Transformation of ammonium nitrogen and response characteristics of nitrifying functional genes in tannery sludge contaminated soil

High concentrations of ammonium nitrogen released from tannery sludge during storage in open air may cause nitrogen pollution to soil and groundwater. To study the transformation mechanism of NH₄⁺-N by nitrifying functional bacteria in tannery sludge contaminated soils, a series of contaminated soil culture experiments were conducted in this study. The contents of ammonium nitrogen (as NH₄⁺-N), nitrite nitrogen (as NO₂^?-N) and nitrate nitrogen (as NO₃^?-N) were analyzed during the culture period under different conditions of pollution load, soil particle and redox environment. Sigmodial equation was used to interpret the change of NO₃^?-N with time in contaminated soils. The abundance variations of nitrifying functional genes (amoA and nxrA) were also detected using the real-time quantitative fluorescence PCR method. The results show that the nitrification of NH₄⁺-N was aggravated in the contaminated silt soil and fine sand under the condition of lower pollution load, finer particle size and more oxidizing environment. The sigmodial equation well fitted the dynamic accumulation curve of the NO₃^?-N content in the tannery sludge contaminated soils. The Cr(III) content increased with increasing pollution load, which inhibited the reproduction and activity of nitrifying bacteria in the soils, especially in coarse-grained soil. The accumulation of NO₂^?-N contents became more obvious with the increase of pollution load in the fine sand, and only 41.5% of the NH₄⁺-N was transformed to NO₃^?-N. The redox environment was the main factor affecting nitrification process in the soil. Compared to the aerobic soil environment, the transformation of NH₄⁺-N was significantly inhibited under anaerobic incubation condition, and the NO₃^?-N contents decreased by 37.2%, 61.9% and 91.9% under low, medium and high pollution loads, respectively. Nitrification was stronger in the silt soil since its copy number of amoA and nxrA genes was two times larger than that of fine sand. Moreover, the copy numbers of amoA and nxrA genes in the silt soil under the aerobic environment were 2.7 times and 2.2 times larger than those in the anaerobic environment. The abundance changes of the amoA and nxrA functional genes have a positive correlation with the nitrification intensity in the tannery sludge-contaminated soil.     

一株H_5N_1亚型禽流感病毒株NA基因的克隆与序列分析

用RT-PCR方法从1个H5N1亚型禽流感病毒分离株A/Chicken/Guangdong/DH/1997扩增NA基因cDNA片段,将其克隆至pMD18-T载体,获得重组质粒pMD-NA,并对其核苷酸序列进行测定和分析。结果表明,该毒株的NA基因长度为1350bp,编码449个氨基酸,与其它H5N1亚型AIV分离株的核苷酸序列同源性为97.0%～99.4%,氨基酸序列同源性为97.7%～99.1%,提示禽流感病毒NA基因保守性较高。NA基因氨基酸序列的聚类分析表明该毒株与来自香港的A/Pheasant/HK/FY155/01和A/Ch/HK/FY150/01两个分离株处于同一进化枝,亲缘关系较近。     

基于转录组测序技术对调控毕加索小丑鱼(Picasso clownfish)白化体征关键基因的研究

毕加索小丑鱼(Picasso clownfish)因其皮肤中的白色斑块分布杂乱抽象而得名,同时也由于其白色斑块的形成无规律性和稀缺性,属于名贵的小丑鱼,因此,解析毕加索小丑鱼的皮肤白斑形成机制,可以为毕加索小丑鱼的人工育种提供理论依据.在本研究中,我们对3种体色毕加索小丑鱼的背鳍和臀鳍之间的身体相同部位3种色块(黑色、...     

倒立水母Cassiopea andromeda 的POU、Hox基因家族的多样性及系统发育分析

与其他钵水母相比,倒立水母具有独特行为：在生命周期的大部分时间里,它都在海底呈倒立附着、“睡眠”的状态。为探索与这一独特行为相关的遗传信息,本研究对安朵仙水母(Cassiopea andromeda)和同属于钵水母纲的海蜇(Rhopilema esculentum)、巴布亚硝水母(Mastigias papua)进行首次全基因组测序、拼接和注释,并重点分析了这3种钵水母与感觉功能、神经系统发育相关的重要转录调节因子Hox和POU基因家族的多样性与系统发育关系。遗传分析显示,刺胞动物门中Hox和POU基因家族具有明显的物种间差异性。对Hox基因的分析首次发现钵水母及水螅(Hydra vulgaris)的“前段Hox基因”产生了部分缺失,并进一步印证了刺胞动物不存在“中段Hox基因”的假说。安朵仙水母和海蜇具有相对全面的ParaHox基因种类,即GSX和XLOX/CDX基因,而巴布亚硝水母只有GSX基因。在POU基因多样性方面,安朵仙水母、海蜇、星状海葵(Nematostella vectensis)的基因组具有全部4类POU,而巴布亚硝水母、海月水母(Aurelia aurita)、水螅只有2类POU。在本研究分析的刺胞动物中,安朵仙水母的POU-1,-6亚型的核苷酸多态性最高;安朵仙水母与指形鹿角珊瑚(Acropora digitifera)、星状海葵的POU-2/3/5亚型的序列多样性较其他钵水母更高。另外,3种钵水母与水螅粘附蛋白的比较结果表明,安朵仙水母具有巴布亚硝水母和海蜇所不具有的粘附相关鼠李糖结合凝集素和一类抗氧化活性物质铁螯合物还原酶。综上所述,安朵仙水母具有更多POU编码基因和复杂POU亚型,以及具有粘附相关凝集素和还原酶的编码基因,可能是与安朵仙水母倒立附着生活方式相关的关键遗传信息。     

合浦珠母贝丝氨酸蛋白酶抑制因子基因pfser1克隆与表达

【目的】克隆合浦珠母贝(Pinctada fucata)丝氨酸蛋白酶抑制因子pfser1基因,探讨该基因的组织表达及其在天然免疫过程中的作用,以及与生物矿化过程的关系。【方法】通过RACE技术获得pfser1基因的全长,通过生物信息学分析其序列结构特征,利用实时荧光定量PCR方法检测pfser1基因在不同组织中的表达,检测健康合浦珠母贝在被大肠杆菌(Escherichia coli)MG1655刺激后和在贝壳损伤修复实验中pfser1基因表达量的变化。【结果】合浦珠母贝pfser1基因cDNA全长为1240 bp,包含1035 bp的开放阅读框(ORF),编码344个氨基酸,氨基酸序列的功能结构域含有丝氨酸蛋白酶抑制因子Serpin家族保守结构域。pfser1基因在合浦珠母贝各个组织中均有表达,在外套膜边缘膜中表达量最高;大肠杆菌MG1655刺激后,该基因表达量显著升高;在贝壳损伤修复过程中,pfser1基因表达先升高后受到抑制。【结论】pfser1基因所表达的蛋白参与了合浦珠母贝的天然免疫应答过程,并与生物矿化过程有一定关系。     

基于景观基因“地域机制”的客家文化保护与传承开发——以湖南省炎陵县为例

根据客家文化景观基因理论及其"地域机制",以客家扩展聚居区炎陵县为例,结合历史文献查阅和实地调研,发掘分析该县客家文化的景观基因。研究结果表明:(1)迁徙由来、地域背景分别是客家文化景观的共同基因、本土基因;(2)由共同基因以"反客为主"的作用方式与本土基因形成了地方客家景观本质特性的主体基因;(3)保护与传承意识形态是决定其主体基因能否在世代遵循并坚守着"原汁原味"文化传统和文化精神传承过程中的人为基因。基于这些独特基因构成内在相关联的地域机制而所表达的特征,提出了应加强客家文化景观基因的数字化保护管理、传承开发政策等对策。     

Morphological and Phylogenetic Studies of a Copepod Species,Irodes parupenei Ho and Lin (2007), Infecting Parupeneus rubescens in Saudi Arabia

Dammam City is one of the gorgeous coastal areas in the Arabian Gulf of Saudi Arabia.The present study aimed to ex-amine one of the copepod species infecting the rosy goatfish that represents a highly consumed fish species by the local population in the Arabian Gulf.The copepod species isolated from the infected fish specimens belong to the family Taeniacanthidae and was iden-tified as Irodes parupenei Ho and Lin(2007),primarily based on its morphological,morphometric,and ultrastructural characteris-tics,especially the structures of the dorsal cephalic area,segmentation of the first antenna,the absence of the maxilliped claw in the fe-male specimens,and the setation and spinulation of the legs 2-4 for the adult females are of great significance in the taxonomic iden-tification.The 18S rRNA gene sequence was analyzed to ensure the precise identity and exact taxonomic status of the copepod species.The result showed that this copepod species belong to Taenicanthidae and closely related to Irodes sauridi(gb|JF781550.1)in the same taxon.More details on the specificity of the goatfish for Irodes species and identifying these parasitic taxa using molecular analysis are given in the present study.     

A Sensitive and Versatile Multiplex PCR System for the Rapid Detection of Enterotoxigenic (ETEC), Enterohaemorrhagic (EHEC) and Enteropathogenic (EPEC) Strains of Escherichia coli

Although Escherichia coli is widely distributed in the marine environment, only a small percentage are pathogenic to humans. Nonetheless, the widespread occurrence of waterborne infections of E. coli origin in humans has become one of the major health problems worldwide. To date, several types of enterovirulent E. coli have been recognized as the aetiologic agents of various gastrointestinal infections in humans. The most commonly encountered are those belonging to the enterotoxigenic (ETEC), enteroinvasive (EIEC), enterohaemorrhagic (EHEC) and enteropathogenic (EPEC) subtypes. In order to better determine the health risks that are associated with exposure to some of these specific subtypes, we have developed a very sensitive multiplex PCR system for the rapid detection and typing of ETEC, EHEC and possibly EPEC strains of E. coli in the aquatic environment. The target genes chosen for this investigation included: the PHO-A housekeeping gene (present in all E. coli); the LT1, LT2 and ST1 genes of ETEC; the VT1 and VT2 verotoxin, and EAE virulence genes of EHEC and EPEC, respectively. Six pairs of oligonucleotide primers were designed to simultaneously amplify internal fragments of these genes by multiplex PCR to generate PCR products that could be analysed and confirmed with relative ease by gel electrophoresis and HincII enzyme digestion. The results showed that the six sets of PCR primers were highly specific for their target genes and produced specific amplimers of the expected size from several control strains of E. coli – ATCC 35401 (LT1+/ST1+); SA53 (LT2+/VT2+); and O157 (VT1+/VT2+/EAE+). The detection sensitivity of the multiplex PCR system for the six target genes in an E. coli cell mixture was optimized and enhanced by preincubating serially diluted cells in Luria-Bertani broth for 6 h prior to PCR analysis. The results obtained indicated a detection sensitivity of 10° CFU (of each strain) per 100 μl reaction. Multiplex PCR analysis of seawater samples collected from four sewage-polluted sites in Hong Kong indicated the presence in all four samples of E. coli bacteria that were positive for LT1, ST1, VT1 and EAE virulence genes. Overall, the data indicated that the multiplex PCR system described in this study is a potentially very useful and powerful method for routine monitoring and risk assessment of water quality.     

狄氏斧蛤(Donax dysoni)线粒体全基因组测定及结构特征分析

为解析狄氏斧蛤(Donax dysoni)线粒体全基因组结构特征以及进化地位,采用二代高通量测序技术获得狄氏斧蛤线粒体基因组全序列,对线粒体基因进行注释并对其序列结构进行分析。结果表明,狄氏斧蛤的线粒体全基因组序列全长为16 908 bp,碱基组成为A (26.81%)、T (41.13%)、G(21.21%)和C (10.85%),A+T含量为67.94%,表现出明显的AT偏向。与其他双壳贝类相似,狄氏斧蛤共编码13个蛋白质编码基因,包含22个t RNA和2个r RNA,且所有基因均位于H链;蛋白质编码基因拥有3种起始密码子(ATG、ATT、ATA),而除了ND3、ND4以及ATP6使用TAG作为终止密码子外,其余所有蛋白质编码基因都使用TAA作为终止密码子。除t RNASer不能折叠成典型的三叶草结构,没有明显的DHU茎环,其余t RNA都能折叠成典型的三叶草结构。狄氏斧蛤与斧蛤科(Donacidae)其他物种具有相同的基因排列,未发现基因重排现象。基于线粒体12个蛋白质编码基因构建62种贝类的进化树,结果显示:Donax semiestriatus和Donax vittatus聚...     

双壳贝类CK1基因家族的鉴定及表达分析

酪蛋白激酶1 (CK1)是一种重要的蛋白激酶家族,其在DNA损伤应答和修复、细胞增殖和凋亡、胚胎发育和稳态等重要的生物学过程中都具有复杂多样的调控作用。为了理解CK1基因家族在双壳贝类中的特征、进化及生物学功能,实验采用比较基因组学及生物信息学的方法对双壳贝类CK1基因家族进行了鉴定分析,通过虾夷扇贝高温应激实验,研究了CK1基因家族在高温应激时的表达规律。结果显示,在虾夷扇贝、栉孔扇贝、长牡蛎与侏儒蛤中均存在CK1α,CK1αlike,CK1δ,CK1γ3,并发现CK1ε,CK1γ1,CK1γ2在双壳贝类中发生了丢失。时空表达分析发现,双壳贝类CK1基因均在胚胎发育早期集中表达,并以多细胞/囊胚期为界呈现出母源/合子特异性表达模式。CK1为关键基因的基因共表达模块显著富集在错配修复、核苷酸剪切修复等相关通路上,暗示了CK1基因在双壳贝类胚胎发育过程中参与调控DNA损伤修复过程,从而维持早期胚胎发育时的基因组稳定性。双壳贝类CK1基因在成体中呈现组织特异的表达模式,主要在鳃和雄性性腺中具有相对较高的表达量。虾夷扇贝受到高温应激后,其鳃中PyCK1α, PyCK1αlike和PyCK1δ...