全氟辛烷磺酸(PFOS)对斑马鱼卵黄蛋白原mRNA水平的影响

为了研究环境低剂量全氟辛烷磺酸（perfluorooctane sulfonate,PFOS）对水生生物的内分泌干扰效应和可能的作用机制,测定了PFOS对斑马鱼（Brachydanio rerio）肝脏中卵黄蛋白原（vitellogenin,VTG）mRNA水平的影响.将斑马鱼暴露于4个PFOS的环境低剂量浓度组（0.1、1、10、100μg.L-1）中进行21d毒性试验,收集肝脏样品,提取RNA,采用荧光定量PCR（qRT-PCR）分别检测VTG1和VTG3的mRNA水平.结果表明：①PFOS暴露引起雄性斑马鱼肝脏VTG1和VTG3 mRNA水平升高,VTG1 mRNA水平升高与剂量呈正相关,在100μg.L-1暴露浓度处与对照组呈现显著性差异;VTG3的mRNA水平变化与剂量呈倒U型曲线,呈现典型的毒物刺激荷尔蒙效应,在10和100μg.L-1暴露浓度处与对照组呈现显著性差异;②PFOS暴露引起雌性斑马鱼肝脏中VTG1 mRNA水平升高,在10μg.L-1暴露浓度处与对照组呈现显著性差异,但在高浓度（10和100μg.L-1）处试验结果误差较大;VTG3 mRNA水平只在10μg.L-1暴露浓度处升高,但相比于对照组均没有显著性差异.试验结果表明PFOS暴露对斑马鱼的内分泌干扰作用明显,其毒性作用机制可能是类雌激素效应,而肝脏中VTG1和VTG3mRNA水平可能作为PFOS内分泌干扰效应评价的敏感生物标志物,但VTG1和VTG3 mRNA水平的响应曲线呈现基因亚型和性别差异.     

Enhanced resistance of yeast mutants deficient in low-affinity iron and zinc transporters to stannous-induced toxicity

Tin or stannous (Sn²⁺) compounds are used as catalysts, stabilizers in plastic industries, wood preservatives, agricultural biocides and nuclear medicine. In order to verify the Sn²⁺ up-take and toxicity in yeast cells we utilized a multi-elemental analysis known as particle-induced X-ray emission (PIXE) along with cell survival assays and quantitative real-time PCR. The detection of Sn²⁺ by PIXE was possible only in yeast cells in stationary phase of growth (STAT cells) that survive at 25 mM Sn²⁺ concentration. Yeast cells in exponential phase of growth (LOG cells) tolerate only micro-molar Sn²⁺ concentrations that result in intracellular concentration below of the method detection limit. Our PIXE analysis showed that STAT XV185-14c yeast cells demonstrate a significant loss of intracellular elements such as Mg, Zn, S, Fe and an increase in P levels after 1 h exposure to SnCl₂. The survival assay showed enhanced tolerance of LOG yeast cells lacking the low-affinity iron and zinc transporters to stannous treatment, suggesting the possible involvement in Sn²⁺ uptake. Moreover, our qRT-PCR data showed that Sn²⁺ treatment could generate reactive oxygen species as it induces activation of many stress-response genes, including SOD1, YAP1, and APN1.     

pH、盐度对微生物还原硫酸盐的影响研究

采用厌氧生物处理工艺，研究了在不同盐度下pH连续降低对硫酸盐还原和有机物去除率的影响。实验结果表明，硫酸盐还原菌有很强的适应pH变化的能力，在pH值达到4以下仍有60％的硫酸盐去除率。NaCl浓度由4g／L增加到50g／L抑制了各厌氧菌的活性，导致硫酸盐和有机物去除率的下降，但硫酸盐还原菌耐受性高于产甲烷菌等其他厌氧菌，在NaCl浓度为50g／L下，硫酸盐去除率能达到50％，而有机物去除率则低于30％。qRT—PCR表明了系统菌落中SRB随着环境的变化情况与化学指标结果相一致，该反应器体系中SRB在整个厌氧菌群落中只占了很小部分。     

Antifungal and anti-aflatoxigenic activity of Heliopsis longipes roots and affinin/spilanthol against Aspergillus parasiticus by downregulating the expression of alfD and aflR genes of the aflatoxins biosynthetic pathway

Abstract

In the present study, ethanolic extract from Heliopsis longipes roots and affinin/spilanthol against Aspergillus parasiticus growth and aflatoxins production were studied in relation to the expression of aflD and aflR, two key genes of aflatoxins biosynthetic pathway. Phytochemical analysis of the ethanolic extract by GC-EIMS identified affinin/spilanthol (7.84?±?0.27?mg g^?1) as the most abundant compounds in H. longipes roots. The antifungal and anti-aflatoxigenic assays showed that affinin/spilanthol at 300?µg mL^?1 produced the higher inhibition of radial growth (95%), as well as, the higher aflatoxins production inhibition (61%) in comparison to H. longipes roots (87% and 48%, respectively). qRT-PCR revealed that the expression of aflD and aflR genes showed a higher downregulation in affinin/spilanthol at 300?µg mL^?1. The expression ratio of alfD was suppressed by affinin/spilanthol in 79% and aflR in 84%, while, a lower expression ratio suppressed by H. longipes was obtained, alfD (55%) and aflR (59%). Affinin/spilanthol possesses higher antifungal and anti-aflatoxigenic activity against A. parasiticus rather than H. longipes roots, and this anti-aflaxotigenic activity occurring via downregulation of the aflD and aflR genes. Thus, H. longipes roots and affinin/spilanthol can be considered potent antifungal agents against aflatoxigenic fungus, especially, affinin/spilanthol.     

The platelet-activating factor acetylhydrolase gene derived from Trichoderma harzianum induces maize resistance to Curvularia lunata through the jasmonic acid signaling pathway

Platelet-activating factor acetylhydrolase (PAF-AH) derived from Trichoderma harzianum was upregulated by the interaction of T. harzianum with maize roots or the foliar pathogen Curvularia lunata. PAF-AH was associated with chitinase and cellulase expressions, but especially with chitinase, because its activity in the KO40 transformant (PAF-AH disruption transformant) was lower, compared with the wild-type strain T28. The result demonstrated that the colonization of maize roots by T. harzianum induced systemic protection of leaves inoculated with C. lunata. Such protection was associated with the expression of inducible jasmonic acid pathway-related genes. Moreover, the data from liquid chromatography-mass spectrometry confirmed that the concentration of jasmonic acid in maize leaves was associated with the expression level of defense-related genes, suggesting that PAF-AH induced resistance to the foliar pathogen. Our findings showed that PAF-AH had an important function in inducing systemic resistance to maize leaf spot pathogen.     

Interactive effects of cadmium and hypoxia on metabolic responses and bacterial loads of eastern oysters Crassostrea virginica Gmelin

Pollution by toxic metals including cadmium (Cd) and hypoxia are important stressors in estuaries and coastal waters which may interactively affect sessile benthic organisms, such as oysters. We studied metabolic responses to prolonged hypoxic acclimation (2 weeks at 5% O₂) in control and Cd-exposed (30 d at 50 μg L⁻¹ Cd) oysters Crassostrea virginica, and analyzed the effects of these stressors on abundance of Vibrio spp. in oysters. Hypoxia-acclimated oysters retained normal standard metabolic rates (SMR) at 5% O₂, in contrast to a decline of SMR observed during acute hypoxia. However, oysters spent more time actively ventilating in hypoxia than normoxia resulting in enhanced Cd uptake and 2.7-fold higher tissue Cd burdens in hypoxia. Cd exposure led to a significant decrease in tissue glycogen stores, increase in free glucose levels and elevated activity of glycolytic enzymes (hexokinase and aldolase) indicating a greater dependence on carbohydrate catabolism. A compensatory increase in activities of two key mitochondrial enzymes (citrate synthase and cytochrome c oxidase) was found during prolonged hypoxia in control oysters but suppressed in Cd-exposed ones. Cd exposure also resulted in a significant increase in abundance of Vibrio parahaemolyticus and Vibrio vulnificus levels during normoxia and hypoxia, respectively. Overall, Cd- and hypoxia-induced changes in metabolic profile, Cd accumulation and bacterial flora of oysters indicate that these stressors can synergistically impact energy homeostasis, performance and survival of oysters in polluted estuaries and have significant consequences for transfer of Cd and bacterial pathogens to the higher levels of the food chain.     

浑河流域野生鲫鱼卵黄蛋白原基因表达

为揭示浑河流域水环境内分泌干扰物对水生态的潜在风险,利用兼并引物扩增获得鲫鱼卵黄蛋白原(Vtg)和核糖体蛋白L-7(RPL-7)基因部分碱基序列(分别为825和450bp),建立以RPL-7为内参基因、定量鲫鱼Vtg基因表达的实时定量逆转录聚合酶链反应(qRT-PCR)方法,并将该方法用于定量浑河野生鲫鱼肝组织Vtg基因表达分析。结果显示:与上游对照点(S1)相比,7月下游各点(S4～S8)雄鱼、S4和S6点雌鱼肝组织的VtgmRNA表达水平皆显著升高(P<0.05);11月在雄鱼中未检出VtgmRNA的有效表达,雌鱼也仅在S2和S3点的表达水平升高(P<0.05)。研究表明,浑河流域野生鲫鱼尤其是在7月明显受到了环境雌激素类物质的影响。另外,qRT-PCR方法能够灵敏检测出鲫鱼Vtg基因表达的时空差异。