中国沿海尖刺拟菱形藻的种下分类学研究*

为了澄清中国沿海尖刺拟菱形藻Pseudo-nitzschia pungens Hasle的种下分类学现状, 从我国沿海分离并建立了32个尖刺拟菱形藻的单克隆培养株系, 结合光学显微镜下的群体特征和透射电镜下的超微形态学特征, 以及基于核糖体转录间隔区(ITS)的分子系统学数据, 鉴定得到尖刺拟菱形藻的两个变种: 尖刺拟菱形藻原变种P. pungens var. pungens和尖刺拟菱形藻阿维罗变种P. pungens var. averiensis。文章对其形态学特征进行了详细描述, 并利用高效液相色谱-质谱联用法(LC-MS/MS)对培养株系的多莫酸(DA)进行了检测, 但未检测到DA。     

核酸指标对桡足类安氏伪镖水蚤摄食率、生长率及产卵率的指示作用

核酸指标可表征桡足类营养及生长状态, 然而多物种和多指标比较研究的相对缺乏却限制了现场海区调查中对核酸标志物的广泛应用。本研究测定了系列饵料浓度(0.2~2.0μg C·mL^-1)喂食的安氏伪镖水蚤Pseudodiaptomus annandalei的桡足幼体Ⅱ期个体的摄食率、生长率及成体产卵率, 并就生化组成, 包括RNA含量、DNA含量、蛋白含量(以下简写为PRO)、RNA:DNA、RNA:PRO、DNA:PRO及元素组成(N含量、P含量、C:N 和C:P比值)进行定量分析和计算。摄食率、生长率、产卵率及桡足幼体的RNA相关的核酸指标随饵料浓度的变化趋势均符合双曲线模型, 而雌性成体中仅RNA:DNA比值符合此模型。对RNA相关指标与各生理参数的相关性分析显示, 桡足幼体的RNA相关指标均与摄食率及生长率呈显著线性相关性, 显著程度的顺序为RNA:DNA>RNA:PRO>RNA含量; 雌性成体中RNA:DNA及RNA:PRO比值与产卵率显著相关, 前者的相关性远较后者显著。此结果表明, 在常用的现场浮游桡足类调查的标志比值(RNA:DNA比值或RNA含量)之外, RNA:PRO比值亦可作为指示桡足类生长状态的标志比值, 但是当指示体生长时, 桡足类必须处于同一生长阶段。RNA相关指标与饵料浓度之间稳固的双曲线相关性提示我们, RNA相关指标可帮助评估桡足类的饵料环境。     

一个新的来自于弧菌QD-5并具有Poly-G内切功能的褐藻胶裂解酶的克隆和性质表征

Seven bacterial clones with alginate-utilizing activity were isolated from rotten kelp. By activity test, the Vibrio sp.QD-5 with the potential alginate-degrading capability was chosen to carry out the draft genome sequencing, and the result showed that the Vibrio sp. QD-5 containing an alginate lyase gene cluster. One of these genes, aly-IV,was cloned and characterized for the first time. After overexpression, Aly-IV, with a molecular mass of about62 kDa and a theoretical isoelectric point(pI) of 5.12, was purified to a specific activity of 1 256.78 U/mg and showed highest activity at 35°C in the Tris-HCl buffer at pH of 8.9. Moreover, the enzyme activity was enhanced by the metal ions of Na~+, K~+ and Mg~(2+) under certain concentration. Aly-IV degraded favorably polyG blocks in an endo-type, yielding monomer and dimer as the main products. Due to its high substrate specificity, Aly-IV could be used as a potential tool for production of polyG oligosaccharides with low degree of polymerization(DP) and for determining the fine structure of alginate.     

重金属锌对海洋微藻藻源氨基酸产出的影响

锌是海洋微藻生长必需的金属元素,低浓度时可促进微藻的生长,而在高浓度时则表现为抑制其生长。受人类活动的影响,海洋中锌有富集增加的趋势,从而会对海洋微藻的生长产生影响。本论文探究了锌离子以及纳米氧化锌对硅藻中肋骨条藻(Skeletonemacostatum)和新月菱形藻(Nitzschia closterium)生长的影响,特别是对藻源氨基酸产生的影响,结果发现:纳米金属颗粒会通过释放离子的形式抑制海洋浮游植物的生长,中肋骨条藻比新月菱形藻对锌的响应更敏感。锌离子和纳米氧化锌可通过抑制浮游植物的生长来减少氨基酸的释放量和改变藻源氨基酸的种类,从而可能会对海洋氮循环过程产生影响。     

几种海洋微藻的碱性磷酸酶性质初步研究

对5种海洋微藻产生的碱性磷酸酶的性质进行了初步研究并确定了碱性磷酸酶的测定条件。结果表明,在pH 8.2的环境中,各藻产生的碱性磷酸酶最佳反应温度在40~50℃内,且存在一定差异;40℃下酶促反应恒速时间及米氏常数也有所不同,酶活恒速时间顺序为东海原甲藻(Prorocentrum donghaiense)<强壮前沟藻(Amphidinium carterae)<旋链角毛藻(Chaetoceros curvisetus)<中肋骨条藻(Skeletonema costatum)<塔玛亚历山大藻(Al-exandrium tamarense);米氏常数大小顺序为塔玛亚历山大藻<东海原甲藻<中肋骨条藻<强壮前沟藻<旋链角毛藻。碱性磷酸酶测定条件为:温度40℃,反应时间90 min,底物浓度260μmol/L。     

矿物培养影响微生物群落的ERIC研究

从江西东乡铜矿、江西银山铅锌矿和广东云浮硫铁矿三地采集了酸性矿坑水样品,用ERIC方法(肠细菌重复内源同源序列方法)分析了经不同矿物培养后的微生物变化情况。结果发现:不同矿山酸性矿坑水样品的化学性质及其微生物组成存在很大差异,同一样品经不同矿粉培养后微生物群落也会发生比较大的变化;样品经组成相似的能源物质培养后,其微生物群落结构也越相似。确定了ERIC方法用于酸性矿坑水样品时的最佳扩增体系和反应程序。     

Effects of Wastewater-borne Heavy Metals on Mangrove Plants and Soil Microbial Activities

Effects of wastewater-borne heavy metals on growth of young plants (9-month-old Bruguiera gymnorrhiza) and soil microbial activities in mangrove microcosms were evaluated. During the 26-week loading period, each mangrove microcosm received 31.2 litres synthetic wastewater of three strengths: normal, medium (5 times of normal strength) and strong (10 times of normal strength). Normal strength wastewater had Cu, Zn, Cd, Cr and Ni concentrations of 3, 5, 0.2, 2 and 3 mg l⁻¹, respectively. Plant growth and total plant biomass in wastewater-treated microcosms were lower than that in the control, and the maximum reduction was found in microcosms receiving strong wastewater. Alkaline phosphatase activity and ATP contents of the mangrove soils receiving wastewater were also reduced. More than 95% reduction in these two parameters was found in soils loaded with strong wastewater. Microtox test demonstrated that soil elutriates obtained from microcosms receiving strong wastewater were of the greatest toxicity (EC₅₀ was 23%). These results show that high concentrations of heavy metals present in strong wastewater were toxic and posed negative effects to both mangrove plants and soil microbial activities. Microbial activities were generally more sensitive to the toxicity of heavy metals than plants.     

鱿鱼营养成分分析

对黑鱿与阿根廷鱿的营养成分作了分析,并评价了它们的营养价值。研究表明：两种鱿所含营养成分大致接近。鱿的蛋白质含量高达７３．３％～７８．８％,而脂肪含量小于４．２０％,这有利于鱿鱼干制品的加工与贮存。两种鱿的氨基酸含量比较丰富,其中谷氨酸、天冬氨酸含量特别高。阿根廷鱿氨基酸含量更为丰富,其亮氨酸稍高于鸡蛋白     

3种海洋养殖贝类体内的脂肪酸组成

分析了华贵栉孔扇贝Ｃｈｌａｍｙｓｎｏｂｉｌｉｓ（Ｒｅｅｖｅ）、合浦珠母贝Ｐｉｎｃｔａｄａｍａｔｅｎｓｉｉ（Ｄｕｕｋｅｒ）、翡翠贻贝Ｐｅｒｎａｖｉｒｌｄｉｓ（Ｌｉｎｎａｅｕｓ）的软体部、肝胰腺和生殖腺的脂肪酸组成及含量。３种贝的各组织中,不饱和脂肪酸含量均较饱和脂肪酸含量高。多不饱和脂肪酸（ＰＵＦＡ）含量丰富,其中二十碳五烯酸（ＥＰＡ）和二十二碳六烯酸（ＤＨＡ）含量相当高,占总脂肪的２５．２６％－２７．２１％。３种贝中,翡翠贻贝的油含量最高（１３．６８％）,软体部的ＥＰＡ和ＤＨＡ含量也较高（２６．９５％）,最具开发利用价值。     

中国对虾(Penaeus chinensis)的白点综合征病毒(WSSV)的提纯和核酸提取

作者从发生白点综合征的中国对虾中,提取了一株白点综合征病毒,用电镜对纯化的病毒进行了形态学观察,并对病毒纯化过程中蛋白酶抑制剂的使用及离心速度的选择进行了探讨。经蔗糖密度梯度离心,分取各带负染后电镜观察,病毒条带位于４０％～５０％蔗糖梯度之间,其完整毒粒长２２５～２７０ｎｍ,直径７５～８８ｎｍ,有的完整毒粒带有很长的尾,病毒衣壳长２５０～３２０ｎｍ,直径６０～８０ｎｍ。在匀浆使用的缓冲液中加入蛋白酶抑制剂ＰＭＳＦ,可得到完整的带包膜的病毒颗粒,比不使用ＰＭＳＦ的提纯效果要好。经３０００ｒ／ｍｉｎ离心５ｍｉｎ,５０００ｒ／ｍｉｎ离心２５ｍｉｎ,去除大部分的组织碎片,经１００００ｒ／ｍｉｎ离心１ｈ,可使大部分病毒沉淀。并采用传统的方法对病毒核酸进行了提取,其基因组经初步分析为ｄｓＤＮＡ。