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 spectr...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.展开更多
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 t...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℃. The SiO2 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 SiO2 morphologies of both biosilica are typical amorphous silica. Besides, IV. 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.展开更多
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 concentr...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, 1 125, 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, 16:1n7 and 20:5n3 ( EPA ) , B13 also has high percentage -of 20:4n6 ( 6.0 ~ 9.1% ).展开更多
A diatom was purified with colony selection and continuous dilution methods. It was identified to Cylindrotheca closterium according to its morphological characteristics and rbc L and 18 s r RNA gene sequences. The al...A diatom was purified with colony selection and continuous dilution methods. It was identified to Cylindrotheca closterium according to its morphological characteristics and rbc L and 18 s r RNA gene sequences. The alga was not sensitive to ampicillin and neomycin, but sensitive to chloramphenicol which inhibited its growth at concentrations ranging from 50 to 150 μg m L-1. The purified alga was easy to culture and its specific growth rate was 0.207 ± 0.002(d-1). It was resistant to pollution and could be harvested in an easy way. It was relatively high in lipid content(20.08% ± 0.67% of dry weight) and the combined amount of its 16:0 and 16:1(n-7), the most suitable resource of biodiesel, was as high as 64% of the total fatty acids, while the amount of polyunsaturated fatty acids reached 19.96%–20% of the total fatty acids. Thus the purified C. closterium can be cultured as a biodiesel producer or a nutrition supplement producer.展开更多
The maximum growth rate ofNitzschia elosterium at various concentrations of petroleum hydrocarbon was studied. The influence of petroleum hydrocarbon on cell size as a function of concentration was discussed. The rela...The maximum growth rate ofNitzschia elosterium 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 experi- mental results showed that the growth rate of Nitzschia elosterium 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 distribu- tion were observed during the growth period ofNitzsehia closterium. The Gaussian function could give a clear description of the cell size distribution ofNitzsehia 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.展开更多
The genus Cylindrotheca consists of a small group of marine diatoms with a few species described. Eleven isolates of diatoms identified as Cylindrotheca closterium morphologically were obtained from Jiaozhou Bay with ...The genus Cylindrotheca consists of a small group of marine diatoms with a few species described. Eleven isolates of diatoms identified as Cylindrotheca closterium morphologically were obtained from Jiaozhou Bay with their nuclear-encoded small-subunit ribosomal RNA （SSU rDNA） and chloroplast-encoded rbcL gene sequences determined in this study. Interestingly, very high sequence divergences of SSU rDNA and rbcL gene were found among these isolates, and numerous nucleotide variation of rbcL gene caused relatively few variation of deduced amino acid sequence. Phylogenetic analyses based on SSU rDNA and rbcL gene, respectively, grouped the isolates into 6 clades. Phylogenetic tree of SSU rDNA placed all the Cylindrotheca isolates together, separating them into two lineages clearly. LineageⅠ was composed of the eleven C. closterium isolates obtained in this study together with another C. closterium isolate, but some clades were not well supported. LineageⅠwas contained two C. closterium isolates and one C. fusiformis isolate. Phylogenetic analysis of rbcL gene also separated the Cylindrotheca isolates into two well-defined lineages. The eleven C. closterium isolates formed a lineage and all clades were supported strongly. Statistical comparisons of SSU rDNA indicated that the average distance within lineageⅠwas significantly higher than that of other microalgae species （P 〈0.01）. These results suggested the existence of cryptic species within C. closterium.展开更多
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 h...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.展开更多
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...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.展开更多
基金Supported by the Program for Changjiang Scholars and Innovative Research Team in University (PCSIRT) (No. IRT0734)Zhejiang Provincial Natural Science Foundation (No. Y506131)+1 种基金National Key Technology Research and Development Program (No. 2007BAD43B09)K. C. Wong Magna Fund of Ningbo University
文摘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.
基金Supported by the Public Science and Technology Research Funds Projects of Ocean,China(No.201305022)the PhD Start-up Fund of Natural Science Foundation of Guangdong Province,China(No.2014A030310326)
文摘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℃. The SiO2 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 SiO2 morphologies of both biosilica are typical amorphous silica. Besides, IV. 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.
基金Project supported by the Hi-Tech "863" programs of the Ministry of Science and Technology China (863-819-02-01)
文摘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, 1 125, 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, 16:1n7 and 20:5n3 ( EPA ) , B13 also has high percentage -of 20:4n6 ( 6.0 ~ 9.1% ).
基金supported by the Major State Basic Research Development Program of China (973 Program) (2011CB200901)National Technical Supporting Project Foundation (2011BAD14B01)Energy Project from State Bureau of Oceanic Administration (Grant No. GHME2011SW03)
文摘A diatom was purified with colony selection and continuous dilution methods. It was identified to Cylindrotheca closterium according to its morphological characteristics and rbc L and 18 s r RNA gene sequences. The alga was not sensitive to ampicillin and neomycin, but sensitive to chloramphenicol which inhibited its growth at concentrations ranging from 50 to 150 μg m L-1. The purified alga was easy to culture and its specific growth rate was 0.207 ± 0.002(d-1). It was resistant to pollution and could be harvested in an easy way. It was relatively high in lipid content(20.08% ± 0.67% of dry weight) and the combined amount of its 16:0 and 16:1(n-7), the most suitable resource of biodiesel, was as high as 64% of the total fatty acids, while the amount of polyunsaturated fatty acids reached 19.96%–20% of the total fatty acids. Thus the purified C. closterium can be cultured as a biodiesel producer or a nutrition supplement producer.
基金supported by the Chinese National Programs for High Technology Research and Development (No.2007AA09Z106).
文摘The maximum growth rate ofNitzschia elosterium 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 experi- mental results showed that the growth rate of Nitzschia elosterium 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 distribu- tion were observed during the growth period ofNitzsehia closterium. The Gaussian function could give a clear description of the cell size distribution ofNitzsehia 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.
基金This research was financially supported by an open project of The Key Laboratory of Mariculture of The Ministry of Education of China affiliated to 0cean umversity of China(200405)by the National Natural Science Foundation of China(30471318,40176028).
文摘The genus Cylindrotheca consists of a small group of marine diatoms with a few species described. Eleven isolates of diatoms identified as Cylindrotheca closterium morphologically were obtained from Jiaozhou Bay with their nuclear-encoded small-subunit ribosomal RNA （SSU rDNA） and chloroplast-encoded rbcL gene sequences determined in this study. Interestingly, very high sequence divergences of SSU rDNA and rbcL gene were found among these isolates, and numerous nucleotide variation of rbcL gene caused relatively few variation of deduced amino acid sequence. Phylogenetic analyses based on SSU rDNA and rbcL gene, respectively, grouped the isolates into 6 clades. Phylogenetic tree of SSU rDNA placed all the Cylindrotheca isolates together, separating them into two lineages clearly. LineageⅠ was composed of the eleven C. closterium isolates obtained in this study together with another C. closterium isolate, but some clades were not well supported. LineageⅠwas contained two C. closterium isolates and one C. fusiformis isolate. Phylogenetic analysis of rbcL gene also separated the Cylindrotheca isolates into two well-defined lineages. The eleven C. closterium isolates formed a lineage and all clades were supported strongly. Statistical comparisons of SSU rDNA indicated that the average distance within lineageⅠwas significantly higher than that of other microalgae species （P 〈0.01）. These results suggested the existence of cryptic species within C. closterium.
基金sponsored by the Ph.D.Start-Up Foundation of Guangdong Ocean University(R19049)Science and Technology Planning Project of Guangdong(2017A030303078)Undergraduate Start-Up Foundation of Guangdong Ocean University(580520135 and 570119011).
文摘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.
基金the National Basic Research Program of China (Grant No. 2005CCA02400)
文摘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.
