Synthetic biology efforts have also led to the development of photosynthetic cyanobacteria as"autotrophic cell factories"for biosynthesis of various biofuels directly from CO_(2).However,the low tolerance to...Synthetic biology efforts have also led to the development of photosynthetic cyanobacteria as"autotrophic cell factories"for biosynthesis of various biofuels directly from CO_(2).However,the low tolerance to toxicity of biofuels has restricted the economic application of cyanobacterial hosts.In this study,RNAseq transcriptomics was employed to reveal stress responses to exogenous n-hexane in Synechocystis sp.PCC 6803.Functional enrichment analysis of the transcriptomic data showed that signal transduction systems were induced significantly.To further identify regulatory genes related to n-hexane tolerance,a library of transcriptional regulators(TRs)deletion mutants was then screened for their roles in nhexane tolerance.The results showed that a knockout mutant of slr0724 that encodes an Hta R suppressor protein was more tolerant to n-hexane than the wild type,indicating the involvement of slr0724 in nhexane tolerance.This study provides the foundation for better understanding the cellular responses to n-hexane in Synechocystis sp.PCC 6803,which could contribute to the further engineering of nhexane tolerance in cyanobacteria.展开更多
Hemolysin produced by various bacteria,may destroy erythrocyte membranes via a pore-forming mechanism,a deter-gent action,or a lipase activity.Previous to this experiment,the mode of action used by cyanobacterial hemo...Hemolysin produced by various bacteria,may destroy erythrocyte membranes via a pore-forming mechanism,a deter-gent action,or a lipase activity.Previous to this experiment,the mode of action used by cyanobacterial hemolysin had not been re-ported.To characterize the action mode of hemolysin produced by the wild-type strain of Synechocystis sp.PCC6803,hemolysis of erythrocytes originating from human,mouse,sheep,rabbit and goldfish was studied.The erythrocytes of mouse,sheep and rabbit were sensitive,while those of human and fish were resistant,to this hemolysin.Using rabbit erythrocytes,it was shown that hemoly-sis occurred in two steps:a binding step within the first 10 min of treatment and a lytic step after 30 min.Both binding and lysis were highly temperature-dependent.Effects of erythrocyte density on hemolysis suggest that the hemolysin might target erythrocytes via a multiple-hit mechanism.In the osmotic protection experiment,all tested osmotic protectants,with molecular diameters ranging from 0.9 ?5.66 nm,failed to effectively inhibit hemolysis.Scanning electron micrographs showed that the hemolysin caused protuberances or echinocytes in rabbit erythrocytes,and then disrupted and ruptured the erythrocytes.Characteristics of hemolysis showed distinct differences from other pore-forming mechanisms,suggesting that this hemolysin might act through a detergent-like or lipase mecha-nism,rather than a pore-forming mechanism.展开更多
Objective:To explore the ability of Synechocystis sp.PCC6803 in transforming 6-deoxypseudoanisatin.Methods:The experiment was performed by incubating 6-deoxypseudoanisatin with the freshwater cyanobacterium Synechocys...Objective:To explore the ability of Synechocystis sp.PCC6803 in transforming 6-deoxypseudoanisatin.Methods:The experiment was performed by incubating 6-deoxypseudoanisatin with the freshwater cyanobacterium Synechocystis sp.PCC6803 under continuous white light at 30C for 5 days.The crude converted product was detected using thin-layer chromatography(TLC)and further analyzed using high-performance liquid chromatography(HPLC)as well as HPLC with electron spray ionization mass spectrometry(HPLC-ESI-MS).Results:TLC results showed that 6-deoxypseudoanisatin was converted into a less polar product.HPLC and MS data indicated that the retention time of the converted product increased in comparison with the standard of 6-deoxypseudoanisatin.Conclusion:Thus,the study appears to demonstrate that Synechocystis sp.PCC6803 can transform 6-deoxypseudoanisatin.The polarity of the converted product is less than that of 6-deoxypseudoanisatin.展开更多
Calcium carbonate(CaCO 3) crystals in their preferred orientation were obtained in BG11 culture media inoculated with Synechocystis sp. PCC6803(inoculated BG11). In this study, the features of calcium carbonate deposi...Calcium carbonate(CaCO 3) crystals in their preferred orientation were obtained in BG11 culture media inoculated with Synechocystis sp. PCC6803(inoculated BG11). In this study, the features of calcium carbonate deposition were investigated. Inoculated BG11 in different calcium ion concentrations was used for the experimental group, while the BG11 culture medium was used for the control group. The surface morphologies of the calcium carbonate deposits in the experimental and control groups were determined by scanning and transmission electron microscopy. The deposits were analyzed by electronic probe micro-analysis, Fourier transform infrared spectrum, X-ray diffraction, thermal gravimetric analysis and differential scanning calorimetry. The results show that the surfaces of the crystals in the experimental group were hexahedral in a scaly pattern. The particle sizes were micrometer-sized and larger than those in the control group. The deposits of the control group contained calcium(Ca), carbon(C), oxygen(O), phosphorus(P), iron(Fe), copper(Cu), zinc(Zn), and other elements. The deposits in the experimental group contained Ca, C, and O only. The deposits of both groups contained calcite. The thermal decomposition temperature of the deposits in the control group was lower than those in the experimental group. It showed that the CaCO 3 deposits of the experimental group had higher thermal stability than those of the control group. This may be due to the secondary metabolites produced by the algae cells, which affect the carbonate crystal structure and result in a close-packed structure. The algae cells that remained after thermal weight loss were heavier in higher calcium concentrations in BG11 culture media. There may be more calciumcontaining crystals inside and outside of these cells. These results shall be benefi cial for understanding the formation mechanism of carbonate minerals.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 Pb2+ and Cd2+ (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 Pb2+) 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 Pb2+), 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 Pb2+), both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents (chlorophyll a, β carotene and phycocyanin) were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb2+, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll a, β carotene and phycocyanin respectively over the control. Corresponding reductions for the same Cd2+concentrations were 57.83, 48.94 and 56.90%. Lethal concentration (96 h LC50) values (3.47 mg/L Cd2+ and 12.11 mg/L Pb2+) indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd2+ than Pb2+.展开更多
Synechocystis sp. PCC 6803 is a model organism widely used in cyanobacterium biology and biotechnology. To know the genetic background of substrains of Synechocystis sp. PCC 6803 is important for further research and ...Synechocystis sp. PCC 6803 is a model organism widely used in cyanobacterium biology and biotechnology. To know the genetic background of substrains of Synechocystis sp. PCC 6803 is important for further research and application. In this study, we reported the genome sequences of two non-motile wild-type substrains of Synechocystis sp. PCC 6803 using whole genome resequencing. 55/56 putative single nucleotide polymorphisms(SNPs) and 8/9 Indels(insertion and deletion) were identified. Among these, 47 SNPs were found in both the GT-AR and GT-CH strains, and 8 were unique to GT-AR and 9 were unique to GT-CH. All of these variations were annotated in metabolism pathway referred to KEGG database. Meanwhile, the deletion in s lr0332 gene was detected in these two strains, which attributed to the non-motile phenotype of them and suggested that the insertion in spkA gene was not essential for non-motile phenotype. These resequencing data provide the genetic background information of these two strains and highlighted the microevolution over decades of laboratory cultivation.展开更多
In this study,cDNA microarrays were developed from 3569 mRNA reads to analyze the expression profi les of the transcriptomes of Synechocystis sp.PCC6803 under low temperature(LT)stress.Among the genes on the cDNA micr...In this study,cDNA microarrays were developed from 3569 mRNA reads to analyze the expression profi les of the transcriptomes of Synechocystis sp.PCC6803 under low temperature(LT)stress.Among the genes on the cDNA microarrays,899 LT-affected genes exhibited a 1.5-fold(or greater)difference in expression compared with the genes from normal unstressed Synechocystis sp.PCC6803.Of the differentially expressed genes,353 were up-regulated and 246 were down-regulated.The results showed that genes involved in photosynthesis were activated at LT(10°C),including genes for photosystem I,photosystem II,photosynthetic electron transport,and cytochrome b6/f complex.Moreover,desB,one of four genes that encode the fatty acid desaturases,was also induced by LT.However,the LT conditions to some degree enhanced the transcription of some genes.In addition,LT(10°C)may reduce cellular motility by regulating the transcription of spkA(sll1575),a serine/threonine protein kinase.The results reported in this study may contribute to a better understanding of the responses of the Synechocystis cell to LT,including pathways involved in photosynthesis and repair.展开更多
基金supported by grants from the National Key Research and Development Program of China(2020YFA0906800,2021YFA0909700,2018YFA0903600 and 2019YFA0904600)。
文摘Synthetic biology efforts have also led to the development of photosynthetic cyanobacteria as"autotrophic cell factories"for biosynthesis of various biofuels directly from CO_(2).However,the low tolerance to toxicity of biofuels has restricted the economic application of cyanobacterial hosts.In this study,RNAseq transcriptomics was employed to reveal stress responses to exogenous n-hexane in Synechocystis sp.PCC 6803.Functional enrichment analysis of the transcriptomic data showed that signal transduction systems were induced significantly.To further identify regulatory genes related to n-hexane tolerance,a library of transcriptional regulators(TRs)deletion mutants was then screened for their roles in nhexane tolerance.The results showed that a knockout mutant of slr0724 that encodes an Hta R suppressor protein was more tolerant to n-hexane than the wild type,indicating the involvement of slr0724 in nhexane tolerance.This study provides the foundation for better understanding the cellular responses to n-hexane in Synechocystis sp.PCC 6803,which could contribute to the further engineering of nhexane tolerance in cyanobacteria.
基金the National Natural Science Fund of China (No. 30870250)Shandong Provincial NaturalScience Fund (No. Q2006D09)
文摘Hemolysin produced by various bacteria,may destroy erythrocyte membranes via a pore-forming mechanism,a deter-gent action,or a lipase activity.Previous to this experiment,the mode of action used by cyanobacterial hemolysin had not been re-ported.To characterize the action mode of hemolysin produced by the wild-type strain of Synechocystis sp.PCC6803,hemolysis of erythrocytes originating from human,mouse,sheep,rabbit and goldfish was studied.The erythrocytes of mouse,sheep and rabbit were sensitive,while those of human and fish were resistant,to this hemolysin.Using rabbit erythrocytes,it was shown that hemoly-sis occurred in two steps:a binding step within the first 10 min of treatment and a lytic step after 30 min.Both binding and lysis were highly temperature-dependent.Effects of erythrocyte density on hemolysis suggest that the hemolysin might target erythrocytes via a multiple-hit mechanism.In the osmotic protection experiment,all tested osmotic protectants,with molecular diameters ranging from 0.9 ?5.66 nm,failed to effectively inhibit hemolysis.Scanning electron micrographs showed that the hemolysin caused protuberances or echinocytes in rabbit erythrocytes,and then disrupted and ruptured the erythrocytes.Characteristics of hemolysis showed distinct differences from other pore-forming mechanisms,suggesting that this hemolysin might act through a detergent-like or lipase mecha-nism,rather than a pore-forming mechanism.
基金Beijing University of Chinese Medicine(2013-JYBZZ-JS-139)Independent Project Topics Foundation.
文摘Objective:To explore the ability of Synechocystis sp.PCC6803 in transforming 6-deoxypseudoanisatin.Methods:The experiment was performed by incubating 6-deoxypseudoanisatin with the freshwater cyanobacterium Synechocystis sp.PCC6803 under continuous white light at 30C for 5 days.The crude converted product was detected using thin-layer chromatography(TLC)and further analyzed using high-performance liquid chromatography(HPLC)as well as HPLC with electron spray ionization mass spectrometry(HPLC-ESI-MS).Results:TLC results showed that 6-deoxypseudoanisatin was converted into a less polar product.HPLC and MS data indicated that the retention time of the converted product increased in comparison with the standard of 6-deoxypseudoanisatin.Conclusion:Thus,the study appears to demonstrate that Synechocystis sp.PCC6803 can transform 6-deoxypseudoanisatin.The polarity of the converted product is less than that of 6-deoxypseudoanisatin.
基金Supported by the National Natural Science Foundation of China(Nos.40972043,41040018,41210104058,21176145,41372108,41302079)the Higher Educational Science and Technology Program of Shandong Province(No.J10LC15)+4 种基金the China Postdoctoral Science Foundation(No.2013M540560)the Program for Scientific Research Innovation Team in Colleges and Universities of Shandong Province,and SDUST Research Fund(No.2010KYTD103)the Open Project of Key Lab of Marine Bioactive Substance and Modern Analytical Technique,State Oceanic Administration,China(No.MBSMAT-2012-03)the Scientific and Technological Program of Qingdao(No.13-1-4-232-jch)the Domestic Visiting Scholar Program for Young Core Teachers in Shandong Universities,Shandong Province,China
文摘Calcium carbonate(CaCO 3) crystals in their preferred orientation were obtained in BG11 culture media inoculated with Synechocystis sp. PCC6803(inoculated BG11). In this study, the features of calcium carbonate deposition were investigated. Inoculated BG11 in different calcium ion concentrations was used for the experimental group, while the BG11 culture medium was used for the control group. The surface morphologies of the calcium carbonate deposits in the experimental and control groups were determined by scanning and transmission electron microscopy. The deposits were analyzed by electronic probe micro-analysis, Fourier transform infrared spectrum, X-ray diffraction, thermal gravimetric analysis and differential scanning calorimetry. The results show that the surfaces of the crystals in the experimental group were hexahedral in a scaly pattern. The particle sizes were micrometer-sized and larger than those in the control group. The deposits of the control group contained calcium(Ca), carbon(C), oxygen(O), phosphorus(P), iron(Fe), copper(Cu), zinc(Zn), and other elements. The deposits in the experimental group contained Ca, C, and O only. The deposits of both groups contained calcite. The thermal decomposition temperature of the deposits in the control group was lower than those in the experimental group. It showed that the CaCO 3 deposits of the experimental group had higher thermal stability than those of the control group. This may be due to the secondary metabolites produced by the algae cells, which affect the carbonate crystal structure and result in a close-packed structure. The algae cells that remained after thermal weight loss were heavier in higher calcium concentrations in BG11 culture media. There may be more calciumcontaining crystals inside and outside of these cells. These results shall be benefi cial for understanding the formation mechanism of carbonate minerals.
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 Pb2+ and Cd2+ (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 Pb2+) 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 Pb2+), 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 Pb2+), both metals inhibited the growth in a dose-dependent manner as incubation progressed. Pigment contents (chlorophyll a, β carotene and phycocyanin) were also decreased with increasing metal concentration. Cells exposed to 6 mg/L Pb2+, resulted in 36.56, 37.39 and 29.34% reductions of chlorophyll a, β carotene and phycocyanin respectively over the control. Corresponding reductions for the same Cd2+concentrations were 57.83, 48.94 and 56.90%. Lethal concentration (96 h LC50) values (3.47 mg/L Cd2+ and 12.11 mg/L Pb2+) indicated that Synechocystis sp. PCC 6803 is more vulnerable to Cd2+ than Pb2+.
基金Supported by the National Key Research and Development Program of China(No.2016YFF0202304)the National Natural Science Foundation of China(No.41376139)+1 种基金the Science and Technology Program of Yantai City(No.2016JHZB007)Natural Science Foundation of Shandong Province,China(No.ZR2016CM48)
文摘Synechocystis sp. PCC 6803 is a model organism widely used in cyanobacterium biology and biotechnology. To know the genetic background of substrains of Synechocystis sp. PCC 6803 is important for further research and application. In this study, we reported the genome sequences of two non-motile wild-type substrains of Synechocystis sp. PCC 6803 using whole genome resequencing. 55/56 putative single nucleotide polymorphisms(SNPs) and 8/9 Indels(insertion and deletion) were identified. Among these, 47 SNPs were found in both the GT-AR and GT-CH strains, and 8 were unique to GT-AR and 9 were unique to GT-CH. All of these variations were annotated in metabolism pathway referred to KEGG database. Meanwhile, the deletion in s lr0332 gene was detected in these two strains, which attributed to the non-motile phenotype of them and suggested that the insertion in spkA gene was not essential for non-motile phenotype. These resequencing data provide the genetic background information of these two strains and highlighted the microevolution over decades of laboratory cultivation.
基金Supported by the National Natural Science Foundation of China(No.40876082)the International Innovation Partnership Program:Typical Environmental Process and Effects on Resources in Coastal Zone Area+3 种基金the Public Science and Technology Research Funds Projects of the Ocean(Nos.200905021,201205027)the Outstanding Young Scholars Fellowship of Shandong Province(Molecular Phycology,No.JQ200914)the Natural Science Foundation of Shandong Province(No.ZR2012DQ015)the Guangdong Province Comprehensive Strategic Cooperation Project of the Chinese Academy of Sciences(No.2011A090100040)
文摘In this study,cDNA microarrays were developed from 3569 mRNA reads to analyze the expression profi les of the transcriptomes of Synechocystis sp.PCC6803 under low temperature(LT)stress.Among the genes on the cDNA microarrays,899 LT-affected genes exhibited a 1.5-fold(or greater)difference in expression compared with the genes from normal unstressed Synechocystis sp.PCC6803.Of the differentially expressed genes,353 were up-regulated and 246 were down-regulated.The results showed that genes involved in photosynthesis were activated at LT(10°C),including genes for photosystem I,photosystem II,photosynthetic electron transport,and cytochrome b6/f complex.Moreover,desB,one of four genes that encode the fatty acid desaturases,was also induced by LT.However,the LT conditions to some degree enhanced the transcription of some genes.In addition,LT(10°C)may reduce cellular motility by regulating the transcription of spkA(sll1575),a serine/threonine protein kinase.The results reported in this study may contribute to a better understanding of the responses of the Synechocystis cell to LT,including pathways involved in photosynthesis and repair.
