The responses of the interspecific competition between Skeletonema costatum and Heterosigma akashiwo to UV-B radiation enhancement were studied by the co-culture method. The results showed that Heterosigma akashiwo ex...The responses of the interspecific competition between Skeletonema costatum and Heterosigma akashiwo to UV-B radiation enhancement were studied by the co-culture method. The results showed that Heterosigma akashiwo exhibited inhibition on the growth of Skeletonema costatum, and with the increase of initial inoculation density of Heterosigma akashiwo, heavier inhibition on Skeletonema costatum appeared. Under different inoculation proportions, Heterosigma akashiwo could always be in predominance in competition with Skeletonema costatum. The UV-B radiation treatment could change the competition relationship between Skeletonema costatum and Heterosigma akashiwo, which could increase the competitive dominance of Skeletonema costatum and decrease the competitive dominance of Heterosigma akashiwo. When the inoculation proportions of Heterosigma akashiwo and Skeletonema costatum were H∶S=1∶4 and H∶S=1∶1,Skeletonema costatum was in predominance in this competition; however, Heterosigma akashiwo was in predominance when the inoculation proportion was H∶S=4∶1.展开更多
Over the past few years, harmful algal blooms （HABs）, such as red tides, have been frequently observed in coastal zones worldwide. The natural allelopathic interactions among macroalgae and red tide microalgae can a...Over the past few years, harmful algal blooms （HABs）, such as red tides, have been frequently observed in coastal zones worldwide. The natural allelopathic interactions among macroalgae and red tide microalgae can alter the structure and succession of aquatic ecosystems. We investigated the influence of four environmental factors （temperature, salinity, light, and pH） on the allelopathic effects of the macroalgae Corallina pilulifera on red-tide forming Heterosigma akashiwo under laboratory conditions. Each of the factors had four levels： temperature （15, 20, 25, and 30℃）, salinity （10, 20, 30, and 40）, light （20, 100, 200 and 400 gmol/（m2.s））, and pH （5.5, 7, 8.5, and 10. Two-factor experiments were designed for each two environmental factors, with six combination treatments （temperature-salinity, temperature-light, temperature-pH, salinity-light, salinity-pH, and light-pH）. Results showed that the allelopathic effect was significantly influenced by temperature, salinity, light, and pH. As single factors, the low temperature （15 ℃）, low salinity （10）, high-intensity light （400umol/（m2.s））, and high pH （10） treatments substantially enhanced the allelopathic effect. The strongest allelopathic effect of C. pilulifera on H. akashiwo was observed under the following treatments： 15℃ and salinity of 40, 25℃ and pH 10, 25℃ with medium- to high-intensity light at 200-400 umol/（m2.s）, 400umol/（m2.s） and salinity of 10, 400 gmol/（m2·s） and pH 10, and pH 10 with a salinity of 40.展开更多
Harmful algal blooms (HABs), caused by the overgrowth of certain phytoplankton species, have negative effects on marine environments and coastal fisheries. In addition to cell-counting methods using phytoplankton nets...Harmful algal blooms (HABs), caused by the overgrowth of certain phytoplankton species, have negative effects on marine environments and coastal fisheries. In addition to cell-counting methods using phytoplankton nets, a hydroacoustic technique based on acoustic backscattering has been proposed for the detection of phytoplankton blooms. However, little is known of the acoustic properties of HAB species. In this study, as essential data to support this technique, we measured the acoustic properties of two HAB species, Akashiwo sanguinea and Alexandrium affine, which occur in the South Sea off the coast of Korea. Due to the small size of the target, we used ultrasound for the measurements. Experiments were conducted under laboratory and field conditions. In the laboratory experiment, the acoustic signal received from each species was directly proportional to the cell abundance. We derived a relationship between the cell abundance and acoustic signal received for each species. The measured signals were compared to predictions of a fluid sphere scattering model. When A. sanguinea blooms appeared at an abundance greater than 3 500 cells/mL, the acoustic signals varied with cell abundance, showing a good correlation. These results confirm that acoustic measurements can be used to detect HAB species.展开更多
Blooms of Akashiwo sanguinea frequently break out around the world,causing huge economic losses to the aquaculture industry and seriously damaging coastal ecosystems.However,the formation mechanisms of A.sanguinea blo...Blooms of Akashiwo sanguinea frequently break out around the world,causing huge economic losses to the aquaculture industry and seriously damaging coastal ecosystems.However,the formation mechanisms of A.sanguinea blooms remain unclear.We investigated the allelopathic effects of A.sanguinea on multiple phytoplankton species,explored the mode of allelochemicals action and the way of nutrient factors regulation of the allelopathic activity.Results show that strains of A.sanguinea could inhibit the growth of co-occurring phytoplankton including Scrippsiella trochoidea,Phaeocystis globosa,and Rhodomonas salina,but inhibition of Prorocentrum micans was not obvious.The inhibition rates on phytoplankton were positively correlated with the cell densities of A.sanguinea.The highest inhibition rate of 94%on R.salina was for A.sanguinea CCMA256 culture of 2000 cells/mL at 72 h.We observed that cells of S.trochoidea,Ph.g lobosa,and R.sali na were lysed when co-cultured with A.sanguinea,with the shortest time for S.trochoidea.Additionally,the growth rates of A.sanguinea were promoted by coculturing with S.trochoidea,Ph.globosa,and R.salina.Four components of A.sanguinea culture were all able to inhibit growth of R.salina:the strongest inhibitory effect was found in the sonicated culture,followed by whole-cell culture,filtrates of sonicated culture,and filtrate culture.The crude extract of A.sanguinea culture also lysed cells of R.salina,and the inhibition rates on R.salina increased with the increasing dose of crude extract.It was shown that both nutrient enrichment and nitrogen:phosphorus ratio imbalance enhanced remarkably the allelopathic activity of A.sanguinea.The highest inhibition rate on R.salina of 70%occurred in A.sanguinea JX13 treatment at 2000 cells/mL under high nutrient condition in 48 h.In JX14 treatment at 2000 cells/mL for N:P of 10:1,the inhibition rate increased by 1.7 times of that for N:P of 20:1.In addition,the allelopathy of A.sanguinea could not only be a competitive strategy but also a nutrition strategy,playing an important role in formation and/or maintenance of blooms of the mixotrophic dinoflagellate A.sanguinea.展开更多
We investigated the effect of CO2 enrichment and initial inoculum density on competition between Skeletonema costatum and Heterosigma akashiwo,two common algae seen in algal blooms.The initial inoculum density(0.2...We investigated the effect of CO2 enrichment and initial inoculum density on competition between Skeletonema costatum and Heterosigma akashiwo,two common algae seen in algal blooms.The initial inoculum density(0.2×104,0.4×104,0.8×104 cells/ml) had a significant effect on population growth and competition between H.akashiwo and S.costatum.The time required for population growth to reach the exponential growth phase and stationary phase decreased significantly as the initial density increased.When the two species were cultured together,S.costatum tended to outcompete H.akashiwo,especially when present at higher initial ratios.CO_2 enrichment(5 000μl/L CO_2) increased the maximum population density and carrying capacity of H.akashiwo but decreased these measures for S.costatum.Thus,CO_2 enrichment favored the growth of H.akashiwo over S.costatum.展开更多
The accurate measurement of gene expression via quantitative real-time reverse transcription PCR（q RT-PCR）heavily relies on the choice of valid reference gene（s） for data normalization. Resting cyst is the dormant...The accurate measurement of gene expression via quantitative real-time reverse transcription PCR（q RT-PCR）heavily relies on the choice of valid reference gene（s） for data normalization. Resting cyst is the dormant stage in the life cycle of dinoflagellate, which plays crucial roles in HAB-forming dinoflagellate ecology. However, only limited investigations have been conducted on the reference gene selection in dinoflagellates. Gap remained in our knowledge about appropriate HKGs for normalizing gene expression in different life stages, which laid obstacles for the application of q RT-PCR to the HAB-forming group. In this study, six candidate reference genes,18 S ribosomal RNA（18S）, glyceraldehyde-3-phosphate dehydrogenase（GAPDH）, α-tubulin（TUA）, β-tubulin（TUB）, actin（ACT） and cytochrome oxidase subunit 1（COX1）, were evaluated for their expression stability with q RT-PCR and three statistical algorithms（Ge Norm, Norm Finder, and Best Keeper） for the cosmopolitan, harmful algal bloom-forming dinoflagellate Akashiwo sanguinea. Expression patterns were observed across 18 biological samples, including cells at resting stages（resting cysts）, different growth stages, in darkness, exposed to abscisic acid（ABA） and exposed to temperature stress. The results indicated that TUA, 18 S and GAPDH were relatively stable across all tested scenarios. While the best-recommended reference genes differed across experimental groups, the pairs of ACT and TUA, 18 S and GAPDH were the most reliable for cells at different growth stages and darkness treatment. The combination of TUA and TUB was the best choice for normalization in resting cysts and in ABA treatment, respectively. The pair of ACT and COX1 was suitable for temperature treatments. This study was the first to investigate the stable internal reference genes in dinoflagellates at different stages of life cycle,particularly in resting cysts. Our results provided useful information for selection of reference genes in dinoflagellates regarding quantification of gene expression at different experimental scenarios, which will facilitate more accurate and widespread use of q RT-PCR in gene analysis of dinoflagellates and help to design primers targeting orthologous genes in other algal species.展开更多
To rapidly detect the harmful algae H.akashiwo qualitatively and quantitatively, sequences of the 18S rDNA deduced from H.akashiwo were used for designing species-specific primers, and a RFQ-PCR (Real-time Fluorescent...To rapidly detect the harmful algae H.akashiwo qualitatively and quantitatively, sequences of the 18S rDNA deduced from H.akashiwo were used for designing species-specific primers, and a RFQ-PCR (Real-time Fluorescent Quantitative Polymerase Chain Reaction) method was developed for quantitative detection of H.akashiwo. Primer H.akashiwo and TaqMan probe were designed, and the specificity of primer was checked with PCR. A calibration curve was constructed with cycle threshold value against visual counted cell number. And the value of the curve was tested with other H.akashiwo samples, which were assayed with both the RFQ-PCR method and visual count under microscope.展开更多
基金supported by the National Natural Science Foundation of China under contract No.30270258Scientific Research Rewards Founda-tion for Outstanding Youth Scientists of Shandong Province under contract No.03BS120.
文摘The responses of the interspecific competition between Skeletonema costatum and Heterosigma akashiwo to UV-B radiation enhancement were studied by the co-culture method. The results showed that Heterosigma akashiwo exhibited inhibition on the growth of Skeletonema costatum, and with the increase of initial inoculation density of Heterosigma akashiwo, heavier inhibition on Skeletonema costatum appeared. Under different inoculation proportions, Heterosigma akashiwo could always be in predominance in competition with Skeletonema costatum. The UV-B radiation treatment could change the competition relationship between Skeletonema costatum and Heterosigma akashiwo, which could increase the competitive dominance of Skeletonema costatum and decrease the competitive dominance of Heterosigma akashiwo. When the inoculation proportions of Heterosigma akashiwo and Skeletonema costatum were H∶S=1∶4 and H∶S=1∶1,Skeletonema costatum was in predominance in this competition; however, Heterosigma akashiwo was in predominance when the inoculation proportion was H∶S=4∶1.
基金Supported by the National Natural Science Foundation of China(No.31200400)
文摘Over the past few years, harmful algal blooms （HABs）, such as red tides, have been frequently observed in coastal zones worldwide. The natural allelopathic interactions among macroalgae and red tide microalgae can alter the structure and succession of aquatic ecosystems. We investigated the influence of four environmental factors （temperature, salinity, light, and pH） on the allelopathic effects of the macroalgae Corallina pilulifera on red-tide forming Heterosigma akashiwo under laboratory conditions. Each of the factors had four levels： temperature （15, 20, 25, and 30℃）, salinity （10, 20, 30, and 40）, light （20, 100, 200 and 400 gmol/（m2.s））, and pH （5.5, 7, 8.5, and 10. Two-factor experiments were designed for each two environmental factors, with six combination treatments （temperature-salinity, temperature-light, temperature-pH, salinity-light, salinity-pH, and light-pH）. Results showed that the allelopathic effect was significantly influenced by temperature, salinity, light, and pH. As single factors, the low temperature （15 ℃）, low salinity （10）, high-intensity light （400umol/（m2.s））, and high pH （10） treatments substantially enhanced the allelopathic effect. The strongest allelopathic effect of C. pilulifera on H. akashiwo was observed under the following treatments： 15℃ and salinity of 40, 25℃ and pH 10, 25℃ with medium- to high-intensity light at 200-400 umol/（m2.s）, 400umol/（m2.s） and salinity of 10, 400 gmol/（m2·s） and pH 10, and pH 10 with a salinity of 40.
基金project titled "Establishment and demonstration of red tide detection and prediction system for minimizing red tide damage" funded by the Ministry of Oceans and Fisheries, Korea (PM61410)
文摘Harmful algal blooms (HABs), caused by the overgrowth of certain phytoplankton species, have negative effects on marine environments and coastal fisheries. In addition to cell-counting methods using phytoplankton nets, a hydroacoustic technique based on acoustic backscattering has been proposed for the detection of phytoplankton blooms. However, little is known of the acoustic properties of HAB species. In this study, as essential data to support this technique, we measured the acoustic properties of two HAB species, Akashiwo sanguinea and Alexandrium affine, which occur in the South Sea off the coast of Korea. Due to the small size of the target, we used ultrasound for the measurements. Experiments were conducted under laboratory and field conditions. In the laboratory experiment, the acoustic signal received from each species was directly proportional to the cell abundance. We derived a relationship between the cell abundance and acoustic signal received for each species. The measured signals were compared to predictions of a fluid sphere scattering model. When A. sanguinea blooms appeared at an abundance greater than 3 500 cells/mL, the acoustic signals varied with cell abundance, showing a good correlation. These results confirm that acoustic measurements can be used to detect HAB species.
基金Supported by the National Natural Science Foundation of China(NSFC)(Nos.41576159,41676099)。
文摘Blooms of Akashiwo sanguinea frequently break out around the world,causing huge economic losses to the aquaculture industry and seriously damaging coastal ecosystems.However,the formation mechanisms of A.sanguinea blooms remain unclear.We investigated the allelopathic effects of A.sanguinea on multiple phytoplankton species,explored the mode of allelochemicals action and the way of nutrient factors regulation of the allelopathic activity.Results show that strains of A.sanguinea could inhibit the growth of co-occurring phytoplankton including Scrippsiella trochoidea,Phaeocystis globosa,and Rhodomonas salina,but inhibition of Prorocentrum micans was not obvious.The inhibition rates on phytoplankton were positively correlated with the cell densities of A.sanguinea.The highest inhibition rate of 94%on R.salina was for A.sanguinea CCMA256 culture of 2000 cells/mL at 72 h.We observed that cells of S.trochoidea,Ph.g lobosa,and R.sali na were lysed when co-cultured with A.sanguinea,with the shortest time for S.trochoidea.Additionally,the growth rates of A.sanguinea were promoted by coculturing with S.trochoidea,Ph.globosa,and R.salina.Four components of A.sanguinea culture were all able to inhibit growth of R.salina:the strongest inhibitory effect was found in the sonicated culture,followed by whole-cell culture,filtrates of sonicated culture,and filtrate culture.The crude extract of A.sanguinea culture also lysed cells of R.salina,and the inhibition rates on R.salina increased with the increasing dose of crude extract.It was shown that both nutrient enrichment and nitrogen:phosphorus ratio imbalance enhanced remarkably the allelopathic activity of A.sanguinea.The highest inhibition rate on R.salina of 70%occurred in A.sanguinea JX13 treatment at 2000 cells/mL under high nutrient condition in 48 h.In JX14 treatment at 2000 cells/mL for N:P of 10:1,the inhibition rate increased by 1.7 times of that for N:P of 20:1.In addition,the allelopathy of A.sanguinea could not only be a competitive strategy but also a nutrition strategy,playing an important role in formation and/or maintenance of blooms of the mixotrophic dinoflagellate A.sanguinea.
基金Supported by the Specialized Research Program for Marine Public Welfare Industry,SOA(No.200805066)
文摘We investigated the effect of CO2 enrichment and initial inoculum density on competition between Skeletonema costatum and Heterosigma akashiwo,two common algae seen in algal blooms.The initial inoculum density(0.2×104,0.4×104,0.8×104 cells/ml) had a significant effect on population growth and competition between H.akashiwo and S.costatum.The time required for population growth to reach the exponential growth phase and stationary phase decreased significantly as the initial density increased.When the two species were cultured together,S.costatum tended to outcompete H.akashiwo,especially when present at higher initial ratios.CO_2 enrichment(5 000μl/L CO_2) increased the maximum population density and carrying capacity of H.akashiwo but decreased these measures for S.costatum.Thus,CO_2 enrichment favored the growth of H.akashiwo over S.costatum.
基金The National Natural Science Foundation of China-Shandong Joint Fund for Marine Science Research Centers under contract No.U1406403the State Key Program of National Natural Science of China under contract No.61533011China Postdoctoral Science Foundation under contract Nos 2014M551969 and 2015T80754
文摘The accurate measurement of gene expression via quantitative real-time reverse transcription PCR（q RT-PCR）heavily relies on the choice of valid reference gene（s） for data normalization. Resting cyst is the dormant stage in the life cycle of dinoflagellate, which plays crucial roles in HAB-forming dinoflagellate ecology. However, only limited investigations have been conducted on the reference gene selection in dinoflagellates. Gap remained in our knowledge about appropriate HKGs for normalizing gene expression in different life stages, which laid obstacles for the application of q RT-PCR to the HAB-forming group. In this study, six candidate reference genes,18 S ribosomal RNA（18S）, glyceraldehyde-3-phosphate dehydrogenase（GAPDH）, α-tubulin（TUA）, β-tubulin（TUB）, actin（ACT） and cytochrome oxidase subunit 1（COX1）, were evaluated for their expression stability with q RT-PCR and three statistical algorithms（Ge Norm, Norm Finder, and Best Keeper） for the cosmopolitan, harmful algal bloom-forming dinoflagellate Akashiwo sanguinea. Expression patterns were observed across 18 biological samples, including cells at resting stages（resting cysts）, different growth stages, in darkness, exposed to abscisic acid（ABA） and exposed to temperature stress. The results indicated that TUA, 18 S and GAPDH were relatively stable across all tested scenarios. While the best-recommended reference genes differed across experimental groups, the pairs of ACT and TUA, 18 S and GAPDH were the most reliable for cells at different growth stages and darkness treatment. The combination of TUA and TUB was the best choice for normalization in resting cysts and in ABA treatment, respectively. The pair of ACT and COX1 was suitable for temperature treatments. This study was the first to investigate the stable internal reference genes in dinoflagellates at different stages of life cycle,particularly in resting cysts. Our results provided useful information for selection of reference genes in dinoflagellates regarding quantification of gene expression at different experimental scenarios, which will facilitate more accurate and widespread use of q RT-PCR in gene analysis of dinoflagellates and help to design primers targeting orthologous genes in other algal species.
文摘To rapidly detect the harmful algae H.akashiwo qualitatively and quantitatively, sequences of the 18S rDNA deduced from H.akashiwo were used for designing species-specific primers, and a RFQ-PCR (Real-time Fluorescent Quantitative Polymerase Chain Reaction) method was developed for quantitative detection of H.akashiwo. Primer H.akashiwo and TaqMan probe were designed, and the specificity of primer was checked with PCR. A calibration curve was constructed with cycle threshold value against visual counted cell number. And the value of the curve was tested with other H.akashiwo samples, which were assayed with both the RFQ-PCR method and visual count under microscope.
