Photoinhibition in benthic diatom assemblages under light stress

Microphytobenthos are frequently subjected to light intensities higher than those required to saturate photosynthesis, which consequently can cause photoinhibition. Photosystem II (PSII) protein D1 (the main target of photoinhibition) and xanthophyll cycle pigments were quantified in epipelic benthic diatom assemblages under high irradiance, in the presence of inhibitors and promoters of photoprotection mechanisms. Levels of D1 protein were significantly lower under high irradiance (1 h, 1500 µmol photons m–2 s–1) than under low light (80 µmol photons m–2 s–1), corresponding to a photoinhibition of 22 to 29%. Photoinhibition increased to 44 and 80% in the presence of lincomycin (inhibitor of chloroplast-protein synthesis) and dithiothreitol (inhibitor of the xanthophyll cycle), respectively. High light treatment had no significant effect on D1 protein concentrations in the presence of added glutathione and ascorbate, scavengers of reactive oxygen species (ROS). In contrast, the ROS promoter methylviologen increased photoinhibition to 63%. Under light stress, the functional stability of PSII reaction centres of the studied epipelic benthic diatoms was more dependent on xanthophyll cycle activation than on D1 protein recycling mechanisms, and our results substantiate the role of antioxidants in photoprotection via ROS scavenging.publishe

Response of the Diatom Phaeodactylum tricornutum to Photooxidative Stress Resulting from High Light Exposure

The response of microalgae to photooxidative stress resulting from high light exposure is a well-studied phenomenon. However, direct analyses of photosystem II (PSII) D1 protein (the main target of photoinhibition) in diatoms are scarce. In this study, the response of the diatom model species Phaeodactylum tricornutum to short-term exposure to high light was examined and the levels of D1 protein determined immunochemically. Low light (LL) acclimated cells (40 µmol photons m−2 s−1) subjected to high light (HL, 1,250 µmol photons m−2 s−1) showed rapid induction of non-photochemical quenching (NPQ) and ca. 20-fold increase in diatoxanthin (DT) concentration. This resulted from the conversion of diadinoxanthin (DD) to DT through the activation of the DD-cycle. D1 protein levels under LL decreased about 30% after 1 h of the addition of lincomycin (LINC), a chloroplast protein synthesis inhibitor, showing significant D1 degradation and repair under low irradiance. Exposure to HL lead to a 3.2-fold increase in D1 degradation rate, whereas average D1 repair rate was 1.3-x higher under HL than LL, leading to decreased levels of D1 protein under HL. There were significant effects of both HL and LINC on P. tricornutum maximum quantum yield of PSII (Fv/Fm), showing a reduction of active PSII reaction centres. Partial recovery of Fv/Fm in the dark demonstrates the photosynthetic resilience of this diatom to changes in the light regime. P. tricornutum showed high allocation of total protein to D1 and an active D1-repair cycle to limit photoinhibition

Photography-based taxonomy is inadequate, unnecessary, and potentially harmful for biological sciences

The question whether taxonomic descriptions naming new animal species without type specimen(s) deposited in collections should be accepted for publication by scientific journals and allowed by the Code has already been discussed in Zootaxa (Dubois & Nemésio 2007; Donegan 2008, 2009; Nemésio 2009a–b; Dubois 2009; Gentile & Snell 2009; Minelli 2009; Cianferoni & Bartolozzi 2016; Amorim et al. 2016). This question was again raised in a letter supported by 35 signatories published in the journal Nature (Pape et al. 2016) on 15 September 2016. On 25 September 2016, the following rebuttal (strictly limited to 300 words as per the editorial rules of Nature) was submitted to Nature, which on 18 October 2016 refused to publish it. As we think this problem is a very important one for zoological taxonomy, this text is published here exactly as submitted to Nature, followed by the list of the 493 taxonomists and collection-based researchers who signed it in the short time span from 20 September to 6 October 2016

N2, N2O and O2profiles in a Tagus estuary salt marsh

Vertical gas profiles of N2, N2O and O2were obtained in intact sediment cores from a Tagus estuary salt marsh using membrane inlet mass spectrometry. This technique allows direct measurements of dissolved gas concentrations with minimal disturbance. O2concentrations decreased sharply with depth, becoming undetectable below 14mm. Denitrification products (N2and N2O) occurred in the surface layer of the sediment where O2was present. Diffusion of N2and N2O from the anaerobic zone, denitrification in anaerobic microsites and aerobic denitrification are possible explanations for this observation. N2was the sole product of denitrification in control sediment cores probably because of the great demand for electron acceptors in this sediment. The addition of NO3 − and CH3CO2 − increased the concentrations of N2and N2O in the sediment. Significantly higher concentrations in treated cores occurred between 1·5 and 2·0cm for N2and between 0·5 and 1·5cm for N2O. The peak in N2concentration occurred in the anaerobic zone of the sediment, close to the aerobic–anaerobic interface while the peak in N2O concentration occurred above this interface where concentrations of O2were approximately 10μM. This is indicative that, in this sediment, production of N2O is less sensitive to the presence of O2than reduction of N2O to N2

N2, N2O and O2profiles in a Tagus estuary salt marsh

Vertical gas profiles of N2, N2O and O2were obtained in intact sediment cores from a Tagus estuary salt marsh using membrane inlet mass spectrometry. This technique allows direct measurements of dissolved gas concentrations with minimal disturbance. O2concentrations decreased sharply with depth, becoming undetectable below 14mm. Denitrification products (N2and N2O) occurred in the surface layer of the sediment where O2was present. Diffusion of N2and N2O from the anaerobic zone, denitrification in anaerobic microsites and aerobic denitrification are possible explanations for this observation. N2was the sole product of denitrification in control sediment cores probably because of the great demand for electron acceptors in this sediment. The addition of NO3 − and CH3CO2 − increased the concentrations of N2and N2O in the sediment. Significantly higher concentrations in treated cores occurred between 1·5 and 2·0cm for N2and between 0·5 and 1·5cm for N2O. The peak in N2concentration occurred in the anaerobic zone of the sediment, close to the aerobic–anaerobic interface while the peak in N2O concentration occurred above this interface where concentrations of O2were approximately 10μM. This is indicative that, in this sediment, production of N2O is less sensitive to the presence of O2than reduction of N2O to N2

Pheophorbide a in Hydrobia ulvae faecal pellets as a measure of microphytobenthos ingestion: variation over season and period of day

The microphytobenthos (MPB) – Hydrobia ulvae trophic interaction is one of the main channels of material transfer to higher trophic levels in intertidal mudflats. A new non-invasive approach to evaluate the grazing activity of H. ulvae on microphytobenthos is proposed. The effects of season and period (combination of tide and day/night) on ingestion rates of H. ulvae (using 14Clabeled MPB) and egested pheopigments a (using HPLC pigment analysis) were also investigated. H. ulvae ingestion rate was found to vary significantly over season and period, being higher in summer and during diurnal low tide periods. This is possibly related to higher growth rates of H. ulvaein summer, as well as to an increase in surface MPB biomass during diurnal low tides. A highly significant relationship was found between ingested chl a and egested pheophorbide a, allowing the estimation of ingestion rate from the amount of egested pheophorbide a on H. ulvae faecal pellets. This new non-invasive methodology may allow the improvement of long-term studies of consumption rates and the evaluation of grazing of H. ulvae on MPB.We thank I. Macário, P. Pochelon and T. Salvaterra for assistance in field work; M. Ruivo for assistance in HPLC and A. Luísa Santos for assistance in the Radioisotope Laboratory; A. Almeida for hosting the radioisotope work in the Radioisotope Laboratory at the Department of Biology, University of Aveiro; and 3 anonymous reviewers for critical comments on the manuscript. H.C. was supported by FCT – Fundação para a Ciência e Tecnologia (SFRH/BD/23720/ 2005). This work is part of the research project Benthic link – trophic links regulated by tidal and daily rhythms: benthic microflora and fauna in estuaries’, funded by FCT (POCI/BIABDE/61977/2004). Both grants were allocated by FCT under the Support Community Framework III, Operational Programme Science, Technology and Innovation. The methods used in this study comply with Portuguese legislation on animal experimentation.publishe

Kleptoplast photosynthesis is nutritionally relevant in the sea slug Elysia viridis

Several sacoglossan sea slug species feed on macroalgae and incorporate chloroplasts into tubular cells of their digestive diverticula. We investigated the role of the "stolen" chloroplasts (kleptoplasts) in the nutrition of the sea slug Elysia viridis and assessed how their abundance, distribution and photosynthetic activity were affected by light and starvation. Elysia viridis individuals feeding on the macroalga Codium tomentosum were compared with starved specimens kept in dark and low light conditions. A combination of variable Chl a fluorescence and hyperspectral imaging, and HPLC pigment analysis was used to evaluate the spatial and temporal variability of photopigments and of the photosynthetic capacity of kleptoplasts. We show increased loss of weight and body length in darkstarved E. viridis as compared to low light-starved sea slugs. A more pronounced decrease in kleptoplast abundance and lower photosynthetic electron transport rates were observed in dark-starved sea slugs than in low light-starved animals. This study presents strong evidence of the importance of kleptoplast photosynthesis for the nutrition of E. viridis in periods of food scarcity. Deprived of photosynthates, E. viridis could accelerate the breakdown of kleptoplasts in the dark to satisfy its' energy requirements

Effects of chlorophyll fluorescence on the estimation of microphytobenthos biomass using spectral reflectance indices

The communities of benthic microalgae that form dense biofilms at the surface of aquatic sediments, or microphytobenthos, are important primary producers in estuarine intertidal flats and shallow coastal waters. The microalgal biomass present in the photic zone of the sediment is a key parameter for ecological and photophysiological studies on microphytobenthos, and has been routinely estimated using hyperspectral reflectance indices based on the chlorophyll (Chl) a red absorption peak at 675 nm, usually the Normalised Difference Vegetation Index (NDVI). This study reports that red region-based biomass indices measured on microphytobenthos biofilms can be significantly affected by the enrichment of reflected light with solar-induced Chl fluorescence emitted by the microalgae. Chl fluorescence emission peaks at 683 nm, counterbalancing the decrease in reflectance centered at 675 nm, thus causing the underestimation of NDVI. The interference of Chl fluorescence was found to be easily identified by a conspicuous double-peak feature in the 670–700 nm region of the second-derivative reflectance spectra. The fluorescence-induced NDVI underestimation was shown to be most pronounced for high surface biomass levels and low incident solar irradiance. Particular aspects of microphytobenthos biofilms, such as the increase in surface Chl fluorescence due the contribution of emission by subsurface layers, and vertical migratory responses by motile microalgae to changes in ambient light, further complicate the effects on biomass estimation using NDVI-like indices. By comparing NDVI with a fluorescence-independent biomass index for a wide range of natural light conditions, it was found that Chl fluorescence interference may cause the underestimation of microalgal biomass to reach over 25%, with errors above 10% being expected for more than half of the measuring occasions. These results indicate that the use of NDVI may compromise the correct assessment of important aspects of microphytobenthos ecology, such as the characterisation of migratory behaviour or the determination of biomass-specific productivity rates, and call for the use of alternative biomass indices, not based on the Chl a red absorption peak