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使用电子微阵列检测来自沿海和微观环境中与有害藻华相关的细菌。

Detection of bacteria associated with harmful algal blooms from coastal and microcosm environments using electronic microarrays.

作者信息

Barlaan Edward A, Furukawa Seiji, Takeuchi Kazuhisa

机构信息

Nagasaki Industrial Promotion Foundation, Ikeda 2-1303-8, Omura City Nagasaki 856-0026, Japan.

出版信息

Environ Microbiol. 2007 Mar;9(3):690-702. doi: 10.1111/j.1462-2920.2006.01188.x.

Abstract

With the global expansion of harmful algal blooms (HABs), several measures, including molecular approaches, have been undertaken to monitor its occurrence. Many reports have indicated the significant roles of bacteria in controlling algal bloom dynamics. Attempts have been made to utilize the bacteria/harmful algae relationship in HAB monitoring. In this study, bacterial assemblages monitored during coastal HABs and bacterial communities in induced microcosm blooms were investigated. Samples were analysed using denaturing gradient gel electrophoresis (DGGE) of the 16S rRNA gene. DGGE bands with peculiar patterns before, during, and after algal blooms were isolated and identified. Probes for six ribotypes representing organisms associated with Chatonella spp., Heterocapsa circularisquama, or Heterosigma akashiwo were used for analysis on NanoChip electronic microarray. In addition, a new approach using cultured bacteria species was developed to detect longer (533 bp) polymerase chain reaction-amplified products on the electronic microarray. The use of fluorescently labelled primers allowed the detection of individual species in single or mixed DNA conditions. The developed approach enabled the detection of the presence or absence and relative abundance of the HAB-related ribotypes in coastal and microcosm blooms. This study indicates the ability of electronic microarray platform to detect or monitor bacteria in natural and induced environments.

摘要

随着有害藻华在全球范围内的扩散,人们采取了包括分子方法在内的多种措施来监测其发生情况。许多报告表明细菌在控制藻华动态方面发挥着重要作用。人们已尝试利用细菌与有害藻类的关系进行有害藻华监测。在本研究中,对沿海有害藻华期间监测到的细菌群落以及诱导微宇宙藻华中的细菌群落进行了调查。使用16S rRNA基因的变性梯度凝胶电泳(DGGE)对样本进行分析。分离并鉴定了藻华之前、期间和之后具有独特模式的DGGE条带。使用代表与卡盾藻属、圆石藻或赤潮异弯藻相关生物的六种核糖型的探针在NanoChip电子微阵列上进行分析。此外,还开发了一种使用培养细菌物种的新方法,以在电子微阵列上检测更长(533 bp)的聚合酶链反应扩增产物。使用荧光标记引物能够在单DNA或混合DNA条件下检测单个物种。所开发的方法能够检测沿海和微宇宙藻华中与有害藻华相关的核糖型的存在与否及其相对丰度。本研究表明电子微阵列平台有能力在自然和诱导环境中检测或监测细菌。

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