the Creative Commons Attribution 4.0 License.
the Creative Commons Attribution 4.0 License.
Variations of polyphenols and carbohydrates of Emiliania huxleyi grown under simulated ocean acidification conditions
Abstract. Global environmental changes strongly affect the growth and biochemical composition of microalgae. Cultures of the coccolithophore Emiliania huxleyi were grown under four different CO2-controlled pH conditions (7.75, 7.90, 8.10, and 8.25) to improve understanding of the adaptive mechanisms of these organisms through changes in phenolic compounds and carbohydrate content and composition under ocean acidification (OA) scenarios. The highest algal biomass peaks, 1.07 (± 0.10) and 1.04 (± 0.06) × 108 cells L−1, were observed in the microcosms with intermediate CO2 levels (pH 8.10 and 7.90 respectively). Intra- and extracellular phenolic compounds were identified and quantified by Reverse Phase-High Performance Liquid Chromatography (RP-HPLC). The highest concentrations of total exuded phenolics were found in cultures with lower cell densities, at pH 8.25 (43±3 nM) and 7.75 (18.0±0.9 nM). Accumulation of intracellular phenolic compounds was observed in cells with decreasing pH, reaching the maximum level (9.24±0.19 attomole cell-1) at the lowest pH (7.75). The total carbohydrate content inside the cells increased with decreasing pH from 8.25 to 8.10, remaining constant at pH 7.90, and decreasing at lower pH. The presence of antioxidants was determined by 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical inhibition and ferric-reducing antioxidant power (FRAP) assays. The highest activity in both tests was exhibited by cells grown at pH 7.75.
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Status: open (until 23 May 2024)
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RC1: 'Comment on bg-2024-1', Anonymous Referee #1, 30 Mar 2024
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The manuscript by M. Rico et al presents data from a cultivation of Emiliania h. under four pH conditions. results concern the growth, phenolic compounds and total carbohydrate in both cells and medium.
The study might be of interest if the authors add some other variables (pigments, coccoliths,...). in its present state, this ms can not be accepted as publication, due to relevant weaknesses.
- The ms totally lacks of statistical analysis. It does seem that for most of the data, no significant variation was revealed among the different treatments. if this was true, the results and discussion section has to be thoroughly re-written.
- the fig. 2 is emblematic: in the legend, "diatoms" are mentioned while results cam from Emiliania h.; dark vs light color is referred to what?; these data correspond to which day ?
- fig.1: no SD bars
- material and methods: no mention on light, photoperiod, temperature...: these factors are probably the most relevant to shape the microalgal physiology
- material and methods: please explain why a first extraction in acetone and then in methanol. what is the role of acetone?
- material and methods: it is not clear: seawater or medium? which was the target of pH manipulation ? (e..
- material and methods: 48h (line 124) ? does it mean that the experiment started after 48 h of cultivation of E.h. under the different conditions? in the fig.1 the day 0 corresponded to this time?. which was the cell concentration at this time?
- carbohydrate concentration measurement: did the protocol setup with acetone and then methanol extraction?
- quantification of phenols: did the authors used some pure standards?
- calibration curve: please explain why there is a "b" factor (y = ax + b; for carbohydrates, frap)?, also especially when it is negative (dpph) ?
- line 165: cell extracts? how they have been done?
- LINE 173: The highest peak?...but NO SIGNIFICANT!
- lines 186-189: re-write!
- table 1: unit of cell and exuded? seems to be different, also from the "levels" (nM).
- legend: three measurements : three replicates ? (different cultures? or technical replicates?)
- lack of significativity tests in all the studies
- I suggest also to the authors to give a look on 10.1080/07388551.2021.1874284, that might help for the discussion
Citation: https://doi.org/10.5194/bg-2024-1-RC1 -
RC2: 'Comment on bg-2024-1', Anonymous Referee #2, 25 Apr 2024
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The manuscript by Rico et al., demonstrated the responses of phenolics and carbohydrates of Emiliania huxleyi to ocean acidification conditions. They found that the intracellular phenolic compounds increased under low pH conditions, and these findings have certain implications for the marine food webs and biogeochemical cycles. However, I do have some concerns need to be addressed before it can be accepted for the publication in BG.
General Comments:
Abstract: The Abstract was not well structed, and is wordy. It should be very concise and highlight the significance of the findings presented in this study. Please rephrase it to be more concise.
Discussion:
1: The discussion could better articulate how the observed changes in polyphenols and carbohydrates could impact broader ecological processes. Specifically, the implications for the marine food web and biogeochemical cycles could be explored in more depth.
2: The manuscript could benefit from a more detailed comparison with existing literature. Specifically, how do these results align or contrast with the findings of similar studies in terms of the magnitude and direction of changes in polyphenol and carbohydrate content?
3: Broader Impacts: The results are discussed primarily in the context of Emiliania huxleyi. Expanding the discussion to consider potential impacts on other phytoplankton species, could provide a more holistic view of ocean acidification impacts.
Specific comments:
1: Line 37-39: Some controversial findings regarding the calcification of coccolithophore to ocean acidification should be mentioned.
2: Line 82-84: There were at least two studies examined the responses of phenolic compounds in marine primary producers to ocean acidification (Arnold et al., 2012, PLOS ONE; Jin et al., 2015, Nature Communications, 6:8714), where are more relevant to the present study should be acknowledged here.
3: Line 115-125: Have you monitored the carbonate chemistry parameters over the 8 days experimental duration? These parameters are important for a typic OA research. Please clarify.
4: Line 170-175: Should it be possible to calculate the specific growth rates for the exponential phase of each growth curve and then compare them under different pH conditions?
5: Line 210: diatoms?
10: Line 255: Typo of the citation “Santana_Casiano et al., 2014”
Citation: https://doi.org/10.5194/bg-2024-1-RC2
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