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首页> 外文期刊>Applied Microbiology >A Double Staining Method Using SYTOX Green and Calcofluor White for Studying Fungal Parasites of Phytoplankton
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A Double Staining Method Using SYTOX Green and Calcofluor White for Studying Fungal Parasites of Phytoplankton

机译：SYTOX Green和Calcofluor white双重染色法研究浮游植物的真菌寄生虫

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We propose a double staining method based on the combination of two fluorochromes, calcofluor white (CFW; specific chitinous fluorochrome) and SYTOX green (nucleic acid stain), coupled to epifluorescence microscopy for counting, identifying, and investigating the fecundity of parasitic fungi of phytoplankton and the putative relationships established between hosts and their chytrid parasites. The method was applied to freshwater samples collected over two successive years during the terminal period of autumnal cyanobacterial blooms in a eutrophic lake. The study focused on the uncultured host-parasite couple Anabaena macrospora (cyanobacterium) and Rhizosiphon akinetum (Chytridiomycota). Our results showed that up to 36.6% of cyanobacterial akinetes could be parasitized by fungi. Simultaneously, we directly investigated the zoosporic content inside the sporangia and found that both the host size and intensity of infection conditioned the final size and hence fecundity of the chytrids. We found that relationships linking host size, final parasite size, and chytrid fecundity were conserved from year to year and seemed to be host-chytrid couple specific. We concluded that our double staining method was a valid procedure for improving our knowledge of uncultured freshwater phytoplankton-chytrid couples and so of the quantitative ecology of chytrids in freshwater ecosystems.
机译：我们提出一种基于两种荧光染料钙荧光白（CFW;特定几丁质荧光染料）和SYTOX绿（核酸染色）的双重染色方法，结合表观荧光显微镜进行计数，鉴定和研究浮游植物寄生真菌的繁殖力以及寄主和其壶菌寄生虫之间建立的推定关系。该方法适用于富营养化湖泊中蓝藻秋季绽放末期连续两年收集的淡水样品。这项研究的重点是未培养的寄主-寄生对虾（Anabaena macrospora）（蓝细菌）和Rhizosiphon akinetum（Chytridiomycota）。我们的研究结果表明，多达36.6％的蓝藻细菌可以被真菌寄生。同时，我们直接调查了孢子囊内部的游动孢子含量，发现宿主的大小和感染强度决定了最终的大小，并因此影响了壶菌的繁殖力。我们发现，关联寄主大小，最终寄生虫大小和壶菌生殖力的关系每年都得到保留，并且似乎是特定于寄主壶虱夫妇的。我们得出的结论是，我们的双重染色方法可提高我们对未养殖的淡水浮游植物-chytrid夫妇以及淡水生态系统中的chytrids定量生态学的了解。

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