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 PCR 检测方法。

IF 4 2区 生物学 Q2 MICROBIOLOGY Frontiers in Microbiology Pub Date : 2024-09-24 eCollection Date: 2024-01-01 DOI:10.3389/fmicb.2024.1421101
Annica Marie Brink, Anke Kremp, Elena Gorokhova
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引用次数: 0

摘要

在波罗的海,甲藻 Apocalathium malmogiense、Biecheleria baltica 和 Gymnodinium corollarium 是春季水华的重要贡献者。然而,由于缺乏对这三个物种独特形态特征的分辨,传统的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,我们介绍了一种基于定量实时聚合酶链式反应(qPCR)引物和探针系统的分子方法,该方法以所有三个物种的 rRNA 基因的 ITS1 和 ITS2 区域为目标,并能对其进行定量。该方法的特异性已在 A.malmogiense、B.baltica、G.corollarium 以及春季在波罗的海共生的其他三种甲藻的单培养物中得到证实,并通过现场采集的浮游植物样本进行了验证。
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Quantitative real-time PCR assays for species-specific detection and quantification of Baltic Sea spring bloom dinoflagellates.

In the Baltic Sea, the dinoflagellates Apocalathium malmogiense, Biecheleria baltica, and Gymnodinium corollarium 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 A. malmogiense, B. baltica, G. corollarium as well as three other dinoflagellate species co-occurring in the Baltic Sea during spring and validated using field-collected phytoplankton samples.

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来源期刊
CiteScore
7.70
自引率
9.60%
发文量
4837
审稿时长
14 weeks
期刊介绍: Frontiers in Microbiology is a leading journal in its field, publishing rigorously peer-reviewed research across the entire spectrum of microbiology. Field Chief Editor Martin G. Klotz at Washington State University is supported by an outstanding Editorial Board of international researchers. This multidisciplinary open-access journal is at the forefront of disseminating and communicating scientific knowledge and impactful discoveries to researchers, academics, clinicians and the public worldwide.
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