Resolving the intra-specific succession within Cochlodinium polykrikoides populations in southern Korean coastal waters via use of quantitative PCR assays

Park, Bum Soo; Wang, Pengbin; Kim, Jin Ho; Kim, Joohwan; Gobler, Christopher J.; Han, Moonsik

doi:10.1016/j.hal.2014.04.019

Cited by 39 publications

(29 citation statements)

References 51 publications

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“…Detection of C. polykrikoides has also been achieved using real-time PCR methods on both natural and cultured samples [82]. This study demonstrated how the application of real-time PCR can be used to look at the variation on a subspecies level, most likely caused by ecological factors and cellular adaptations.…”

Section: Real-time Pcr Applicationsmentioning

confidence: 92%

Approaches for the detection of harmful algal blooms using oligonucleotide interactions

et al. 2014

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Blooms of microscopic algae in our waterways are becoming an increasingly important environmental concern. Many are sources of harmful biotoxins that can lead to death in humans, marine life and birds. Additionally, their biomass can cause damage to ecosystems such as oxygen depletion, displacement of species and habitat alteration. Globally, the number and frequency of harmful algal blooms has increased over the last few decades, and monitoring and detection strategies have become essential for managing these events. This review discusses developments in the use of oligonucleotide-based 'molecular probes' for the selective monitoring of algal cell numbers. Specifically, hybridisation techniques will be a focus.

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Section: Real-time Pcr Applicationsmentioning

confidence: 92%

Approaches for the detection of harmful algal blooms using oligonucleotide interactions

et al. 2014

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“…PCR amplification of LSU rDNA (28S) using primers specific to C. polykrikoides American/Malaysian ribotype was performed using the primers developed by Park et al (53) (forward primer, AMCPSF, CTC AAT CGC CTT TCG CCT GAT, and reverse primer, AMCPSR, ACC GGA CAC CTC GGA TAT GAT), which were originally designed to positively identify New York clones of C. polykrikoides. PCR conditions were as described by Park et al (53) with minor modifications, specifically using an initial denaturation at 95°C for 10 min, 60 cycles at 95°C for 20 s, 56°C for 20 s, and 72°C for 50 s, followed by a 5-min extension step at 72°C.…”

Section: Methodsmentioning

confidence: 99%

“…PCR conditions were as described by Park et al (53) with minor modifications, specifically using an initial denaturation at 95°C for 10 min, 60 cycles at 95°C for 20 s, 56°C for 20 s, and 72°C for 50 s, followed by a 5-min extension step at 72°C. Reaction mixtures (25 l) were made with 12.5 l of Go Taq Green (Promega), 10 l of sample DNA (Ͻ20 ng), and 2.5 l of forward and reverse primer mixture (4 M).…”

Section: Methodsmentioning

confidence: 99%

Mapping the Distribution of Cysts from the Toxic Dinoflagellate Cochlodinium polykrikoides in Bloom-Prone Estuaries by a Novel Fluorescence In Situ Hybridization Assay

Hattenrath-Lehmann

Zhen

Wallace

et al. 2016

Appl Environ Microbiol

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Cochlodinium polykrikoides is a cosmopolitan dinoflagellate that is notorious for causing fish-killing harmful algal blooms (HABs) across North America and Asia. While recent laboratory and ecosystem studies have definitively demonstrated that Cochlodinium forms resting cysts that may play a key role in the dynamics of its HABs, uncertainties regarding cyst morphology and detection have prohibited even a rudimentary understanding of the distribution of C. polykrikoides cysts in coastal ecosystems. Here, we report on the development of a fluorescence in situ hybridization (FISH) assay using oligonucleotide probes specific for the large subunit (LSU) ribosomal DNA (rDNA) of C. polykrikoides. The LSU rDNA-targeted FISH assay was used with epifluorescence microscopy and was iteratively refined to maximize the fluorescent reaction with C. polykrikoides and minimize cross-reactivity. The final LSU rDNA-targeted FISH assay was found to quantitatively recover cysts made by North American isolates of C. polykrikoides but not cysts formed by other common cyst-forming dinoflagellates. The method was then applied to identify and map C. polykrikoides cysts across bloom-prone estuaries. Annual cyst and vegetative cell surveys revealed that elevated densities of C. polykrikoides cysts (>100 cm ؊3 ) during the spring of a given year were spatially consistent with regions of dense blooms the prior summer. The identity of cysts in sediments was confirmed via independent amplification of C. polykrikoides rDNA. This study mapped C. polykrikoides cysts in a natural marine setting and indicates that the excystment of cysts formed by this harmful alga may play a key role in the development of HABs of this species. Resting cysts of dinoflagellates can be associated with genetic recombination, maintenance of blooms, termination of blooms, recurrence of annual blooms, resistance against unfavorable environmental conditions, protection from viruses, grazers, or parasite attacks, and geographical expansion of populations (1-10). Resting cysts, therefore, play an important role in the ecology of harmful algal blooms (HABs) caused by dinoflagellates (8, 11) and have been considered a fundamental attribute of dinoflagellate life cycles (12). About 200 marine and freshwater dinoflagellates are known to produce resting cysts, a small number relative to the ϳ2,300 extant dinoflagellate species (5, 11, 13). More than 20 of these cyst-producing dinoflagellates are known to cause harmful algal blooms (HABs) (5), including Cochlodinium polykrikoides Margalef (10).Cochlodinium polykrikoides Margalef is an unarmored dinoflagellate that has caused fish-killing HABs in locations across much of Asia and North America (14-17). The initiation and development of C. polykrikoides blooms have been shown to be related to multiple factors, including stimulation by nutrients such as nitrogen and vitamins (18, 19), mixotrophy (20), the production of extracellular toxins lethal to grazers (21-23), bacterial mutualism (24), and allelopathic effects on competing phy...

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“…The ribotype "Philippines" was fi rst reported including samples collected from Omura Bay, Japan, in July 2003 and Manila Bay, the Philippines, in October 2004 . Cells of C. polykrikoides assigned to this ribotype were later reported from the southern coast of Korea, by specifi c primer sets of qPCR, and L'Ametlla Harbour, Spain (Reñé et al 2013a ;Park et al 2014 ). Considering the occurrences in the Western Pacifi c and Mediterranean, the ribotype is likely to distribute widely, from tropical to temperate regions.…”

Section: Intraspecifi C Relationships In Cochlodinium Polykrikoidesmentioning

confidence: 97%

Taxonomy and Distribution of the Unarmored Dinoflagellates Cochlodinium polykrikoides and C. fulvescens

Iwataki

Takayama²,

Takahashi

et al. 2015

Marine Protists

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Red tides of the unarmored dinofl agellate Cochlodinium polykrikoides and related chain-forming Cochlodinium species have been responsible for the mass mortality of fi sh in coastal areas of East Asia, and in Central and North America over the past few decades. Since 2000, C. polykrikoides has been reported for the fi rst time from Southeast Asia, India, the Middle East and the Mediterranean, so its distribution has seemingly expanded. Recent molecular phylogenetic works have detected intraspecifi c populations of C. polykrikoides , which presumably indicates their original distribution area. Reported occurrences of the chain-forming Cochlodinium spp., Cochlodinium fulvescens and C. polykrikoides are summarized with reference to their ribotypes, in order to consider the expansion of their distribution area and recent migration. The ultrastructure of C. polykrikoides was recently analyzed and showed the characteristic features seen in all other dinofl agellates. The apical groove structure of C. fulvescens is also provided in this chapter, for morphological comparison with C. polykrikoides and further characterization of chain-forming Cochlodinium spp. that produce fi sh-killing red tides.

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Resolving the intra-specific succession within Cochlodinium polykrikoides populations in southern Korean coastal waters via use of quantitative PCR assays

Cited by 39 publications

References 51 publications

Approaches for the detection of harmful algal blooms using oligonucleotide interactions

Approaches for the detection of harmful algal blooms using oligonucleotide interactions

Mapping the Distribution of Cysts from the Toxic Dinoflagellate Cochlodinium polykrikoides in Bloom-Prone Estuaries by a Novel Fluorescence In Situ Hybridization Assay

Taxonomy and Distribution of the Unarmored Dinoflagellates Cochlodinium polykrikoides and C. fulvescens

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