL,其次为吡唑醚菌酯、代森锰锌和嘧菌酯。多菌灵的抑菌效果最差,EC₅₀为15.03 μg/mL。生防菌发酵上清液和发酵上清粗提物对病菌的EC₅₀分别为2.38 μL/mL和0.53 μL/mL。发酵上清液粗提物和代森锰锌、吡唑醚菌酯的复配均表现为增效作用,其中体积比6∶4和7∶3达最大毒性比,分别为1.39和1.41。结论 戊唑醇、苯醚甲环唑和丙环唑对苹果斑点落叶病菌的毒力最大。解淀粉芽孢杆菌SJ1606所产粗提物对化学杀菌剂代森锰锌和吡唑醚菌酯具有增效作用,为后续农药新制剂研发提供了参考数据。     

Lipids of sulfate-reducing bacteria and sulfur-oxidizing bacteria found in the Dongsheng uranium deposit

U-bearing sandstones from the Dongsheng deposit in Ordos Basin contain abundant C15-C18 fatty acids. The fatty acids may have been derived from modern and ancient organisms including organisms from the intervals of U mineralization. A certain amount of i15:0, a15:0, a17:0 fatty acids coexist with small amounts of i17:1ω7c and 10me16:0, characteristic biomarkers of Desulfovibrio and Desulfobacter sp., respectively. This indicates the existence of sulfate-reducing bacteria (SRB) in the sandstones. The presence of sulfur-oxidizing bacteria (SOB), such as Beggiatoa and Thioploca, is indicated by significant amounts of 16:1ω7c and 18:1ω7c fatty acids. The existence of the SRB in the deposit, as inferred from the fatty acids, is consistent with results from fossilized microorganisms and isotopic compositions of ore-stage pyrite. This suggests that the environment may have been favorable for the SRB to grow since ore formation (9.8-22 Ma). The bacteria may have degraded hydrocarbons directly, or indirectly utilized hydrocarbons degraded by oxic microbes in the deposits. This process may have produced 12C-rich calcite and prominent baseline humps of unresolved complex mixtures (UCM), and 25-demethylated hopanes and tricyclic terpanes. The existence of sulfur-oxidizing bacteria and sulfate-reducing bacteria in the deposit may have resulted in bacterial sulfate reduction to sulfide, re-oxidization of the sulfide to sulfate and subsequent reduction of the sulfate to sulfide. This assertion is supported by ore-stage pyrite with δ34S values as low as-39.2‰, and the lightest sulfate (about 11‰) measured during the Phanerozoic, a difference of more than 46‰.     

窖泥己酸菌的分离培养与诱变选育

泥窖己酸菌产生的己酸是浓香型大曲酒主体香成分的前体物质之一。从老窖泥中分离纯化出一株产酸能力较高的己酸菌GS5,以其为亲株,通过紫外诱变(照射功率15W,照射距离30cm,时间为3m in),筛选到1株产酸量提高12%的突变株GSUT3,多次继代培养结果表明其产量稳定。研究还发现GSUT3与亲株之间,GSUT3第1代与第8代之间,其生长变化规律和己酸产量的变化规律表现一致。