[Objective] The aim was to study the effects of L-carnitine on population growth of Nannochloropsis oculata and Tetrahymena sp..[Method] When the concentration of L-carnitine was 0,50,100 and 1 000 mg/L,population den...[Objective] The aim was to study the effects of L-carnitine on population growth of Nannochloropsis oculata and Tetrahymena sp..[Method] When the concentration of L-carnitine was 0,50,100 and 1 000 mg/L,population densities of Nannochloropsis oculata and Tetrahymena were determined respectively.[Result] Adding high-dose L-carnitine had a significant inhibition effect on the population growth of Nannochloropsis oculata (P0.05).Adding L-carnitine had a significant proliferation promoting effect on the population growth of Tetrahymena (P0.05).[Conclusion] The research provides theoretical basis for the application of L-carnitine as feed additive in aquaculture.展开更多
The aim of this study was to explore the potency of N. oculata extracts as antibacterial, antioxidant and antiviral on grouper Cromileptes altivelis (C. altivelis) infected by Vibrio alginolyticus (V. alginolyticus...The aim of this study was to explore the potency of N. oculata extracts as antibacterial, antioxidant and antiviral on grouper Cromileptes altivelis (C. altivelis) infected by Vibrio alginolyticus (V. alginolyticus) and Viral Nervous Necrotic (VNN). Dilution test was used to measure antibacterial activity of N. oculata extracts. Antioxidant activity of extracts was expressed by levels of MDA (Malondialdehyde) and SOD (superoxide dismutase) in the brain and kidneys of fish. Antiviral capability of N. oculata was determined by the expression of cellular immune cells Major Histocompatibility Complex (MHC) Class I for proliferation and inhibition of VNN in blood cells with immunocytochemistry. The results showed that N. oculata extract was able to suppress the growth of V. alginolyticus at a concentration of 40%. Increasing levels of SOD and reducing level of MDA indicated that N. oculata extracts may serve as an antibacterial and antioxidant. Providing cellular response of MHC class I cell expressed on C. altive/is blood cells, N. oculata demonstrated its antiviral activity.展开更多
A gene (NANOC-D6D) encoding a desaturase that removes two hydrogen atoms from fatty acids at delta 6 position was isolated from a cDNA library of Nannochloropsis oculata (Droop) D. J. Hibberd (Eustigmatophyceae)...A gene (NANOC-D6D) encoding a desaturase that removes two hydrogen atoms from fatty acids at delta 6 position was isolated from a cDNA library of Nannochloropsis oculata (Droop) D. J. Hibberd (Eustigmatophyceae). The unicellular marine microalga N. oculata synthesizes rich long chain polyunsaturated fatty acids (LCPUFAs), including eicosapentaenoic acid (20:5n-3, EPA). The deduced protein contains 474 amino acids that fold into 4 trans-membrane domains. The neighbor-joining phylogenetic tree indicates that NANOC-D6D is phylogenetically close to the delta-6 fatty acid desaturase of marine microalgae such as Glossomastix chrysoplasta, Thalassiosira pseudonana, and Phaeodactylum tricornutum. The gene was expressed in Saccharomyces cerevisiae INVScl to verify the substrate specificity of NAN OC-D6D. Our results suggest that the recombinant NANOC-D6D simultaneously desaturates linoleic acid (LA) and a-linolenic acid (ALA).展开更多
Nannochloropsis oculata CS179, a unicellular marine microalga, is rich in long-chain polyunsaturated fatty acids (LCPUFAs). Elongase and desaturase play a key role in the biosynthesis of PUFAs. A new elongase gene, ...Nannochloropsis oculata CS179, a unicellular marine microalga, is rich in long-chain polyunsaturated fatty acids (LCPUFAs). Elongase and desaturase play a key role in the biosynthesis of PUFAs. A new elongase gene, which encodes 322 amino acids, was identified via RT-PCR and 5' and 3' RACE. The sequence of the elongase gene was blast-searched in the NCBI GenBank and showed a similarity to those of the coptosporidium. But the NJ-tree revealed that the N. oculata CS 179 elongase clustered with those of the microalgae Phaeodac^lum tricornutum, Ostreocoecus tauri and Thalassiosira pseudonana.展开更多
We constructed and characterized a normalized cDNA library of Nannochloropsis oculata CS-179,and obtained 905 nonredundant sequences(NRSs) ranging from 431-1 756 bp in length.Among them,496 were very similar to nonred...We constructed and characterized a normalized cDNA library of Nannochloropsis oculata CS-179,and obtained 905 nonredundant sequences(NRSs) ranging from 431-1 756 bp in length.Among them,496 were very similar to nonredundant ones in the GenBank(E ≤1.0e-05),and 349 ESTs had significant hits with the clusters of eukaryotic orthologous groups(KOG).Bases G and/or C at the third position of codons of 14 amino acid residues suggested a strong bias in the conserved domain of 362 NRSs(>60%).We also identified the unigenes encoding phosphorus and nitrogen transporters,suggesting that N.oculata could efficiently transport and metabolize phosphorus and nitrogen,and recognized the unigenes that involved in biosynthesis and storage of both fatty acids and polyunsaturated fatty acids(PUFAs),which will facilitate the demonstration of eicosapentaenoic acid(EPA) biosynthesis pathway of N.oculata.In comparison with the original cDNA library,the normalized library significantly increased the efficiencies of random sequencing and rarely expressed genes discovering,and decreased the frequency of abundant gene sequences.展开更多
Illuminating conditions are crucial factors affecting the microalgal growth and biosynthesis.The effects of illuminating spectral quality on the growth and bio-component production of Nannochloris oculata were investi...Illuminating conditions are crucial factors affecting the microalgal growth and biosynthesis.The effects of illuminating spectral quality on the growth and bio-component production of Nannochloris oculata were investigated.The results indicated that a high proportion of Red-light enhanced the pigments and carbohydrate production but reduced those of the biomass and lipid.Mono-chromatic Blue-light has advantageous effects on lipid production compared with Red-and White-light.The optimal light spectrum for the protein production was the combination of Red-and Blue-light at a ratio of 4:3 or 5:2.Among the seven fatty acids identified from N.oculata,the contents of C16:0,C18:0,and C18:3(n-3)in the lipid were inhibited by the increased proportion of Red-light while those of C18:2(n-9),C16:2(n-6),and C20:0 were inhibited by Blue-light.Monochromatic Red-and Blue-light and their com-binations were proposed as a promising illuminating strategy for the large-scale cultivation aiming for various bio-components.展开更多
Nannochloropsis oculata CS-179, a marine eukaryotic unicellular microalga, is rich in long-chain polyunsaturated fatty acids (LC-PUFAs). Culture temperature affected cell growth and the composition of LC-PUFAs. At an ...Nannochloropsis oculata CS-179, a marine eukaryotic unicellular microalga, is rich in long-chain polyunsaturated fatty acids (LC-PUFAs). Culture temperature affected cell growth and the composition of LC-PUFAs. At an initial cell density of 1.5 × 106 cell mL-1, the highest growth was observed at 25℃ and the cell density reached 3 × 107 cell mL-1 at the beginning of loga-rithmic phase. The content of LC-PUFAs varied with culture temperature.The highest content of LC-PUFAs (43.96%) and EPA (36.6%) was gained at 20℃. Real-time PCR showed that the abundance of Δ6-desaturase gene transcripts was significantly different among 5 culture temperatures and the highest transcript level (15℃) of Nanoc-D6D took off at cycle 21.45. The gene transcript of C20-elongase gene was higher at lower temperatures (10, 15, and 20℃), and the highest transcript level (20℃) of Nanoc-E took off at cycle 21.18. The highest conversion rate (39.3%) of Δ6-desaturase was also gained at 20℃.But the conversion rate of Nanoc-E was not detected. The higher content of LC-PUFAs was a result of higher gene transcript level and higher enzyme activity. Compared with C20-elongase gene, Δ6-desaturase gene transcript and enzyme activity varied significantly with temperature. It will be useful to study the mechanism of how the content of LC-PUFAs is affected by temperature.展开更多
Microalgae Tetraselmis subcordiformis and Nannochloropsis oculata were cultured at 15,20,25,30,and 35℃ and their properties as potential biofuel resources were examined.The results indicate that T.subcordiformis and ...Microalgae Tetraselmis subcordiformis and Nannochloropsis oculata were cultured at 15,20,25,30,and 35℃ and their properties as potential biofuel resources were examined.The results indicate that T.subcordiformis and N.oculata grew best at 20℃ and 25℃ and yielded the highest total lipids at 20℃and 30℃,respectively.With increased temperature,neutral lipid and polyunsaturated fatty acids(FAs)decreased while saturated FAs increased,accompanied by increased monounsaturated FAs(MUFAs) in T.subcordiformis and decreased MUFAs in N.oculata;meanwhile,the predicted cetane number of FA methyl esters increased from 45.3 to 47.6 in T.subcordiformis and from 52.3 to 60.3 in N.oculata.Therefore,optimizing culture temperatures is important for improving microalgal biodiesel production.展开更多
The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker. It can control the phleomycin resistance, and significantly increase the tolerance of hosts to zeocin. The unicellular ...The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker. It can control the phleomycin resistance, and significantly increase the tolerance of hosts to zeocin. The unicellular marine microalga Nannochloropsis oculata is extremely sensitive to zeocin. We selected ble as the selective marker for the genetic transformation of N. oculata. After the algal cells at a density of 2× 10^7 cells mL-1 was digested with 4% hemicellulase and 2% driselase for 1 h, the protoplasts accounted for 90% of the total. The ble was placed at the downstream of promoter HSP70A-RUBS2 isolated from Chlamydomonas reinhardtii, yielding a recombinant expression construct pMS 188. The construct was transferred into the protoplasts through electroporation (1 kV, 15 μS). The transformed protoplasts were cultured in fresh f/2 liquid medium, and selected on solid f/2 medium supplemented with 500 ng mL-t zeoein. The PCR result proved that ble existed in the transformants. Three transformants had been cultured for at least 5 generations without losing ble. Southern blotting analysis showed that the ble has been integrated into the genome of N. oculata. The ble will serve as a new dominant selective marker in genetic engineering N. oculata.展开更多
Hydroesterification process has been presented biodiesel production from oil the green microalga Nannochloropsis oculata raw materials. Biodiesel studied in this work is the main product got from the hydroesterificati...Hydroesterification process has been presented biodiesel production from oil the green microalga Nannochloropsis oculata raw materials. Biodiesel studied in this work is the main product got from the hydroesterification of biomass the Nannochloropsis oculata and was obtained from esterification of fatty acid (product of a hydrolysis reaction) with methanol. It was used as catalyst the niobic acid pure and supported on δ-aluminum. The product was evaluated by gas chromatography and other analyses. The optimum conditions found in the conversion (%) for the hydrolysis reactions of the biomass (92.3%). Better results were observed in the algae concentration 20%, lead at 300?C with 20% of catalyst. For esterification of fatty acids of Nannochloropsis oculata (92.24%), were observed the molar ratio methanol: fat acid 3, lead at 200°C with 20% of catalyst supported.展开更多
Ultraviolet mutagenesis was applied to Nannochloropsis oculata and three mutants resistant to 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) were isolated. The cellular chlorophyll a and total lipid content of the wil...Ultraviolet mutagenesis was applied to Nannochloropsis oculata and three mutants resistant to 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) were isolated. The cellular chlorophyll a and total lipid content of the wild are higher in the medium supplemented with DCMU than in the control without DCMU. Without DCMU, the growth rates and chlorophyll a contents of the mutants are similar to those of the wild. Significant changes of fatty acid content and composition have occurred in DCMU-resistant mutants growing in the medium supplemented with DCMU. The total lipid, palmitic acid (16∶0), palmitoleic acid (16∶1ω9) and oleic (18∶1ω9) contents decrease significantly, while the vaccenic acid (18∶1ω11) increases significantly and the EPA content of dried powder increases slightly in the mutants. The study may provide a basis to improve EPA content in Nannochloropsis oculata in the future.展开更多
基金Supported by National Natural Science Foundation of China(30671621)~~
文摘[Objective] The aim was to study the effects of L-carnitine on population growth of Nannochloropsis oculata and Tetrahymena sp..[Method] When the concentration of L-carnitine was 0,50,100 and 1 000 mg/L,population densities of Nannochloropsis oculata and Tetrahymena were determined respectively.[Result] Adding high-dose L-carnitine had a significant inhibition effect on the population growth of Nannochloropsis oculata (P0.05).Adding L-carnitine had a significant proliferation promoting effect on the population growth of Tetrahymena (P0.05).[Conclusion] The research provides theoretical basis for the application of L-carnitine as feed additive in aquaculture.
文摘The aim of this study was to explore the potency of N. oculata extracts as antibacterial, antioxidant and antiviral on grouper Cromileptes altivelis (C. altivelis) infected by Vibrio alginolyticus (V. alginolyticus) and Viral Nervous Necrotic (VNN). Dilution test was used to measure antibacterial activity of N. oculata extracts. Antioxidant activity of extracts was expressed by levels of MDA (Malondialdehyde) and SOD (superoxide dismutase) in the brain and kidneys of fish. Antiviral capability of N. oculata was determined by the expression of cellular immune cells Major Histocompatibility Complex (MHC) Class I for proliferation and inhibition of VNN in blood cells with immunocytochemistry. The results showed that N. oculata extract was able to suppress the growth of V. alginolyticus at a concentration of 40%. Increasing levels of SOD and reducing level of MDA indicated that N. oculata extracts may serve as an antibacterial and antioxidant. Providing cellular response of MHC class I cell expressed on C. altive/is blood cells, N. oculata demonstrated its antiviral activity.
基金Supported by the National Key Technology R&D Program in the 11th Five-Year Plan of China (No. 2006BAD09A03-2)the National High-tech Research and Development Program of China(863 Program) (No. 2007AA09Z427)
文摘A gene (NANOC-D6D) encoding a desaturase that removes two hydrogen atoms from fatty acids at delta 6 position was isolated from a cDNA library of Nannochloropsis oculata (Droop) D. J. Hibberd (Eustigmatophyceae). The unicellular marine microalga N. oculata synthesizes rich long chain polyunsaturated fatty acids (LCPUFAs), including eicosapentaenoic acid (20:5n-3, EPA). The deduced protein contains 474 amino acids that fold into 4 trans-membrane domains. The neighbor-joining phylogenetic tree indicates that NANOC-D6D is phylogenetically close to the delta-6 fatty acid desaturase of marine microalgae such as Glossomastix chrysoplasta, Thalassiosira pseudonana, and Phaeodactylum tricornutum. The gene was expressed in Saccharomyces cerevisiae INVScl to verify the substrate specificity of NAN OC-D6D. Our results suggest that the recombinant NANOC-D6D simultaneously desaturates linoleic acid (LA) and a-linolenic acid (ALA).
基金supported by the Project for Supporting the National Development (No. 2006BAD09A03)National 863 Program (No. 2007AA09Z427)
文摘Nannochloropsis oculata CS179, a unicellular marine microalga, is rich in long-chain polyunsaturated fatty acids (LCPUFAs). Elongase and desaturase play a key role in the biosynthesis of PUFAs. A new elongase gene, which encodes 322 amino acids, was identified via RT-PCR and 5' and 3' RACE. The sequence of the elongase gene was blast-searched in the NCBI GenBank and showed a similarity to those of the coptosporidium. But the NJ-tree revealed that the N. oculata CS 179 elongase clustered with those of the microalgae Phaeodac^lum tricornutum, Ostreocoecus tauri and Thalassiosira pseudonana.
基金Supported by National Key Technology R&D Program in the 11th-Five-Year Plan of China(No.2006BAD09A03-2)National High Technology Research and Development of China(863 Program)(No.2007AA09Z427)
文摘We constructed and characterized a normalized cDNA library of Nannochloropsis oculata CS-179,and obtained 905 nonredundant sequences(NRSs) ranging from 431-1 756 bp in length.Among them,496 were very similar to nonredundant ones in the GenBank(E ≤1.0e-05),and 349 ESTs had significant hits with the clusters of eukaryotic orthologous groups(KOG).Bases G and/or C at the third position of codons of 14 amino acid residues suggested a strong bias in the conserved domain of 362 NRSs(>60%).We also identified the unigenes encoding phosphorus and nitrogen transporters,suggesting that N.oculata could efficiently transport and metabolize phosphorus and nitrogen,and recognized the unigenes that involved in biosynthesis and storage of both fatty acids and polyunsaturated fatty acids(PUFAs),which will facilitate the demonstration of eicosapentaenoic acid(EPA) biosynthesis pathway of N.oculata.In comparison with the original cDNA library,the normalized library significantly increased the efficiencies of random sequencing and rarely expressed genes discovering,and decreased the frequency of abundant gene sequences.
基金supported by the National Natural Sci-ence Foundation of China(Nos.41776156 and 42177459)the Open-End Funds of Jiangsu Key Laboratory of Ma-rine Bioresources and Environment(No.SH20201206).
文摘Illuminating conditions are crucial factors affecting the microalgal growth and biosynthesis.The effects of illuminating spectral quality on the growth and bio-component production of Nannochloris oculata were investigated.The results indicated that a high proportion of Red-light enhanced the pigments and carbohydrate production but reduced those of the biomass and lipid.Mono-chromatic Blue-light has advantageous effects on lipid production compared with Red-and White-light.The optimal light spectrum for the protein production was the combination of Red-and Blue-light at a ratio of 4:3 or 5:2.Among the seven fatty acids identified from N.oculata,the contents of C16:0,C18:0,and C18:3(n-3)in the lipid were inhibited by the increased proportion of Red-light while those of C18:2(n-9),C16:2(n-6),and C20:0 were inhibited by Blue-light.Monochromatic Red-and Blue-light and their com-binations were proposed as a promising illuminating strategy for the large-scale cultivation aiming for various bio-components.
基金supported by the grants from the National Key Technologies Research and Development Program of China during the Eleventh Five-Year Plan Period(No.2006BAD09A03-2)National High Technology Research and Development Program of China(863 Program)(No.2007AA09Z427)Basic Research Program of Municipal Bureau of Science and Technology of Qingdao(No.09-1-3-22-jch)
文摘Nannochloropsis oculata CS-179, a marine eukaryotic unicellular microalga, is rich in long-chain polyunsaturated fatty acids (LC-PUFAs). Culture temperature affected cell growth and the composition of LC-PUFAs. At an initial cell density of 1.5 × 106 cell mL-1, the highest growth was observed at 25℃ and the cell density reached 3 × 107 cell mL-1 at the beginning of loga-rithmic phase. The content of LC-PUFAs varied with culture temperature.The highest content of LC-PUFAs (43.96%) and EPA (36.6%) was gained at 20℃. Real-time PCR showed that the abundance of Δ6-desaturase gene transcripts was significantly different among 5 culture temperatures and the highest transcript level (15℃) of Nanoc-D6D took off at cycle 21.45. The gene transcript of C20-elongase gene was higher at lower temperatures (10, 15, and 20℃), and the highest transcript level (20℃) of Nanoc-E took off at cycle 21.18. The highest conversion rate (39.3%) of Δ6-desaturase was also gained at 20℃.But the conversion rate of Nanoc-E was not detected. The higher content of LC-PUFAs was a result of higher gene transcript level and higher enzyme activity. Compared with C20-elongase gene, Δ6-desaturase gene transcript and enzyme activity varied significantly with temperature. It will be useful to study the mechanism of how the content of LC-PUFAs is affected by temperature.
基金Supported by the National High Technology Research and Development Program of China(863 Program)(No.2009AA064401)the Special Fundation for Marine Renewable Energy from the State Oceanic Administration of China(No.SHME2011SW02)the Shanghai Universities First-class Disciplines Project of Fisheries(A)
文摘Microalgae Tetraselmis subcordiformis and Nannochloropsis oculata were cultured at 15,20,25,30,and 35℃ and their properties as potential biofuel resources were examined.The results indicate that T.subcordiformis and N.oculata grew best at 20℃ and 25℃ and yielded the highest total lipids at 20℃and 30℃,respectively.With increased temperature,neutral lipid and polyunsaturated fatty acids(FAs)decreased while saturated FAs increased,accompanied by increased monounsaturated FAs(MUFAs) in T.subcordiformis and decreased MUFAs in N.oculata;meanwhile,the predicted cetane number of FA methyl esters increased from 45.3 to 47.6 in T.subcordiformis and from 52.3 to 60.3 in N.oculata.Therefore,optimizing culture temperatures is important for improving microalgal biodiesel production.
基金supported by the Provincial Natural Science Foundation of Shandong Province,China (2013ZRB14055)and the National Natural Science Foundation of China (Grant No.31372518)
文摘The gene ble from Streptoalloteichus hindustanus is widely used as a selective antibiotic marker. It can control the phleomycin resistance, and significantly increase the tolerance of hosts to zeocin. The unicellular marine microalga Nannochloropsis oculata is extremely sensitive to zeocin. We selected ble as the selective marker for the genetic transformation of N. oculata. After the algal cells at a density of 2× 10^7 cells mL-1 was digested with 4% hemicellulase and 2% driselase for 1 h, the protoplasts accounted for 90% of the total. The ble was placed at the downstream of promoter HSP70A-RUBS2 isolated from Chlamydomonas reinhardtii, yielding a recombinant expression construct pMS 188. The construct was transferred into the protoplasts through electroporation (1 kV, 15 μS). The transformed protoplasts were cultured in fresh f/2 liquid medium, and selected on solid f/2 medium supplemented with 500 ng mL-t zeoein. The PCR result proved that ble existed in the transformants. Three transformants had been cultured for at least 5 generations without losing ble. Southern blotting analysis showed that the ble has been integrated into the genome of N. oculata. The ble will serve as a new dominant selective marker in genetic engineering N. oculata.
文摘Hydroesterification process has been presented biodiesel production from oil the green microalga Nannochloropsis oculata raw materials. Biodiesel studied in this work is the main product got from the hydroesterification of biomass the Nannochloropsis oculata and was obtained from esterification of fatty acid (product of a hydrolysis reaction) with methanol. It was used as catalyst the niobic acid pure and supported on δ-aluminum. The product was evaluated by gas chromatography and other analyses. The optimum conditions found in the conversion (%) for the hydrolysis reactions of the biomass (92.3%). Better results were observed in the algae concentration 20%, lead at 300?C with 20% of catalyst. For esterification of fatty acids of Nannochloropsis oculata (92.24%), were observed the molar ratio methanol: fat acid 3, lead at 200°C with 20% of catalyst supported.
基金support of the National Advanced Technology‘863’Project Foundation(819-02-01).
文摘Ultraviolet mutagenesis was applied to Nannochloropsis oculata and three mutants resistant to 3-(3,4-dichlorophenyl)-1,1-dimethylurea (DCMU) were isolated. The cellular chlorophyll a and total lipid content of the wild are higher in the medium supplemented with DCMU than in the control without DCMU. Without DCMU, the growth rates and chlorophyll a contents of the mutants are similar to those of the wild. Significant changes of fatty acid content and composition have occurred in DCMU-resistant mutants growing in the medium supplemented with DCMU. The total lipid, palmitic acid (16∶0), palmitoleic acid (16∶1ω9) and oleic (18∶1ω9) contents decrease significantly, while the vaccenic acid (18∶1ω11) increases significantly and the EPA content of dried powder increases slightly in the mutants. The study may provide a basis to improve EPA content in Nannochloropsis oculata in the future.