重金属铅与两种淡水藻的相互作用

为了研究重金属铅与淡水藻类之间的相互作用,采用不同Pb2+浓度处理铜绿微囊藻(Microcysis aeruginosa Kutz.)和斜生栅藻[Scenedesmus obliquus(Turp.)Kutz.],分别对两种藻的生物量、藻液电导率、O-·2含量、过氧化物酶(POD)、过氧化氢酶(CAT)活性以及藻对Pb2+的吸收作用等进行了测定,并通过扫描电镜观察了不同Pb2+浓度处理下两种藻细胞的表面结构。结果显示:(1)Pb2+浓度低于3 mg/L促进铜绿微囊藻生长,高于9 mg/L抑制其生长;但在3—12 mg/L范围内,Pb2+均明显抑制了斜生栅藻的生长,说明斜生栅藻对Pb2+毒性的敏感程度要高于铜绿微囊藻。(2)受到铅离子的胁迫,两种藻细胞膜通透性均有一定改变,扫描电子显微镜的照片观察,两种藻细胞表面的絮状物随着Pb2+的升高而增多,尤其是斜生栅藻细胞结构改变明显,多数细胞变形破裂;同时,O-·2含量升高,POD、CAT活性早期均可随Pb2+的增加而上升,表明氧自由基的产生增多以及由其引起的细胞生理生化改变可能是铅离子作用于藻细胞的主要机制。(3)两种淡水藻对Pb2+均有吸收作用,单位量藻细胞内,斜生栅藻对Pb2+的吸收能力好于铜绿微囊藻。所有结果提示:斜生栅藻不仅可以作为对重金属敏感的指示生物来监测水体Pb2+污染程度;同时由于斜生栅藻比铜绿微囊藻具有更好的Pb2+吸收能力,因此还可以利用斜生栅藻作为处理水体Pb2+的生物材料。     

斜生栅藻在不同磷条件下氮生态幅的研究

为获得斜生栅藻（Scendesmus obliquus）的氮生态幅,研究根据中华人民共和国地表水环境质量标准磷浓度界定,利用谢尔福德（Shelford）耐受定律进行曲线拟合对斜生栅藻在低磷（0.02 mg/L）、中磷（0.2 mg/L）和高磷（0.4 mg/L）三种不同磷浓度下氮的生态幅进行定量表达,获得三种磷起始条件下斜生栅藻生长的最佳氮浓度、氮适宜生长范围和氮耐受范围。研究表明,在三种磷条件下斜生栅藻生长的最佳氮浓度分别为1.02、8.91和18.05 mg/L,对应的最大比生长速率分别为（0.1420.006）、（0.3140.002）和（0.3460.007） /d,氮适宜生长范围分别为（0.521.52）、（4.4813.34）和（11.7224.38） mg/L,氮耐受限度分别为（0.022.02）、（0.0517.77）和（5.3930.71） mg/L。这表明富营养化水体可能引起斜生栅藻的大量生长、繁殖,也暗示了斜生栅藻能作为高氮水环境的一个良好指示生物。       

斜生栅藻对振荡和磷胁迫的生理生化响应

为了探讨水文条件的变化对常见绿藻水华发生的潜在影响,实验以斜生栅藻为材料,研究了不同水流状态及磷浓度对其生长及磷利用策略的影响。实验分别设置静止条件(0 r/min)、低振荡条件(90 r/min)和高振荡条件(120 r/min),同时设置磷限制(0.2 mg/L)和磷充足(2 mg/L)两组磷浓度,整个实验过程为3周。实验过程测定的指标为:比生长速率、胞外碱性磷酸酶活性、叶绿素a含量以及磷吸收动力学参数。实验结果表明:(1)同静止条件相比,低振荡和高振荡条件均能显著降低斜生栅藻的比生长速率,但是两种不同磷浓度下其比生长速率却无显著性差异。(2)在实验第21天时,振荡条件培养下,斜生栅藻的胞外碱性磷酸酶活性显著高于静止条件培养,与此相反,胞内磷浓度却显著高于静止条件。(3)在磷限制条件下斜生栅藻的叶绿素a含量显著降低,同时两种振荡条件培养均使其在磷充足条件下叶绿素a含量降低。(4)通过磷吸收动力学参数的比较,静止条件培养的斜生栅藻对磷的亲和力高于其在振荡条件下。由此可见,斜生栅藻适应于静止无扰动且磷营养丰富的水体,随着水体中磷浓度进一步升高,静止和水流缓慢的水体存在导致像斜生栅藻这一类绿藻发生水华的风险。       

斜生栅藻对不同形态锑胁迫的响应及抗性研究

该研究以斜生栅藻(Scenedesmus obliquus)为研究对象,研究不同浓度的Sb(Ⅲ)、Sb(Ⅴ)胁迫下对斜生栅藻生长速率、叶绿素a、抗氧化酶活性和细胞表面结构的影响,以探讨重金属胁迫下斜生栅藻的抗性机理。结果表明:(1)在高浓度Sb(Ⅲ)、Sb(Ⅴ)胁迫处理中,斜生栅藻生长对Sb(Ⅲ)胁迫的响应更加迅速;随着Sb(Ⅲ)、Sb(Ⅴ)胁迫浓度增加,斜生栅藻体内叶绿素a含量呈下降趋势,且对Sb(Ⅲ)胁迫浓度的变化反应更为敏感。(2)在800μmol/L Sb(Ⅲ)和Sb(Ⅴ)胁迫下,斜生栅藻体内丙二醛(MDA)含量及超氧化物歧化酶(SOD)活性分别在胁迫第7~8天和第6~7天时变化更为明显;过氧化氢酶(CAT)活性在胁迫处理的第4~5天就迅速增加,对Sb胁迫响应比SOD更为迅速。(3)在高浓度Sb(Ⅲ)、Sb(Ⅴ)胁迫下,斜生栅藻细胞结构受到严重损伤,而且Sb(Ⅲ)对斜生栅藻细胞的损伤更加明显。研究发现,Sb(Ⅲ)、Sb(Ⅴ)对斜生栅藻生长、叶绿素a及细胞结构表现出不同程度的抑制和破坏作用,其细胞受到的影响程度与Sb(Ⅲ)、Sb(Ⅴ)的处理浓度及处理时间均密切相关,但斜生栅藻依靠自身抗氧化酶系统在一定程度上能够缓解Sb胁迫的不利影响。     

基于感光色素吸收信号的太湖藻类识别

铜绿微囊藻和斜生栅藻是太湖"水华"的主要藻种,基于室内纯铜绿微囊藻、斜生栅藻组成色素的吸收系数,通过四阶微分、标准化系数等方法对太湖水体浮游植物中铜绿微囊藻和斜生栅藻进行识别,并确定其组成比例。结果表明,纯藻组成色素的吸收系数应用于其在太湖水体浮游植物中比例的确定和识别中,能够全面考虑辅助色素的识别作用,较好地避免非色素物质吸收信号的干扰,具有较好的识别效果。太湖水体浮游色素中铜绿微囊藻比例最高,斜生栅藻次之,由夏季向冬季过度中,铜绿微囊藻比例不断减小,斜生栅藻的比例逐渐升高,但铜绿微囊藻比例仍略高于斜生栅藻;铜绿微囊藻的区域分布差异性较小,但时间差异性相对较大,而斜生栅藻恰恰相反,空间分布差异性较大而时间分布差异性较小。       

大豆主要株型和产量指标对大气CO2和温度升高的响应

针对当前气候变暖和大气CO_2浓度升高同步发生现实,以高光效大豆品种黑农41(HN41)和3个常规对照品种周豆16号(ZD16)、中豆35号(ZD35)和桂黄豆2号(GHD2)为研究对象,通过开顶式气室模拟高CO_2浓度(650μL/L)和温度升高(±0.5—0.6℃)研究了大气CO_2和温度升高对大豆的生长发育与产量影响。结果表明,CO_2浓度升高对株高、茎粗、单株干重和单株籽粒重影响极显著;温度、CO_2与品种互作极显著地影响单株籽粒重。CO_2浓度升高有增加大豆株高、茎粗、干重和单株籽粒重的趋势,且高温下CO_2浓度升高对株高和茎粗的促进作用更大,而正常温度水平下高CO_2浓度升高更有利于干物质积累。与对照CO_2浓度比,高CO_2浓度显著促进了高温下HN41、ZD16和GHD2的株高,并显著提高了正常温度下HN41、ZD16、ZD35和GHD2的单株干重。与正常温度相比,高温仅显著提高了高CO_2处理下HN41的茎粗,并显著提高了对照CO_2处理下HN41的单株籽粒重。此外,同一CO_2浓度和温度处理下,高光效大豆HN41的茎粗、根冠比和单株籽粒重等都显著高于ZD16、ZD35和GHD2;而仅在正常温度与高CO_2浓度处理下HN41的单株干重显著高于ZD16和GHD2。CO_2浓度和温度升高显著影响了高光效大豆的生长,其中,高温下CO_2浓度升高有利于其生长势,正常温度下CO_2浓度升高有利于其光合产物积累。     

梭鱼草化感物质丁二酸对微囊藻和栅藻生长及竞争的影响

为了解梭鱼草(Pontederia cordata)根茎有机酸类化感物质对铜绿微囊藻(Micro-cystis aeruginosa,M)和斜生栅藻(Scenedesmus obliquus,S)生长和竞争关系的影响,将不同浓度丁二酸作用于两种藻的单种培养和共培养,即铜绿微囊藻与斜生栅藻初始比例为1∶0(100%M)、1∶1(50%M+50%S)、2∶3(40%M+60%S)、3∶2(60%M+40%S)、0∶1(100%S)。结果表明:丁二酸对铜绿微囊藻和斜生栅藻均有抑制作用,抑制能力与浓度呈正相关;无化感物质作用下,单种培养和共培养中铜绿微囊藻生长能力均强于斜生栅藻; 60mg·L-1丁二酸处理下,40%M+60%S、50%M+50%S培养组中β(斜生栅藻对铜绿微囊藻的抑制参数)0,60%M+40%S培养组中β0,表明藻类初始比例影响蓝绿藻之间竞争;丁二酸可改变蓝绿藻之间竞争关系,低浓度(20、40 mg·L-1)作用下,表现为铜绿微囊藻抑制斜生栅藻,斜生栅藻促进铜绿微囊藻,高浓度(60、80 mg·L-1)作用下,表现为两者相互抑制,表明两种藻之间竞争加强。     

三聚氰胺对藻类的毒性效应研究

以近头状伪蹄形藻(Pseudokirchneriella subcapitata)和斜生栅藻(Scenedesmus obliquus)为材料,利用藻类生长抑制试验测定了三聚氰胺对藻类的毒性效应。结果表明,三聚氰胺对两种藻的生长存在不同程度的抑制效应,浓度越高,抑制效应越强;对斜生栅藻的毒性效应还随处理时间的延长而增强。以生物量计,三聚氰胺抑制近头状伪蹄形藻和斜生栅藻生长的72h EbC50值分别是537.67和485.17mg/L,均为较低的毒性效应。两种藻的生长率均随处理浓度的增加而逐渐降低;依据估算的EC20和EC10值推断,斜生栅藻对三聚氰胺较为敏感,而且还具有明显的慢性毒性效应。       

铜绿微囊藻、斜生栅藻对氮磷饥饿的耐受能力研究

在太湖微囊藻水华暴发期间 , 监测了梅梁湾水体中叶绿素 a 和氮磷营养盐的动态变化, 并通过批量培养实验研究了氮磷缺乏对铜绿微囊藻和斜生栅藻生长的影响 , 以探讨两种藻对氮磷饥饿的耐受能力对微囊藻水华形成的影响。结果表明 , 在水华期间 , 水体的氮磷营养盐浓度并不高, 并处于波动状态; 铜绿微囊藻和斜生栅藻对氮磷饥饿的耐受能力存在很大差异 : 在无磷培养基中 , 铜绿微囊藻能保持 8 天的指数生长, 而斜生栅藻在无氮或无磷培养基中生长都非常缓慢。两种藻对磷饥饿的耐受能力均高于氮。通过生长速率的比较, 发现铜绿微囊藻对磷饥饿的耐受能力明显高于斜生栅藻, 因此更能够适应波动的营养供给条件。对营养盐饥饿耐受能力的不同导致两种藻在营养盐波动的情况下微囊藻更易取得优势, 成为水华期间浮游植物的优势种。     

大气CO_2增加对不同生长光强下龙须菜光合生理特性的影响

为了探讨未来大气CO2升高对不同生长光强下大型海藻的影响,选取经济红藻龙须菜为实验材料,研究了其生长速率、光合作用、呼吸作用、叶绿素荧光参数以及光合色素对CO2和光强的响应。实验设置两个CO2浓度,正常空气水平CO2浓度(390μL/L)和高CO2浓度(1000μL/L);两个光强梯度,高光(300μmol m-2s-1)和低光(100μmol m-2s-1)。结果表明,CO2和光强对龙须菜的生长和光合作用有明显的交互作用。大气CO2升高并没有显著影响龙须菜的生长速率,但在不同CO2处理下,龙须菜对光强的响应不同。在空气水平下,光强的变化对其生长速率影响不显著。而在高CO2作用下,高光处理下的藻体有更高的生长速率。CO2显著促进高光生长下龙须菜的呼吸作用速率,但是在低光下作用不明显。而对于光合作用速率来说,低光培养下的藻体CO2表现为负面效应,但对高光下生长的藻体作用不明显。CO2增加没有改变龙须菜生长状态下的电子传递速率,但在高光下,CO2表现为一定的抑制作用。CO2显著降低了龙须菜天线色素藻红蛋白和叶绿素a的含量。这些CO2与光强的结合效应表明,大气CO2的升高对龙须菜光合生理特性的影响随着光强的变化而呈现不同的效应,在未来评估CO2的增加对大型海藻的影响时,要充分考虑其他环境因子的耦合效应。     

Characterization of imipenem-resistant Serratia marcescens producing IMP-type and TEM-type beta-lactamases encoded on a single plasmid

To evaluate the roles of blaIMP and blaTEM genes in the resistance of Serratia marcescens against beta-lactams and to find the spreading ways of these genes, 19 clinical isolates of imipenem-resistant Serratia marcescens were analyzed. Six strains bore blaIMP and blaTEM genes on a single plasmid, as confirmed by transferring resistance determinants via conjugation and transformation, and by detecting bla genes with PCR analysis. The six strains showed two different genomic patterns on pulsed-field gel electrophoresis. All the transconjugants and transformants gained high-level resistance to ampicillin, cephalexin, cefoxitin and cefotaxime, and showed a reduced susceptibility to imipenem, but maintained full susceptibility to aztreonam. In addition, the expressions of blaIMP and blaTEM genes were constitutive, either in Serratia marcescens clinical isolates or in their transconjugants and transformants. These findings may explain the rapid spread of the above resistance determinants among Enterobacteriaceae via transmissible plasmids in the clinical setting.     

On the integration of subthreshold inputs from Perforant Path and Schaffer Collaterals in hippocampal CA1 pyramidal neurons

Using a realistic model of a CA1 hippocampal pyramidal neuron, we make experimentally testable predictions on the roles of the non-specific cation current, I _h , and the A-type Potassium current, I _A , in modulating the temporal window for the integration of the two main excitatory afferent pathways of a CA1 neuron, the Schaffer Collaterals and the Perforant Path. The model shows that the experimentally observed increase in the dendritic density of I _h and I _A could have a major role in constraining the temporal integration window for these inputs, in such a way that a somatic action potential (AP) is elicited only when they are activated with a relative latency consistent with the anatomical arrangement of the hippocampal circuitry.     

Low levels of quantitative and molecular genetic differentiation among natural populations of Medicago ciliaris Kroch. (Fabaceae) of different Tunisian eco-geographical origin

In this paper, we analyze the genetic variability in four Tunisian natural populations of Medicago ciliaris using 19 quantitative traits and six polymorphic microsatellite loci. We investigated the amplification transferability of 30 microsatellites developed in the model legume M. truncatula to M. ciliaris. Results revealed that about 56.66% of analyzed markers are valuable genetic markers for M. ciliaris. The most genetic diversity at quantitative traits and microsatellite loci was found to occur within populations (>80%). Low differentiations among populations at quantitative traits Q _ST  = 0.146 and molecular markers F _ST  = 0.18 were found. The majority of measured traits exhibited no significant difference in the level of Q _ST and F _ST . Furthermore, significant correlations established between these traits and eco-geographical factors suggested that natural selection should be invoked to explain the level of phenotypic divergence among populations rather than drift. There was no significant correlation between population differentiation at quantitative traits and molecular markers. Significant spatial genetic structure consistent with models of isolation by distance was detected within all studied populations. The site-of-origin environmental factors explain about 9.07% of total phenotypic genetic variation among populations. The eco-geographical factors that influence more the variation of measured traits among populations are the soil texture and altitude. Nevertheless, there were no consistent pattern of associations between gene diversity (He) and environmental factors.     

Gas Exchange Responses to CO2 Concentration Instantaneously Elevated in Flag Leaves of Winter Wheat Cultivars Released in Different Years

Three winter wheat (Triticum aestivum L.) cultivars, representatives of those widely cultivated in Beijing over the past six decades, were grown in the same environmental conditions. Net photosynthetic rate (P _N) per unit leaf area and instantaneous water use efficiency (WUE) of flag leaves increased with elevated CO₂ concentration. With an increase in CO₂ concentration from 360 to 720 µmol mol^–1, P _N and WUE of Jingdong 8 (released in 1990s and having the highest yield) increased by 173 and 81 %, while those of Nongda 139 (released in 1970s) increased by 88 and 66 %, and Yanda 1817 (released in 1945, with lowest yield) by 76 and 65 %. Jingdong 8 had the highest P _N and WUE values under high CO₂ concentration, but Yanda 1817 showed the lowest P _N. Stomatal conductance (g _s) of Nongda 139 and Yanda 1817 declined with increasing CO₂ concentration, but g _s of Jingdong 8 firstly went down and then up as the CO₂ concentration further increased. Intercellular CO₂ concentration (C _i) of Jingdong 8 and Nongda 139 increased when CO₂ concentration elevated, while that of Yanda 139 increased at the first stage and then declined. Jingdong 8 had the lowest C _i of the three wheat cultivars, and Yanda 1817 had the highest C _i value under lower CO₂ concentrations. However, Jingdong 8 had the highest P _N and lowest C _i at the highest CO₂ concentration which indicates that its photosynthetic potential may be high.     

黄花棘豆脱水素OoY_2K_4的鉴定及表达分析

该研究以黄花棘豆cDNA为模板,采用同源克隆法,从黄花棘豆转录组数据库中克隆获得1个响应逆境胁迫的胚胎发育晚期丰富蛋白基因,命名为OoY_2K_4;OoY_2K_4基因ORF为786bp,编码261个氨基酸,含有2个保守的Y片段和4个K片段,为典型的Y_2K_4类脱水蛋白亚家族成员;OoY_2K_4蛋白不具有跨膜结构域,不存在信号肽,亲水性极强,含有1个糖基化位点和17个磷酸化位点;亚细胞定位显示,OoY_2K_4蛋白定位于细胞质中。多序列比对发现,OoY_2K_4蛋白与其他物种第二组LEA蛋白(脱水素)序列高度保守;进化树分析显示,该序列与三叶草、蒺藜苜蓿和紫花苜蓿相似度最高,亲缘关系最近。采用qRT-PCR对OoY_2K_4基因在干旱、高盐、低温以及脱落酸、乙烯、赤霉素处理下的表达分析显示,干旱和高盐胁迫可显著诱导OoY_2K_4基因表达,而低温胁迫下基本无变化;激素处理均可诱导OoY_2K_4基因高效表达,其中脱落酸诱导下OoY_2K_4基因表达最显著。研究推测,OoY_2K_4基因可能通过依赖ABA的信号途径参与黄花棘豆对干旱和高盐逆境胁迫的应答反应。     

Genomics of the<Emphasis Type="Italic"> ccoNOQP</Emphasis>-encoded<Emphasis Type="Italic"> cbb</Emphasis><Subscript>3</Subscript> oxidase complex in bacteria

Many bacteria adapt to microoxic conditions by synthesizing a particular cytochrome c oxidase (cbb ₃) complex with a high affinity for O₂, encoded by the ccoNOQP operon. A survey of genome databases indicates that ccoNOQP sequences are widespread in all sub-branches of Proteobacteria but otherwise are found only in bacteria of the CFB group (Cytophaga, Flexibacter, Bacteroides). Our analysis of available genome sequences suggests four major strategies of regulating ccoNOQP expression in response to O₂. The most widespread strategy involves direct regulation by the O₂-responsive protein Fnr. The second strategy involves an O₂-insensitive paralogue of Fnr, FixK, whose expression is regulated by the O₂-responding FixLJ two-component system. A third strategy of mixed regulation operates in bacteria carrying both fnr and fixLJ-fixK genes. Another, not yet identified, strategy is likely to operate in the -Proteobacteria Helicobacter pylori and Campylobacter jejuni which lack fnr and fixLJ-fixK genes. The FixLJ strategy appears specific for the -subclass of Proteobacteria but is not restricted to rhizobia in which it was originally discovered.     

Genes determining leucine aminopeptidase and mildew resistance from the ornamental apple, ‘White Angel’

Mildew resistance in the ornamental apple White Angel was found to be determined by complementary genes. The gene R _w was found to be necessary for the expression of resistance controlled by the resistance gene Pl _w . The close linkage between the isoenzyme gene, Lap-2, for leucine aminopeptidase and P1 _w was confirmed. The efficiency of Lap-2 as a marker in screening for mildew resistance is limited, as it cannot account for susceptible plants with the r _w r _w P1 _w p1 _w genotype. It has, however, an important role to play in combining resistance genes from different sources. The genotypes of White Angel (R _w r _w , Pl _w pl _w , Lap-2an), Jester (R _w r _w , p1 _w p _w , Lap-2an) Katja (R _w r _w ,p1 _w p1 _w , Lap-2an) and Gloster 69 (r _w r _w , p1 _w p1 _w , Lap-2an) were determined. It also appeared that R _w might influence Lap-2 activity in young seedlings.     

Carbon dioxide and methane fluxes in boreal peatland microcosms with different vegetation cover—effects of ozone or ultraviolet-B exposure

O₃ concentrations in the troposphere are rising and those in the stratosphere decreasing, the latter resulting in higher fluxes of solar ultraviolet-B (UV-B) radiation to the earth's surface. We assessed whether the fluxes of CO₂ and CH₄ are altered by enhanced UV-B radiation or elevated tropospheric O₃ concentrations in boreal peatland microcosms (core depth 40 cm, diameter 10.5 cm) with different vegetation cover. At the end of the UV-B experiment which lasted for a growing season, net CO₂ exchange (NEE) and dark ecosystem respiration (R _TOT) were sevenfold higher, and CH₄ efflux 12-fold higher, in microcosms with intact vegetation dominated by Eriophorum vaginatum L. and Sphagnum spp., compared to microcosms from which we removed E. vaginatum. Vegetation treatment had minor effects on CH₄ production and consumption potentials in the peat, suggesting that the large difference in CH₄ efflux is mainly due to efficient CH₄ transport via the aerenchyma of E. vaginatum. Ambient UV-B supplemented with 30% and elevated O₃ concentrations (100 and 200 ppb, for 7 weeks) significantly increased R _TOT in both vegetation treatments. Elevated O₃ concentrations reduced NEE over time, while UV-B had no clear effects on the fluxes of CO₂ or CH₄ in the cloudy summer of the study. Field experiments are needed to assess the significance of increasing UV-B radiation and elevated tropospheric O₃ concentration on peatland gas exchange in the long-term.     

A novel phospholipase A2/esterase from hyperthermophilic archaeon Aeropyrum pernix K1

An open reading frame of the hyperthermophilic archaeon Aeropyrum pernix K1 APE2325, which composed of 474 bases, was cloned and expressed in Escherichia coli BL21 (DE3) Codon Plus-RIL. The recombinant protein was purified by Ni-chelation affinity chromatography. It showed a single band with a molecular mass of 18kDa in SDS-PAGE. The purified enzyme exhibited both phospholipase A(2) and esterase activities with the optimal catalytic temperature at 90 degrees C. The enzyme activity was Ca(2+)-independent. Kinetic analysis revealed its Km, k cat, and Vm for the p-nitrophenyl propionate substrate were 103microM, 39s(-1), and 249micromol/min/mg, respectively. The recombinant protein was thermostable and its half-life at 100 degrees C was about 1h.