Quantification and distinction of aplastidic and plastidic marine nanoplankton by fluorescence in situ hybridization
Quantification and distinction of aplastidic and plastidic marine nanoplankton by fluorescence in situ hybridization
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DOI：
10.3354/ame041163
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发表时间：
2005-11-25
影响因子：
1.4
通讯作者：
Pernthaler, J
中科院分区：
文献类型：
--
作者：
Beardsley, C;Knittell, K;Pernthaler, J
本研究建立了一种利用荧光原位杂交（FISH）与18S rDNA靶向寡核苷酸探针结合酪胺信号扩增（TSA）技术对纳米浮游生物进行显微检测的方法。利用可紫外激发荧光染料标记的酪酰胺，可以根据杂交生物的营养模式同时进行鉴定和分类。该方案最初在纯培养物上进行了验证，随后与北海水域时间序列中原生生物计数的标准技术进行了比较。新方案对再生和塑性纳米浮游生物的细胞计数都明显更高，这主要是由于对2 - 5 μ m之间的细胞进行了更好的检测。FISH-TSA对针藻和一组新型层水母的特异性探针显示，在调查时，这些细菌捕食者在北海沿海地表水中并不丰富。结合特异的18S rRNA靶向寡核苷酸探针，新方案可能为同时快速分析环境样品中纳米浮游生物的身份和营养模式提供有价值的工具。
We developed a protocol for the microscopic detection of nanoplankton by fluorescence in situ hybridization (FISH) with 18S rDNA targeting oligonucleotide probes in combination with tyramide signal amplification (TSA). The use of tyramides labeled with a UV-excitable fluorochrome allowed simultaneous identification and classification of the hybridized organisms according to their trophic modes. The protocol was initially validated on pure cultures and was subsequently compared with a standard technique for the enumeration of protists in a time series from North Sea waters. Cell counts with the new protocol were significantly higher for both aplastidic and plastidic nanoplankton, mainly due to superior detection of cells between 2 and 5 mu m. FISH-TSA with specific probes for Pedinellales and a group of novel stramenopiles revealed that these lineages of bacterivores were not abundant in coastal North Sea surface waters at the time of investigation. In combination with specific 18S rRNA targeted oligonucleotide probes the new protocol may provide a valuable tool for a simultaneous rapid analysis of the identity and trophic mode of nanoplankton in environmental samples.