Protein kinase C (PKC) has a crucial role in signal transduction for a variety of biologically active substances which activate cellular functions and proliferation. We previously isolated the full-length PKC gene fro...Protein kinase C (PKC) has a crucial role in signal transduction for a variety of biologically active substances which activate cellular functions and proliferation. We previously isolated the full-length PKC gene from Dunaliella salina (DsPKC) using rapid amplification of cDNA ends (RACE) and RT-PCR methods. And we submitted the mRNA sequence of DsPKC gene to NCBI (Genbank No. JN625213). In the present paper, the DsPKC gene open reading frame obtained by PCR was cloned into pGS-21a vector and transformed into Escherichia coli to generate the fusion protein. Bioinformatics analysis revealed that DsPKC gene was a member of serine/threonine kinase with two conserved domains and highly conserved motifs. The DsPKC was highly expressed upon induction with isopropyl-β-d-thiogalactoside (IPTG) at a final concentration of 0.2 mmol L 1 at 37℃. Under salt stress, the fu- sion protein Green Fluorescent Protein (GFP)-DsPKC was transferred from the cytoplasm to the cell membrane. The expression pat- tern of DsPKC gene was analyzed using real-time quantitative PCR, and indicated that DsPKC gene was up-regulated by 3.0 mol L 1 NaCl at 12 h, which was significantly higher than in control values (P < 0.05). These results suggest that the DsPKC gene plays an important role in response to salt stress in D. salina.展开更多
The research status and the future study prospect of salt lake organisms of Chinese salt lakes were elaborated in this paper.Many saline lakes occur widely in arid and semiarid areas of China,and more than half of the...The research status and the future study prospect of salt lake organisms of Chinese salt lakes were elaborated in this paper.Many saline lakes occur widely in arid and semiarid areas of China,and more than half of the total展开更多
The seasonal variations in biomass, abundance, and species composition of plankton in relation to hydrography were studied in the saline Bange Lake, northern Tibet, China. Sampling was carried out between one to three...The seasonal variations in biomass, abundance, and species composition of plankton in relation to hydrography were studied in the saline Bange Lake, northern Tibet, China. Sampling was carried out between one to three times per month from May 2001 to July 2002. Salinity ranged from 14 to 146. The air and water temperature exhibited a clear seasonal pattern, and mean annual temperatures were approximately 4.8℃ and 7.3℃, respectively. The lowest water temperature occurred in winter from December to March at-2℃ and the highest in June and July at 17.7℃. Forty-one phytoplankton taxa, 21 zooplankton, and 5 benthic or facultative zooplankton were identifi ed. The predominant phytoplankton species were Gloeothece linearis, Oscillatoria tenuis, Gloeocapsa punctata, Ctenocladus circinnatus, Dunaliella salina, and Spirulina major. The predominant zooplankton species included H olophrya actra, Brachionus plicatilis, Daphniopsis tibetana, Cletocamptus dertersi, and A rctodiaptomus salinus. The mean annual total phytoplankton density and biomass for the entire lake were 4.52×10^7 cells/L and 1.60 mg/L, respectively. The annual mean zooplankton abundance was 52, 162, 322, and 57, 144 ind./L, in the three sublakes. The annual mean total zooplankton biomass in Lakes 1–3 was 1.23, 9.98, and 2.13 mg/L, respectively. The annual mean tychoplankton abundances in Bg1, 2, and 3 were 47, 67, and 654 ind./L. The annual mean tychoplankton biomass was 2.36, 0.16, and 2.03 mg/L, respectively. The zooplankton biomass（including tychoplankton） in the lake was 9.11 mg/L. The total number of plankton species in the salt lake was signifi cantly negatively correlated with salinity.展开更多
Plants possess effective mechanisms to respond quickly to the external environment. Rapid activation of phosphatidylinositol-specific phospholipase C (PLC) enzymes occurs after a stimulus. The PLC in Dunaliella salina...Plants possess effective mechanisms to respond quickly to the external environment. Rapid activation of phosphatidylinositol-specific phospholipase C (PLC) enzymes occurs after a stimulus. The PLC in Dunaliella salina plays important roles in growth and stress responses. However, the molecular basis of PLC action in D. salina remains little understood. To gain insight into the potential biological functions of this enzyme, we cloned a phospholipase C gene from D. salina in a previous study, named DsPLC (GenBank No. KF573428). Here, we present the prokaryotic expression, purification, and characterization of the DsPLC gene. The entire coding region of DsPLC was inserted into an expression vector pET32a, and the DsPLC gene was successfully expressed in Escherichia coli. The DsPLC protein was purified and identified using a polyclonal antibody and western blotting. Expressing DsPLC fused with a green fluorescent protein (GFP) in onion showed that DsPLC-GFP was localized to the intracellular membrane. Quantitative real-time PCR analysis revealed that the relative expression of the DsPLC gene was induced significantly by 3.0-mol/L NaCl at 4 h. Our results support the importance of PLC enzymes in plant defense signaling. This study provides a basis for further functional studies of the DsPLC gene and for additional analysis of the potential roles of PLC enzymes in response to abiotic stress.展开更多
We investigated the eff ects of temperature（14,20,and 22℃）,salinity（5,10,15,20,and 25）,and body length（0.83,1.17,1.49,and 2.33 mm）on the energy budget of Daphniopsis tibetana Sars in the laboratory.The results ...We investigated the eff ects of temperature（14,20,and 22℃）,salinity（5,10,15,20,and 25）,and body length（0.83,1.17,1.49,and 2.33 mm）on the energy budget of Daphniopsis tibetana Sars in the laboratory.The results demonstrated no diff erence in D.tibetana assimilation efficiency（AE）with values of 51.59%–56.22% at the trial temperatures.Gross growth efficiency（K1）and net growth efficiency（K2）were 28.78% and 55.71%,respectively,at 14℃.Daphniopsis tibetana energy conversion efficiency（AE,K1,and K2）was higher at salinities of 10–20,with values of 72.80%–77.31%,42.50%–50.42%,and 58.43%–65.02%,respectively.Daphniopsis tibetana energy conversion efficiency was highest when body length was 1.17–1.19 mm,with values of 83.70%–84.73%,50.11%–50.81%,and 59.13%–60.71%,respectively.The results of this study can provide a reference for large-scale cultivation use of D.tibetana in the further.展开更多
With the total RNA of Dunaliella salina as a template,the cD NA sequence of D. salina small GTP-binding protein gene was amplified by RT-PCR technique,and cloned onto pM Dl8-T simple vector,the recon was subjected to ...With the total RNA of Dunaliella salina as a template,the cD NA sequence of D. salina small GTP-binding protein gene was amplified by RT-PCR technique,and cloned onto pM Dl8-T simple vector,the recon was subjected to PCR detection and restriction endonuclease analysis,and the total sequence of DNA was determined. The results showed that the cloned fragment was 612 bp,and shared 100% homology with reported D. salina DsRab gene（ GenB ank： JN989548）. The target gene fragment was inserted downstream of pM DCG 35 S promoter,constructing subcellular localization recombinant vector pM DCG-DsRab. The successfully constructed subcellular localization recombinant vector pM DCG-DsRab was transformed into Agrobacterium tumefaciens LBA4404,and positive single clones were screened and used for transinfection of onion epidemical cells by Agrobacterium-mediated method,and the instant expression of DsRab was observed under fluorescence microscope. The results showed that the fusion protein GFP-DsRab was successfully expressed in onion epidemical cells,and mainly distributed on cytomembrane. This study will provide reference for further illumination of the function and action mechanism of D. salina small GTP-binding protein DsRab.展开更多
Stable carbon and nitrogen isotopes were used to investigate nutrient pathways and trophic relationships from the rice-crab system in Panjin, Liaoning Province, China. Values of 313C ranged from -27.38%±0.44%o to...Stable carbon and nitrogen isotopes were used to investigate nutrient pathways and trophic relationships from the rice-crab system in Panjin, Liaoning Province, China. Values of 313C ranged from -27.38%±0.44%o to -18.34%±0.26%o and δ^15N ranged from 1.10%o4-0.88%o to 9.33%±0.57%. Pseudorasbora parva （Stone moroko） had the highest δ^13C and 8tSN values. The lowest δ^13C values were obtained for the macrophytes and the lowest δ^15N value was found in sediments. Stable carbon and nitrogen isotopes were used to determine the contribution of different food items to the diets of crabs. The δ^13C results indicated that the Pseudorasbora parva made the greatest contribution to the diet of Eriocheir sinensis （Chinese mitten crab）, while the δ^15N results indicated that most food items contributed more than 10% to the diet of the crab. There were three trophic levels identified in the system （Levels 0-2）. The crab Eriocheir sinensis, fish Pseudorasbora parva and Misgurnus anguillicaudatus （Oriental weatherfish）, and the oligochaete Limnodrilus hoffmeisteri （Limnodrilus worm）, were at the second level, zooplankton were at the first level and suspended particulate matter and macrophytes were at trophic position 0.展开更多
We evaluated the effects of lithium on the survival, growth, and reproduction ofD. tibetana in the laboratory. The safe concentration value was 69.3 mg/L. The time to first brood was significant lower for individuals ...We evaluated the effects of lithium on the survival, growth, and reproduction ofD. tibetana in the laboratory. The safe concentration value was 69.3 mg/L. The time to first brood was significant lower for individuals reared in 5 and 10 mg/L （24.4 and 24.0 d, respectively） compared with individuals reared in 20, 40, or 60 mg/L. Females reared in 5 mg/L lithium produced a mean of 16.5 neonates/brood and had the highest number of broods （3.0±1.95）. The rate of egg production （a）, the intrinsic rate of increase （rm）, net reproduction rate （R0）, and finite rate of increase （2） were highest for D. tibetana reared at 5 and 10 mg/L lithium, and the duration of development was shorter than for the remaining groups. The results indicated that rearing in 5 10 mg/L lithium can accelerate the growth and reproduction ofD. tibetana.展开更多
The food sources of aquacultured Apostichopus japonicus and the trophic levels of organisms in a sea cucumber（A. japonicus） and prawn（Penaeus japonica） polyculture system in a saltwater pond in Zhuanghe, Liaoning ...The food sources of aquacultured Apostichopus japonicus and the trophic levels of organisms in a sea cucumber（A. japonicus） and prawn（Penaeus japonica） polyculture system in a saltwater pond in Zhuanghe, Liaoning Province were examined using carbon and nitrogen stable isotopes. Across organisms, δ13C ranged from（–25.47±0.20）‰ to（–16.48±0.17）‰（mean±SD）, and δ15N ranged from（4.23±0.49）‰ to（12.44±0.09）‰. The δ13C and δ15N contents of A. japonicus, P. japonica and Fenneropenaeus chinensis were comparatively higher than those of other organisms. Values of δ13C and δ15N revealed that P. japonica, Hemigrapsus sanguineus and Neomysis japonica comprised the largest component of the diet of A. japonicus. The mean trophic level of the organisms in this saltwater pond polyculture system was（2.75±0.08）. P. japonica, A. japonicus, F. chinensis,Synechogobius hasta and Neomysis japonica were in the 3rd trophic level（2–3）; jellyfish, H. sanguineus and zooplankton were in the 2nd trophic level（1–2）; and Enteromorpha prolifera, benthic microalgae, periphyton and suspended matter primarily consisting of phytoplankton, bacteria and humus were in the primary trophic level（0–1）.展开更多
In order to study the effect of temperature on the growth of juvenile rock scallop(Crassadoma gigantea),learn more about its growth with temperature changes under the conditions of indoor circulating water temperature...In order to study the effect of temperature on the growth of juvenile rock scallop(Crassadoma gigantea),learn more about its growth with temperature changes under the conditions of indoor circulating water temperature-controlled culture.Four temperature gradients of10℃,12.5℃,15℃and17.5℃were set,observed the growth of juvenile rock scallop at different test temperatures during two month.The results show:With the increase of test temperature,the growth rate first increased and then decreased.One-way analysis of variance showed that temperature had a significant effect on the growth rate(P<0.05),the optimum temperature was15℃,under this condition,the shell height average daily growth was significantly higher than the other three groups,the average daily growth of shell length and wet weight had no significant difference with17.5℃experimental group but significantly higher than the previous two groups.Within the experimental range,the change of temperature had no significant effect on the survival rate of juvenile rock scallop(P>0.05).展开更多
基金the functional analysis of PKC signaling pathway involved in response to salt stress of Dunaliella salinathe National Natural Science Foundation of China (No. 31472260)
文摘Protein kinase C (PKC) has a crucial role in signal transduction for a variety of biologically active substances which activate cellular functions and proliferation. We previously isolated the full-length PKC gene from Dunaliella salina (DsPKC) using rapid amplification of cDNA ends (RACE) and RT-PCR methods. And we submitted the mRNA sequence of DsPKC gene to NCBI (Genbank No. JN625213). In the present paper, the DsPKC gene open reading frame obtained by PCR was cloned into pGS-21a vector and transformed into Escherichia coli to generate the fusion protein. Bioinformatics analysis revealed that DsPKC gene was a member of serine/threonine kinase with two conserved domains and highly conserved motifs. The DsPKC was highly expressed upon induction with isopropyl-β-d-thiogalactoside (IPTG) at a final concentration of 0.2 mmol L 1 at 37℃. Under salt stress, the fu- sion protein Green Fluorescent Protein (GFP)-DsPKC was transferred from the cytoplasm to the cell membrane. The expression pat- tern of DsPKC gene was analyzed using real-time quantitative PCR, and indicated that DsPKC gene was up-regulated by 3.0 mol L 1 NaCl at 12 h, which was significantly higher than in control values (P < 0.05). These results suggest that the DsPKC gene plays an important role in response to salt stress in D. salina.
文摘The research status and the future study prospect of salt lake organisms of Chinese salt lakes were elaborated in this paper.Many saline lakes occur widely in arid and semiarid areas of China,and more than half of the total
基金Supported by the China Geological Survey(Resources No.[2002]004)the National Natural Science Foundation of China(No.30371112)+1 种基金the Liaoning Science and Technology Foundation(Nos.002119,20022100)the Special Program for Key Basic Research of Ministry of Science and Technology,China(No.2014FY210700)
文摘The seasonal variations in biomass, abundance, and species composition of plankton in relation to hydrography were studied in the saline Bange Lake, northern Tibet, China. Sampling was carried out between one to three times per month from May 2001 to July 2002. Salinity ranged from 14 to 146. The air and water temperature exhibited a clear seasonal pattern, and mean annual temperatures were approximately 4.8℃ and 7.3℃, respectively. The lowest water temperature occurred in winter from December to March at-2℃ and the highest in June and July at 17.7℃. Forty-one phytoplankton taxa, 21 zooplankton, and 5 benthic or facultative zooplankton were identifi ed. The predominant phytoplankton species were Gloeothece linearis, Oscillatoria tenuis, Gloeocapsa punctata, Ctenocladus circinnatus, Dunaliella salina, and Spirulina major. The predominant zooplankton species included H olophrya actra, Brachionus plicatilis, Daphniopsis tibetana, Cletocamptus dertersi, and A rctodiaptomus salinus. The mean annual total phytoplankton density and biomass for the entire lake were 4.52×10^7 cells/L and 1.60 mg/L, respectively. The annual mean zooplankton abundance was 52, 162, 322, and 57, 144 ind./L, in the three sublakes. The annual mean total zooplankton biomass in Lakes 1–3 was 1.23, 9.98, and 2.13 mg/L, respectively. The annual mean tychoplankton abundances in Bg1, 2, and 3 were 47, 67, and 654 ind./L. The annual mean tychoplankton biomass was 2.36, 0.16, and 2.03 mg/L, respectively. The zooplankton biomass（including tychoplankton） in the lake was 9.11 mg/L. The total number of plankton species in the salt lake was signifi cantly negatively correlated with salinity.
基金Supported by the National Natural Science Foundation of China(No.31472260)the Key Laboratory of Hydrobiology in Liaoning Province,College of Fisheries and Life Science,Dalian Ocean University
文摘Plants possess effective mechanisms to respond quickly to the external environment. Rapid activation of phosphatidylinositol-specific phospholipase C (PLC) enzymes occurs after a stimulus. The PLC in Dunaliella salina plays important roles in growth and stress responses. However, the molecular basis of PLC action in D. salina remains little understood. To gain insight into the potential biological functions of this enzyme, we cloned a phospholipase C gene from D. salina in a previous study, named DsPLC (GenBank No. KF573428). Here, we present the prokaryotic expression, purification, and characterization of the DsPLC gene. The entire coding region of DsPLC was inserted into an expression vector pET32a, and the DsPLC gene was successfully expressed in Escherichia coli. The DsPLC protein was purified and identified using a polyclonal antibody and western blotting. Expressing DsPLC fused with a green fluorescent protein (GFP) in onion showed that DsPLC-GFP was localized to the intracellular membrane. Quantitative real-time PCR analysis revealed that the relative expression of the DsPLC gene was induced significantly by 3.0-mol/L NaCl at 4 h. Our results support the importance of PLC enzymes in plant defense signaling. This study provides a basis for further functional studies of the DsPLC gene and for additional analysis of the potential roles of PLC enzymes in response to abiotic stress.
基金Supported by the National Natural Science Foundation of China(No.41501535)the Special Program for Key Basic Research of the Ministry of Science and Technology,China(No.2014FY210700)
文摘We investigated the eff ects of temperature（14,20,and 22℃）,salinity（5,10,15,20,and 25）,and body length（0.83,1.17,1.49,and 2.33 mm）on the energy budget of Daphniopsis tibetana Sars in the laboratory.The results demonstrated no diff erence in D.tibetana assimilation efficiency（AE）with values of 51.59%–56.22% at the trial temperatures.Gross growth efficiency（K1）and net growth efficiency（K2）were 28.78% and 55.71%,respectively,at 14℃.Daphniopsis tibetana energy conversion efficiency（AE,K1,and K2）was higher at salinities of 10–20,with values of 72.80%–77.31%,42.50%–50.42%,and 58.43%–65.02%,respectively.Daphniopsis tibetana energy conversion efficiency was highest when body length was 1.17–1.19 mm,with values of 83.70%–84.73%,50.11%–50.81%,and 59.13%–60.71%,respectively.The results of this study can provide a reference for large-scale cultivation use of D.tibetana in the further.
基金Supported by National Natural Science Foundation of China(31472260)
文摘With the total RNA of Dunaliella salina as a template,the cD NA sequence of D. salina small GTP-binding protein gene was amplified by RT-PCR technique,and cloned onto pM Dl8-T simple vector,the recon was subjected to PCR detection and restriction endonuclease analysis,and the total sequence of DNA was determined. The results showed that the cloned fragment was 612 bp,and shared 100% homology with reported D. salina DsRab gene（ GenB ank： JN989548）. The target gene fragment was inserted downstream of pM DCG 35 S promoter,constructing subcellular localization recombinant vector pM DCG-DsRab. The successfully constructed subcellular localization recombinant vector pM DCG-DsRab was transformed into Agrobacterium tumefaciens LBA4404,and positive single clones were screened and used for transinfection of onion epidemical cells by Agrobacterium-mediated method,and the instant expression of DsRab was observed under fluorescence microscope. The results showed that the fusion protein GFP-DsRab was successfully expressed in onion epidemical cells,and mainly distributed on cytomembrane. This study will provide reference for further illumination of the function and action mechanism of D. salina small GTP-binding protein DsRab.
基金Supported by the National Special Research Fund for Non-Profit Sector(Agriculture)(No.201203O81-7)
文摘Stable carbon and nitrogen isotopes were used to investigate nutrient pathways and trophic relationships from the rice-crab system in Panjin, Liaoning Province, China. Values of 313C ranged from -27.38%±0.44%o to -18.34%±0.26%o and δ^15N ranged from 1.10%o4-0.88%o to 9.33%±0.57%. Pseudorasbora parva （Stone moroko） had the highest δ^13C and 8tSN values. The lowest δ^13C values were obtained for the macrophytes and the lowest δ^15N value was found in sediments. Stable carbon and nitrogen isotopes were used to determine the contribution of different food items to the diets of crabs. The δ^13C results indicated that the Pseudorasbora parva made the greatest contribution to the diet of Eriocheir sinensis （Chinese mitten crab）, while the δ^15N results indicated that most food items contributed more than 10% to the diet of the crab. There were three trophic levels identified in the system （Levels 0-2）. The crab Eriocheir sinensis, fish Pseudorasbora parva and Misgurnus anguillicaudatus （Oriental weatherfish）, and the oligochaete Limnodrilus hoffmeisteri （Limnodrilus worm）, were at the second level, zooplankton were at the first level and suspended particulate matter and macrophytes were at trophic position 0.
基金Supported by the National Natural Science Foundation of China(Nos.30371112,30671625,40776065)
文摘We evaluated the effects of lithium on the survival, growth, and reproduction ofD. tibetana in the laboratory. The safe concentration value was 69.3 mg/L. The time to first brood was significant lower for individuals reared in 5 and 10 mg/L （24.4 and 24.0 d, respectively） compared with individuals reared in 20, 40, or 60 mg/L. Females reared in 5 mg/L lithium produced a mean of 16.5 neonates/brood and had the highest number of broods （3.0±1.95）. The rate of egg production （a）, the intrinsic rate of increase （rm）, net reproduction rate （R0）, and finite rate of increase （2） were highest for D. tibetana reared at 5 and 10 mg/L lithium, and the duration of development was shorter than for the remaining groups. The results indicated that rearing in 5 10 mg/L lithium can accelerate the growth and reproduction ofD. tibetana.
基金The National Marine Public Welfare Project of China under contract No.201305005
文摘The food sources of aquacultured Apostichopus japonicus and the trophic levels of organisms in a sea cucumber（A. japonicus） and prawn（Penaeus japonica） polyculture system in a saltwater pond in Zhuanghe, Liaoning Province were examined using carbon and nitrogen stable isotopes. Across organisms, δ13C ranged from（–25.47±0.20）‰ to（–16.48±0.17）‰（mean±SD）, and δ15N ranged from（4.23±0.49）‰ to（12.44±0.09）‰. The δ13C and δ15N contents of A. japonicus, P. japonica and Fenneropenaeus chinensis were comparatively higher than those of other organisms. Values of δ13C and δ15N revealed that P. japonica, Hemigrapsus sanguineus and Neomysis japonica comprised the largest component of the diet of A. japonicus. The mean trophic level of the organisms in this saltwater pond polyculture system was（2.75±0.08）. P. japonica, A. japonicus, F. chinensis,Synechogobius hasta and Neomysis japonica were in the 3rd trophic level（2–3）; jellyfish, H. sanguineus and zooplankton were in the 2nd trophic level（1–2）; and Enteromorpha prolifera, benthic microalgae, periphyton and suspended matter primarily consisting of phytoplankton, bacteria and humus were in the primary trophic level（0–1）.
基金supported by Liaoning Provincial Education Department Project(NO.L201624)
文摘In order to study the effect of temperature on the growth of juvenile rock scallop(Crassadoma gigantea),learn more about its growth with temperature changes under the conditions of indoor circulating water temperature-controlled culture.Four temperature gradients of10℃,12.5℃,15℃and17.5℃were set,observed the growth of juvenile rock scallop at different test temperatures during two month.The results show:With the increase of test temperature,the growth rate first increased and then decreased.One-way analysis of variance showed that temperature had a significant effect on the growth rate(P<0.05),the optimum temperature was15℃,under this condition,the shell height average daily growth was significantly higher than the other three groups,the average daily growth of shell length and wet weight had no significant difference with17.5℃experimental group but significantly higher than the previous two groups.Within the experimental range,the change of temperature had no significant effect on the survival rate of juvenile rock scallop(P>0.05).
