A model for carbohydrate metabolism in the diatom Phaeodactylum tricornutum deduced from comparative whole genome analysis

Background

Diatoms are unicellular algae responsible for approximately 20% of global carbon fixation. Their evolution by secondary endocytobiosis resulted in a complex cellular structure and metabolism compared to algae with primary plastids.

Methodology/Principal Findings

The whole genome sequence of the diatom Phaeodactylum tricornutum has recently been completed. We identified and annotated genes for enzymes involved in carbohydrate pathways based on extensive EST support and comparison to the whole genome sequence of a second diatom, Thalassiosira pseudonana. Protein localization to mitochondria was predicted based on identified similarities to mitochondrial localization motifs in other eukaryotes, whereas protein localization to plastids was based on the presence of signal peptide motifs in combination with plastid localization motifs previously shown to be required in diatoms. We identified genes potentially involved in a C4-like photosynthesis in P. tricornutum and, on the basis of sequence-based putative localization of relevant proteins, discuss possible differences in carbon concentrating mechanisms and CO₂ fixation between the two diatoms. We also identified genes encoding enzymes involved in photorespiration with one interesting exception: glycerate kinase was not found in either P. tricornutum or T. pseudonana. Various Calvin cycle enzymes were found in up to five different isoforms, distributed between plastids, mitochondria and the cytosol. Diatoms store energy either as lipids or as chrysolaminaran (a β-1,3-glucan) outside of the plastids. We identified various β-glucanases and large membrane-bound glucan synthases. Interestingly most of the glucanases appear to contain C-terminal anchor domains that may attach the enzymes to membranes.

Conclusions/Significance

Here we present a detailed synthesis of carbohydrate metabolism in diatoms based on the genome sequences of Thalassiosira pseudonana and Phaeodactylum tricornutum. This model provides novel insights into acquisition of dissolved inorganic carbon and primary metabolic pathways of carbon in two different diatoms, which is of significance for an improved understanding of global carbon cycles.     

Unique configuration of genes of silicon transporter in the freshwater pennate diatom <Emphasis Type="Italic">Synedra acus</Emphasis> subsp. <Emphasis Type="Italic">radians</Emphasis>

The existence of the cluster of duplicated sit silicon transporter genes in the chromosome of the diatom Synedra acus subsp. radians was shown for the first time. Earlier, the localization of sit genes in the same chromosome and cluster formation caused by gene duplication was shown only for the marine raphid pennate diatom P. tricornutum. Only non-clustered sit genes were found in the genomes of other diatoms. It is reasonable to assume that sit tandem (sit-td) and sit triplet (sit-tri) genes of S. acus subsp. radians occurred as a result of gene duplication followed by divergence of gene copies.     

The ancestors of diatoms evolved a unique mitochondrial dehydrogenase to oxidize photorespiratory glycolate

Like other oxygenic photosynthetic organisms, diatoms produce glycolate, a toxic intermediate, as a consequence of the oxygenase activity of Rubisco. Diatoms can remove glycolate through excretion and through oxidation as part of the photorespiratory pathway. The diatom Phaeodactylum tricornutum encodes two proteins suggested to be involved in glycolate metabolism: PtGO1 and PtGO2. We found that these proteins differ substantially from the sequences of experimentally characterized proteins responsible for glycolate oxidation in other species, glycolate oxidase (GOX) and glycolate dehydrogenase. We show that PtGO1 and PtGO2 are the only sequences of P. tricornutum homologous to GOX. Our phylogenetic analyses indicate that the ancestors of diatoms acquired PtGO1 during the proposed first secondary endosymbiosis with a chlorophyte alga, which may have previously obtained this gene from proteobacteria. In contrast, PtGO2 is orthologous to an uncharacterized protein in Galdieria sulphuraria, consistent with its acquisition during the secondary endosymbiosis with a red alga that gave rise to the current plastid. The analysis of amino acid residues at conserved positions suggests that PtGO2, which localizes to peroxisomes, may use substrates other than glycolate, explaining the lack of GOX activity we observe in vitro. Instead, PtGO1, while only very distantly related to previously characterized GOX proteins, evolved glycolate-oxidizing activity, as demonstrated by in gel activity assays and mass spectrometry analysis. PtGO1 localizes to mitochondria, consistent with previous suggestions that photorespiration in diatoms proceeds in these organelles. We conclude that the ancestors of diatoms evolved a unique alternative to oxidize photorespiratory glycolate: a mitochondrial dehydrogenase homologous to GOX able to use electron acceptors other than O₂.     

Uncertainty of using habitat fidelity in biomonitoring based on benthic diatoms - the Raška River case study

Epilithic diatoms are frequently recommended for river biomonitoring, while much less emphasis is placed on epiphytic communities. This study considers use of epiphytic and epilithic diatom communities from the Ra?ka River in biomonitoring. A total of 212 diatom taxa were recorded in both communities. Dominant diatoms were Achnanthidium minutissimum (Kützing) Czarnecki, A. affine (Grunow) Czarnecki, A. subatomus (Hustedt) Lange-Bertalot, Amphora pediculus (Kützing) Grunow, Diatoma vulgaris Bory, Gomphonema tergestinum (Grunow) Fricke, Cocconeis placentula var. lineata (Ehrenberg) Van Heurck, Melosira varians Agardh and Navicula tripunctata (Müller) Bory. Redundancy analysis (RDA) was used to detect the relationships between diatoms, some environmental factors and sampling sites. Diatoms of the epiphytic community showed a clearly grouping in relation to the sampling sites as compared with diatoms of the epilithic community. Species common to both communities showed a similar correlation with some environmental variables (BOD, pH, NH₄⁺, TP, NO₃^? and conductivity), with three exceptions (Diatoma vulgaris, Cymbella compacta, and Encyonema silesiacum). Pearson’s correlation coefficient showed correlation between selected environmental variables and diatom indices calculated using OMNIDIA 6.2 software (TDI, IPS, CEE, GENRE, TID, SID, SHE and IDSE/5). Diatoms of the epiphytic community are more clearly clustered in relation to the sampling sites as compared with diatoms of the epilithic community. According to our results there is still “place” in biomonitoring for epiphytic community. The present study highlights the necessity of sampling both epiphytic and epilithic communities because substrate specificity could play important role in biomonitoring.     

Changes in growth and composition of the marine microalgae <Emphasis Type="Italic">Phaeodactylum tricornutum</Emphasis> and <Emphasis Type="Italic">Nannochloropsis salina</Emphasis> in response to changing sodium bicarbonate concentrations

Oils, carbohydrates, and fats generated by microalgae are being refined in an effort to produce biofuels. The research presented here examines two marine microalgae, Nannochloropsis salina (green alga) and Phaeodactylum tricornutum (diatom), when grown with 0 (no addition), 0.5, 1.0, 2.0, and 5.0 g L^?1 NaHCO₃ added to an f/2 medium during the growth phase (GP) and a nutrient induced (nitrate limitation) lipid formation phase (LP). We hypothesize that the addition of NaHCO₃ is a sustainable and practical strategy to increase cellular density and concentrations of lipids in microalgae as well as the rate of lipid accumulation. In N. salina, final cell densities were significantly (p?<?0.05) higher in the NaHCO₃-treated cells than the control while in P. tricornutum the cell densities were higher with >[NaHCO₃] during the GP. During the LP, cell densities were generally higher in the NaHCO₃-treated cells compared with controls. F _V/F _M (efficiency of photosystem II) patterns paralleled those for cell density with generally higher values with higher concentrations of NaHCO₃ and significantly different values between controls and 5.0 g L^?1 NaHCO₃ at the end of the GP (p?<?0.05). F _V/F _M was variable between treatments in P. tricornutum (0.3–0.65) but less so in N. salina for (0.5–0.7) regardless of [NaHCO₃]. The lipid index (measured with Nile red), used as a proxy for triacylglycerides (TAGs), was 10.2?±?6.5 and 4.4?±?2.9 (fluorescence units/OD cells ×1000) for N. salina and P. tricornutum, respectively, at the end of the GP. At the end of the LP, the lipid index was eight and four times higher than during the GP in the corresponding 5.0 g L^?1 NaHCO₃ treatments, revealing that N. salina was accumulating more lipid than P. tricornutum. Dry weights essentially doubled during LP compared with GP for N. salina; this was not the case for P. tricornutum. In general, the percentage of ash in dry weights was significantly higher in the LP relative to the corresponding GP treatments for P. tricornutum; this was not the case for N. salina. During the LP, there was also less soluble protein in N. salina compared to GP; differences were not significant in cells growing with 2.0 or 5.0 g L^?1 NaHCO₃. In P. tricornutum, faster growing cells had more soluble protein during the GP and LP; differences between treatments were significant. P. tricornutum generally accumulated significantly more crude protein than N. salina at higher [NaHCO₃]; there was three times more crude protein in the highest NaHCO₃ (5.0 g L^?1) treatment compared with the controls. C:N ratios (mol:mol) were similar across treatments during GP: 7.03?±?0.12 and 10.16?±?0.41 for N. salina and P. tricornutum, respectively. Further, C:N ratios increased with increasing [NaHCO₃] during LP. Species-specific fatty acid methyl ester (FAMEs) profiles were observed. While C16:0 was lower in P. tricornutum compared to N. salina, the diatom produced more C16:1 and C14 but not C18:3. Monounsaturated fatty acids (MUFA) significantly increased in N. salina in the LP compared to GP and in response to increasing [NaHCO₃] (t tests; p?<?0.05). Saturated fatty acids (SFA) responded similarly but to a lesser degree. There were more polyunsaturated fatty acids (PUFA) in N. salina than MUFAs or SFAs. In P. tricornutum, there were generally more SFAs, MUFAs and PUFAs in P. tricornutum during LP than GP in the corresponding NaHCO₃ treatments. These findings reveal the importance of considering NaHCO₃ as a supplemental carbon source in the culturing marine phytoplankton in large-scale production for biofuels.     

Bacterial diketopiperazines stimulate diatom growth and lipid accumulation

Diatoms are photosynthetic microalgae that fix a significant fraction of the world’s carbon. Because of their photosynthetic efficiency and high-lipid content, diatoms are priority candidates for biofuel production. Here, we report that sporulating Bacillus thuringiensis and other members of the Bacillus cereus group, when in co-culture with the marine diatom Phaeodactylum tricornutum, significantly increase diatom cell count. Bioassay-guided purification of the mother cell lysate of B. thuringiensis led to the identification of two diketopiperazines (DKPs) that stimulate both P. tricornutum growth and increase its lipid content. These findings may be exploited to enhance P. tricornutum growth and microalgae-based biofuel production. As increasing numbers of DKPs are isolated from marine microbes, the work gives potential clues to bacterial-produced growth factors for marine microalgae.     

Diatom Vacuolar 1,6‐β‐Transglycosylases can Functionally Complement the Respective Yeast Mutants

Diatoms are unicellular photoautotrophic algae, which can be found in any aquatic habitat. The main storage carbohydrate of diatoms is chrysolaminarin, a nonlinear β‐glucan, consisting of a linear 1,3‐β‐chain with 1,6‐β‐branches, which is stored in cytoplasmic vacuoles. The metabolic pathways of chrysolaminarin synthesis in diatoms are poorly investigated, therefore we studied two potential 1,6‐β‐transglycosylases (TGS) of the diatom Phaeodactylum tricornutum which are similar to yeast Kre6 proteins and which potentially are involved in the branching of 1,3‐β‐glucan chains by adding d ‐glucose as 1,6‐side chains. We genetically fused the full‐length diatom TGS proteins to GFP and expressed these constructs in P. tricornutum, demonstrating that the enzymes are apparently located in the vacuoles, which indicates that branching of chrysolaminarin may occur in these organelles. Furthermore, we demonstrated the functionality of the diatom enzymes by expressing TGS1 and 2 proteins in yeast, which resulted in a partial complementation of growth deficiencies of a transglycosylase‐deficient ?kre6 yeast strain.     

Production characteristics of <Emphasis Type="Italic">Phaeodactylum tricornutum</Emphasis> Bohlin grown on medium with artificial sea water

The growth of marine diatoms Phaeodactylum tricornutum was investigated on a medium with artificial sea water under artificial and natural light. The maximum specific growth rate was 0.7 day^–1, the productivity was 0.8 g/L day, and the maximum biomass was 3.86 g/L under artificial light in laboratory conditions. In the conditions of Crimea, the maximum productivity of P. tricornutum amounted to 6 g/m² day under natural light in an outdoor photobioreactor (pool). The results of cultivation of P. tricornutum in a pool with artificial seawater under natural light may serve as a basis for developing technologies for the industrial cultivation of algae.     

Fast Fourier infrared spectroscopy to characterize the biochemical composition in diatoms

Diatoms are photosynthetic unicellular microalgae and are nature’s hidden source of several biosynthetic metabolites with their use in biofuel, food and drug industries. They mainly contain various lipids, sterols, isoprenoids and toxins with their use in apoptotic, fertility controlling and cancer drugs. Chemical studies on diatoms are limited due to various limitations such as variation of nutrients, contaminants and change in seasonal factors in the environment. To overcome these limitations, we obtained axenic cultures of 12 fresh-water diatom strains on the 22nd day of inoculation having a dry weight of 1 mg each and performed their Fourier transform infrared (FTIR) study for the detection of functional groups responsible for their chemical moiety. The spectral mapping showed a varied level of polyunsaturated fatty acids, amides, amines, ketone bodies and esters for their applications in various pharmacological, food and biofuel industries in the exponential phase of their growth in f/2 media. The FTIR study of the 12 diatom strains showed various similarities in the form of some common peak patterns ranging from 3000 to 3600 cm^?1 for ν_O–H absorption. The symmetric stretching vibration frequency of Diadesmis confervaceae (V2) type species showed different behaviour than others in the spectral region starting from 1600 to 1700 cm^?1. The absorption between 1500 and 1575 cm^?1 reflects the presence of the –N–H group. Infrared (IR) absorptions falling between 1600 and 1700 cm^?1 reflect the presence of amide’s ν_C=O in all species. Placoneis elginensis (V8) type species showed an additional absorption band which is centred around 1735–1750 cm^?1 which perhaps reflects the presence of ester’s ν_C=O. Diadesmis confervaceae (V2), Nitzschia palea (V4), Placoneis elginensis (V8), Nitzschia palea var. debilis (V6), Nitzschia inconspicua (V10), Gomphonema parvulum (V11) and Sellaphora (V12) showed distinct structural features with important key functionalities that can make them essential drug markers in the pharmaceutical industry.     

Impacts of enhanced UVB radiation on photosynthetic characteristics of the marine diatom <Emphasis Type="Italic">Phaeodactylum tricornutum</Emphasis> (Bacillariophyceae,Heterokontophyta)

Solar ultraviolet B (UVB) irradiance at the Earth’s surface is increasing due to anthropogenic influences. To evaluate the effects of enhanced UVB radiation on photosynthetic characteristics of the marine diatom Phaeodactylum tricornutum, the species was exposed to four levels of UVB radiation, 0, 0.25, 0.75, and 1.50 KJ m^?2 day^?1 for 7 days. Effects of UVB stress on net photosynthetic rate, net respiration rate, variable chlorophyll (Chl) fluorescence parameters, Chl a and carotenoid contents, and UV-absorbing compounds (UVACs) were investigated. Results showed that there were no significant differences in terms of net respiration rate or maximal photochemical efficiency of photosystem II (F_v/F_m) between the treatments in the short or long term. However, enhanced UVB radiation at an intensity of 0.16 W m^?2 had a negative effect on the net photosynthetic rate, electron transport rate, and on the pathway of excess energy dissipation over the short term (1 to 5 days). Carotenoid and UVACs content increased under UVB radiation. Photosynthetic parameters were unaffected by UVB radiation on the seventh day indicating that P. tricornutum can adapt to UVB radiation in the long term. Results of the present study indicate that there is a dynamic balance between damage and adaptation in microalgae that enables them to adapt to UVB-induced photosystem alterations during both short-term and long-term exposure.     

Paul Henry Latimer (1925–2011): discoverer of selective scattering in photosynthetic systems

The response of marine phytoplankton to the ongoing increase in atmospheric pCO₂ reflects the consequences of both increased CO₂ concentration and decreased pH in surface seawater. In the model diatom Thalassiosira weissflogii, we explored the effects of varying pCO₂ and pH, independently and in concert, on photosynthesis and respiration by incubating samples in water enriched in H₂ ¹⁸O. In long-term experiments (~6-h) at saturating light intensity, we observed no effects of pH or pCO₂ on growth rate, photosynthesis or respiration. This absence of a measurable response reflects the very small change in energy used by the carbon concentrating mechanism (CCM) compared to the energy used in carbon fixation. In short-term experiments (~3 min), we also observed no effects of pCO₂ or pH, even under limiting light intensity. We surmise that in T. weissflogii, it is the photosynthetic production of NADPH and ATP, rather than the CO₂-saturation of Rubisco that controls the rate of photosynthesis at low irradiance. In short-term experiments, we observed a slightly higher respiration rate at low pH at the onset of the dark period, possibly reflecting the energy used for exporting H⁺ and maintaining pH homeostasis. Based on what is known of the biochemistry of marine phytoplankton, our results are likely generalizable to other diatoms and a number of other eukaryotic species. The direct effects of ocean acidification on growth, photosynthesis and respiration in these organisms should be small over the range of atmospheric pCO₂ predicted for the twenty-first century.     

The effect of constant darkness and short light periods on the survival and physiological fitness of two phytoplankton species and their growth potential after re-illumination

We tested the survival potential and fitness of two different algae strains (the diatom Thalassiosira weissflogii and the cryptophyceae Rhodomonas sp.) under different growth conditions (complete darkness and short light intervals, simulating conditions in a deep mixed water column) at different temperatures, plus the effect of these conditions on the physiological fitness and growth after re-illumination was examined. Both species survived the experimental conditions without significant cell loss or physiological damage. Two different survival strategies were observed: (1) the diatom T. weissflogii immediately reduced its metabolic rate and stopped cell division. The effect on chlorophyll a (chl-a) content and photosynthetic capacity was negligible. At 10 °C, T. weissflogii used the short light windows to metabolize carbohydrates and growth. (2) The cryptophyte Rhodomonas sp. initially continued to grow after transfer into all trials. However, the cell number decreased after day 6. Carbohydrate and chl-a content went on to decrease dramatically (70 and 50%, respectively). After 3 days of re-illumination, T. weissflogii grew faster than of Rhodomonas sp.. The diatom seemed to benefit from better start conditions and would out-compete the cryptophyte during a spring bloom. Our results highlight that these algae groups have different strategies in dealing with darkness, which potentially endow diatoms with a competitive advantage in deep mixed waters and in the season of early spring.     

Productivity and biochemical composition of <Emphasis Type="Italic">Tetradesmus obliquus</Emphasis> and <Emphasis Type="Italic">Phaeodactylum tricornutum</Emphasis>: effects of different cultivation approaches

The present work evaluated biomass productivity, carbon dioxide fixation rate, and biochemical composition of two microalgal species, Phaeodactylum tricornutum (Bacillariophyta) and Tetradesmus obliquus (Chlorophyta), cultivated indoors in high-technology photobioreactors (HT-PBR) and outdoors both in pilot ponds and low-technology photobioreactors in a greenhouse in southern Italy. Microalgae were grown in standard media, under nitrogen starvation, and in two liquid digestates obtained from anaerobic digestion of agro-zootechnical and vegetable biomass. P. tricornutum, cultivated in semi-continuous mode in indoor HT-PBRs with standard medium, showed a biomass productivity of 21.0?±?2.3 g m^?2 d^?1. Applying nitrogen starvation, the lipid productivity increased from 2.3 up to 4.5?±?0.5 g m^?2 d^?1, with a 24 % decrease of biomass productivity. For T. obliquus, a biomass productivity of 9.1?±?0.9 g m^?2 d^?1 in indoor HT-PBR was obtained using standard medium. Applying liquid digestates as fertilizers in open ponds, T. obliquus gave a biomass productivity (10.8?±?2.0 g m^?2 d^?1) not statistically different from complete medium such as P. tricornutum (6.5?±?2.2 g m^?2 d^?1). The biochemical data showed that the fatty acid composition of the microalgal biomass was affected by the different cultivation conditions for both microalgae. In conclusion, it was found that the microalgal productivity in standard medium was about doubled in HT-PBR compared to open ponds for P. tricornutum and was about 20 % higher for T. obliquus.     

Genome-wide analysis of the diatom cell cycle unveils a novel type of cyclins involved in environmental signaling

Background

Despite the enormous importance of diatoms in aquatic ecosystems and their broad industrial potential, little is known about their life cycle control. Diatoms typically inhabit rapidly changing and unstable environments, suggesting that cell cycle regulation in diatoms must have evolved to adequately integrate various environmental signals. The recent genome sequencing of Thalassiosira pseudonana and Phaeodactylum tricornutum allows us to explore the molecular conservation of cell cycle regulation in diatoms.

Results

By profile-based annotation of cell cycle genes, counterparts of conserved as well as new regulators were identified in T. pseudonana and P. tricornutum. In particular, the cyclin gene family was found to be expanded extensively compared to that of other eukaryotes and a novel type of cyclins was discovered, the diatom-specific cyclins. We established a synchronization method for P. tricornutum that enabled assignment of the different annotated genes to specific cell cycle phase transitions. The diatom-specific cyclins are predominantly expressed at the G1-to-S transition and some respond to phosphate availability, hinting at a role in connecting cell division to environmental stimuli.

Conclusion

The discovery of highly conserved and new cell cycle regulators suggests the evolution of unique control mechanisms for diatom cell division, probably contributing to their ability to adapt and survive under highly fluctuating environmental conditions.     

Occurrence of the planktonic bloom-forming marine diatom <Emphasis Type="Italic">Chaetoceros tenuissimus</Emphasis> Meunier and its infectious viruses in western Japan

The genus Chaetoceros is among the most species-rich marine planktonic diatoms. Most Chaetoceros are considered important primary producers in various marine environments, but because of their small size, we know little about their ecology and distribution. Therefore, from 2008 to 2012, we examined the occurrence of C. tenuissimus Meunier, one of the smallest members in the genus, and its infectious viruses in western Japanese coastal waters. Using real-time quantitative PCR, we found that C. tenuissimus was widely detected throughout our study sites, with a maximum concentration of 2.4 × 10⁷ cells/l in May 2012. Sediment analysis revealed that C. tenuissimus resting-stage cells were present at potentially high levels, despite its infectious viruses being detected in the same region. The present study suggests that C. tenuissimus remains highly productive even when surrounded by its infectious viruses. This tolerance to viral infection, along with the diatom’s fast growth rate, suggests that C. tenuissimus might play an important role in maintaining the growth of important filter feeders.     

Does allelopathy affect co-culturing <Emphasis Type="Italic">Haslea ostrearia</Emphasis> with other microalgae relevant to aquaculture?

Haslea ostrearia is a marine diatom known to produce marennine, a water-soluble blue-green pigment responsible for the greening of oysters in ponds along the French Atlantic coast. This phenomenon occurs seasonally when H. ostrearia blooms in oyster ponds, and it increases the economic value of cultured oysters. From an ecological perspective, H. ostrearia blooms are accompanied by a decrease in the abundance of other microalgae, suggesting that this diatom produces allelochemicals. Recent studies showed that purified marennine has other biological activities, for instance antioxidant, antibacterial, and antiviral activities, which could be used in aquaculture to promote this pigment as a natural antipathogen agent. One important issue regarding the possible use of H. ostrearia in aquaculture as a mixed algal diet, however, is the importance of marennine allelopathy. In this study, we investigated the allelopathic effect of H. ostrearia on the growth of five microalgal species relevant to aquaculture: Chaetoceros calcitrans, Skeletonema costatum, Phaeodactylum tricornutum, Tetraselmis suecica, and Tisochrysis lutea. Allelopathic tests were realized by co-culturing these microalgae with H. ostrearia in batch and in semi-continuous mode, based on initial biovolume ratios. Our findings showed that inhibition of the growth of microalgae due to the presence of H. ostrearia and marennine was species dependent. Skeletonema costatum, C. calcitrans, and T. lutea were significantly more sensitive, whereas T. suecica and P. tricornutum appeared to be more resistant. Growth irradiance significantly influenced the allelopathic effect against the sensitive species S. costatum, and the H. ostrearia production of marennine increases with irradiance. Data presented in this study partly support the hypothesis that marennine released into the culture medium possibly acts as an allelochemical compound, thus explaining the dominance of H. ostrearia and the loss of sensitive algae in oyster ponds, but also that some species are insensitive, which allows co-culturing and use in a mixed algal diet in aquaculture.     

Electron requirements for carbon incorporation along a diel light cycle in three marine diatom species

Diatoms account for about 40% of primary production in highly productive ecosystems. The development of a new generation of fluorometers has made it possible to improve estimation of the electron transport rate from photosystem II, which, when coupled with the carbon incorporation rate enables estimation of the electrons required for carbon fixation. The aim of this study was to investigate the daily dynamics of these electron requirements as a function of the diel light cycle in three relevant diatom species and to apprehend if the method of estimating the electron transport rate can lead to different pictures of the dynamics. The results confirmed the species-dependent capacity for photoacclimation under increasing light levels. Despite daily variations in the photosynthetic parameters, the results of this study underline the low daily variability of the electron requirements estimated using functional absorption of the photosystem II compared to an estimation based on a specific absorption cross section of chlorophyll a. The stability of the electron requirements throughout the day would suggest it is potentially possible to estimate high-frequency primary production by using autonomous variable fluorescence measurements from ships-of-opportunity or moorings, without taking potential daily variation in this parameter into consideration, but this result has to be confirmed on natural phytoplankton assemblages. The results obtained in this study confirm the low electron requirements of diatoms to perform photosynthesis, and suggest a potential additional source of energy for carbon fixation, as recently described in the literature for this class.