为研究我国浮丝藻(Planktothrix Anagnostidis et Komárek)的毒素相关基因,选取分离自我国不同省份水体的13株阿氏浮丝藻,通过PCR检测其微囊藻毒素合成酶基因mcyA、mcyE及mcyT研究其毒素基因特性。PCR结果表明除mcyT之外其他引物...为研究我国浮丝藻(Planktothrix Anagnostidis et Komárek)的毒素相关基因,选取分离自我国不同省份水体的13株阿氏浮丝藻,通过PCR检测其微囊藻毒素合成酶基因mcyA、mcyE及mcyT研究其毒素基因特性。PCR结果表明除mcyT之外其他引物检测均无扩增产物,说明这13株浮丝藻不具备产微囊藻毒素的能力。通过克隆测序得到mcyT序列,并进行分子系统分析,构建了关于mcyT序列的Neighbor-Joining系统树,结果表明mcyT序列可以将产毒与不产毒浮丝藻分为两大独立的分支,两个分支之间的最低序列相似度分别为98.5%和99.1%。研究结果可为后续研究我国浮丝藻的微囊藻毒素合成相关基因的多样性以及分子监测提供参考。展开更多
The glutathione S-transferases(GSTs)play important roles in detoxification of microcystins(MCs),but the responses of GSTs to MC-LR have not been well characterized in freshwater gastropod,Cipangopaludina cahayensis.In...The glutathione S-transferases(GSTs)play important roles in detoxification of microcystins(MCs),but the responses of GSTs to MC-LR have not been well characterized in freshwater gastropod,Cipangopaludina cahayensis.In the present study,we cloned full-length cDNAs of mu-and pi-class GSTs(GSTM and GSTP)and partial cDNA of omega-class GST(GSTO),and determined the transcriptional responses of the three GST genes to different concentrations of MC-LR(0,1,10 and 100 lg/L)in C.cahayensis.The full-length cDNAs of GSTM and GSTP of C.cahayensis were 813 and 820 bp,containing an open reading frame(ORF)of 648 bp(encoding 215 amino acids)and 624 bp(encoding 207 amino acids),respectively.The mRNA expression of GSTM and GSTO significantly decreased after exposure to 10 lg/L MC-LR,and the mRNA expression of GSTP significantly decreased after100 lg/L MC-LR exposure.This might contribute to the detoxication of MCs in C.cahayensis,which is consistent with its sedentary life and filter-feeder status.The mRNA expression of the three GST isoforms in C.cahayensis could be used as biomarkers for water contamination.展开更多
文摘微囊藻毒素(Microcystin,MC)的产生受微囊藻毒素合成酶基因簇(microcystin biosynthesis gene,mcy)调控,常用PCR扩增mcy基因检测产毒微囊藻。采集江苏省5大淡水湖泊——太湖、滆湖、高宝-邵伯湖、洪泽湖和骆马湖的水样,测定水体营养盐浓度和叶绿素a浓度,根据叶绿素a浓度计算5个湖泊的富营养化指数(Trophic state index,TSI),同时应用单一和多重PCR扩增mcy基因。结果表明,太湖和滆湖处于富营养和超富营养化水平,洪泽湖和骆马湖处于中营养和富营养化水平,高宝-邵伯湖处于寡营养水平。太湖、滆湖、洪泽湖和骆马湖的所有水样均检出mcy基因,4个湖泊水体受到微囊藻毒素的潜在威胁,高宝-邵伯湖没有检测出mcy基因的存在,尚未受微囊藻毒素的污染。
文摘为研究我国浮丝藻(Planktothrix Anagnostidis et Komárek)的毒素相关基因,选取分离自我国不同省份水体的13株阿氏浮丝藻,通过PCR检测其微囊藻毒素合成酶基因mcyA、mcyE及mcyT研究其毒素基因特性。PCR结果表明除mcyT之外其他引物检测均无扩增产物,说明这13株浮丝藻不具备产微囊藻毒素的能力。通过克隆测序得到mcyT序列,并进行分子系统分析,构建了关于mcyT序列的Neighbor-Joining系统树,结果表明mcyT序列可以将产毒与不产毒浮丝藻分为两大独立的分支,两个分支之间的最低序列相似度分别为98.5%和99.1%。研究结果可为后续研究我国浮丝藻的微囊藻毒素合成相关基因的多样性以及分子监测提供参考。
基金supported by the Key Projects in the National Science&Technology Pillar Program during the Twelfth Five-year Plan Period(2012BAD25B04)the National Natural Science Foundation of China(31272641,31172420)the Fundamental Research Funds for the Central Universities(2013SC13)
文摘The glutathione S-transferases(GSTs)play important roles in detoxification of microcystins(MCs),but the responses of GSTs to MC-LR have not been well characterized in freshwater gastropod,Cipangopaludina cahayensis.In the present study,we cloned full-length cDNAs of mu-and pi-class GSTs(GSTM and GSTP)and partial cDNA of omega-class GST(GSTO),and determined the transcriptional responses of the three GST genes to different concentrations of MC-LR(0,1,10 and 100 lg/L)in C.cahayensis.The full-length cDNAs of GSTM and GSTP of C.cahayensis were 813 and 820 bp,containing an open reading frame(ORF)of 648 bp(encoding 215 amino acids)and 624 bp(encoding 207 amino acids),respectively.The mRNA expression of GSTM and GSTO significantly decreased after exposure to 10 lg/L MC-LR,and the mRNA expression of GSTP significantly decreased after100 lg/L MC-LR exposure.This might contribute to the detoxication of MCs in C.cahayensis,which is consistent with its sedentary life and filter-feeder status.The mRNA expression of the three GST isoforms in C.cahayensis could be used as biomarkers for water contamination.
