节杆菌AD26的分离鉴定及其与假单胞菌ADP对阿特拉津的联合降解

用富集培养法,从农药厂的工业废水中分离到高效降解除草剂阿特拉津的AD26菌株,通过16SrRNA基因序列分析,该菌株被鉴定为节杆菌（Arthrobacter sp.）。降解基因的PCR分析表明,AD26含有阿特拉津降解基因trzN和atzBC,它能以阿特拉津为唯一氮源、蔗糖或柠檬酸钠为碳源生长,将阿特拉津降解成氰尿酸,降解速度快但降解不完全。假单胞菌（Pseudomonas sp.）ADP是Wackett实验室分离的阿特拉津降解菌株,含有阿特拉津降解基因atzABCDEF,能以阿特拉津为唯一氮源、柠檬酸钠为碳源（不能以蔗糖为碳源）生长,将阿特拉津降解成NH3和CO2,降解完全但降解速度慢。在阿特拉津浓度为200mg·L^-1的无机盐培养基中进行的AD26和ADP混合培养表明,它们对阿特拉津的降解发生了互补和增强作用,两个菌株能在以阿特拉津为唯一氮源、蔗糖为碳源的培养基中生长,而且生长和降解速率都好于单个菌株,培养72h后阿特拉津去除率达到99.9%,其中76.7%的阿特拉津被降解成NH3和CO2。这表明由节杆菌AD26和假单胞菌ADP组成的混合菌株在阿特拉津废水处理和污染土壤的生物修复中有很好的应用潜力。     

琉璃苣种子发育过程中脂肪酸成分的变化

为探讨琉璃苣种子发育过程中脂肪酸合成的变化规律,采用脂肪酸甲酯气相色谱法(GC-FAME)对琉璃苣种子发育过程中脂肪酸成分的相对含量进行测定。结果表明,琉璃苣种子主要含有棕榈酸、硬脂酸、油酸、亚油酸、γ-亚麻酸、α-亚麻酸、花生一烯酸和芥子酸等8种脂肪酸成分;成熟种子不饱和脂肪酸成分占总脂肪酸的84.05%,其中亚油酸最高(38.20%),γ-亚麻酸次之(24.59%);随着种子发育,棕榈酸、硬脂酸和α-亚麻酸的相对含量不断下降,其它不饱和脂肪酸成分的相对含量不断增加;棕榈酸与α-亚麻酸呈显著正相关,二者与其它脂肪酸成分呈负相关。本研究结果为进一步揭示琉璃苣种子脂肪酸的代谢及其调控机制提供了数据支持。     

摘除卵巢对母山羊肾周脂肪中脂肪酸组成及相关基因mRNA表达量的影响

公羊去势(摘除睾丸)作为一项十分成熟的生产技术,在畜牧生产上得到了非常广泛的应用,但是有关母山羊(Capra hircus)去势的研究还不多。本研究旨在探讨去势对母山羊血清中血糖、甘油三酯、总胆固醇水平,肾周脂肪组织中脂肪酸的组成以及对乙酰Co A羧化酶(acetly-Co A carboxylase,ACC)、脂肪酸合成酶(fatty acid synthase,FAS)、脂蛋白酯酶(lipoprotein lipase,LPL)和激素敏感酯酶(hormone sensitive lipase,HSL)基因表达的影响。将体重相近的5月龄母山羊摘除卵巢后测定其血糖、甘油三酯、胆固醇水平;采用Agilent-6890N气相色谱仪测定其肾周脂肪组织中脂肪酸的组成;利用实时荧光定量PCR(q RT-PCR)相对定量方法检测FAS、ACC、LPL和HSL m RNA的相对表达量。结果表明,摘除卵巢后,母山羊血清中血糖、甘油三酯和总胆固醇水平较对照组均无显著性差异(P0.05),而且在整个实验过程中保持稳定;肾周脂肪组织中总饱和脂肪酸和4种饱和脂肪酸(SFA)(癸酸(C10∶0)、十五碳酸(C15∶0)、棕榈酸(C16∶0)和木质素酸(C24∶0))的比例(P0.05或P0.01)显著降低;肾周脂肪中总单不饱和脂肪酸(MUFA)和总多不饱和脂肪酸(PUFA)的比例与对照组差异不显著(P0.05),但是表现出了一种上升的趋势,油酸(C18∶1)比例显著上升(P0.05);脂肪酸代谢相关基因中LPL m RNA表达量显著提高(P0.05),ACC和FAS基因的表达量极显著提高(P0.01),HSL基因的表达量显著降低(P0.05)。摘除卵巢不会打破母山羊血清中血糖、甘油三酯、总胆固醇的平衡,可能通过影响肾周脂肪组织中脂肪酸代谢相关基因(FAS,ACC,LPL和SHL)m RNA的表达,降低肾周脂肪中饱和脂肪酸的比例,有利于改善羊肉的风味。本研究结果为揭示摘除母山羊卵巢改善肾周脂肪中脂肪酸的构成提供基础资料。     

Changes in the Amino Acid Composition of Decomposing Plant Materials in Soil: Species and Depth Effects

Abstract

This project studied the amino acid composition of samples taken from an experiment of in situ incubation of plant debris, mixed with mineral earth, to identify changes during decomposition. Medicago sativa debris, and Eucalyptus globulus and Pinus halepensis ground litter, were mixed with a OM‐poor mineral earth, and buried in the soil at 5, 20, and 40 cm depth, for 2 years. The total amino acid composition of the final samples was compared with that of the initial ones. The amino acid composition of the several initial mixtures was quite similar. Also, the differences between initial and incubated samples were small, but still large enough to detect consistent changes. The pattern of change was consistent for only nine amino acids: Gly consistently increased, whereas Lys, His, Leu, and (to a lesser extent) Arg, Tyr, Phe, Ser, and Tau consistently decreased. The ratio Gly/(Lys+His+Leu+Arg+Tyr+Phe+Ser) is taken as the amino acid signature (AaS). This ratio increased from about 0.30–0.35 in the undecomposed mixtures, to about 0.55–0.60 after 2 years of field decomposition, and the comparison with data from other authors suggests that it could be a useful indicator of the degree of biochemical evolution of nitrogen compounds in soil samples. In contrast, changes in the traditional groups of amino acids, i.e., acid, basic, neutral, and sulfur (S)‐containing, were hardly detectable.     

脂肪酸合成酶基因在皮下脂肪中表达及其与血清Leptin的关系

动物的体脂沉积所需要的脂肪酸大多是来自脂肪酸的全程合成,即由脂肪酸合成酶(FAS)催化乙酰辅酶A和丙二酸单酰辅酶A合成甘油三脂(TAG)(Simith et al.,2003)。FAS表达水平的升高显著增加甘油三脂在体内的沉积而导致肥胖(Semenkovich,1997)。leptin是ob基因表达产物,是一种能调节     

Bioremediation of Pulp and Paper Mill Effluent by Tannic Acid Degrading Enterobacter sp.

Wastewaters from pulp and paper mills are highly toxic and around 250 xenobiotic compounds have been reported in the effluents. Tannic acid degrading bacterium, Enterobacter sp. was isolated from soil by tannic acid enrichment. This isolate was used for bioremediation of pulp and paper mill effluents. Parameters like temperature, agitation, inoculum size and treatment duration were optimized by using Qualiteck-4 software. Reduction in lignin 73% and colour up to 82% was also observed. Encouraging results were observed is reduction of COD, BOD with 16-h retention time in batch culture.     

哈夫尼菌降解毒死蜱条件的优化及动力学模型建立

利用响应面法优化哈夫尼菌降解毒死蜱的条件,并建立了降解模型。采用Box-Behnken设计试验,以响应面法进行优化,结果表明,哈夫尼菌降解毒死蜱的最优条件为：蔗糖浓度0.31%,毒死蜱初浓度51.33mg·L-1,培养温度30.7℃,在该条件下理论预测毒死蜱降解率可达74.7%。通过假设和验证,得出哈夫尼菌降解毒死蜱为一级动力学模型,哈夫尼菌能水解毒死蜱P-O键,属一级酶促反应。     

Breakdown Products in the Metabolic Pathway of Anthracene Degradation by a Ligninolytic Fungus Polyporus sp. S133

Polyporus sp. S133 fungi were selected based on their ability to degrade anthracene in liquid media. The degradation efficiency of anthracene increased by adding 0.5% Tween 80 to reach 71%; agitation at 120 rev/min increased the degradation to 92% after 30?days of incubation. Enzymes such as manganese peroxidase (MnP), lignin peroxidase (LiP), laccase, 1,2-dioxygenase and 2,3-dioxgenase were produced by Polyporus sp. S133 during incubation, and the highest enzyme activity was 182.3 U l^?1 by 1,2-dioxygenase after 20?days of incubation. These results indicate that ligninolytic and dioxygenase enzymes secreted from Polyporus sp. S133 could play an important role in anthracene degradation efficiency. The metabolites detected through the degradation pathway were anthraquinone, phthalic acid, benzoic acid and catechol.     

Biodegradation of the Anionic Surfactant Linear Alkylbenzene Sulfonate (LAS) by Autochthonous Pseudomonas sp.

Anionic surfactants, the earliest and the most common surfactants in detergent and cosmetic product formulations contribute significantly to the pollution profile of the ecosystem. Linear alkylbenzene sulfonates (LAS), a major chemical constituent of detergents, forms an imperative group of anionic surfactants. Bioremediation of LAS by conventional processes such as activated sludge is ineffective due to the low kinetics of degradation by unsuitable organisms and foam production. Hence this study was focused on isolating and characterizing indigenous LAS-degrading bacteria from soil. Twenty different LAS-degrading bacteria were isolated from detergent-contaminated soil by enrichment culture technique and degradation efficiency was assessed by Methylene Blue Active Substances (MBAS) assay and by reverse-phase high-performance liquid chromatography (HPLC) analysis. The most efficient LAS-degrading isolates, L9 (81.33?±?0.7) and L12 (81.81?±?0.8), were selected and identified as Pseudomonas nitroreducens (MTCC 10463) and Pseudomonas aeruginosa (MTCC 10462). The 16S rDNA sequences of the isolates were deposited in NCBI GenBank under the accession numbers HQ 271083 (L9) and HQ 271084 (L12). The isolates were capable of degrading 0.05?g/l LAS at 25?°C and pH 7.0?C7.5. Presence of a solid support caused biofilm formation which in turn enhanced LAS degradation. The isolates tend to display diauxic growth with alternate carbon source such as dextrose. These isolates also have the capability to degrade other xenobiotics like hydrocarbons and pesticides. Since xenobiotic pollutants in nature occur as a mixture of compounds rather than single pollutants, the potential of these two indigenous LAS-degrading isolates to degrade multiple xenobiotics gains relevance.     

普施特降解菌Bacillus sp.zx2和zx7生长及降解特性

芽孢杆菌zx2和zx7是普施特高效降解菌,研究其生长和降解特性旨在为普施特污染土壤的生物修复提供科学依据。采用瓶培养法,对芽孢杆菌zx2和zx7的生长特性及单菌和复合菌对普施特的降解特性进行了研究。结果表明,zx2和zx7均可在普施特初始浓度≤200mg·L-1的无机盐培养液中生长良好,zx2在温度25～35℃和pH4.0～7.0时生长良好,而zx7适宜在温度30～35℃和pH5.0～8.0时生长,可见在适应性上二者互补。在最佳条件（温度32℃、pH6.0和普施特初始浓度为200mg·L-1）下,zx2和zx7在无机盐培养液中对普施特降解动态均符合阻滞动力学,半衰期分别为3.8d和2.8d,培养6d时普施特降解率分别为85.81%和90.27%。在培养过程中,zx2的pH是降低的,而zx7的pH基本不变,可初步表明二者降解机理不同;zx2和zx7复合菌（1∶1）对普施特降解率比单菌低,为82.70%,这可能是因为zx2或zx7降解普施特的过程中利用了对方产生的降解产物。     

禾长蠕孢菌的紫外诱变改良和除草活性评价

为进一步提高稗草生防潜力菌禾长蠕孢菌(Helminthosporium gramineum Rabenh f.sp.echinochloae,HGE)的杀草活性,采用紫外诱变的方法对该菌种进行改良,经筛选获得1株适合液体发酵的突变菌株M1。与出发菌株HGE菌落墨绿色相比,突变株M1菌落呈灰白色至白色,表现出明显差异,并且M1生长速度快,其发酵液对稗草(Echinochloa crusgalli(L.)Beauv)根芽的抑制效果均优于出发菌株HGE。连续7次传代培养发现M1仍具有良好的遗传稳定性。采用RAPD-PCR方法对出发菌株和突变菌株M1同时进行PCR扩增,结果表明,突变菌株的遗传物质已经发生了改变。同时对其液体发酵工艺进行了研究,初步确定了最佳发酵工艺。所用培养基马铃薯200gL,葡萄糖30gL,麸皮20gL,酵母膏1gL,KH2PO42gL,MgSO4·7H2O1gL,初始pH7,发酵温度24℃和150rmin振荡培养6d的发酵方案,突变菌株M1菌丝体产量最高,达14.37gL。液体培养获得毒素的最佳发酵工艺为培养基蔗糖30gL,(NH4))2C4H4O65gL,NH4NO31gL,酵母膏1gL,KH2PO41gL,MgSO4·7H2O0.5gL,NaCl0.1gL,CaCl20.1gL,水解酪蛋白0.5gL;初始pH7.25,发酵温度为24℃和静置培养14d。     

Restriction Fragment Length Analysis of the Cytochrome b Gene and Muscle Fatty Acid Composition Differentiate the Cryptic Flatfish Species Solea solea and Solea aegyptiaca

Overlapping external morphometric characters easily confound the flatfishes Solea aegyptiaca and Solea solea (Soleidae) in areas of the Mediterranean Sea where both species live in sympatry. This leads to uncertainties in the fisheries and marketing of the species, in addition to misinterpretations in biogeography and conservation studies. This paper describes a simple restriction fragment length-based diagnostic test that differentiates S. solea from S. aegyptiaca, as well as from other species of the Soleidae family. Furthermore, the two species living in sympatry in the Gulf of Kavala (North Aegean Sea, Greece) present significant qualitative differences in muscle fatty acid composition, a property that can also be used to distinguish the two cryptic species.     

乙草胺降解菌株Rhodococcus sp.AC-1的分离及降解特性研究

为解决乙草胺残留问题,采用连续传代富集培养的方法,从长期生产乙草胺的农药厂污染土壤中分离筛选到1株乙草胺降解菌株AC-1。经过形态学特征、生理生化特征和16S rRNA基因序列系统发育分析,菌株AC-1被鉴定为红球菌属(Rhodococcus)。菌株AC-1在48h内能将0.2 m M的乙草胺完全降解,但不能矿化乙草胺。借助LC-MS,确定乙草胺降解终产物为2-氯-N-(2-乙基-6-甲基苯基)乙酰胺(CMEPA)。菌株AC-1降解乙草胺的最适温度为30℃,最适p H值为7.5。0.1 m M的Cu2+和Hg2+对菌株AC-1降解乙草胺具有很强的抑制作用,而0.1 m M的Fe2+则对菌株AC-1降解乙草胺具有微弱的促进作用。菌株AC-1对乙草胺的降解效果与起始接种量呈正相关。菌株AC-1对甲草胺和丁草胺表现出良好的降解效果,但对异丙甲草胺的降解能力非常微弱。土壤降解试验表明,投加菌株AC-1可以明显促进土壤中乙草胺的降解。本研究为乙草胺的有效降解提供了数据和技术支持。     

PchsA驱动的烟草orf25基因表达载体构建及遗传转化

为研究orf25基因的功能及其与烟草雄性不育性的相关性,通过PCR方法从质粒p UC57-Pchs A中扩增了花特异性启动子Pchs A,并以其取代植物表达载体p BI121中的组成型启动子Ca MV35S,然后将orf25基因插入到Pchs A启动子下游,构建由Pchs A启动子驱动的orf25基因表达载体,测序结果显示植物表达载体p BI121-Pchs A-orf25构建成功。将该表达载体导入根癌农杆菌LBA4404中,采用根癌农杆菌介导法遗传转化雄性不育烟草MS革新3号,共获得76株转基因烟草植株,经PCR鉴定,转基因阳性植株为18株,转化率为23.7%。研究结果证明目的基因orf25已整合到烟草基因组中,为进一步探究该基因的功能及其与烟草雄性不育性的关系奠定了基础。     

脂肪酸单体碳同位素的沉积学记录及其环境意义

湖泊/水库沉积物是有机化学领域的重要研究载体,与其他自然记录比较,它具有信息丰富、时间分辨率高、记录连续性好和覆盖面广等优点。自20世纪80年代以来,在古环境/古气候的研究中,沉积物有机质分子的标志物研究发挥了无可取代的作用。脂肪酸及其单体碳同位素的分布特征与环境意义是目前研究热点。以沉积物为研究载体,采用文献荟萃的方法,探讨了脂肪酸及其单体碳同位素技术在有机质溯源、流域内植被的恢复和气候的反演等方面的研究进展。结果表明:脂类化合物在来源上存在重叠,脂肪酸的研究可以作为正构烷烃分子标志物研究的补充。与单体碳同位素组成特征的联合应用,能够准确地对沉积有机质进行溯源,可以充分揭示保存在陆源脂肪酸中的气候环境信息,为指示气候和环境变化提供重要的线索。在重建区域性的气候和环境状况方面具有广阔的应用前景。     

Degradation of cyhalofop-butyl (CyB) by Pseudomonas azotoformans strain QDZ-1 and cloning of a novel gene encoding CyB-hydrolyzing esterase

Cyhalofop-butyl (CyB) is a widely used aryloxyphenoxy propanoate (AOPP) herbicide for control of grasses in rice fields. Five CyB-degrading strains were isolated from rice field soil and identified as Agromyces sp., Stenotrophomonas sp., Aquamicrobium sp., Microbacterium sp., and Pseudomonas azotoformans; the results revealed high biodiversity of CyB-degrading bacteria in rice soil. One strain, P. azotoformans QDZ-1, degraded 84.5% of 100 mg L(-1) CyB in 5 days of incubation in a flask and utilized CyB as carbon source for growth. Strain QDZ-1 could also degrade a wide range of other AOPP herbicides. An esterase gene, chbH, which hydrolyzes CyB to cyhalofop acid (CyA), was cloned from strain QDZ-1 and functionally expressed. A chbH-disrupted mutant dchbH was constructed by insertion mutation. Mutant dchbH could not degrade and utilize CyB, suggesting that chbH was the only esterase gene responsible for CyB degradation in strain QDZ-1. ChbH hydrolyzed all AOPP herbicides tested as well as permethrin. The catalytic efficiency of ChbH toward different AOPP herbicides followed the order quizalofop-P-ethyl ≈ fenoxaprop-P-ethyl > CyB ≈ fluazifop-P-butyl > diclofop-methyl ≈ haloxyfop-P-methyl; the results indicated that the chain length of the alcohol moiety strongly affected the biodegradability of the AOPP herbicides, whereas the substitutions in the aromatic ring had only a slight influence.     

一种新的精子膜蛋白sp18基因的克隆及其在E.coli 中的表达

摘要：利用sp18 mAb筛选小鼠睾丸λgt11-cDNA表达文库,命名为sp18（GenBank登录号：AF129872）的阳性克隆由 1 468 bp组成,开放阅读框（open reading frame, ORF）长约429 bp,编码142个氨基酸。起始密码ATG位于第694 bp,终止密码位于第1 122 bp,polyA位于1 245~1 250 bp。核苷酸同源性分析表明,sp18 cDNA的10~651 bp与人视神经Hr44的532~ 1 173 bp、sp18 cDNA的10~816 bp、864~1 444 bp与牛视神经Hr44的1 408~2 208 bp、2 250~2 309 bp的cDNA序列有很高的同源性, 高达98%。推测sp18基因可能是生殖系统存在的一种新基因。sp18编码蛋白的理论分子量约为15.7 kD,有8个N-糖基化位点和12个O-糖基化位点,亲水性分析表明sp18多肽是一种精子跨膜蛋白,跨膜区位于第17~33氨基酸处。将sp18 cDNA亚克隆到原核表达载体pET-30a(+),在E. coli BL21（DE3）中得到诱导表达,其表观分子量约为16.5 kD,表达量约占菌体总蛋白的32%。Western blot分析表明sp18 mAb能识别重组的sp18膜蛋白,表明重组蛋白具有免疫原性,为进一步研究sp18基因的功能打下了基础。     

不同种植年限设施菜地土壤有机质组成与结构变化

试验采集不同种植年限设施菜地土壤样品,测定土壤有机质(SOM)、可溶性有机碳(DOC)、胡敏酸(HA)、富里酸(FA)、微生物量碳(MBC)、微生物商(C_(mic)-to-C_(org))、C矿化和E4/E6值等指标,结合傅立叶变换红外光谱技术解析不同种植年限土壤有机质组成与结构变化的规律。结果显示,随着种植年限的增加,SOM、DOC、HA、HA/FA、MBC、C_(mic)-to-C_(org)、E4/E6值、C矿化率和CO_2释放速率均出现明显下降趋势,其中MBC、C_(mic)-to-C_(org)、C矿化率、CO_2释放速率和HA/FA在种植6 a时下降最明显,之后变化缓慢。与露天农田土壤(CK)相比,设施菜地呈现较高的SOM、DOC、Humus、HA、FA、HA/FA和E4/E6。胡敏酸红外光谱图谱显示设施菜地种植20 a处理土壤芳构化程度高,结构稳定,难以降解,其他三种种植年限(3 a、6 a和10 a)的芳构化程度相对较低,脂肪族成分相对较高。综上可知,设施菜地土壤有机质组成与结构随着种植年限的增加具有规律性变化,结果加深了对设施菜地土壤有机质演变规律的认知。