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Flow Cytometry for Rapid Enumeration and Biomass Quantification of Thraustochytrids in Coastal Seawaters

机译:流式细胞仪用于沿海海水中破囊壶菌的快速计数和生物量定量

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摘要

Marine fungus-like eukaryotic unicellular protists (thraustochytrids) are considered to play an important role in the marine microbial food web. However, their abundance, distribution, and relative biomass in coastal waters have not yet been examined in detail. By using a flow cytometry method (FCM) for the rapid enumeration of thraustochytrids in nearshore and offshore stations along the Gulf of Bohai, China, we herein expanded current knowledge on their ecological significance. The FCM method allows for the rapid detection and quantification of prokaryotic and eukaryotic cells, but is rarely applied to the enumeration of small eukaryotic protists. Epifluorescence microscopy (EpiM) has been commonly used for the direct detection and enumeration of thraustochytrids; however, this method is time-consuming and inapplicable to a large-scale analysis of complex seawater samples. There is no available FCM method to track the abundance and biomass of thraustochytrids in marine habitats. The FCM enumeration of thraustochytrids in seawater samples ranged between 400 and 4,080 cells mL−1 with a biomass range of 8.15–83.96 μg C L−1. The thraustochytrid biomass contributed 10.9% to 98.1% of the total biomass of the heterotrophic microbial community comprising bacterioplankton and thraustochytrids. Their overall abundance in nearshore stations was significantly different from that in offshore stations (P<0.5). The present results provide an optimized method for the rapid detection and enumeration of thraustochytrids in seawater and facilitate large-scale studies of the ecological role of thraustochytrids in the microbial food web of coastal waters.
机译:海洋真菌样的真核单细胞生物(破囊壶菌)被认为在海洋微生物食物网中起着重要的作用。但是,尚未详细研究其在沿海水域的丰度,分布和相对生物量。通过使用流式细胞仪方法(FCM)快速枚举中国渤海沿岸近海站和近海站的破囊壶菌,我们在此扩展了有关其生态意义的最新知识。 FCM方法可以快速检测和定量原核和真核细胞,但很少用于枚举小的真核生物。表层荧光显微镜(EpiM)已普遍用于直接检测和计数破囊壶菌。然而,该方法耗时且不适用于复杂海水样品的大规模分析。没有可用的FCM方法来追踪海洋生境中破囊壶菌的丰度和生物量。海水样品中破囊壶菌的FCM计数范围为400至4,080个细胞mL -1 ,生物量范围为8.15-83.96μgC L -1 。在包括浮游细菌和破囊壶菌的异养微生物群落中,破囊壶菌的生物量占总生物量的10.9%至98.1%。它们在近海站的总体丰度与近海站的差异很大(P <0.5)。目前的结果提供了一种快速检测和计数海水中破囊壶菌的优化方法,并有助于大规模研究破囊壶菌在沿海水域微生物食物网中的生态作用。

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