Detection of Heterosigma akashiwo (Hada) using specific RNA probes: Variability of RNA content with environmental conditions

Blanco, Eva Perez; Hagström, Johannes A.; Salomon, Paulo S.; Granéli, Edna

doi:10.1016/j.hal.2013.02.001

Cited by 9 publications

(3 citation statements)

References 32 publications

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“…Like gapdh , 18S is commonly used as a reference gene [ 52 ], and it has been used to design probes for quantifying the abundance of H . akshiwo in field samples [ 53 ]. In the present study we found that 18S was a moderately stable gene across all conditions we examined, although it appeared to be a suitable reference gene under light treatment ( Fig 2 ).…”

Section: Discussionmentioning

confidence: 99%

Screening for Suitable Reference Genes for Quantitative Real-Time PCR in Heterosigma akashiwo (Raphidophyceae)

Lin

et al. 2015

PLoS ONE

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The raphidophyte Heterosigma akashiwo is a globally distributed harmful alga that has been associated with fish kills in coastal waters. To understand the mechanisms of H. akashiwo bloom formation, gene expression analysis is often required. To accurately characterize the expression levels of a gene of interest, proper reference genes are essential. In this study, we assessed ten of the previously reported algal candidate genes (rpL17-2, rpL23, cox2, cal, tua, tub, ef1, 18S, gapdh, and mdh) for their suitability as reference genes in this species. We used qRT-PCR to quantify the expression levels of these genes in H. akashiwo grown under different temperatures, light intensities, nutrient concentrations, and time points over a diel cycle. The expression stability of these genes was evaluated using geNorm and NormFinder algorithms. Although none of these genes exhibited invariable expression levels, cal, tub, rpL17-2 and rpL23 expression levels were the most stable across the different conditions tested. For further validation, these selected genes were used to normalize the expression levels of ribulose-1, 5-bisphosphate carboxylase/oxygenase large unite (HrbcL) over a diel cycle. Results showed that the expression of HrbcL normalized against each of these reference genes was the highest at midday and lowest at midnight, similar to the diel patterns typically documented for this gene in algae. While the validated reference genes will be useful for future gene expression studies on H. akashiwo, we expect that the procedure used in this study may be helpful to future efforts to screen reference genes for other algae.

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Section: Discussionmentioning

confidence: 99%

Screening for Suitable Reference Genes for Quantitative Real-Time PCR in Heterosigma akashiwo (Raphidophyceae)

Lin

et al. 2015

PLoS ONE

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“…This species can be difficult to identify, especially once preserved in Lugol’s. Two species-specific probes were designed from the 18S region (SSHaka) and six more from the 28S region (LSHaka) for H. akashiwo ( Table 2 , [ 25 ]). Their calibration curves show the sensitivity of each probe and point out a low affinity with the H. akashiwo RNA.…”

Section: Resultsmentioning

confidence: 99%

Testing a Microarray to Detect and Monitor Toxic Microalgae in Arcachon Bay in France

et al. 2013

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Harmful algal blooms (HABs) occur worldwide, causing health problems and economic damages to fisheries and tourism. Monitoring agencies are therefore essential, yet monitoring is based only on time-consuming light microscopy, a level at which a correct identification can be limited by insufficient morphological characters. The project MIDTAL (Microarray Detection of Toxic Algae)—an FP7-funded EU project—used rRNA genes (SSU and LSU) as a target on microarrays to identify toxic species. Furthermore, toxins were detected with a newly developed multiplex optical Surface Plasmon Resonance biosensor (Multi SPR) and compared with an enzyme-linked immunosorbent assay (ELISA). In this study, we demonstrate the latest generation of MIDTAL microarrays (version 3) and show the correlation between cell counts, detected toxin and microarray signals from field samples taken in Arcachon Bay in France in 2011. The MIDTAL microarray always detected more potentially toxic species than those detected by microscopic counts. The toxin detection was even more sensitive than both methods. Because of the universal nature of both toxin and species microarrays, they can be used to detect invasive species. Nevertheless, the MIDTAL microarray is not completely universal: first, because not all toxic species are on the chip, and second, because invasive species, such as Ostreopsis, already influence European coasts.

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“…In order to produce such a predictive tool, it was necessary to test how Similarly, this was also determined to be the case for toxic species Heterosigma akashiwo and Alexandrium species (Blanco et al 2013;2014).…”

Section: Environmental Stressesmentioning

confidence: 99%

An evaluation of the applicability of microarrays for monitoring toxic algae in Irish coastal waters

McCoy

Touzet

Fleming

et al. 2012

Environ Sci Pollut Res

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Detection of Heterosigma akashiwo (Hada) using specific RNA probes: Variability of RNA content with environmental conditions

Cited by 9 publications

References 32 publications

Screening for Suitable Reference Genes for Quantitative Real-Time PCR in Heterosigma akashiwo (Raphidophyceae)

Screening for Suitable Reference Genes for Quantitative Real-Time PCR in Heterosigma akashiwo (Raphidophyceae)

Testing a Microarray to Detect and Monitor Toxic Microalgae in Arcachon Bay in France

An evaluation of the applicability of microarrays for monitoring toxic algae in Irish coastal waters

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