微囊藻毒素对大鳞副泥鳅胚胎和幼鱼的毒性效应

微囊藻毒素(Microcystins,MC)是一类单环七肽的生物毒素,它主要由淡水藻类铜绿微囊藻(Microcystisaeruginosa)产生,由于MC的作用靶器官为肝脏,故又称肝毒素。       

南四湖泥鳅和大鳞副泥鳅随机扩增多态DNA分析初报

采用随机扩增多态性DNA(RAPD)技术对南四湖的泥鳅（Misgurnus anguillicaudatus）和大鳞副泥鳅（Paramisgurnus dabryanus）进行了初步分析。共用10个10bp的OPG组寡核苷酸片段作为引物,筛选出结果好的5个引物,从泥鳅4个体基因组DNA中扩增出142个片段,共有片段12个,特有片段5个;从大鳞副泥鳅3个体基因组DNA中扩增出101个片段,共有片段18个,特有片段5个,在总扩增出的243个片段中,共有片段仅4个。所有扩增片段大小介于0.2-3kb之间。用PoPGen32软件计算出个体间的遗传距离,依此进行UPGMA聚类得到的分子系统树与形态分类学的结果一致,这7个体被聚成2大类,正对应着形态学的2个种：泥鳅和大鳞副泥鳅。     

PCR扩增泥鳅和大鳞副泥鳅SRY盒基因

以特异扩增人ＳＲＹ基因保守区的一对引物,研究了泥鳅和大鳞副泥鳅基因组中ＳＲＹ盒基因的扩增。结果表明,该引物可以在泥鳅中扩增出四条带,其长度分别为２００,５５０、９４０和１０００ｂｐ。在大鳞副泥鳞中扩增出三条带,大小为２００,５５０和９００ｂｐ。经Ｓｏｕｔｈｅｒｎ杂交显示出二者的阳性带为２００和５５０ｂｐ。阳性带在雌雄个体间和两个物种间无差异。     

大鳞副泥鳅精子结构研究

用光学显微镜和透射电子显微镜对大鳞副泥鳅精子的结构进行研究。结果表明,大鳞副泥鳅的精子主要分为头部、中段和尾部;头部无顶体,在光学显微镜下近圆形,在透射电子显微镜下纵切亦近圆形,主要由细胞核组成,核内染色质致密,核内有核空泡,核外可见清晰核膜,核外可见细胞质,细胞质很少且紧贴细胞核,细胞质外是质膜,质膜在细胞质外呈波浪状;头部后端有一较浅的植入窝,约占核的1/4,植入窝的长轴几乎与细胞核的长轴平行,植入窝内有中心粒复合体;精子的中片与头部无明显分割,位于头部的后方,由中心粒复合体和袖套组成,袖套两边不对称,中心粒复合体由近端中心粒和远端中心粒组成,近端中心粒与远端中心粒之间呈一钝角;精子尾部无侧鳍,可分为主段和末段,尾部主段具有典型的＂9＋2＂的轴丝结构。精子头长为（1.79±0.28）μm,中片长为（1.86±0.42）μm,尾长为（28.06±2.78）μm,全长为（31.65±2.82）μm。     

大鳞副泥鳅雌雄个体的形态分析

本文利用数理统计手段研究南四湖大鳞副泥鳅雌雄个体形态差异,结果表明大鳞副泥鳅雌雄个体形态整体差异不显著,但体长/吻腹长指标差异显著;1龄鱼、3龄鱼雌性个体体长、体重大于雄性个体,2龄鱼则呈相反趋势;t检验表明,仅3龄鱼雌雄个体间体长、体重差异显著。     

大鳞副泥鳅、泥鳅和北方泥鳅肉质比较分析

文章采用组织切片、生化组分分析以及实时荧光定量PCR等方法,研究了大鳞副泥鳅(Paramisgurnus dabryanus)、泥鳅(Misgurnus anguillicaudatus)和北方泥鳅(Misgurnus bipartitus)肉质差异。结果显示:大鳞副泥鳅、泥鳅和北方泥鳅的肌纤维横截面积分别为(3589.17±2326.01)、(2809.7±1818.69)和(2511.93±1949.03)μm2。粗蛋白和粗脂肪含量均是大鳞副泥鳅最高[分别为(17.07±0.31)%和(2.57±0.38)%],依次为泥鳅[分别为(14.57±0.59)%和(1.37±0.12)%]和北方泥鳅[分别为(12.33±0.15%和0.57±0.06%]。必需氨基酸指数(EAAI)依次为泥鳅(74.38)、大鳞副泥鳅(65.11)和北方泥鳅(60.14);呈味氨基酸含量依次为泥鳅(32.60±1.64)%、大鳞副泥鳅(27.75±2.13)%和北方泥鳅(24.86±1.00)%;除亚油酸以外的总多不饱和脂肪酸含量依次为北方泥鳅(24.43±0.26)%、泥鳅(24.18±1.99)%和大...     

泥鳅和大鳞副泥鳅卵雌核的紫外辐射灭活条件筛选

采用紫外线对泥鳅和大鳞副泥鳅卵雌性原核进行干法灭活以及在合成卵巢液、TC-199溶液、Ringer氏溶液和Holtfreter氏溶液中灭活,结果表明:当辐射剂量为90-180mJ/cm~2时,卵子在四种溶液中的单倍体诱导率均较高。合成卵巢液组的最佳辐射剂量为120-180mJ/cm~2;TC-199和Ringer氏溶液组的最佳辐射剂量为120mJ/cm~2;原液使用Holtfreter氏溶液效果欠佳,干法灭活效果最差。在上述各辐射剂量下,畸形苗经染色体组鉴定92％为雄核发育单倍体。综合评价,灭活效果依次是:人工合成卵巢液>TC-199溶液>Ringer氏溶液>Holtfreter氏溶液>干法。     

大鳞副泥鳅鳞片表面结构的扫描电镜观察

对大鳞副泥鳅的鳞片作了扫描电镜观察。结果表明,大鳞副泥鳅具圆鳞;基区、侧区和顶区均具有辐射沟及环沟,二者交织成网状,将鳞片分割成块状,可增加鳞片的柔软度;次级辐射沟发出部位可作为确定年轮的主要依据。     

泥鳅与大鳞副泥鳅正反交子代遗传变异的RAPD分析

我国利用远缘杂交方法在鱼类杂交优势利用上已经取得了不少成果1.不仅如此,不少学者还通过胚胎发育形态学、染色体组型、同工酶等生化表型的研究对远缘杂交中的核质关系、基因表达、不亲和性原理进行了广泛探索2-5.       

铜对鱼类慢性毒性研究进展

重金属铜既是生物体自身所必需的元素,也可造成污染。       

TOXICITY OF COPPER TO THALASSIOSIRA PSEUDONANA IN UNENRICHED INSHORE SEAWATER1

Toxicity of copper to T. pseudonana (formerly Cyclotella nana, clone 13-1) wax examined in inshore seawater using a 96-hr bioassay method. Raw unenriched seawater wax filtered through a 0.22-μ membrane filter and then pasteurized, for 30 min at 60 C. Following this treatment, samples contained 0.68–1.14 μg Cu/liter. Copper was added as the chloride in 5-μg increments over the range of 5 to 30 μg/liter (about 0.1–0.5 μM). Population densities, mean cell, volume, and ¹⁴C bicarbonate uptake were measured. Population growth, and ¹⁴C uptake by T. pseudonana displayed inhibition over the entire range of added copper. Growth rate constant (k) of T. pseudonana decreased with increasing copper concentration and during the course of growth at each concentration. Correspondingly, mean cell volumes increased with copper concentration and time. Copper toxicity varied in different water samples. The presence of decomposed natural plankton and detritus decreased toxicity. In the absence of enrichment, bacteria had little effect on copper toxicity. H exults were influenced by glassware treatment, collection and storage of seawater, and absence of enrichments.     

两种泥鳅不同群体遗传变异的RAPD分析

应用RAPD技术,分析了采自于我国黄河、长江和珠江三大水系中游的大鳞副泥鳅和泥鳅不同群体间的遗传变异。结果表明：种内不同群体间带纹相似度在0．730－0．938之间;遗传距离为0．089－0．245;种间不同群体间的带纹相似度为0．392－0．505,遗传距离为0．620－0．800。两种泥鳞采自武汉的群体,其不同个体间的相似度较之其它群体为低,表明其群体内遗传变异程度较高。     

COPPER TOXICITY TO SKELETONEMA COSTATUM (BACILLARIOPHYCEAE)1,2

Copper toxicity to Skeletonema costatum (Grev.) Cleve has been studied in batch cultures of chemically defined culture media. The alga is relatively insensitive to cupric ion activity, demonstrating no effect on growth up to (Cu²⁺) = 10^?8.5 M. Cultures inoculated from stationary phase stocks exhibit a prolongation of the lag phase with increasing copper concentrations near and above the point of precipitation of the copper. The toxicity of copper is a function of the silicic acid concentration in the medium. This effect is observed in a range of Si(OH)₄ concentrations (10^?5 M to 10^?4 M) above known values for the saturation of silicon uptake kinetics, thus suggesting an influence of copper on silicate metabolism.     

THE ROLE OF REACTIVE OXYGEN SPECIES IN COPPER TOXICITY TO TWO FRESHWATER GREEN ALGAE1

The role of reactive oxygen species (ROS) in copper (Cu) toxicity to two freshwater green algal species, Pseudokirchneriella subcapitata (Korshikov) Hindák and Chlorella vulgaris Beij., was assessed to gain a better mechanistic understanding of this toxicity. Cu‐induced formation of ROS was investigated in the two algal species and linked to short‐term effects on photosynthetic activity and to long‐term effects on cell growth. A light‐ and time‐dependent increase in ROS concentrations was observed upon exposure to environmentally relevant Cu concentrations of 50 and 250 nM and was comparable in both algal species. However, effects of 250 nM Cu on photosynthesis were different, leading to a 12% reduction in photosynthetic activity in P. subcapitata, but not in C. vulgaris. These results indicate that differences in species‐specific sensitivities measured as photosynthetic activity were not caused by differences in the cellular ROS content of the algae, but probably by different species‐specific ROS defense systems. To investigate the role of ROS in Cu‐mediated inhibition of photosynthesis, the ROS scavenger N‐tert‐butyl‐α‐phenylnitrone (BPN) was used, resulting in a reduction of Cu‐induced ROS production up to control level and a complete restoration of photosynthetic activity of Cu‐exposed P. subcapitata. This finding implied that ROS play a primary role in Cu toxicity to algae. Furthermore, we observed a time‐dependent ROS release process across the plasma membrane. More than 90% of total ROS were determined to be extracellular in P. subcapitata, indicating an efficient method of cellular protection against oxidative stress.     

INFLUENCE OF PHOSPHORUS NUTRITION ON COPPER TOXICITY TO THREE STRAINS OF SCENEDESMUS ACUTUS (CHLOROPHYCEAE)1

Three strains of Scenedesmus acutus f. alternans Hortobagyi with markedly different sensitivities to copper were examined to determine the relative importance of cellular polyphosphate content on acute copper toxicity to photosynthesis. By manipulating the phosphate concentration in semicontinuous cultures, the response of each strain was assessed at three cellular phosphorus states: P-loaded, P-sufficient, and P-deficient. The results demonstrated the importance of cellular polyphosphate content in reducing the toxic effect of copper on photosynthesis; the greater the cellular P content, the less inhibition of photosynthesis occurred during copper exposure. This relationship was evident for both Cu-tolerant strains (XCu and B-4) and a Cu-sensitive strain (X72). The ranges of response to 9.9 μM Cu (measured as the percentage of control rate of photosynthesis within strains) were, from P-loaded to P-deficient cells, X72, 78–54%; XCu, 95–77%; and B-4, 99–94%. The data suggest that polyphosphate plays a passive role in protecting cells from copper; however, with respect to the mechanism of Cu tolerance, polyphosphate appeared to be relatively unimportant because the sensitivity of the Cu-tolerant strains showed less dependence on cellular polyphosphate than did the Cu-sensitive strain.     

大肠杆菌抗铜启动子的亚克隆

大肠杆菌的抗铜质粒中含有两个抗铜启动子,PpcoA和PpcoE,它们均含有copperbox。为了证实copper box在抗铜作用中的重要性以及研究抗铜启动子的特性,以质粒pUCD615为报告载体进行了这两个启动子不同长度片段的亚克隆。限制性酶切和DNA序列分析的结果表明,这些片段的亚克隆都是成功的;此外报告基因lux-荧光酶活性测定的结果表明,两个不含有copper box的Ppco short-lux亚克隆均不表达荧光酶活性,而其他启动子片段的亚克隆则都具有较高的酶活性,说明这两个亚克隆不具有启动子功能,初步证明在抗铜启动子中copper box是不可少的。