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Microorganisms
Volume 9
Issue 3
10.3390/microorganisms9030510
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Article

Assessing the Use of Molecular Barcoding and qPCR for Investigating the Ecology of Prorocentrum minimum (Dinophyceae), a Harmful Algal Species

by
Kate McLennan
,
Rendy Ruvindy
,
Martin Ostrowski
and
Shauna Murray
*
Faculty of Science, University of Technology Sydney, Ultimo, NSW 2007, Australia
*
Author to whom correspondence should be addressed.
Academic Editor: Thomas Mock
Microorganisms 2021, 9(3), 510; https://doi.org/10.3390/microorganisms9030510
Received: 20 January 2021 / Revised: 12 February 2021 / Accepted: 19 February 2021 / Published: 28 February 2021
(This article belongs to the Special Issue Dinoflagellate Biology: Using Molecular Approaches to Unlock Their Ecology and Evolution)
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Abstract

Prorocentrum minimum is a species of marine dinoflagellate that occurs worldwide and can be responsible for harmful algal blooms (HABs). Some studies have reported it to produce tetrodotoxin; however, results have been inconsistent. qPCR and molecular barcoding (amplicon sequencing) using high-throughput sequencing have been increasingly applied to quantify HAB species for ecological analyses and monitoring. Here, we isolated a strain of P. minimum from eastern Australian waters, where it commonly occurs, and developed and validated a qPCR assay for this species based on a region of ITS rRNA in relation to abundance estimates from the cultured strain as determined using light microscopy. We used this tool to quantify and examine ecological drivers of P. minimum in Botany Bay, an estuary in southeast Australia, for over ~14 months in 2016–2017. We compared abundance estimates using qPCR with those obtained using molecular barcoding based on an 18S rRNA amplicon. There was a significant correlation between the abundance estimates from amplicon sequencing and qPCR, but the estimates from light microscopy were not significantly correlated, likely due to the counting method applied. Using amplicon sequencing, ~600 unique actual sequence variants (ASVs) were found, much larger than the known phytoplankton diversity from this region. P. minimum abundance in Botany Bay was found to be significantly associated with lower salinities and higher dissolved CO2 levels. View Full-Text
Keywords: Prorocentrum minimum; harmful algae; next-generation sequencing Prorocentrum minimum; harmful algae; next-generation sequencing
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MDPI and ACS Style

McLennan, K.; Ruvindy, R.; Ostrowski, M.; Murray, S. Assessing the Use of Molecular Barcoding and qPCR for Investigating the Ecology of Prorocentrum minimum (Dinophyceae), a Harmful Algal Species. Microorganisms 2021, 9, 510. https://doi.org/10.3390/microorganisms9030510

AMA Style

McLennan K, Ruvindy R, Ostrowski M, Murray S. Assessing the Use of Molecular Barcoding and qPCR for Investigating the Ecology of Prorocentrum minimum (Dinophyceae), a Harmful Algal Species. Microorganisms. 2021; 9(3):510. https://doi.org/10.3390/microorganisms9030510

Chicago/Turabian Style

McLennan, Kate, Rendy Ruvindy, Martin Ostrowski, and Shauna Murray. 2021. "Assessing the Use of Molecular Barcoding and qPCR for Investigating the Ecology of Prorocentrum minimum (Dinophyceae), a Harmful Algal Species" Microorganisms 9, no. 3: 510. https://doi.org/10.3390/microorganisms9030510

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