利用ND1、ITS1序列研究三种背角无齿蚌（Anodontawoodiana）分类学问题

利用ND1、ITS1序列差异对采自太湖、在形态上被认为是背角无齿蚌、圆背角无齿蚌、椭圆背角无齿蚌的三种个体进行鉴定,并与从NCBI中下载到的同属不同种褶纹冠蚌的ND1、ITS1序列进行对比,分析了基于Kimura2-parameter模型参数得到的遗传距离,并构建了NJ树。结果表明,本研究中所有背角无齿蚌种间遗传距离变...     

稀土元素镧对三角帆蚌(Hyriopsis cumingii)早期珍珠色泽的影响

采用酶学、电镜扫描以及X射线衍射等方法,研究不同浓度稀土元素镧对三角帆蚌(Hyriopsis cumingii)早期珍珠色泽的影响。实验共分五组,I至V组稀土元素镧终浓度分别为0、0.7、1.0、1.3、1.6mg/L。结果显示,稀土元素镧对酸性磷酸酶、碱性磷酸酶具有Hormesis效应,当镧浓度为1.0mg/L时,三角帆蚌外套膜组织中的酸性磷酸酶、碱性磷酸酶比活力较高,分别为(29.3±19.4)U/g、(212.3±61.6)U/g,此浓度下三角帆蚌白色珍珠圆润度、结晶度最好,且生长比较均匀,XRD衍射显示珍珠中只含有少量的方解石和球文石结晶。提示,适量的稀土元素镧(1.0mg/L)能有效增加珍珠的本色,促进白色珍珠CaCO3文石规则有序地结晶,较好地提高三角帆蚌早期珍珠的色泽度。     

三角帆蚌(Hyriopsis cumingii)珍珠钙代谢的周年性变化研究

采用生态调查和生理生化方法,对植片后的1龄三角帆蚌(Hyriopsis cumingii)进行了为期一年的调查研究,拟探讨珍珠形成和钙代谢的相关性。定期测定珍珠囊和珍珠的重量、直径及水温,同时取其鳃、围心腔和外套膜组织进行酸、碱性磷酸酶活性检测及钙含量等测定。结果表明,1龄植片后的三角帆其蚌珍珠囊和珍珠在一周年之中出现三次快速生长期。外套膜中的酸、碱性磷酸酶均在165d时升至最高。鳃组织中钙含量显著高于围心腔和外套膜。经Pearson相关分析显示,珍珠囊和珍珠的重量、直径分别与三种组织中的碱性磷酸酶活性及外套膜中的钙含量呈显著性正相关(P<0.05);同时,外套膜中的钙含量与三种组织中的碱性磷酸酶呈显著性正相关(P<0.05);水温与外套膜、围心腔中的钙含量呈显著性负相关(P<0.05),与三种组织中的酸性磷酸酶呈显著性正相关(P<0.05)。提示:鳃是三角帆蚌钙吸收、贮存和钙调节的重要器官,形成珍珠的钙一部分来源于循环系统,另一部分可由外套膜直接从环境中吸收,碱性磷酸酶对珍珠生长起着重要的调节作用。     

四种营养盐对舟形藻(Navicula)BT001生长速率的影响

以I.A.M.收集的培养基Ⅱ为基本培养基,采用单因子和L16(45)正交设计法,进行了舟形藻BT001对N、P、Fe、Si四种营养盐最适需求的研究,并在此基础上,研究了尿素对正交优化组合的影响以及以尿素作为氮源对正交优化组合的影响。结果表明,N、P、Fe、Si四种营养盐最佳单因子水平为:KNO3,300mg/L;Na2HPO4·12H2O,40mg/L;FeCl3,4mg/L;Na2SiO3·9H2O,200mg/L。四种营养盐正交组合水平为:KNO3,150mg/L;Na2HPO4·12H2O,40mg/L;FeCl3,4mg/L;Na2SiO3·9H2O,200mg/L。在正交组合水平基础上,添加16mg/LCO(NH2)2,可更好的促进该藻的生长和繁殖。在8天的培养中,最大细胞相对生长率可达0.1577。在等摩尔氮源的条件下,以CO(NH2)2代替正交优化水平组合中的KNO3对舟形藻BT001进行11天的培养。结果表明,以CO(NH2)2为氮源的藻细胞最大生长密度可达2.69×105cell/ml,明显地高于硝酸钾作为氮源培养的藻细胞密度。     

人工育珠对三角帆蚌(Hyriopsis cumingii)遗传基因影响的初步研究

采用酚-氯仿和试剂盒两种提取法对三角帆蚌怀珠群与非怀珠群各6个个体进行基因组DNA的提取,然后在优化RAPD(随机扩增多态性DNA)检测条件基础上,从80个随机引物中筛选出12个扩增较好且多态性强的引物进行RAPD扩增,产物通过水平板琼脂糖凝胶电泳和垂直板聚丙烯酰胺凝胶电泳两种方法进行检验并对DNA的多态性进行分析。结果显示在怀珠群和非怀珠群检测到的位点数、多态位点比例、平均Shannon多态性信息指数、平均Nei's基因多样性指数、群体内个体间平均遗传相似率和平均遗传距离分别为100、33%、0.1927、0.1324、0.904、0.096,95、47.37%、0.2711、0.1879、0.861、0.139,群体间的平均遗传距离为0.821,非怀珠群的变异性大于怀珠群;本研究还获得了两群体各自的特异扩增谱带及两群体间表达差异较大谱带,这些位点很可能是由人工育珠所引起。根据MEGA4.0软件的UPGMA和NJ程序构建的分子系统树可直观地将两群体分开。     

三角帆蚌(Hyriopsis cumingii)外套膜无核珍珠颜色成因的育珠实验

用紫色蚌和非紫色蚌制取细胞小片,分别插入紫色蚌和非紫色蚌外套膜内,探究三角帆蚌(Hyriopsis cumingii)无核珍珠颜色与制片蚌、育珠蚌的关系;用不同类型制片蚌的不同部位制取细胞小片,分别插入育珠蚌外套膜内,进一步探究珍珠颜色与制片蚌珍珠质颜色的关系。结果表明,以紫色蚌为制片蚌,所产的珍珠为紫色系,珍珠紫色深浅与细胞小片所对应部位的制片蚌珍珠质紫色深浅呈正相关。以非紫色蚌为制片蚌,所产的珍珠有白色系和黄色系,珍珠黄色深浅与细胞小片所对应部位的制片蚌珍珠质黄色深浅呈正相关。三角帆蚌外套膜无核珍珠颜色是由提供细胞小片的制片蚌珍珠质颜色所决定,而与育珠蚌无关。该结论支持珍珠囊表皮细胞来自于移植细胞小片的观点。该结果表明,通过定向选育纯紫、纯白色贝壳珍珠质三角帆蚌新品系,即可培育出纯紫、纯白色无核珍珠。     

滇池螺蛳(Margarya melanioides)分布和底泥营养元素调查

研究了滇池螺蛳(Margarya melanioides)分布和底泥营养成分之间的关系。2013年在滇池87个采样点采集螺蛳和底泥样本,图示分析所有采样点的活的螺蛳数量,确定活螺在滇池中的分布位点及分布密度;选择8个采样点的底泥样本,进行16项营养元素(有机质、磷、氮、锌、铁、锰、镁、钙、铜、钠、钾、砷、汞、铅、镉、铬)含量测定,分析这些样本营养元素的差异。根据这8个采样点中有无螺蛳生长,将8个底泥样本分为两组,采用完全随机设计两样本比较的t检验,结果两组样本间仅铜的含量有统计学差异(P0.05),这可能是由于滇池螺蛳的生长有效富集了重金属铜元素,使有螺区域比无螺区域底泥中铜的含量明显减少。这在一定程度上说明了螺蛳生长与底泥的关系,可为滇池污染防治及螺蛳种群保护提供相关的科学依据。     

三角帆蚌(Hyriopsis cumingii)叉头蛋白2(foxl2)基因和性别决定框14(sox14)基因克隆与表达分析

三角帆蚌(Hyriopsis cumingii)生殖发育研究相对较少,相关基因信息缺乏。本文以三角帆蚌卵巢和精巢为研究对象,对转录组测序获得的叉头蛋白2(foxl2)基因和性别决定框14(sox14)基因保守区进行克隆和表达分析。定量PCR显示foxl2主要在卵巢中表达,而sox14主要在精巢中表达。在不同发育时期,foxl2和sox14在受精卵和卵裂期基本不表达,从原肠期开始表达,钩介幼虫期表达量比较高。原位杂交染色证明foxl2基因主要表达在卵巢的间质细胞和颗粒细胞中,sox14在精巢的间质细胞和一些精原细胞中表达。三角帆蚌foxl2的表达和脊椎动物有类似的模式,foxl2可能在河蚌卵巢发育中起到重要作用,可以作为卵巢的一个标志分子;Sox14可能对精巢发育和功能维持有重要作用,可以作为精巢的一个标记分子。Foxl2和sox14在胚胎发育中也会发挥一定的功能。本文结果丰富了河蚌的基因资源,可为三角帆蚌的生殖与发育生物学研究提供基础数据。     

自然环境下虾夷扇贝(Patinopecten yessoensis)体内麻痹性贝毒的净化研究

为探究在自然环境中虾夷扇贝Patinopecten yessoensis体内麻痹性贝毒(PSTs)的净化过程,从黄海北部大窑湾海域采集了受PSTs污染的虾夷扇贝,将其转移至黄海的棋盘磨水域,该水域未曾有PSTs污染的报道。本研究采用高效液相色谱-荧光检测法(HPLC-FLD)检测30天净化过程中虾夷扇贝体内PSTs的组分和水平。研究结果发现,在第9天,虾夷扇贝软体中的PSTs的含量下降到一个相对较低的水平。而且,初期扇贝体内PSTs的净化率高于后期。扇贝各组织器官中PSTs含量分析发现,消化腺中的PSTs含量最高,这对于人类健康和扇贝养殖具有重要的意义。在本研究中还发现,虾夷扇贝的死亡率与PSTs的水平有一定的关系。     

黄河三角洲关键种泥螺Bullacta exarata对沉积物-水体界面溶解氧和营养盐通量的影响

泥螺自2001因养殖目的引入黄河三角洲以来,随着分布范围的扩大和种群数量的增长,已成为当地潮间带的关键种。泥螺给当地民众带来较高收入,但其对底栖生态系统的潜在影响还不清楚。为了明确该种群的生物扰动对底栖微藻和微生物的初级生产,以及对水体-沉积物界面营养盐交换的影响,开展了该项实验。结果表明,在光照条件下,泥螺能显著影响沉积物-水体界面的溶解氧DO通量;在黑暗及无生物处理组的恢复期,泥螺能显著增加两界面间氨氮的通量。底栖微藻和小型生物在黑暗条件下也能增加氨氮的通量;但在有光照条件下,上述微藻和小型底栖动物对沉积物-水体界面间溶解氧和氨氮通量的影响不明显。实验结果为更深入理解泥螺对生态系统的影响提供重要理论支持。     

栉孔扇贝体内寄生的病毒的分离纯化及其形态学观察

栉孔扇贝(Chlamys farreri)是我国传统海水养殖贝种,养殖范围主要分布于黄海、渤海沿岸的山东、河北、辽宁等地,但是,近年来因病害发生,造成了巨大的经济损失.扇贝大规模死亡流行病学调查研究已进行多年,流行病学调查、病理学、病原感染实验及病原分离纯化等研究初步认为类立克次体是扇贝(包括栉孔扇贝和海湾扇贝Argopecten irradians)的病原生物[1～4].在用泛影葡胺密度梯度离心纯化栉孔扇贝类立克次体时发现一种具囊膜的病毒粒子,本文首次报道栉孔扇贝体内该球形病毒纯化的形态学特征.     

Distribution of nitrogenous nutrients and denitrifiers strains in estuarine sediment profiles of the Tanshui River, northern Taiwan

Chemical profiles of both oxidized (nitrate and sulfate) and reduced (ammonium, sulfide, acid-volatile sulfide [AVS], and pyrite) materials and the corresponding distribution of denitrifier microbial communities were measured at low tide in sediments at Guandu in the estuary of the Tanshui River, northern Taiwan in August 2002. Denitrifier strains were isolated for physiological and phylogenic analyses. Based on the distribution of nitrogenous compounds and denitrifier abundances, the vertical profile of Guandu sediments could be separated into four layers: a mixed layer (the top 1 cm of depth, respectively containing 0.82–2.37 and 535.9–475.0 μM of nitrate and ammonium), a nitrate-concentrated layer (1–5 cm in depth, 2.37–0.53 and 475.0–1192.1 μM, respectively), a denitrifier-aggregation layer (5–7 cm in depth, 0.53–0.72 and 1192.1–1430.1 μM, respectively), and an ammonium-enriched layer (7–12 cm in depth, 0.72–0.78 and 1430.1–2196.6 μM, respectively). Denitrifier strains were detected in all layers except for the mixed layer. A variety of metabolic processes by these strains may occur in different layers. Bacillus jeotgali-, Bacillus sphaericus-, and Bacillus firmus-related strains isolated from the nitrate-concentrated layer may be involved in the nitrification-denitrification coupling process due to the relatively low nitrate concentrations (maximum = 2.37 μM), and may contribute to denitrification not nitrification. Bacillus bataviensis- and B. jeotgali-related strains isolated from the denitrifier-aggregation layer comprised the predominant denitrifier population (3.64 × 10⁴ cells/g of denitrifier abundance). They possess the ability of dissimilatory nitrate reduction to ammonium (DNRA). Bacillus jeotgali-related strains and two newly identified strains of GD0705 and GD0706 isolated from the ammonium-enriched layer possibly use fermentative processes as the main metabolic pathway instead of denitrification when nitrate is scarce, and this further supports the high ammonium concentrations (up to 2.20 mM) found in the Guandu sediments. In addition, spore formation also enhances the chance of survival of these strains in the face with such a nitrate-deficient environment.     

在流水系统中浒苔对营养盐吸收和转化的模拟实验

在实验室内选用不进行营养盐加富的流水系统模拟研究了浒苔（Ulva prolifera）对海水营养盐的吸收,测定了浒苔体内不同形态的碳、氮、磷的含量变化,探讨了浒苔对海水中碳、氮、磷3种生源要素的转化作用。结果显示,流动海水的实验体系是一个能给藻体创造稳定环境的模拟装置,可以不断地给藻体更新海水和补充营养盐。浒苔对海水中溶解无机氮（DIN）、溶解有机氮（DON）、溶解无机磷（DIP）和溶解有机磷（DOP）都有吸收作用,其平均吸收速率分别为10.87 μmol·g^-1·d^-1、2.41 μmol·g^-1·d^-1、0.183 μmol·g^-1·d^-1和0.023 μmol·g^-1·d^-1。光照时段的DTN和DTP的平均吸收速率与无光照时段的平均吸收速率没有显著差异（P>0.05）。浒苔对海水中无机形态的营养盐的吸收量高于对该元素的有机形态的吸收量;而浒苔的体内有机形态的成分均远高于无机形态的成分。浒苔对无机形态的氮、磷和碳转化为自身有机形态的转化效率分别为：97.33%,99.99%和96.84%。以上结果表明,浒苔能快速吸收无机形态的生源要素并转化为有机形态,能够加快生源物质进入生态系统的物质循环。     

异养蛋白核小球藻净化对虾池塘养殖尾水的实验

为研究异养培养的蛋白核小球藻(Chlorellapyrenoidosa)去除对虾池塘养殖尾水氮、磷营养盐的效果,本实验通过对异养藻种的光自养转换、盐度驯化,并以不同初始密度接种入养殖尾水,定期检测水体中氨氮、磷酸盐、硝酸盐氮和亚硝酸盐氮等指标,分析其对营养盐的吸收利用规律。结果表明:光照强度6 600 lx、光照周期16 h∶8 h(L/D)、温度25±0.5℃条件下,接种细胞密度为1.0×105个/mL、5.0×105个/mL、1.0×106个/mL的蛋白核小球藻均在接种初期快速进入指数生长期, 10 d后进入生长平台期, 20 d藻细胞生物量分别增加39.25、7.98和4.07倍;蛋白核小球藻能明显降低养殖水体中氮、磷营养盐浓度,起到去氮除磷的净化作用,磷酸盐的去除率随藻细胞的生长持续上升, 21 d去除率分别为58.8%±0.72%、72.9%±1.7%、81.4%±9.86%;对氮盐的利用规律依次为氨氮、硝酸氮和亚硝酸氮,氨氮6 d的去除率达81.9%±6.0%, 96.2%±1.16%, 95.4%±1.24%,硝酸氮21d去除率达82%±1.35%、93.3%±4.41%...     

2013年夏季琼东海域营养盐与叶绿素a的周日波动及其影响因素

基于2013年8月琼东海域两个连续站的观测资料,对比分析了近岸站位和陆架站位的营养盐和叶绿素a浓度周日变化特征及其影响因素。结果表明:在垂直分布上,近岸站位S1与陆架站位S2的温跃层减弱了营养盐的向上输运;在时间变化上,S1站底层硅酸盐、硝酸盐和磷酸盐浓度具有半日波动的特点,而S2站的营养盐则不具有周日波动的现象,这说明近岸站位的营养盐受到潮汐作用的影响更显著。S1站的叶绿素a分层不明显,夜间的低值可能体现浮游动物的摄食作用,S2站位的叶绿素a分层明显,夜间没体现浮游动物的摄食作用。总体上,琼东海域近岸站位S1和陆架区站位S2叶绿素a和营养盐周日波动都受到温跃层、潮汐、生物作用和光照的影响,但S1站受潮汐作用影响更显著,且S1站叶绿素a浓度还受到浮游动物摄食作用影响。由于叶绿素a和营养盐受到多种环境要素的影响,使得两者相关性不显著。     

浒苔多糖的分级纯化及保湿活性研究

用DEAE-Sepharose FF离子交换凝胶层析柱对浒苔(Enteromorpha)多糖进行分级纯化,利用HPLC色谱分析了单糖组成,并以甘油和透明质酸为对照,研究其吸湿和保湿活性。结果表明,浒苔多糖主要为葡萄糖醛酸-鼠李糖-木糖-甘露糖聚合物;浒苔多糖主要由两种组分组成,其一组分木糖和鼠李糖含量摩尔比为1.11:1,另一组分木糖和鼠李糖含量比为1:3.28;浒苔多糖具有与透明质酸相似的吸湿和保湿能力。     

刚毛藻对静态模拟刺参养殖池塘上覆水营养盐含量的影响

利用室内装置模拟的刺参养殖池塘上覆水,在温度为25℃,光照为(4000±200)lx条件下静态培养刚毛藻48 h,定时测定上覆水中总氮(TN)、氨态氮(NH_4~+-N)、硝态氮(NO_3~–-N)、亚硝态氮(NO_2~–-N)、总磷(TP)、活性磷(PO_4~(3–)-P)等营养盐含量,分析了氮磷营养盐含量的动态变化规律,探讨了刚毛藻生物量对沉积物-水界面营养盐含量变化的影响。结果表明,刚毛藻生物量对各营养盐含量的影响存在显著性差异(P0.05)。随着培养时间延长,上覆水中营养盐含量均呈现先增大后减小的趋势。上覆水中各营养盐含量在培养6～24 h时分别达到最大值,其中当刚毛藻生物量为0.5～4.5 g/L时,TN、NO_3~–-N、NO_2~–-N、NH_4~+-N、TP、PO_4~(3–)-P含量达到最大值。继续培养,上覆水中各营养盐的含量逐渐减小,其中生物量为8.5 g/L时刚毛藻对NO_3~–-N、NO_2~–-N的吸收效果最好;生物量为0.5 g/L时刚毛藻对NH_4~+-N、PO_4~(3–)-P吸收效果最好。因此,高生物量(8.5 g/L)刚毛藻可降低上覆水中NO_3~–-N、NO_2~–-N含量,但低生物量(0.5 g/L)刚毛藻能有效吸收上覆水中的NH_4~+-N、PO_4~(3–)-P。     

Effects of temperature and organic and inorganic nutrients on the growth of Chattonella marina (Raphidophyceae) from the Daya Bay, South China Sea

The effects of temperature and different forms of nutrients on Chattonella marina growth have been investigated in strains isolated from the Daya Bay,the South China Sea.The strain of C.marina preferred high temperatures, with an optimal temperature of 25°C,and 18 °C was the minimum for its survival.Higher cell number and growth rate were obtained in high nitrogen and phosphorus concentrations (500 μ g/L, 74 μ g/L) than under nutrient limitation.Nitrogen influenced the growth most, as the specific growth rate and maximum cell density were lower in nitrogen-limited cultures than noted under phosphorus limitation or under limitation from both.C.marina was capable of using many kinds of organic nitrogen sources including L-serine (L-Ser),glycine (Gly),alanine (Ala),L-threonine (L-Thr), glutamic acid (Glu) and urea, but could not utilize uric acid.Various forms of organic phosphorus compound such as glucose-6-phosphate (G6P),sodium glycerophosphate (GYP),adenosine triphosphate (ATP),adenosine monophosphate (AMP),cytidine monophosphate (CMP),guanosine monophosphate (GMP),uridine monophosphate (UMP),4-nitrophenylphosphate (NPP) and triethyl phosphate (TEP) supported the growth as well. Algal cells had the ability to sustain growth under nitrogen-and/or phosphorus-free conditions particularly under phosphorus depleted condition.These results led to the hypothesis that high loading of nitrogen has played an important role in frequent C.marina blooms in the past decade,and its capability for utilization of diverse forms of organic nutrients and growth in low nutrient conditions make this species a likely recurrent dominant in the Daya Bay phytoplankton assemblages,visible as     

水母(沙海蜇)降解与营养盐(氮、磷)释放的模拟与实证

The aim of this study was to investigate nitrogen and phosphorus released in the process of the decomposition of giant jellyfish in the laboratory and found the evidence to verify the influence of nutrients released by the decomposition of jellyfish on the ecosystem in the field. The release of nitrogen and phosphorus from the decomposition of Nemopilema nomurai was examined in a series of experiments under different incubation conditions such as different p H values, salinity values, temperatures and nitrogen and phosphorus concentrations. The results showed that the complete decomposition of Nemopilema nomurai generally took about 4–8 d. The release of nitrogen and phosphorus from the decomposition of Nemopilema nomurai could be divided into two stages: the early stage and the later stage, although the efflux rate of nitrogen was one order more than phosphorus. In the early stage of the decomposition of Nemopilema nomurai, the concentrations of dissolved nitrogen, dissolved phosphorus, total nitrogen and total phosphorus in seawater increased rapidly, and the concentration of nitrogen could reach the highest level in the whole degradation process. In the later stage of the decomposition, the concentrations of dissolved nitrogen and total nitrogen declined slowly, while the concentration of phosphorus in water could reach a maximum in the degradation process. High p H, low salinity,high temperature and N/P will promote the release of nitrogen; low p H is unfavorable to the release of nitrogen but favorable to the release of phosphorus. In addition, we found the concentrations of ammonium and phosphate in the bottom water were higher than those in the surface water during the period of jellyfish bloom in the Jiaozhou Bay, proving that nutrients released by the decomposition of jellyfish have significant influence on nitrogen and phosphorus in the field. For the whole Yellow Sea, nutrients released by jellyfish carcasses may reach up to(2.63±2.98)×107 mol/d of dissolved nitrogen(DN) and(0.74±0.84)×106 mol/d of dissolved phosphorus(DP) during the period of jellyfish bloom. The values are comparable to riverine inputs in a day, but much higher than sediment–water exchange flux in the Yellow Sea. The great amounts of nutrients must have significant influence on the nutrients balance of the Yellow Sea during the period of jellyfish dead and decomposition. Both the experimental data and field observations proved that the decomposition of jellyfish may release a great amount of nutrient to the surrounding environment during the period of jellyfish decomposition.