家猪基因指纹图的初步研究

寡核苷酸探针（CAC）5／（GTG）5检测了五指山猪、枫泾猪、枫泾猪与长白山猪杂交子一代的基因指纹图．各个体的可分辨谱带，分布于0．7～21．2kb之间．在约1．3kb处的一条共有谱带，可能是家猪的特征带．     

常乐稀土复合肥对玉米（Zea may L.）的细胞毒性和遗传毒性效应

为了研究常乐稀土多元复合肥对主要农作物--玉米(Zea mays L.)是否产生细胞毒性和遗传毒性效应,采用不同梯度质量浓度的稀土复合肥溶液对玉米进行处理,观察其根尖生长状况,统计分析细胞死亡率、微核率、有丝分裂指数以及对细胞DNA的损伤情况.结果表明,质量浓度为10mg·L-1以下时,常乐稀土复合肥能够促进玉米根尖的生长,100mg·L-1以上时抑制根尖生长;随着试验浓度的递增,细胞死亡率、微核率、DNA损伤率逐渐上升并具有正相关性,表现出明显的剂量-效应关系,而有丝分裂指数却随着浓度的增加逐渐下降.以上结果表明,常乐稀土复合肥对玉米确有一定的细胞毒性和遗传毒性效应,其细胞毒性阈值和遗传毒性阈值皆为100mg·L-1.     

Population structure and historical demography of the thorny skate (<Emphasis Type="Italic">Amblyraja radiata</Emphasis>, Rajidae) in the North Atlantic

Population genetic structure of the thorny skate (Amblyraja radiata) was surveyed in >300 individuals sampled from Newfoundland, Iceland, Norway, the Kattegat and the central North Sea. A 290-bp fragment of the mt cytochrome-b gene was first screened by SSCP. Sequences of SSCP haplotypes revealed 34 haplotypes, 14 of which were unique to Iceland, 3 to Newfoundland, 1 to Norway and 3 to the Kattegat. The global F _ST was weak but significant. Removal of the two Kattegat locations, which were the most differentiated, resulted in no significant genetic differentiation. Haplotype diversity was high and evenly distributed across the entire Atlantic (h = 0.8) with the exception of the North Sea (h = 0.48). Statistical parsimony revealed a star-like genealogy with a central widespread haplotype. A subsequent nested clade analysis led to the inference of contiguous expansion with evidence for long distance dispersal between Newfoundland and Iceland. Historical demographic analysis showed that thorny skates have undergone exponential population expansion that started between 1.1 million and 690,000 years ago; and that the Last Glacial Maximum apparently had little effect. These results strongly differ from those of a parallel study of the thornback ray (Raja clavata) in which clear structure and former refugial areas could be identified. Although both species have similar life history traits and overlapping ranges, the continental shelf edge apparently does not present a barrier to migration in A. radiata, as it does for R. clavata.     

RAPD扩增中商品酶体系对扩增产物的影响

随机选用了10条RAPD引物，采用3种商品TapDNA聚合酶体系，以5个不同物种的动植物总DNA为模板，进行RAPD扩增，实验发现，在对同一模板、采用同一引物进行的RAPD扩增中，仅仅因为使用了不同的商品TaqDNA聚合酶，获得的扩增产物差别极大；不同商品来源的PCR反应缓冲体系，对扩增产物也有一定影响，但相对较小，这说明，不同的商品酶体系对RAPD扩增产物影响很大，因此，影响RAPD扩增产物稳定性的一个关键因素是商品TaqDNA聚合酶本身；研究中前后一致地使用同一商品TaqDNA聚合酶体系，是成功进行RAPD分析的基础。     

用改良彗星试验检测工业废水的DNA损伤作用

采用改良彗星试验直接检测了多种工业废水不所引起的Ｖ７９细胞ＤＮＡ损伤。结果表明,所测的种各工业废话水均含有ＤＮＡ损伤剂,能够诱发培养的Ｖ７９细胞ＤＮＡ链断裂,其中以制革厂不的ＤＮＡ损伤作用最强。本研究显示出改良彗星试验在综合评价各种废水遗传毒性方面具有广阔的应用前景。     

分层型水库藻类垂向演替的水质与细菌种群调控

浮游藻类和细菌是水生食物网的重要组成部分,在水源水库生态系统物质循环和能量流动方面发挥关键作用,藻类的空间演替与水体细菌种群代谢和结构演变关系密切.因此,本文以李家河水库为研究对象,在分析水库水质指标基础上,采用高通量DNA测序技术和Biolog技术,研究水库藻类暴发期,浮游藻类和细菌群落结构垂向演替特征及其与水质的偶联关系.结果表明,李家河水库在8月处于热分层期,水体pH、DO和NH~+_4-N随深度变化均逐渐降低(P0.001).藻密度和Chla呈现同步变化趋势(P0.001),表层最大分别为3 364.33×10~4 cells·L~(-1)和7.03μg·L~(-1).藻类群落结构在水深0 m和3 m处以微囊藻为主,而在水深6 m处,小环藻取代微囊藻成为最优藻属,相对丰度达57.28%.Biolog结果表明,微囊藻的暴发对细菌代谢活性及其相对丰度产生较大影响,但细菌种群代谢活性多样性变化不显著.高通量测序共发现1 420个OTUs,隶属于10个细菌门类,其中放线菌门(Actinobacteria)和变形菌门(Proteobacteria)在不同水层均为最优门类,其相对丰度之和达50%以上;绿菌门(Chlorobi)和浮霉菌门(Planctomycetes)的相对丰度随着水深增加发生显著变化,均在6 m深度达到最大值,分别为10.29%和6.78%,且与水层藻密度呈显著负相关(P0.05);厚壁菌门(Firmicutes)和芽单胞菌门(Gemmatimonadetes)与藻密度呈显著正相关(P0.05).热图(Heat map)指纹图谱表明,李家河水库的细菌群落结构垂向分布差异显著,且随着水深的增加,细菌群落分布更均匀并趋于多样性.冗余分析(RDA)表明,细菌和藻类的群落结构的垂向分布受不同水质指标调控且差异显著.对水源水库藻类暴发期藻类与细菌群落的偶联机制进行探究,为水源藻华消涨的分子微生态驱动机制研究提供科学依据.     

质粒pJP4水平转移介导生物膜系统强化降解2,4-D效应

以携带质粒pJP4[其上含编码2,4-二氯苯氧基乙酸(2,4-dichlorophenoxyacetic acid,2,4-D)降解功能的基因簇(tfd)]的基因工程菌Pseudomonas putida SM1443::gfp2x(pJP4::dsRed)为供体菌,以生物膜系统为对象,通过半连续流实验研究了质粒pJP4水平转移介导基因强化降解2,4-D效应,考察了目标基因在系统中存在状况及基因强化对系统菌群结构的影响.结果表明,以2,4-D(初始浓度为170 mg/L±10 mg/L)为唯一碳源,向生物膜系统加入携pJP4质粒的基因工程菌对2,4-D的降解具有促进作用,运行初期,促进作用较弱,随着半连续流反应的进行,促进作用显著增强,基因强化系统较对照系统对2,4-D的平均降解速率之差达13.3 mg/(L.h).通过对基因强化系统功能基因片段tfdB基因及报告基因gfp的跟踪检测,证实了在pJP4质粒介导下生物膜系统基因水平转移的发生.PCR-DGGE结果表明基因强化的生物膜系统较对照系统在受到2,4-D冲击条件下保持了相对更加稳定的菌群结构.     

一株苯酚降解菌的筛选鉴定及响应面法优化其降解

从某化工厂污水处理车间活性污泥中分离、筛选到一株能以苯酚为唯一碳源和能源生长的菌株YH8.基于形态特征、生理生化特性、BIOLOG细菌自动鉴定系统、16S rDNA和gyrB基因序列同源性分析鉴定菌株YH8,鉴定菌株YH8为Acinetobacter guillouiae.在苯酚浓度低于1200 mg·L-1,温度为26~34℃,pH为7.0~10.0时,菌株YH8培养60 h对苯酚的降解率达70%以上.运用单因素实验初步确定苯酚降解的最适外加碳源和氮源分别为山梨醇和NaNO3,最适温度为30℃,最适初始pH为9.0,最适接种量为5%.为了提高菌株YH8的降解率,首先利用Plackett-Burman实验设计评估并筛选出影响苯酚降解的3个关键因素为初始pH、苯酚浓度、山梨醇浓度.用最陡爬坡实验逼近以上3个因子的最大响应区域,采用Box-Behnken实验设计及响应面法分析,确定其最优降解条件为初始pH 9.26、苯酚浓度1163.63 mg·L-1、山梨醇浓度7.81%、接种量5%、NaNO_3浓度2%、温度30℃、培养时间96 h,在此条件下苯酚降解率可达98.95%.苯酚降解酶活性及酶定域实验表明,菌株YH8相关降解酶为胞内酶,且苯酚可诱导苯酚羟化酶(LmPH)和邻苯二酚1,2-双加氧酶(C_(12)O)的合成.通过降解酶特异性引物从菌株YH8扩增得到LmPH和C12O基因片段,经质粒检测和消除实验发现菌株YH8相关降解基因位于质粒上.此外,菌株YH8能耐受高浓度NaCl和多种重金属离子,对多种抗生素具有抗性.     

大亚湾浮游细菌的变性梯度凝胶电泳分析

于2010年12月以及2011年6月采集了大亚湾9个站点的表层水样,利用PCR-变性梯度凝胶电泳(PCR-DGGE)技术对浮游细菌DNA指纹进行了分析.结果显示,大亚湾海域浮游细菌DNA指纹比较丰富,冬季和夏季样品中分别共得到32条和33条条带,每个样品得到的条带数为18~25条.DNA指纹的Shannon-Weaver多样性指数(H')和Pielou均匀度(J)的变化范围分别为2.48~2.88和0.80~0.93.DNA指纹条带数与温度和盐度明显正相关,而与p H、DO、透明度呈明显负相关关系.结果说明PCR-DGGE技术可以在一定程度上反映细菌的群落结构和多样性,但需要与其他技术相结合方能较全面反映细菌的结构与功能.     

Genotoxicity of ferric oxide nanoparticles in Raphanus sativus: Deciphering the role of signaling factors, oxidative stress and cell death

We have studied the genotoxic and apoptotic potential of ferric oxide nanoparticles(Fe_2O_3-NPs) in Raphanus sativus(radish).Fe_2O_3-NPs retarded the root length and seed germination in radish.Ultrathin sections of treated roots showed subcellular localization of Fe_2O_3-NPs,along with the appearance of damaged mitochondria and excessive vacuolization.Flow cytometric analysis of Fe_2O_3-NPs(1.0 mg/m L) treated groups exhibited 219.5%,161%,120.4% and 161.4% increase in intracellular reactive oxygen species(ROS),mitochondrial membrane potential(ΔΨm),nitric oxide(NO) and Ca2+influx in radish protoplasts.A concentration dependent increase in the antioxidative enzymes glutathione(GSH),catalase(CAT),superoxide dismutase(SOD) and lipid peroxidation(LPO) has been recorded.Comet assay showed a concentration dependent increase in deoxyribonucleic acid(DNA) strand breaks in Fe_2O_3-NPs treated groups.Cell cycle analysis revealed 88.4% of cells in sub-G1 apoptotic phase,suggesting cell death in Fe_2O_3-NPs(2.0 mg/m L) treated group.Taking together,the genotoxicity induced by Fe_2O_3-NPs highlights the importance of environmental risk associated with improper disposal of nanoparticles(NPs) and radish can serve as a good indicator for measuring the phytotoxicity of NPs grown in NP-polluted environment.