机构地区:[1]中国科学院南京地理与湖泊研究所,南京210008 [2]中国科学院研究生院,北京100049
出 处:《生态学报》2011年第21期6551-6560,共10页Acta Ecologica Sinica
基 金:973计划"大中型浅水湖泊蓝藻水华暴发机理研究"(2008CB418000);中国科学院南京地理与湖泊研究所所长基金(07SL021001)
摘 要:应用荧光定量PCR对冬季太湖不同湖区底泥表面有毒微囊藻和总微囊藻种群丰度进行调查,同时基于PCR-DGGE技术对底泥中有毒微囊藻群落结构进行分析。结果表明:微囊藻在太湖底泥表面分布广泛,所有采样点都检测到有毒微囊藻存在,且不同湖区有毒微囊藻和总微囊藻种群丰度存在显著差异,有毒微囊藻和微囊藻基因型丰度范围分别为1.23×104—3.75×106拷贝数/g干重和2.56×104—1.07×107拷贝数/g干重,有毒微囊藻与微囊种群丰度的比例为4.8%—35.2%;DGGE指纹图谱显示,冬季太湖不同湖区表层底泥中有毒微囊藻群落结构相似性较高,相似性系数为70.2%—96.0%。虽然不同湖区基因型组成存在差异,但所有样品中占优势的基因型是一致的。同时发现,优势基因型所占的比例与样品的香农多样性指数呈负相关。序列分析表明,mcyA序列长度为291bp,序列相似性超过97%。综合定量PCR结果和底泥中叶绿素a和藻蓝素浓度的测定结果,可以得出2010年冬季太湖蓝藻越冬主要集中在梅梁湾、竺山湾、贡湖湾和湖心。通过建立荧光定量PCR分析方法,为研究湖泊底泥中蓝藻种群丰度动态变化奠定了基础。Lake Taihu is a large,shallow,eutrophic freshwater lake,which has frequently experienced cyanobacterial bloom in summer in the past decades.In winter,a large quantity of cyanobacterial cells sink on the sediment surface to overwinter.Therefore,it is important to quantify the biomass of cyanobacteria on the sediment to predict the location and the extent of water bloom in summer.However it is difficult to identify and to count cyanobacterial cells in sediment using microscopy method,and microscopic identification cannot differentiate between the toxigenic and non-toxigenic genotypes.In recent years,this problem has been resolved by using molecular techniques such as PCR and quantitative real-time PCR(qPCR) with primers targeting toxin-producing genes.The microcystin gene cluster which contains 10 genes,namely from mcyA to mcyJ has been widely used to reveal microcystin-producing species in genera Anabaena,Microcystis and Planktothrix.Detection of cyanobacteria and their toxin-producing ability using molecular methods has been successfully applied to a diverse range of water and sediment environments.In this study,we use quantitative real time PCR to quantify the abundance of microcystin-producing cyanobacteria and Microcystis,and PCR-denaturing gradient gel electrophoresis technique to investigate the community structure of microcystin-producing cyanobacteria in surface sediment of Lake Taihu in winter.qPCR data showed that Microcystis and microcystin-producing cyanobacteria were present in all of sediment samples and their abundance varied significantly in different lake areas,ranging from 1.23×104copies/g dry weight to 3.75×106copies/g dry weight and from 2.56×104 to 1.07×107 copies/g dry weight,respectively.The proportion of microcystin-producing cyanobacteria to Microcystis ranged from 4.8% to 35.2%.DGGE patterns indicated that the composition of microcystin-producing cyanobacteria were very similar in surface sediments collected from different lake areas,and the similarity values were between 70.2% an
关 键 词:荧光定量PCR 有毒蓝藻 群落结构 变性梯度凝胶电泳 太湖
分 类 号:X524[环境科学与工程—环境工程] X173
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