五氯酚对土壤跳虫代谢转化酶基因和蜕皮相关基因表达的影响

五氯酚(PCP)是一种广泛存在于环境中的有机污染物。利用土壤跳虫(Folsomia candida)作为受试生物,研究了不同浓度的PCP暴露下细胞色素P450(CYP450)基因、谷胱甘肽转移酶(GST)基因和蜕皮相关基因表达水平的变化。结果显示,PCP暴露下,跳虫CYP450基因Fcc01651、Fcc02155、Fcc03650和GST基因Fcc04073、Fcc05260的表达水平未发生显著变化。在PCP浓度为240 mg·kg-1时,跳虫GST基因Fcc00494的表达显著上调。在PCP浓度为120 mg·kg-1和240 mg·kg-1时,几丁质酶基因Fcc00881和几丁质结合域基因Fcc01306的表达受到显著抑制。研究结果可为评价PCP暴露对跳虫的毒性提供一定依据。     

非生物胁迫诱导青蒿素生物合成基因的表达

对离体培养青蒿试管苗中3个青蒿素合成相关基因的非生物胁迫诱导表达模式进行了初步研究.半定量RT-PCR测定结果表明,当暴露于冷、热和紫外光后,青篙植株的紫穗槐-4,11-二烯合酶基因(ADS)和细胞色素P450单加氧酶基因(CYP71AV1)转录上调;相反,在未经胁迫处理的情况下,ADS和CYP71AV1基因的表达水平较低,而在胁迫处理前后细胞色素P450还原酶基因(CPR)转录所产生的mRNA均保持恒定.同时,冷胁迫的诱导效果也得到实时荧光定量RT-PCR测定数据的支持.经低温锻炼的青蒿试管苗,其ADS和CYP71AV1基因的转录产物拷贝数比对照分别提高11倍和7倍,而CPR基因的mRNA拷贝数与对照基本持平.短暂预冷处理还可显著提高青蒿试管苗的青蒿素含量,达到7.5～8.8 mg/g干重,比对照提高66.7%～95.6%,为进一步探索利用环境胁迫促进青蒿素高产的新途径提供了可能性.图2表3参21     

农药对秀丽隐杆线虫毒性效应及其机制的研究进展

随着大量农药被广泛的使用并最终汇聚到环境介质中,对生态环境和人体健康产生潜在影响。秀丽隐杆线虫(Caenorhabditis elegans)是土壤中最丰富的后生动物,在土壤生态系统中具有重要地位,并作为一种重要的模式生物广泛应用于环境毒理学研究。本文从秀丽隐杆线虫常用的毒理学研究方法入手,以致死率、生长发育、运动行为、繁殖、活性氧自由基(ROS)水平、细胞凋亡水平、相关基因表达量和蛋白水平等作为测试指标,归纳总结农药对秀丽隐杆线虫的衰老性、神经及生殖系统的毒性效应,分析探索其毒性机理,并展望了未来的研究重点。     

The meta-hydroxylation step in the phenylpropanoid pathway: a new level of complexity in the pathway and its regulation

Together in silico and genetic mining approaches have recently designated the CYP98 family of plant cytochromes P450 as the family of enzymes that catalyzes the meta-hydroxylation step in the phenylpropanoid pathway. This meta-hydroxylation is not catalyzed on the free p-coumaric acid as anticipated, but on its conjugates with shikimic, quinic, or phenyllactic acids. While all CYP98s have in common phenol meta-hydroxylase activity, p-coumaroylshikimate remains their preferred substrate. High expression of CYP98s is detected in lignifying tissues in stems, roots, and siliques. The CYP98A3 gene disruption in Arabidopsis thaliana leads to a drastic inhibition of lignin synthesis, cell growth, and plant development. The meta-hydroxylation of phenolic precursors is thus essential for higher plant development. Isolation of coding sequences belonging to the CYP98 family from basil, wheat, and extensive functional analysis of the recombinant enzymes, together with CYP98s from other plant taxa, helps shedding some light on mechanisms of P450s evolution. Most importantly, the occurrence of the meta-hydroxylation on esters of shikimic or quinic acids introduces a new biochemical regulation mechanism in the phenylpropanoid pathway.     

Bedeutung der Expositionswege auf Bioakkumulation und Wirkung in C. elegans

Different uptake routes of contaminants were analysed with focus on particle-bound pollutants. Bioavailability of cadmium was determined with quantitative mRNA-analysis using the expression of the “cadmium responsive gene” cdr- in Caenorhabditis elegans. Comparable experiments were performed with fluoranthene as organic pollutant. The exposure in medium was compared with and without bacteria as food particles. Both bioaccumulation and toxic effects were most influenced via the food exposure. Inorganic particles increased also the uptake of cadmium, but significantly less than food or mixtures of food and inorganic particles. The exposure of fluoranthene reduced the reproduction of the nematodes, if a gene expression threshold was exceeded. However, bioavailabilty of fluoranthene via food particles caused a faster response of the cyp-expression. Thus means, the internal availability is crucial influenced by the uptake route.     

AzaC预处理增加TCDD对特殊细胞P450基因的诱导

利用RT-PCR检测不同物质诱导细胞的CYP基因 mRNA的表达水平.在HepG2细胞,TCDD能诱导CYP1A1、CYP1B1及CYP1A2基因表达,CYP1A1、CYP1B1基因比CYP1A2基因更容易被诱导,用AzaC处理后CYP1B1基因表达无改变;在A549细胞和SPC-A1细胞,Azac预处理后增加了TCDD对CYP1家族的诱导.也就是AzaC增加了CYP1A1、CYP1A2和CYP1B1基因表达水平.图1表1参10     

Chaotic genetic patchiness and high relatedness of a poecilogonous polychaete in a heterogeneous estuarine landscape

The genetic structure of benthic marine invertebrates is often described as “chaotic” when genetic structure cannot be explained and barriers to dispersal and gene flow cannot be identified. Here, chaotic patterns of genetic structure for the polychaete Pygospio elegans (Claparède) sampled at 16 locations from the heterogeneous Isefjord–Roskilde Fjord estuary complex in Denmark were found. There was no isolation by distance, and the geography of the estuary complex did not seem to pose a barrier to dispersal and gene flow in this species. We investigated whether characteristics of the environment could be related to the genetic structure and possibly restrict gene flow in this species. Additionally, since P. elegans is poecilogonous, producing larvae with different pelagic developmental periods, we investigated whether observed developmental modes in the samples might clarify the genetic patterns. None of the tested factors explained the population genetic structure. However, a high degree of relatedness among individuals in almost all samples was found. Samples with a larger percentage of young individuals had more related individuals, suggesting that different cohorts could be comprised of individuals with different degrees of relatedness. Relatedness within a site could be increased by limited larval dispersal, collective dispersal of related larvae, sweepstakes reproductive success, or asexual reproduction, but distinguishing between these requires further study. Using a “seascape genetics” approach allowed us to investigate some of the numerous potential factors that could influence population genetic structure in a poecilogonous species.     

纳米二氧化硅对秀丽线虫的毒性作用研究

为探讨二氧化硅纳米颗粒(SiNP)的毒性作用.以微米SiO2和空白做对照,将尺寸为20和60 nm的SiNP分别以0.2,1.0和5.0 g·L-13个浓度对模式生物秀丽隐杆线虫进行染毒.通过第一代和第二代线虫体长、身体弯曲频率、头部摆动频率以及第一代线虫后代数目、世代时间等发育、运动和生殖的相关评价指标,对SiNP毒...     

Role of bacteria in larval settlement and metamorphosis of the polychaete Hydroides elegans

The serpulid polychaete Hydroides elegans Haswell, 1883 is an early colonist of new substrata in Pearl Harbor, Oahu, Hawaii. When metamorphically competent, larvae of H. elegans will settle rapidly upon an acceptably biofilmed surface, but not on a clean surface. In this study we found the ability of larvae to respond selectively to inductive surfaces to be retained for at least 3 wk. Of a series of bacterial strains isolated from Hawaiian marine biofilms, 13 induced larval settlement, 11 gave moderate or mixed results, and 10 others did not stimulate the settlement of H. elegans. The amount of settlement induced by monospecific strains was rarely as great as with natural, multispecies films. Most of the isolated bacteria were motile Gram-negative rods, but Gram-positive strains were also present, and diverse metabolic types were represented in the study. Biofilms killed by treatment with heat, ultraviolet radiation or chemical fixatives were no longer inductive. Soluble, dialyzable, heat-stable bacterial products induced settlement and metamorphosis more slowly. The range of bacteria producing an inductive signal suggests either that there are multiple cues, or that the cue is common to many bacteria. Chemical signals characteristic of early microbial biofilms may indicate freshly available substrata with optimal potential for the growth and survival of H. elegans. Received: 30 January 1998 / Accepted: 12 September 1998     

Thermal tolerance of larvae of Pollicipes elegans, a marine species with an antitropical distribution

For the antitropical gooseneck barnacle Pollicipes elegans, population-specific physiological temperature tolerance of larvae may serve as a barrier to larval dispersal across the warmest regions of the tropical Pacific Ocean. Thermal tolerance ranges of larvae of three different populations of P. elegans sampled in 2011 and 2012 (Mexico [MX], El Salvador [ES], and Peru [PE]) were investigated by measuring three indicators of physiological performance: swimming activity, oxygen consumption, and lethality or LT₅₀. The thermal tolerance profiles, which include measurable optimum (maximum aerobic performance), pejus (“getting worse”) and pessimum (worst aerobic performance) ranges, of larvae from the three populations were consistent with their characteristic environmental temperatures. In MX, larvae live close to the upper border of their optimum during warm months and so have a limited capacity to tolerate higher-than-normal temperatures. Larvae from the ES population likewise appear to live within their optimum temperature range, but these larvae lack a detectable pessimum range, suggesting they would be unable to cope with temperatures above their pejus range. Larvae from PE have a broad optimum but no pejus range. Different thermal tolerance ranges provide strong evidence for population-dependent physiological adaptations in P. elegans. For the southern (PE) and northern (MX) P. elegans populations, high tropical temperatures are likely to be a strong direct physiological barrier to larval survival and dispersal, which is in contrast to the more thermally tolerant ES population.     

真鲷去毒相关基因cDNA序列的克隆与肝脏组成型表达分析

赤潮发生时产生的一些海洋生物毒素对人类和海洋动物的安全形成潜在的威胁甚至导致死亡.为从分子水平探讨鱼类中海洋藻毒素的去毒分子机理,采用RT-PCR法克隆了真鲷(Pagrus major)肝脏芳香烃受体核转位蛋白(ARNT)和I时相异生素代谢酶细胞色素P4501A(CYP1A)基因cDNA核心序列,同时,应用半定量RT-PCR方法,以β-肌动蛋白作为外参照,在其指数期增长的范围内研究了芳香烃受体(AHR)、ARNT、CYP1A、II时相异生素代谢酶alpha型谷胱甘肽S-转移酶(GSTA1、GSTA2)、rho型谷胱甘肽S-转移酶(GSTR)、热休克蛋白70(HSP70)基因组成型表达水平.结果发现,真鲷ARNT、CYP1A基因cDNA核心序列片段分别长438bp和908bp,分别编码146和302个氨基酸.序列同源性分析发现,真鲷与门齿鲷(Stenotomus chrysops)、石首鱼(Micropogon undulatus)ARNT基因氨基酸序列同源性高达97.2%、95.2%,与斑马鱼(Brachydanio rerio)、人、大鼠、小鼠ARNT同源性较低(77.2%～79.3%).真鲷与门齿鲷、金头鲷(Sparus auratus)、欧洲川鲽(Rhombus maximus)、欧洲海鲈(Dicentrachus labrax)CYP1A基因氨基酸序列同源性较高,为84.8%～94.0%,与斑马鱼、人、小鼠CYP1A同源性较低,为59.6%～77.8%.真鲷肝脏AHR、ARNT、CYP1A、GSTA1、GSTA2、GSTR和HSP70基因组成型表达水平分别为(25.32±6.56)%、(26.22±4.24)%、(146.5±16.06)%、(55.42±3.75)%、(48.82±10.89)%、(79.47±3.13)%、(107.42±14.34)%.     

Species-specific variation in cuttlebone δ13C and δ18O for three species of Mediterranean cuttlefish

Stable carbon (δ¹³C) and oxygen (δ¹⁸O) isotopes in cuttlebones of three species of Mediterranean cuttlefish (Sepia elegans, S. officinalis, and S. orbignyana) with different life histories were contrasted. Cuttlebone δ¹³C and δ¹⁸O were quantified at both the core and edge (representing early life and recent deposition, respectively) for all three species sampled from the southern Adriatic Sea in 2010 (n = 28). For S. officinalis, cuttlebone δ¹³C and δ¹⁸O values were both lower relative to S. elegans and S. orbignyana at the core by approximately 1.0–2.0 and 3.0 ‰, respectively. Differences between core and edge in cuttlebone δ¹³C and δ¹⁸O were also observed for S. officinalis with observed values at the cuttlebone edge (recent) exceeding core (early life) values by 2.5 ‰ for δ¹³C and 1.4 ‰ for δ¹⁸O. Differences in isotopic composition across S. officinalis cuttlebones are possibly reflective of ontogenetic migrations from nearshore nurseries (lower seawater δ¹³C and δ¹⁸O values) to offshore overwintering habitats (higher seawater δ¹³C and δ¹⁸O values). Overall, results from this study suggest that cuttlebone δ¹³C and δ¹⁸O hold promise as natural tags for determining the degree of spatial connectivity between nearshore and offshore environments used by cuttlefish.     

Isolation of a Pseudomonas Stutzeri strain that degrades 1,2,4-trichlorobenzene and characterization of its degradative plasmid

The genetic information encoding metabolic pathways for xenobiotic compounds in bacteria often resides on catabolic plasmids. The aim of the present work was to know the location of the genes for degrading 1,2,4-trichlorobenzen. In this paper a 1,2,4-trichlorobenzene-degrading strain THSL-1 was isolated from the soil of Tianjin Chemical Plant using 1,2,4-trichlorobenzene as the sole carbon source. The strain was identified as Pseudomonas stutzeri through morphologic survey and 16S rDNA sequence determination. A plasmid was discovered from strain THSL-1 by using the alkali lysis method. When the plasmid was transformed into E. coli. JM109 by the CaCl₂ method, the transformant could grow using 1,2,4-trichlorobenzene as the sole carbon source and had the degradation function of 1,2,4-trichlorobenzene. Therefore, it could be deemed that the plasmid carried the degradative genes of 1,2,4-trichlorobenzene. The average size of the plasmid was finally determined to be 40.2 Kb using selectively three kinds of restricted inscribed enzymes (HindIII, BamHI, and XholI) for single cutting and double cutting the plasmid pTHSL-1, respectively.     

Gene expression profiles of the swimming crab <Emphasis Type="Italic">Portunus trituberculatus</Emphasis> exposed to salinity stress

The swimming crab, Portunus trituberculatus, is an important marine fishery and aquaculture species. Although P. trituberculatus is a euryhaline species, water salinity condition influenced its distribution, migration route, and artificial propagations. To investigate gene expression in the P. trituberculatus exposed to different salinity stresses, 2426 expressed sequence tags (ESTs) from gill cDNA library were selected to spot on a cDNA microarray chip. In total, 417 differentially expressed genes were identified and grouped into eight clusters by hierarchical clustering analysis. Approximately 71.5% of grouped genes belonged to three independent expression patterns, indicating that these three expression patterns may represent three important stress tolerance pathways or networks in P. trituberculatus. Moreover, our cDNA microarray data suggested that there were differences in gene expression patterns of P. trituberculatus for low salinity and high salinity acclimation, suggesting that two salinity challenges resulted in a wide variation of gene expression in P. trituberculatus. In addition, a series of genes such as CCAAT/enhancer-binding protein, Na/K ATPase β-subunit, and heat shock proteins (HSPs) genes were suggested to be key elements during salinity acclimation process. Overall, this work represented an important step toward understanding the molecular processes and mechanisms involved in salinity acclimation of the swimming crab.     

GO通过氧化损伤激活秀丽线虫未折叠蛋白反应

以秀丽线虫为受试生物,探讨氧化石墨烯(GO)对秀丽线虫未折叠蛋白应答的激活及其与氧化应激的关系。将L4幼虫暴露于GO中24 h,通过检测活性氧(ROS)水平、乳酸脱氢酶(LDH)、丙二醛(MDA)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)和谷胱甘肽过氧化物酶(GSH-Px)等指标评估GO对秀丽线虫所致氧化损伤;以hsp-4和hsp-6分别作为秀丽线虫内质网和线粒体未折叠蛋白反应(UPR)的报告基因评价GO对秀丽线虫UPR应答的激活;同时以谷胱甘肽(GSH)作为抗氧化剂探讨GO所致UPR应答与氧化应激的关系。研究结果显示,GO暴露后秀丽线虫体内ROS、LDH和MDA水平升高,抗氧化酶活性上升;内质网和线粒体UPR反应增强,GSH预处理可以降低内质网和线粒体UPR应答。研究表明,GO短期暴露可以诱导秀丽线虫机体氧化应激进而激活UPR应答,抗氧化保护可以降低UPR应答反应。     

Ocean acidification reduces biomineralization-related gene expression in the sea urchin, Hemicentrotus pulcherrimus

Although recent studies have demonstrated that calcification in a wide range of marine organisms is profoundly affected by CO₂-induced ocean acidification, the mechanism of this phenomenon is still unclear. To clarify the effects of ocean acidification on the calcification process at the molecular level, we evaluated the expression of three biomineralization-related genes in the sea urchin Hemicentrotus pulcherrimus exposed under control, 1,000, and 2,000?ppm CO₂ from egg to pluteus larval stage. We found that the expression of the gene msp130, which is proposed to transport Ca²⁺ to the calcification site, is suppressed by increased CO₂ at pluteus larval stage. Meanwhile, expression of the spicule protein matrix genes SM30 and SM50 was apparently not affected. The results suggest that the combined effects of ocean acidification on the expression of skeletogenesis-related genes as well as the change in seawater carbonate chemistry affect the biomineralization ability of sea urchins.     

PFOS对多齿围沙蚕CYPs、GST基因转录及酶活性的毒性效应

多毛类沙蚕已经广泛应用于海洋环境污染的生物监测,但其对新型持久性有机污染物全氟辛烷磺酰基化合物PFOS的毒理学研究尚无报道。本研究以潮间带优势种多齿围沙蚕(Perinereis nuntia)为研究对象,以细胞色素P450(CYP)、谷胱甘肽硫转移酶(GST)的基因和酶作为联合指标,研究了在PFOS亚致死浓度(4、8、16 mg·L-1)暴露第1、4、7、14天及净水恢复5 d后多齿围沙蚕CYP431A1、CYP424A1基因转录水平和EROD酶活性、GST omega基因转录水平和GST酶活性的响应情况。结果表明,PFOS暴露对EROD的抑制具有明显的时间-效应关系;CYP2系成员CYP431A1基因转录水平对PFOS的响应具有良好的剂量-效应关系并在胁迫第14天表现出最高的可诱导性;CYP4系基因CYP424A1的转录在4、8 mg·L-1处理组中与PFOS暴露时间正相关。II相解毒系统成员GST酶活和GST omega基因的响应均表现出随着PFOS胁迫时间的延长先下降,后上升的规律;多齿围沙蚕在高强度PFOS胁迫下仍可加速新陈代谢,表现出对PFOS的耐受性;净水恢复阶段,各指标都有向对照组水平恢复的趋势。总之,基因和蛋白的响应表明CYPs和GST在多齿围沙蚕PFOS新陈代谢中发挥重要作用,这些基因和酶具有作为生物标志物监测海洋潮间带PFOS污染效应的潜力。     

Biosynthesis of lipds in zooplankton from Saanich Inlet,British Columbia,Canada

Lipid analyses were carried out on various species of zooplankton captured in Saanich Inlet, British Columbia, Canada, during September, 1972. The amphipod Cyphocaris challengeri had the highest level of lipid, consisting mainly of wax esters. The copepod Calanus pacificus had moderate amounts of lipid, with triglyceride as the major neutral lipid. The euphausiid Thysanoessa raschii contained mainly triglyceride in its moderate levels of neutral lipid, while wax esters and, to a lesser extent, triglycerides were present in the very small levels of neutral lipid in the chaetognath Sagitta elegans. The major fatty alcohols in wax esters of both Cyphocaris challengeri and Calanus pacificus were 20:1 and 22:1, although notable differences were found in the major fatty acids. Biosynthetic studies showed that phospholipids were labelled faster than neutral lipids in all species with both (U-¹⁴C) glucose and (1-¹⁴C) palmitic acid as precursors. Only species containing significant amounts of wax esters in their neutral lipids incorporated substantial amounts of radio-activity from (1-¹⁴C) palmitic acid into wax esters in (i) living animals, (ii) preparations containing fragments of tissue, (iii) cell-free systems. All species incorporated added fatty alcohols into wax esters in preparations containing tissue fragments and in cell-free preparations. Both the fatty acid and fatty alcohols of the wax esters of both Calanus pacificus and S. elegans were labelled from (1-¹⁴C) palmitic acid, consistent with de novo biosynthesis of the esters. (1-¹⁴C) hexadecanol was incorporated into wax esters almost entirely in the fatty alcohol moiety. It is concluded that all species examined formed was esters when presented with preformed fatty alcohols, but only those species that had wax esters as a major component of the neutral lipids were capable of de novo biosynthesis of the lipids.     

五氯酚(PCP)对鸡肝癌细胞(LMH)毒性效应的机制研究

五氯酚(pentachlorophenol,PCP)是一种持久性有机污染物,广泛用于灭钉螺、木材防腐、除草剂等方面,由于PCP在环境中的持久性和生物累积性,其对生态环境和人类健康造成潜在危害。本文以鸡肝癌细胞系(chicken hepatoma cells,LMH)为受试对象,探讨了PCP对细胞色素P450(CYP450)和抗氧化系统的影响。MTT结果显示LMH细胞经不同浓度PCP暴露后,呈现出先促进细胞增殖后抑制的J-型曲线,PCP对LMH细胞24 h的半数效应浓度(24 h-EC50)为427.52μmol·L~(-1)。LMH细胞在1.56、6.25、25、100μmol·L~(-1)PCP染毒条件下可增加细胞EROD、MROD、PROD和BFC活性,并可使CYP1A、1B、1C、2H及3A家族基因mRNA表达水平升高。LMH细胞在0.4~100μmol·L~(-1)PCP染毒下可显著降低硫酸基转移酶(SULT1B1和SULT 1C1)基因mRNA水平。此外,LMH细胞在6.25、25、100μmol·L~(-1)PCP染毒下可引起细胞内ROS升高,同时PCP(1.56~100μmol·L~(-1))可显著增加细胞内MDA含量和降低GSH/GSSH比值。这些结果表明细胞色素P450(CYP450)基因及酶活性的变化、细胞内ROS和MDA含量及GSH/GSSH可作为评价LMH细胞PCP毒性效应的敏感性生物标志物。此研究在细胞水平上利用多个评价指标研究PCP对细胞的毒性效应,为PCP环境风险评价提供依据。