Irene Olivé

Establishing research strategies, methodologies and technologies to link genomics and proteomics to seagrass productivity, community metabolism, and ecosystem carbon fluxes

A complete understanding of the mechanistic basis of marine ecosystem functioning is only possible through integrative and interdisciplinary research. This enables the prediction of change and possibly the mitigation of the consequences of anthropogenic impacts. One major aim of the European Cooperation in Science and Technology (COST) Action ES0609 “Seagrasses productivity. From genes to ecosystem management,” is the calibration and synthesis of various methods and the development of innovative techniques and protocols for studying seagrass ecosystems. During 10 days, 20 researchers representing a range of disciplines (molecular biology, physiology, botany, ecology, oceanography, and underwater acoustics) gathered at The Station de Recherches Sous-marines et Océanographiques (STARESO, Corsica) to study together the nearby Posidonia oceanica meadow. STARESO is located in an oligotrophic area classified as “pristine site” where environmental disturbances caused by anthropogenic pressure are exceptionally low. The healthy P. oceanica meadow, which grows in front of the research station, colonizes the sea bottom from the surface to 37 m depth. During the study, genomic and proteomic approaches were integrated with ecophysiological and physical approaches with the aim of understanding changes in seagrass productivity and metabolism at different depths and along daily cycles. In this paper we report details on the approaches utilized and we forecast the potential of the data that will come from this synergistic approach not only for P. oceanica but for seagrasses in general.

Interactions of light and organic matter under contrasting resource simulated environments: the importance of clonal traits in the seagrass Zostera noltii

Light reduction in the water column and enhanced organic matter (OM) load into the sediments are two main consequences of eutrophication in marine coastal areas. This study addresses the combined effects of light, OM, and clonal traits in the seagrass Zostera noltii. Large Z. noltii plants were grown in sand with or without the addition of OM and under two light levels (high light and low light). Whereas some complete plant replicates were grown under homogeneous light and/or OM conditions, other replicates were grown under contrasting light and/or OM levels between the apical and the distal parts of the same plant. The three-way factorial design (light, OM load, and apex position) allowed us to determine the harmful effect of light reduction and OM enrichment on the growth, photosynthetic performance, and biochemical composition of Z. noltii. The addition of OM to the sediment promoted a decrease, or even an inhibition, in net plant growth regardless of the light level when the whole plants were grown under homogeneous light conditions. However, the results differed when plants were grown under contrasting light and/or OM conditions between apical and distal parts. In this case, the harmful effect of OM load was alleviated when apical parts were grown under high light conditions. OM loads also negatively affected the photosynthetic performance, evaluated as leaf fluorescence. The results indicate the importance of clonal traits in the response of Z. noltii growth to light conditions and OM enrichment.

Depth-specific fluctuations of gene expression and protein abundance modulate the photophysiology in the seagrass Posidonia oceanica

Here we present the results of a multiple organizational level analysis conceived to identify acclimative/adaptive strategies exhibited by the seagrass Posidonia oceanica to the daily fluctuations in the light environment, at contrasting depths. We assessed changes in photophysiological parameters, leaf respiration, pigments, and protein and mRNA expression levels. The results show that the diel oscillations of P. oceanica photophysiological and respiratory responses were related to transcripts and proteins expression of the genes involved in those processes and that there was a response asynchrony between shallow and deep plants probably caused by the strong differences in the light environment. The photochemical pathway of energy use was more effective in shallow plants due to higher light availability, but these plants needed more investment in photoprotection and photorepair, requiring higher translation and protein synthesis than deep plants. The genetic differentiation between deep and shallow stands suggests the existence of locally adapted genotypes to contrasting light environments. The depth-specific diel rhythms of photosynthetic and respiratory processes, from molecular to physiological levels, must be considered in the management and conservation of these key coastal ecosystems.

Linking gene expression to productivity to unravel long- and short-term responses of seagrasses exposed to CO2 in volcanic vents

Ocean acidification is a major threat for marine life but seagrasses are expected to benefit from high CO2. In situ (long-term) and transplanted (short-term) plant incubations of the seagrass Cymodocea nodosa were performed near and away the influence of volcanic CO2 vents at Vulcano Island to test the hypothesis of beneficial effects of CO2 on plant productivity. We relate, for the first time, the expression of photosynthetic, antioxidant and metal detoxification-related genes to net plant productivity (NPP). Results revealed a consistent pattern between gene expression and productivity indicating water origin as the main source of variability. However, the hypothesised beneficial effect of high CO2 around vents was not supported. We observed a consistent long- and short-term pattern of gene down-regulation and 2.5-fold NPP decrease in plants incubated in water from the vents and a generalized up-regulation and NPP increase in plants from the vent site incubated with water from the Reference site. Contrastingly, NPP of specimens experimentally exposed to a CO2 range significantly correlated with CO2 availability. The down-regulation of metal-related genes in C. nodosa leaves exposed to water from the venting site suggests that other factors than heavy metals, may be at play at Vulcano confounding the CO2 effects.

Genome-wide transcriptional reprogramming in the seagrass Cymodocea nodosa under experimental ocean acidification.

Here, we report the first use of massive‐scale RNA‐sequencing to explore seagrass response to CO2‐driven ocean acidification (OA). Large‐scale gene expression changes in the seagrass Cymodocea nodosa occurred at CO2 levels projected by the end of the century. C. nodosa transcriptome was obtained using Illumina RNA‐Seq technology and de novo assembly, and differential gene expression was explored in plants exposed to short‐term high CO2/low pH conditions. At high pCO2, there was a significant increased expression of transcripts associated with photosynthesis, including light reaction functions and CO2 fixation, and also to respiratory pathways, specifically for enzymes involved in glycolysis, in the tricarboxylic acid cycle and in the energy metabolism of the mitochondrial electron transport. The upregulation of respiratory metabolism is probably supported by the increased availability of photosynthates and increased energy demand for biosynthesis and stress‐related processes under elevated CO2 and low pH. The upregulation of several chaperones resembling heat stress‐induced changes in gene expression highlighted the positive role these proteins play in tolerance to intracellular acid stress in seagrasses. OA further modifies C. nodosa secondary metabolism inducing the transcription of enzymes related to biosynthesis of carbon‐based secondary compounds, in particular the synthesis of polyphenols and isoprenoid compounds that have a variety of biological functions including plant defence. By demonstrating which physiological processes are most sensitive to OA, this research provides a major advance in the understanding of seagrass metabolism in the context of altered seawater chemistry from global climate change.

Epiphytes Modulate Posidonia oceanica Photosynthetic Production, Energetic Balance, Antioxidant Mechanisms, and Oxidative Damage

Epiphytes impose physical barriers to light penetration into seagrass leaves causing shading, which may decrease the production of oxygen reactive species (ROS), but also constitute a physical aggression that may trigger the production of ROS, leading to oxidative damage. Here we investigate the effects of epiphytes on Posidonia oceanica under both interactive perspectives, light attenuation and oxidative stress. Specifically the role of epiphytes in net photosynthesis, chlorophyll a and b, photoprotection (Violaxanthin+Anteraxanthin+Zeaxanthin cycle), soluble sugar and starch contents, enzymatic (ascorbate peroxidase (APX) and dehydroascorbate reductase (DHAR)) and global (trolox equivalent antioxidant capacity (TEAC)) and oxygen radical antioxidant capacity (ORAC)) antioxidant responses, phenolics and oxidative damage (malondialdehyde) are tested. Leaves with epiphytes showed higher chlorophyll b and lower content in VAZ cycle carotenoids. Epiphyte shading was the probable reason for the lower VAZ de-epoxidation-ratio of leaves with epiphytes. In spite of being shaded, leaves with epiphytes showed higher antioxidant levels, indicating that epiphytes trigger the production of ROS. Both ORAC and TEAC and also APX and DHAR activities were higher in leaves with epiphytes, indicating that this response was related with its presence. Malondialdehyde concentrations also suggest oxidative damage caused by epiphytes. We conclude that the epiphyte load causes oxidative stress in P. oceanica and the mechanisms to scavenge ROS were not completely effective to avoid cell damage.

Acoustic detection of bubbles in a pond covered by the seagrass Cymodocea nodosa

This paper describes two experiments conducted in a pond covered by the seagrass Cymodocea nodosa at the Aquaculture Research Station of the Portuguese Institute for the Sea and Atmosphere in Olhao, Portugal, aiming at developing acoustic methods to assess oxygen production of seagrasses. The first experiment was carried out in July covering two days, when warm water and high photosynthetic rates give a high probability of oxygen supersaturation in water. The second experiment was carried out in late October, covering a period of 10 days, when seagrass productivity was expected to be lower than in July given the low irradiance and photoperiod. In the July experiment the high attenuation of low frequency pulses and broadband water pump noise (<20 kHz) in the afternoon is ascribed to bubbles formation during oxygen supersaturation conditions. This hypothesis is coherent with the significant increase of the backscattering level, as measured by an acoustic backscatter system operating at 0.5,1,2,4 MHz. Both, the attenuation of low frequency signals and backscattering level are correlated with oxygen supersaturation in water as measured by an optode. In the October experiment, when only water pump noise was acquired, the acoustic variability that can be related to photosynthetic activity was much weaker, nevertheless the attenuation shows a diurnal pattern correlated with the dissolved oxygen. The results suggest a significant release of oxygen as bubbles during photosynthesis, and therefore the potential contribution of acoustic methods to assess oxygen production of seagrass ecosystems.

Monitoring bubble production in a seagrass meadow using a source of opportunity

Under high irradiance, the photosynthetic activity of dense seagrass meadows saturates the water forming oxygen bubbles. The diel cycle of bubble production peaks at mid-day, following light intensity pattern. It is well known that bubbles strongly affect the acoustic propagation, increasing signal attenuation and decreasing the effective water sound speed, noticeable at low frequencies. Thus, the diurnal variability of bubbles may show an interference pattern in the spectrograms of low frequency acoustic signals. In an experiment conducted in July 2016 at the Aquaculture Research Station of the Portuguese Institute for the Sea and Atmosphere in Olhao, Portugal, the spectrograms of low frequency (<20kHz) broadband noise produced by water pumps in a pond of 0.48ha covered by the seagrass Cymodocea nodosa showed interference patterns that can be ascribed to the variability of the sound speed in the water. Preliminary analysis suggests that the daily cycle of bubble concentration can be inferred from these i...