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用于波罗的海春季水华甲藻物种特异性检测和定量的定量实时聚合酶链式反应分析

Quantitative real-time PCR assays for species-specific detection and quantification of Baltic Sea spring bloom dinoflagellates.

作者信息

Brink Annica Marie, Kremp Anke, Gorokhova Elena

机构信息

Department of Ecology, Environment and Plant Sciences, Stockholm University, Stockholm, Sweden.

Biological Oceanography, Institute for Baltic Sea Research Warnemünde, Rostock, Germany.

出版信息

Front Microbiol. 2024 Sep 24;15:1421101. doi: 10.3389/fmicb.2024.1421101. eCollection 2024.

DOI:10.3389/fmicb.2024.1421101
PMID:39380673
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11458424/
Abstract

In the Baltic Sea, the dinoflagellates , , and are important contributors to the spring bloom. However, their relative contribution to the bloom community cannot be unambiguously determined by conventional light microscopy due to a lack of resolution of distinctive morphological features of the three species. Here, we describe a molecular approach based on a quantitative real-time polymerase chain reaction (qPCR) primer and probe system, targeting the ITS1 and ITS2 regions of the rRNA gene for all three species and enabling their quantification. The specificity of the method was demonstrated using monocultures of , , as well as three other dinoflagellate species co-occurring in the Baltic Sea during spring and validated using field-collected phytoplankton samples.

摘要

在波罗的海，甲藻、和是春季水华的重要贡献者。然而，由于这三个物种独特形态特征的分辨率不足，通过传统光学显微镜无法明确确定它们对水华群落的相对贡献。在此，我们描述了一种基于定量实时聚合酶链反应（qPCR）引物和探针系统的分子方法，该方法针对这三个物种的rRNA基因的ITS1和ITS2区域，能够对它们进行定量分析。使用、和的单培养物，以及春季在波罗的海同时出现的其他三种甲藻物种，证明了该方法的特异性，并使用现场采集的浮游植物样本进行了验证。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/8350bdde6369/fmicb-15-1421101-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/bfffad0ea56a/fmicb-15-1421101-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/0680dfd8c6ab/fmicb-15-1421101-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/ce895df13285/fmicb-15-1421101-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/c6bc75261806/fmicb-15-1421101-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/903d04f83c87/fmicb-15-1421101-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/8587951c373f/fmicb-15-1421101-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/8350bdde6369/fmicb-15-1421101-g007.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/bfffad0ea56a/fmicb-15-1421101-g001.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/0680dfd8c6ab/fmicb-15-1421101-g002.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/ce895df13285/fmicb-15-1421101-g003.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/c6bc75261806/fmicb-15-1421101-g004.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/903d04f83c87/fmicb-15-1421101-g005.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/8587951c373f/fmicb-15-1421101-g006.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/774c/11458424/8350bdde6369/fmicb-15-1421101-g007.jpg

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Evaluation of sxtA and rDNA qPCR assays through monitoring of an inshore bloom of Alexandrium catenella Group 1.
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Using synthetic oligonucleotides as standards in probe-based qPCR.在基于探针的定量聚合酶链反应中使用合成寡核苷酸作为标准品。
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