在从武汉东湖水样中培养分离水华蓝藻噬藻体(Planktothrix agardhii Virus from Lake Donghu,PaV-LD)的基础上,对在不同条件培养的宿主蓝藻细胞中,PaV-LD增殖效率及裂解作用进行了测定分析。分别将PaV-LD接种到生长期、半连续培养更新...在从武汉东湖水样中培养分离水华蓝藻噬藻体(Planktothrix agardhii Virus from Lake Donghu,PaV-LD)的基础上,对在不同条件培养的宿主蓝藻细胞中,PaV-LD增殖效率及裂解作用进行了测定分析。分别将PaV-LD接种到生长期、半连续培养更新率或光照不同的宿主蓝藻液中,并采用稀释培养计数(Mostprobable number,MPN)方法与电镜观察,测定子代PaV-LD释放量及宿主细胞的裂解作用。结果显示:对数生长期宿主蓝藻单个细胞中子代PaV-LD的平均释放量为350感染单位(Infectious Units,IU/cell),显著高于稳定生长期的平均释放量110 IU/cell。在用新鲜培养基更新率为0%、35%、50%和65%的半连续培养宿主蓝藻中,接种PaV-LD 5d之后,噬藻体的释放量分别约为50 IU/cell、70 IU/cell、220 IU/cell或310 IU/cell,表明子代PaV-LD释放率随培养基更新率的增加而显著提高。在光照条件下感染3—4d后,宿主蓝藻细胞充分裂解,并释放大量子代PaV-LD,滴度可由初始7.00×103IU/mL快速增加到8.56×107IU/mL;但在遮光条件下,同样感染的蓝藻细胞未见裂解,也检测不到释放的子代噬藻体。电镜观察显示,在光照条件下感染的蓝藻细胞类囊体膜结构消失,而大量子代PaV-LD颗粒主要分布在原有类囊体的部位。显然,宿主蓝藻细胞的培养条件和状态可能对获得噬藻体纯培养有决定性影响。展开更多
The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the ...The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the present study. The species was cultured in BG11 liquid medium contained various initial concentrations of Pb^2＋ and Cd〉 （0, 0.5, 1, 2, 4, 6 and 8 mg/L）. The experiment was conducted for six days and the metal induced alterations in the ultrastructure, growth and pigment contents were assessed. Alterations in the ultrastructure of the Synechocystis sp. PCC 6803 cells became evident with the increased （〉4 mg/L Pb^2＋） metal concentration. The photosynthetic apparatus （thylakoid membranes） were found to be the worst affected. Deteriorated or completely destroyed thylakoid membranes have made large empty spaces in the cell interior. In addition, at the highest concentration （8 mg/L pb^2＋）, the polyphosphate granules became more prominent both in size and number. Despite the initial slight stimulations （0.2, 3.8 and 6.5% respectively at 0.5, 1 and 2 mg/L pb^2＋）, both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents （chlorophyll a, 13 carotene and phycocyanin） were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb^2＋, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll a, 13 carotene and phycocyanin respectively over the control. Corresponding reductions for the same CdZ＋concentrations were 57.83, 48.94 and 56.90%. Lethal concentration （96 h LC50） values （3.47 mg/L Cd^2＋ and 12.11 mg/L Pb^2＋） indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd^2＋ than Pb^2＋.展开更多
Gas vesicles are hollow, air-filled polyprotein structures that provide the buoyancy to cells. They are found in a variety of prokaryotes. In this study, we isolated a partial gas vesicle protein gene cluster containi...Gas vesicles are hollow, air-filled polyprotein structures that provide the buoyancy to cells. They are found in a variety of prokaryotes. In this study, we isolated a partial gas vesicle protein gene cluster containing gvpA and gvpC20ψ from Planktothrix rubescens, and inserted it into an expression vector and expressed it in E. coli. The gas vesicle was developed in bacterial cells, which made bacterial cells to float on medium surface. We also amplified gvpA and gvpC20ψ separately and synthesized an artificial operon by fusing these two genes with the standardized gene expression controlling elements of E. coli. The artificial operon was expressed in E. coli, forming gas vesicles and floating bacteria cells. Our findings verified that the whole set of genes and the overall structure of gas vesicle gene cluster are not necessary for developing gas vesicles in bacteria cells. Two genes, gvpA and gvpC20ψ, of the gas vesicle gene cluster are sufficient for synthesizing an artificial operon that can develop gas vesicles in bacteria cells. Our findings provided a wide range of applications including easing the harvest of cultured microalgae and bacteria, as well as enriching and remediating aquatic pollutants by constructing gas vesicles in their ceils.展开更多
The biosorption mechanism of Cr （Ⅳ） ions on Synechococcus sp. biosorbent was studied by analyzing the biosorption kinetics as well as speciation change and bond formation during the biosorption process. The kinetic...The biosorption mechanism of Cr （Ⅳ） ions on Synechococcus sp. biosorbent was studied by analyzing the biosorption kinetics as well as speciation change and bond formation during the biosorption process. The kinetics study shows that the adsorption process of Cr （Ⅳ） consists of a very fast stage in the first several minutes, in which more than half of the saturation adsorption is attained, and a slower stage that approximately follows the first order kinetic model, basically Freundlich isotherm models were observed. Comparative studies of FT-LR spectra of K2Cr2O7, free cells of Synechococcus sp., and Cr-bound cells of Synechococcus sp show that the speciation of chromium that binds to the cells ofSynechococcus sp. is Cr （Ⅲ）, instead of Cr （Ⅳ）, and the carboxylic, alcoholic, amido and amino groups may be involved in the binding of Cr （Ⅲ）. Integrative analyses of the surface electric potential, the effect of pH value on adsorption behavior of Cr （Ⅵ）, and the results of FT-IR show that the biosorption of Cr （Ⅵ） follows two subsequent steps, biosorption of Cr2O7 ^2- by electrostatical force at the protonated active sites and reduction of Cr2O7^2- to Cr^3＋ by the reductive groups on the surface of the biosorbents.展开更多
The freshwater cyanobacterium, Cylindrospermopsis raciborskii （Woloszyflska） Seenayya and Subba Raju is a common species in lakes and reservoirs globally. In some areas of the world it can produce cytoand hepatotoxi...The freshwater cyanobacterium, Cylindrospermopsis raciborskii （Woloszyflska） Seenayya and Subba Raju is a common species in lakes and reservoirs globally. In some areas of the world it can produce cytoand hepatotoxins （cylindrospermopsins, saxitoxins）, making blooms of this species a serious health concern for humans. In the last 10 15 years, there has been a considerable body of research conducted on the ecology, physiology and toxin production of this species and this paper reviews these studies with a focus on the cylindrospermopsin （CYN）-producing strains. C. raciborskii has low light requirements, close to neutral buoyancy, and a wide temperature tolerance, giving it the capacity to grow in many lentic waterbodies. It also has a flexible strategy with respect to nitrogen （N） utilisation; being able to switch between utilising fixed and atmospheric N as sources of N fluctuate. Additionally this species has a high phosphate （DIP） affinity and storage capacity. Like many cyanobacteria, it also has the capacity to use dissolved organic phosphorus （DOP）. Changes in nutrient concentrations, light levels and temperature have also been found to affect production of the toxin CYN by this species. However, optimal toxin production does not necessarily occur when growth rates are optimal. Additionally, different strains of C. raciborskii vary in their cell quota of CYN, making it difficult to predict toxin concentrations, based on C. raciborskii cell densities. In summary, the ecological flexibility of this organism means that controlling blooms of C. raciborskii is a difficult undertaking. However, improved understanding of factors promoting the species and toxin production by genetically capable strains will lead to improved predictive models of blooms.展开更多
Eutrophication has become a serious concern in many lakes, resulting in cyanobacterial blooms. However, the mechanism and pathways of cyanobacteria decline are less understood. To identify and define the growth and de...Eutrophication has become a serious concern in many lakes, resulting in cyanobacterial blooms. However, the mechanism and pathways of cyanobacteria decline are less understood. To identify and define the growth and decline of Microcystis blooms in Taihu Lake of China, and to illuminate the destination of surface floating blooms, we investigated the biomass distribution and variations in colony size, morphology, and floating velocity from October 2008 to September 2009. The results showed that the Microcystis bloom declined in response to biomass decrease, colony disaggregation, buoyancy reduction, and increased phytoplankton biodiversity, and these indicative parameters could be applied for recognition of the development phases of the bloom. Three major decline pathways were proposed to describe the bloom decline process, colony disaggregation （Pathway I）, colony settlement （Pathway II）, and cell lysis in colonies （Pathway III）. We proposed a strategy to define the occurrence and decline of Microcystis blooms, to evaluate the survival state under different stress conditions, and to indicate the efficiency of controlling countermeasures against algal blooms.展开更多
文摘The unicellular cyanobacterium Synechocystis sp. PCC 6803, a model organism known for its unique combination of highly desirable molecular genetic, physiological and morphological characteristics, was employed in the present study. The species was cultured in BG11 liquid medium contained various initial concentrations of Pb^2＋ and Cd〉 （0, 0.5, 1, 2, 4, 6 and 8 mg/L）. The experiment was conducted for six days and the metal induced alterations in the ultrastructure, growth and pigment contents were assessed. Alterations in the ultrastructure of the Synechocystis sp. PCC 6803 cells became evident with the increased （〉4 mg/L Pb^2＋） metal concentration. The photosynthetic apparatus （thylakoid membranes） were found to be the worst affected. Deteriorated or completely destroyed thylakoid membranes have made large empty spaces in the cell interior. In addition, at the highest concentration （8 mg/L pb^2＋）, the polyphosphate granules became more prominent both in size and number. Despite the initial slight stimulations （0.2, 3.8 and 6.5% respectively at 0.5, 1 and 2 mg/L pb^2＋）, both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents （chlorophyll a, 13 carotene and phycocyanin） were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb^2＋, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll a, 13 carotene and phycocyanin respectively over the control. Corresponding reductions for the same CdZ＋concentrations were 57.83, 48.94 and 56.90%. Lethal concentration （96 h LC50） values （3.47 mg/L Cd^2＋ and 12.11 mg/L Pb^2＋） indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd^2＋ than Pb^2＋.
基金supported by Undergraduate Institution of Marine Biological Science, Ocean University of China (OUC)
文摘Gas vesicles are hollow, air-filled polyprotein structures that provide the buoyancy to cells. They are found in a variety of prokaryotes. In this study, we isolated a partial gas vesicle protein gene cluster containing gvpA and gvpC20ψ from Planktothrix rubescens, and inserted it into an expression vector and expressed it in E. coli. The gas vesicle was developed in bacterial cells, which made bacterial cells to float on medium surface. We also amplified gvpA and gvpC20ψ separately and synthesized an artificial operon by fusing these two genes with the standardized gene expression controlling elements of E. coli. The artificial operon was expressed in E. coli, forming gas vesicles and floating bacteria cells. Our findings verified that the whole set of genes and the overall structure of gas vesicle gene cluster are not necessary for developing gas vesicles in bacteria cells. Two genes, gvpA and gvpC20ψ, of the gas vesicle gene cluster are sufficient for synthesizing an artificial operon that can develop gas vesicles in bacteria cells. Our findings provided a wide range of applications including easing the harvest of cultured microalgae and bacteria, as well as enriching and remediating aquatic pollutants by constructing gas vesicles in their ceils.
基金Project(50321402) supported by the National Natural Science Foundation of China
文摘The biosorption mechanism of Cr （Ⅳ） ions on Synechococcus sp. biosorbent was studied by analyzing the biosorption kinetics as well as speciation change and bond formation during the biosorption process. The kinetics study shows that the adsorption process of Cr （Ⅳ） consists of a very fast stage in the first several minutes, in which more than half of the saturation adsorption is attained, and a slower stage that approximately follows the first order kinetic model, basically Freundlich isotherm models were observed. Comparative studies of FT-LR spectra of K2Cr2O7, free cells of Synechococcus sp., and Cr-bound cells of Synechococcus sp show that the speciation of chromium that binds to the cells ofSynechococcus sp. is Cr （Ⅲ）, instead of Cr （Ⅳ）, and the carboxylic, alcoholic, amido and amino groups may be involved in the binding of Cr （Ⅲ）. Integrative analyses of the surface electric potential, the effect of pH value on adsorption behavior of Cr （Ⅵ）, and the results of FT-IR show that the biosorption of Cr （Ⅵ） follows two subsequent steps, biosorption of Cr2O7 ^2- by electrostatical force at the protonated active sites and reduction of Cr2O7^2- to Cr^3＋ by the reductive groups on the surface of the biosorbents.
文摘The freshwater cyanobacterium, Cylindrospermopsis raciborskii （Woloszyflska） Seenayya and Subba Raju is a common species in lakes and reservoirs globally. In some areas of the world it can produce cytoand hepatotoxins （cylindrospermopsins, saxitoxins）, making blooms of this species a serious health concern for humans. In the last 10 15 years, there has been a considerable body of research conducted on the ecology, physiology and toxin production of this species and this paper reviews these studies with a focus on the cylindrospermopsin （CYN）-producing strains. C. raciborskii has low light requirements, close to neutral buoyancy, and a wide temperature tolerance, giving it the capacity to grow in many lentic waterbodies. It also has a flexible strategy with respect to nitrogen （N） utilisation; being able to switch between utilising fixed and atmospheric N as sources of N fluctuate. Additionally this species has a high phosphate （DIP） affinity and storage capacity. Like many cyanobacteria, it also has the capacity to use dissolved organic phosphorus （DOP）. Changes in nutrient concentrations, light levels and temperature have also been found to affect production of the toxin CYN by this species. However, optimal toxin production does not necessarily occur when growth rates are optimal. Additionally, different strains of C. raciborskii vary in their cell quota of CYN, making it difficult to predict toxin concentrations, based on C. raciborskii cell densities. In summary, the ecological flexibility of this organism means that controlling blooms of C. raciborskii is a difficult undertaking. However, improved understanding of factors promoting the species and toxin production by genetically capable strains will lead to improved predictive models of blooms.
基金Supported by the National Basic Research Program (973 Program)(No. 2008CB418002)the National Major Programs of Water Body Pollution Control and Remediation (Nos. 2009ZX07104-005,2009ZX07106-001)
文摘Eutrophication has become a serious concern in many lakes, resulting in cyanobacterial blooms. However, the mechanism and pathways of cyanobacteria decline are less understood. To identify and define the growth and decline of Microcystis blooms in Taihu Lake of China, and to illuminate the destination of surface floating blooms, we investigated the biomass distribution and variations in colony size, morphology, and floating velocity from October 2008 to September 2009. The results showed that the Microcystis bloom declined in response to biomass decrease, colony disaggregation, buoyancy reduction, and increased phytoplankton biodiversity, and these indicative parameters could be applied for recognition of the development phases of the bloom. Three major decline pathways were proposed to describe the bloom decline process, colony disaggregation （Pathway I）, colony settlement （Pathway II）, and cell lysis in colonies （Pathway III）. We proposed a strategy to define the occurrence and decline of Microcystis blooms, to evaluate the survival state under different stress conditions, and to indicate the efficiency of controlling countermeasures against algal blooms.
