Effects of Two Potential Allelochemicals on the Photosystem II of Nitzschia closterium and Monostroma nitidum

In aquaculture,high-density seaweed farming brings higher economic benefits but also increases outbreaks of diatom felt.The effective control of diatom felt in high-density seaweed farming has always been a research hotspot.This study selected two potential allelochemicals 2-hydroxycinnamic acid and quinic acid to explore their effects on a diatom Nitzschia closterium and an economic seaweed Monostroma nitidum.The results showed that 2-hydroxycinnamic acid had better inhibitory effects than quinic acid on the growth,pigment content and photosynthetic efficiency of N.closterium.Their half-maximal inhibitory concentrations at 120 h(IC_(50–120 h))were 0.9000 and 1.278 mM,respectively.Additionally,these allelochemicals had limited inhibitory effects on the growth,pigment content and photosynthetic efficiency of M.nitidum before 24 h.To further explore the allelopathic effect of these chemicals,this study focused on the photosystem II energy fluxes of N.closterium.It was found that 3 mM 2-hydroxycinnamic acid could destroy the whole photosynthetic system by devastating the PSII reaction centre(RC)before 24 h;however,the same concentration of quinic acid could only down-regulate the electron transport efficiency by changing the effective antenna size of an active RC and downregulating the PSII reaction centre density.These experimental results are expected to provide a new strategy to control diatom felt blooms on the high-density seaweed farming areas.

Pb^(2+)对淡水硅藻谷皮菱形藻(Nitzschia palea)生长的影响

通过在自制培养基中加入不同浓度的Pb2+,研究水环境中重金属Pb2+污染对谷皮菱形藻生长和叶绿素a含量的影响,并在光学显微镜下观察细胞形态变化,研究该藻对铅离子(Pb2+)的耐受性。试验结果表明,谷皮菱形藻对Pb2+的耐受性较强。Pb2+浓度低于5 mg/L,促进其生长;Pb2+浓度为10 mg/L时,对其无明显影响;Pb2+浓度大于50 mg/L时,明显抑制其生长,且藻细胞出现异常现象;96 h EC50值为27.26 mg/L。Pb2+浓度小于10 mg/L时,藻叶绿素a含量均较高,与正常生长情况相似;Pb2+浓度大于50 mg/L时,叶绿素a含量明显降低。Pb2+浓度较低时,藻细胞体形态正常,且运动能力很强,Pb2+浓度大于50 mg/L时,藻细胞体出现畸形,颜色加深,两端由圆形变成方形,且运动能力很弱。

Cd^(2+)对淡水底栖硅藻Nitzschia amplectens生长的影响

对不同浓度Cd2+中淡水底栖硅藻Nitzschia amplectens的存活数、叶绿素a含量及运动速度进行测定,以探索硅藻对重金属毒性的响应规律,为硅藻监测水体污染提供科学依据。纯化培养Nitzschia amplectens,生长数量达到7万个/m L时,加入不同浓度的Cd2+进行急性毒性和亚急性毒性胁迫培养。在急性毒性试验中,Nitzschia amplectens的存活数在96 h的EC50值为4.989 mg/L。在亚急性毒性试验中,培养15 d后Cd2+浓度为0.625、0 mg/L的Nitzschia amplectens运动平均速度为2.5、5.56μm/s,其余试验组无运动迹象;培养13 d后,Cd2+浓度与叶绿素a含量呈负相关(P<0.05)。Cd2+浓度对Nitzschia amplectens的生长、运动速度和叶绿素a含量有明显影响,所以Nitzschia amplectens具监测水体重金属Cd2+污染潜力。

长菱形藻(Nitzschia longissima)和新月细柱藻(Cylindrotheca closterium)分子分类研究

本研究自胶州湾分离了两种底栖硅藻,形态学初步鉴定为长菱形藻和新月细柱藻。对其18S rDNA和rbcL基因进行了测序并用邻接法构建了系统树。结果表明,两藻在18S rD-NA和rbcL基因序列上均存在较大的差异,基于DNA序列的分类结果与形态学分类结果一致。在依据形态指标难以确定藻类分类地位的情况下,18S rDNA和rbcL基因序列是有用的鉴定工具。

福氏拟菱形藻(Pseudo-nitzschia fukuyoi)——中国产毒拟菱形藻的新记录

为了提高对我国海域拟菱形藻属(Pseudo-nitzschia)物种多样性的认识,并明确其是否能够产生多莫酸(domoic acid,DA)。本文从中国沿海分离和建立了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、透射电镜下的超微形态学特征、基于核糖体转录间隔区(internal transcribed spacer,ITS)的分子系统学数据,以及基于ITS2转录RNA的二级结构分析,鉴定到我国拟菱形藻属的1个新记录种:福氏拟菱形藻(P.fukuyoi Lim,Teng,Leaw&Lim)。本文对其形态学特征进行描述,与相似物种进行了对比分析;对其ITS2-RNA序列进行了分析;利用高效液相色谱-串联质谱联用法(liquid chromatography tandem mass spectrometry,LC-MS/MS)对培养藻株的产毒特征进行了分析,多个株系中均检测到DA,单细胞产毒水平为0.02—0.23pg,这是我国报道的第二种产毒拟菱形藻。利用卤虫(Artemia salina)和褶皱臂尾轮虫(Brachionus plicatilis)对福氏拟菱形藻的产毒水平进行了诱导,发现浮游动物能够提高福氏拟菱形藻的DA含量,增强幅度在4.7—28.5倍之间。本文提高了对我国拟菱形藻属物种多样性的认识,明确了福氏拟菱形藻的产毒特征和水平,为后续深入研究提供了基础数据。

Photo-induced transformations of Hg(Ⅱ) species in the presence of Nitzschia hantzschiana,ferric ion,and humic acid

Effects of algae Nitzschia hantzschiana, Fe（Ⅲ） ions, humic acid, and pH on the photochemical reduction of Hg（Ⅱ） using the irradiation of metal halide lamps （λ〉 365 nm, 250 W） were investigated. The photoreduction rate of Hg（Ⅱ） was found to increase with increasing concentrations of algae, Fe（Ⅲ） ions, and humic acid. Alteration of pH value affected the photoreduction of Hg（Ⅱ） in aqueous solution with or without algae. The photoreduction rate of Hg（Ⅱ） decreased with increasing initial Hg（Ⅱ） concentration in aqueous solution in the presence of algae. The photochemical kinetics of initial Hg（Ⅱ） and algae concentrations on the photoreduction of Hg（Ⅱ） were studied at pH 7.0. The study on the total Hg mass balance in terms of photochemical process revealed that more than 42% of Hg（Ⅱ） from the algal suspension was reduced to volatile metallic Hg under the conditions investigated.

固定化菱形藻(Nitzschia sp.)的生长及营养成分变化

选用底栖硅藻菱形藻(Nitzschiasp.)为实验对象,分别研究了CaCl2浓度、胶珠直径、接种密度及胶珠密度对其生长的影响及后两因素对其主要营养成分含量变化的影响。结果表明:(1)随培养时间的延长,CaCl2浓度对菱形藻生长有极显著影响,在2%CaCl2溶液中形成的胶珠里菱形藻比生长速率最高;(2)胶珠直径对菱形藻在初期和指数期的生长影响显著,且较小直径(2.5～3.0mm)的胶珠较适宜菱形藻生长;(3)随接种密度和胶珠密度的提高,菱形藻的比生长速率都有先增高后下降的变化趋势,接种密度较高(2.50×104～5.50×104mL-1)时,固定化组的比生长速率是对照组的2倍左右。胶珠密度为220粒/(100mL培养液)左右较适宜(胶珠直径3.0mm);(4)初始接种密度和胶珠密度试验中,蛋白质、总糖及粗脂肪的含量固定化组低于或相当于对照组,而叶绿素的含量固定化组高于对照组。

Photodegradation of 17α-Ethynylestradiol in Aqueous Solution with Nitzschiahantzschiana or Chlorella vulgaris and Fe^(3+)

The photodegradation of 17α-ethynylestradiol (EE2) induced by highpressure mercury lamp (λ≥313 nm, 250 W) in aqueous solution with algae (e. g.Nitzschia hantzschiana andChlorella vulgaris) and Fe3+ was ineestigated initially. The affecting factors on the photodegradation were studied and described in details, such as algae concentration, Fe3+, exposure time, and so on. The concentration of EE2 in distilled water was determined using fluorescence spectrophotometer. The photodegradation of EE2 in aqueous solution exposed to 250 W high-pressure mercury lamp was evident in the presence of algae and Fe3+. With the algae concentration increasing, photodegradation rate increased. Fe3+ could accelerate the photocatalytic degradation of EE2 in aqueous solution with algae. The colloidal ferric hydroxide particles that might have absorbed on the algae cells could enhance the photocatalytic degradation of EE2 by algae. The catalysis in photocatalytic degradation reaction mainly resulted from the active oxygen (H2O2,1O2 and ·OH) that was caused by algae and Fe3+ under 250 W HPML. In this paper, the mechanism of photocatalytic degradation of EE2 by algae and Fe3+ is discussed theoretically in details. Key words 17α-ethynylestradiol - photodegradation - high-pressure mercury lamp - Nitzschia hantzschiana - Chlorella vulgaris - Fe3+ CLC number X 131. 2 Foundation item: Supported by the Scientific Research Foundation of Wuhan Environmental Protection Bureau and the National Natural Science Foundation of China (20177017)Biography: Liu Xian-li (1965-), male, Ph. D candidate, Associate professor, research direction: environmental chemistry

新月菱形藻(Nitzschia closterium)制备参数对藻球性质及去氮磷效果的影响

为了制备出高效的固定化藻球,完善固定化藻球的制备工艺,以新月菱形藻为藻种,采用褐藻胶包埋技术和单因子试验,研究了不同褐藻酸钠质量分数(2.0%、2.5%、3.0%)和不同溶剂(海水、蒸馏水、NaCl溶液)对藻球形状、强度、传质性、去除氮磷效果及藻细胞生长的影响.结果表明:不同褐藻酸钠质量分数对藻球形状影响显著(P<0.05),褐藻酸钠质量分数为2.0%和2.5%时藻球呈球形,为3.0%时藻球呈不规则的水滴形;不同溶剂种类对藻球形状则无影响.不同褐藻酸钠质量分数对藻球强度影响显著(P<0.05),随着褐藻酸钠浓度的增加,藻球强度逐渐增强;不同溶剂对藻球强度的影响显著(P<0.05),影响大小的排序为海水>NaCl>蒸馏水.不同褐藻酸钠质量分数和不同溶剂对藻球传质性影响显著(P<0.05),随着褐藻酸钠浓度的增加,藻球传质性也逐渐增强;溶剂对传质性影响的强弱排序为海水>NaCl>蒸馏水;不同褐藻酸钠质量分数和不同溶剂对固定化藻球藻细胞生长及去氮磷效率影响显著(P<0.05),其中以3.0%褐藻酸钠+海水的藻细胞生长最快,去除氮磷效率最佳.

Physiological Characteristics of Nitzschia hantzschia in Response to Nitrobenzene Stress

This study was conducted to investigate the effects of different mass concentrations of nitrobeneze on the growth, soluble sugar content, soluble protein content, chlorophyll a content and algal cell conductivity of Nitzschia hantzschia. The results showed that as the concentration of nitrobenzene increased, the growth of N. hantzschia was inhibited, and the algal culture liquids gradually changed from dark yellow to light yellow after 5 d of treatment;the soluble sugar content increased after 2 d;and the soluble protein content of the 100 mg/L nitrobenzene treatment group was 89.1% of the control group on day 1, which was the lowest value, and then showed a gradual upward trend. The low-mass concentration of nitrobenzene promoted the chlorophyll a content of algal cells, the medium and high mass concentrations had an inhibitory effect, and the chlorophyll a content of the 50 mg/L treatment gradually recovered after 3 d. The electrical conductivity of algal cells gradually increased with the increase of the mass concentration of nitrobenzene. The electrical conductivity gradually recovered after 3 d of the low mass concentration treatment, while the high mass concentration harmed the algae cells, causing N. hantzschia to gradually die.

Effect of NaNO_3 Concentrations on the Growth and Fatty Acid Compositions of Nitzschia closterium and Chaetoceros gracilis*

Fatty acid compositions of two strains of marine diatoms Nitzschia closterium ( MACC B222 ) and Chaetoceros gracilis ( MACC B13 ) have been examined. The microalgae have been grown at different initial NaNO3 concentrations (75,375, 750, 1125,1 500, 1 875 mg/L ) and harvested at the late exponential phase. The results by one factor analysis of variance show that the NaNO3 concentrations have no significant influence on the relative growth rate of two strains of marine diatoms; the influence of NaNO3 concentrations on the fatty acids differs from species to species. The major fatty acids of two diatom strains are 14:0, 16:0, 16:ln- 7 and 20:5n- 3 ( EPA ) , B13 also has high percentage of20:4n-6 (6.0～9.1%).

Analysis of phospholipids in microalga Nitzschia closterium by UPLC-Q-TOF-MS

Precise structural identification of phospholipids in the microalga Nitzschia closterium has been established using ultra performance liquid chromatography-electrospray ionization-quadrupole-time of flight-mass spectrometry (UPLC-ESI-Q-TOF-MS) for direct analysis of total lipid extracts.Mass spectrometry was performed in reflective time-of-flight using electron spraying ionization in negative mode.Phospholipid molecular species identification was based on the characteristic product ions and neutral loss yielded by different phospholipids under ESI-MS/MS mode.The molecular species were confirmed by the carboxylate anions produced by phospholipids in negative mode;the regiospecificity of the two acyl chains was determined from the ratio of sn-1 to sn-2 carboxylate anion abundances.As a result,18 lipid molecular species were identified for the first time in this microalga,comprising seven phosphatidylcholines (PC),two phosphatidylethanolamines (PE),two phosphatidylinositols (PI),and seven phosphatidylglycerols (PG).Lipid standards of PC,PE,PI,and PG were added to the total lipids as internal standards for semiquantitative analysis,revealing concentrations of phospholipids in this species between 0.09 and 3.37 nmol/mg.This method can produce a full structural profile of intact phospholipid molecular species and can be used for study of the physiological and ecological functions of lipids by monitoring their individual changes over time.

Morphological and toxicity characteristics of Pseudo-nitzschia pungens strain PP0201-01 isolated from the East China Sea

The morphology and toxicity of Pseudo-nitzschia strain PP0201-01, isolated near the estuary of Changjiang (Yangtze) River in Year 2002 and tentatively identified as Pseudo-nitzschia pungens, were studied. The strain was identified morphologically as P. pungens using scanning electronic microscope (SEM). The ana- lytical results of LC-PDA (liquid chromatography with photodiode array detector), LC-MS (liquid chromatogra- phy with mass detector), and the toxicity data from mouse bioassay, showed that no domoic acid was produced by the strain. However, the mechanism of domoic acid production by Pseudo-nitzschia species is complicated. More works are needed to assess the toxic species and to evaluate the potential risk of amnesic shellfish poison- ing (ASP) in China.

Effects of Nutritional Factors on the Growth and Heterotrophic Eicosapentaenoic Acid Production of Diatom Nitzschia laevis

The effects of several nutritional factors on the growth and eicosapentaenoic acid(EPA) production of diatom Nitzschia laevis were studied. 4 LDM(quadrupled concentration of the nutrient salt) was the optimal concentration of nutrient salt for the growth and EPA production of N. laevis. The growth of N. laevis was inhibited when the glucose concentration was either lower than 10 gL-1 or higher than 15 gL-1. Both sodium nitrate and urea were good nitrogen sources for the growth and EPA production,while ammonium chloride seriously decreased the dry cell weight(DW) and the EPA content. Silicate seriously influenced the growth of N. laevis. The maximum DW of 2.34 gL-1 was obtained in the presence of 150 mgL-1 Na2SiO3· 9H2O. The EPA content remained almost the same when the silicate concentration was lower than 150 mgL-1;however,higher silicate concentrations resulted in a steady de-crease of EPA content. Low medium salinity(≤29) did not seem to influence the DW of N. laevis,and high salinity resulted in a decrease of DW. The highest EPA content(4.08%) and yield(110 mgL-1) were observed at the salinity of 36 and 29,respectively.

Preparation and characteristics of biosilica derived from marine diatom biomass of Nitzschia closterium and Thalassiosira

In this study, biosilica of high purity was successfully prepared from marine diatom( Nitzschia closterium and Thalassiosira) biomass using an optimized novel method with acid washing treatment followed by thermal treatment of the biomass. The optimal condition of the method was 2% diluted HCl washing and baking at 600°C. The SiO_2 contents of N. closterium biosilica and Thalassiosira biosilica were 92.23% and 91.52%, respectively, which were both higher than that of diatomite biosilica. The SiO_2 morphologies of both biosilica are typical amorphous silica. Besides, N. closterium biosilica possessed micropores and fibers with a surface area of 59.81 m^2/g. And Thalassiosira biosilica possessed a mesoporous hierarchical skeleton with a surface area of 9.91 m^2/g. The results suggest that the biosilica samples obtained in this study present highly porous structures. The prepared porous biosilica material possesses great potential to be used as drug delivery carrier, biosensor, biocatalyst as well as adsorbent in the future.

Size-Dependent Growth Rate of Nitzschia closterium at Different Concentrations of Petroleum Hydrocarbon

The maximum growth rate of Nitzschia closterium at various concentrations of petroleum hydrocarbon was studied. The influence of petroleum hydrocarbon on cell size as a function of concentration was discussed. The relationship between maximum growth rate of Nitzschia closterium and cell median equivalent spherical diameter (MESD) was also carefully studied. The experimental results showed that the growth rate of Nitzschia closterium was generally suppressed by petroleum hydrocarbon, which had greater effects at both low and high concentrations than at intermediate concentrations. No significant changes in cell size distribution were observed during the growth period of Nitzschia closterium. The Gaussian function could give a clear description of the cell size distribution of Nitzschia closterium, and the MESD value ranged from 2.71 to 6.82 μm. The MESD decreased when the cell was exposed to petroleum hydrocarbon, and the reduction of cell MESD was much more significant at both relatively high and low hydrocarbon concentrations. The presence of petroleum hydrocarbon changed the relationship between μmax and MESD from an allometric function to a U-shaped curve. When the MESD was below 5.07 μm, μmax decreased along with increased MESD, whereas when MESD was above 5.07 μm, μmax increased along with MESD, which deviated from the allometric model.

Effect of nitrogen/phosphorus concentration on algal organic matter generation of the diatom Nitzschia palea:Total indicators and spectroscopic characterization

Critical algal blooms in great lakes increase the level of algal organic matters（AOMs）,significantly altering the composition of natural organic matters（NOMs） in freshwater of lake.This study examined the AOM＇s characteristics of Nitzschia palea（N.palea）,one kind of the predominant diatom and an important biomarker of water quality in the great lakes of China,to investigate the effect of AOMs on the variation of NOMs in lakes and the process of algal energy.Excitation–emission matrix fluorescence（EEM） spectroscopy,synchronous fluorescence（SF） spectroscopy and deconvolution UV–vis（D-UV） spectroscopy were utilized to characterize AOMs to study the effects of nutrient loading on the composition change of AOMs.From results,it was revealed that the phosphorus is the limiting factor for N.palea＇s growth and the generation of both total organic carbon and amino acids but the nitrogen is more important for the generation of carbohydrates and proteins.EEM spectra revealed differences in the composition of extracellular organic matter and intracellular organic matter.Regardless of the nitrogen and phosphorus concentrations,aromatic proteins and soluble microbial products were the main components,but the nitrogen concentration had a significant impact on their composition.The SF spectra were used to study the AOMs for the first time and identified that the protein-like substances were the major component of AOMs,creating as a result of aromatic group condensation.The D-UV spectra showed carboxylic acid and esters were the main functional groups in the EOMs,with –OCH_3,–SO_2NH_2,–CN,–NH_2,–O– and –COCH_3functional groups substituting into benzene rings.

Hierarchical recognition on the taxonomy of Nitzschia closterium f. minutissima

Nitzschia closterium f. minutissima, a marine eukaryotic unicellular diatom, originally classified as Bacillariophyta/Bacillariophyceae/Bacillariales/Bacillariaceae/Nitzschia, is one of the most important feed sources in mariculture. In this study, its morphological features were examined under DIC Microscopy (differential interference contrast microscope); its pigments and fatty acids composition were analyzed by using High Performance Liquid Chromatography (HPLC) and gas chromatography (GC); the complete Actin cDNA, part 18S rDNA, complete ITS1 and ITS2 sequences, part 28S rDNA se- quences, and a putatively encoding ?5 fatty acid desaturase gene were cloned respectively and further functioned in transgenic yeast. The sequence alignments were separately conducted using the related sequences from Nitzschia closterium f. minutissima, Cylindrotheca closterium (Bacillariophyta/Baci- llariales/Bacillariaceae/Cylindrotheca) and Phaeodactylum tricornutum (Naviculales) with ClustalX 1.83. No distinct difference was discovered between N. closterium f. minutissima and P. tricornutum in both biochemical and molecular level. Their identity was more than 99.6% among 18S rDNA, 5.8S rDNA and actin-gene sequences, and is up to 98.6% even among ITS1 and ITS2 sequences. Their ?5 desaturase similarity was 99.4%. However, the lower similarity was disclosured between N. closterium f. minutis- sima and Cylindrotheca closterium, which shared less than 40% identity in the ITS1 and ITS2 se- quences. So, N. closterium f. minutissima should not be placed in Bacillariales, Bacillariaceae, Nitzschia, but in Naviculales, Phaeodactylaceae, Phaeodactylum, and it was actually a strain of P. tri- cornutum.

丹江口水库一株硅藻的分子鉴定及氮素对其生长的影响

采用稀释分离法和小液滴分离法,从丹江口水库水样中分离了一株硅藻。通过形态学观察和18S rDNA、SSU rDNA、rbcL的序列分析,将其鉴定为谷皮菱形藻。以NaNO_(3)、NH_(4)Cl和尿素为氮源,研究了不同氮素对其生长特性的影响。结果表明,谷皮菱形藻在铵态氮、硝态氮和有机氮中均可正常生长,但生长特性对不同氮素形态的响应有所不同。在NaNO_(3)和尿素的培养条件下,谷皮菱形藻的生长速率随氮素浓度的增加而相应增加,而在NH_(4)Cl培养基中,生长速率随氮素浓度的增加表现出先增加后降低的趋势。在NH_(4)Cl 350μmol·L^(-1)时达到最高,最大比生长速率和细胞密度分别为0.2920 d^(-1)和189×10^(4)个·mL^(-1),表明高浓度铵态氮对其生长有抑制作用。本研究可为谷皮菱形藻的分子鉴定提供依据,也可为保护丹江口水库的水质安全提供基础资料。

光质对新月菱形藻生长、总脂含量和脂肪酸组成的影响

【目的】研究不同光质对新月菱形藻(Nitzschia closterium)生长、总脂含量和脂肪酸组成的影响,确定新月菱形藻生长及油脂产量的最适光质。【方法】在智能光质培养箱的静置培养条件下,采用单因子试验分别研究不同光质(LED白光、LED红光、LED蓝光、LED绿光和LED黄光)条件下新月菱形藻的生长、总脂含量和脂肪酸组成。【结果】光质对新月菱形藻的生长、总脂累积和脂肪酸合成有显著影响(P<0.05):LED绿光和黄光培养下新月菱形藻比生长速率显著高于白光、蓝光和红光组;LED白光、绿光和黄光培养下新月菱形藻的叶绿素a含量显著高于红光和蓝光组;LED白光、绿光和黄光培养下新月菱形藻的总脂含量显著高于红光和蓝光组;LED蓝光、绿光和黄光培养下新月菱形藻的多不饱和脂肪酸(PUFAs)含量没有显著差异(P>0.05),显著高于白光和红光组;LED蓝光培养下新月菱形藻的二十碳五烯酸(EPA)含量最高,显著高于其他组;LED红光培养下新月菱形藻的二十二碳六烯酸(DHA)含量最高,显著高于其他组。【结论】LED绿光和黄光能提高新月菱形藻的生长速率,LED蓝光有利于提高新月菱形藻的PUFAs和EPA的含量,LED红光有利于提高新月菱形藻的DHA含量。