Abstract:

Oil exposure has been suggested to induce reactive oxygen species (ROS) in phytoplankton, that in turn can lead to cellular damage. To test this hypothesis, we measured the reactive oxygen species in oil resistant Phaeodactylum tricornutum and oil sensitive Thalassiosira pseudonana diatom species.

Suggested Citation:

Kamalanathan, Manoj. 2020. Reactive oxygen diatom species Phaeodactylum tricornutum and Thalassiosira pseudonana in response to oil exposure. Distributed by: GRIIDC, Harte Research Institute, Texas A&M University–Corpus Christi. doi:10.7266/ZBJY1NQM

Publications:

Quigg, A., Parsons, M., Bargu, S., Ozhan, K., Daly, K. L., Chakraborty, S., … Buskey, E. J. (2021). Marine phytoplankton responses to oil and dispersant exposures: Knowledge gained since the Deepwater Horizon oil spill. Marine Pollution Bulletin, 164, 112074. doi:10.1016/j.marpolbul.2021.112074

Purpose:

To understand how oil affects phytoplankton species intracellularly.

Data Parameters and Units:

The headers are Species, Treatment, Replicate and Value. Reactive oxygen species per cell: Relative fluorescence per cell.

Methods:

The WAF was made using CORSSERF method (Singer et al., 2001) Briefly, 400 µL of Macondo surrogate oil was added per L sterile f/2 growth media and stirred overnight in the dark. Afterwards, it is filtered using 20 µm Teflon sieve to obtain water accommodated fraction (WAF) free of large oil droplets. The cellular concentration for both Phaeodactylum tricornutum and Thalassiosira pseudonanacells cells (cell count) at the beginning of the experiment were 10^5 cells/ml. ROS levels were quantified using modifications of methods described by Rijstenbil (2002). Briefly, 1 mL of samples were stained with 10 mM of 2',7'-dichlorofluorescien diacetate (DCFH-DA) for 60 mins in complete darkness. The samples were then centrifuged at 10,000 g for 5 mins, following which the supernatant was discarded, and the stained cells were re-suspended with fresh f/2 media. The fluorescence was then measured at 488/520 nm excitation/emission using BioTek CytationTM 5 imaging reader controlled by Gen52.09 software (USA).

Instruments:

Gen52.09 software; BioTek CytationTM 5 imaging reader

Provenance and Historical References:

Rijstenbil, J.W. 2002. Assessment of oxidative stress in the planktonic diatom Thalassiosira pseudonana in response to UVA and UVB radiation. Journal Plankton Research 24(12):1277–1288. DOI: 10.1093/plankt/24.12.1277 Singer, Michael M., Don V. Aurand, Gina M. Coelho, Gail E. Bragin, James R. Clark, Michael Sowby, and Ronald S. Tjeerdema. 2001. Making, measuring, and using water-accommodated fractions of petroleum for toxicity testing. International Oil Spill Conference 2001(2): 1269-1274. DOI: 10.7901/2169-3358-2001-2-1269

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