Coralline photosynthetic physiology across a steep light gradient.

Coralline algae (CA) are globally distributed and fulfil many important roles within coastal ecosystems. In this study, photosynthetically active radiation (PAR) measured for 616 days at 2 and 10 m in a temperate subtidal kelp forest in southern New Zealand provided context to photosynthesis vs. irradiance relationships for, and pigment concentrations of, an articulated coralline alga, Arthrocardia sp. and a crustose coralline species assemblage within the Hapalidiales order. The maximum photosynthetic rate Pmax of the Arthrocardia sp. (20.38 ± 2.38 µmol O2. gDW-1 h-1) was significantly higher than the Pmax of crustose coralline spp. (3.72 ± 0.74 µmol O2. gDW-1 h-1) at the same 2 m stratum. Pigment concentration of Arthrocardia sp. was significantly higher than that of crustose coralline spp. at the same depth, while pigment concentration of crustose coralline spp. at 2 and 10 m were not significantly affected by depth. The photosynthetic characteristics of these coralline algae represent a shade acclimated organism with low saturation irradiance (all Ek < 100 µmol photons m-2 s-1). Despite sevenfold difference in average daily dose between 2 and 10 m there was no significant effect of depth on the photosynthetic performance of crustose coralline algae measured. The lack of evidence for acclimation to low light could be because periods of clear water provide enough light to maintain photosynthesis, lower energetic requirements of species found at depth or constraints on the synthesis of photosynthetic pigments at greater depth.

Revealing hydrogen peroxide as an external stressor in macrophyte-dominated coastal ecosystems.

Benthic primary producers in coastal ecosystems provide important habitat for marine organisms through the provision of complex 3D habitat. Primary producers produce organic matter, while simultaneously producing reactive oxygen species, including hydrogen peroxide (H2O2), a driver of oxidative stress. Through their high biomass, productivity and effect on local hydrodynamics, benthic primary producers can potentially increase H2O2 concentrations surrounding the biogenic structures they form. The aim of this study was to identify the potential role of H2O2 produced by benthic primary producers as an external stressor in coastal ecosystems. This was achieved by measuring H2O2 concentrations within sea lettuce blooms (Ulva sp.), giant kelp forests (Macrocystis pyrifera), and seagrass meadows (Zostera muelleri); quantifying H2O2 production rates of these species; and testing heterotrophic bacterial response to relevant H2O2 concentrations. Ulva sp. produced five times more H2O2 than other species. At in situ concentrations, H2O2 inhibited bacterial production and carbon flow through the microbial loop by 75%. This study reveals H2O2 as an additional stressor in bloom-forming Ulva sp. with higher H2O2 production compared to the ecosystem engineers M. pyrifera and Z. muelleri. H2O2 production by benthic primary producers can affect carbon flow through the microbial loop, with the potential to propagate a stress signal up the food web.

Whole community estimates of macroalgal pigment concentration within two southern New Zealand kelp forests1.

Light availability is a fundamental factor that controls the productivity and distribution of macroalgae and is highly variable, both spatially and temporally, in subtidal coastal systems. Our comprehension of how macroalgae respond to such variability is a significant knowledge gap that limits our understanding of how light influences the structure and productivity of these environments. Here, we examined the pigment characteristics of individual species, and for the first time the whole community, within one low-light, and one high-light kelp-forest system in southern New Zealand. The aim was to quantify the range of pigmentation seen within the two kelp-forests which differed in irradiance regime. Light availability was 33% and 64% greater at the high-light compared to the low-light site at 2 and 10 m depth, respectively. Results suggested Phaeophyceae species at deeper depths in the low-light site may be living at the edge of their photosynthetic ability and pigment synthesis appeared significantly restricted. Even with greater investment in the pigment fucoxanthin, biomass of Phaeophyceae species was significantly lower in the low-light site. Highly pigmented Rhodophyceae species made a greater proportional contribution to community biomass within the low-light site where they likely possessed a photosynthetic advantage. This work helps explain discrepancies in community structure between the two study sites and explores the complex relationship between irradiance and photoacclimation. The comparison of community pigment concentration holds potential as a tool for assessing the relative degree of photoacclimation occurring between sites and provides a proxy of photosynthetic cost under a specific light regime.

Light dose versus rate of delivery: implications for macroalgal productivity.

The role of how light is delivered over time is an area of macroalgal photosynthesis that has been overlooked but may play a significant role in controlling rates of productivity and the structure and persistence of communities. Here we present data that quantify the relative influence of total quantum dose and delivery rate on the photosynthetic productivity of five ecologically important Phaeophyceae species from southern New Zealand. Results suggested that greater net oxygen production occurs when light is delivered at a lower photon flux density (PFD) over a longer period compared to a greater PFD over a shorter period, given the same total dose. This was due to greater efficiency (α) at a lower PFD which, for some species, meant a compensatory effect can occur. This resulted in equal or greater productivity even when the total quantum dose of the lower PFD was significantly reduced. It was also shown that light limitation at Huriawa Peninsula, where macroaglae were sourced, may be restricting the acclimation potential of species at greater depths, and that even at shallow depth periods of significant light limitation are likely to occur. This research is of particular interest as the variability of light delivery to coastal reef systems increases as a result of anthropogenic disturbances, and as the value of in situ community primary productivity estimates is recognised.

A kelp with integrity: Macrocystis pyrifera prioritises tissue maintenance in response to nitrogen fertilisation.

Our understanding of the response of vascular, terrestrial plants to nitrogen (N) addition is advanced and provides the foundation for modern agriculture. In comparison, information on responses of marine macroalgae to increased nitrogen is far less developed. We investigated how in situ pulses of nitrate (NO3 (-)) affected the growth and N physiology of Macrocystis pyrifera by adding N using potassium nitrate dissolution blocks during a period of low seawater N concentration. Multiple parameters (e.g. growth, pigments, soluble NO3 (-)) were measured in distinct tissues throughout entire fronds (apical meristem, stipe, adult blade, mature blade, sporophyll, and holdfast). Unexpectedly, N fertilisation did not enhance elongation rates within the frond, but instead thickness (biomass per unit area) increased in adult blades. Increased blade thickness may have enhanced tissue integrity as fertilised kelp had lower rates of blade erosion. Tissue chemistry also responded to enrichment; pigmentation, soluble NO3 (-), and % N were higher throughout fertilised fronds. Labelled (15)N traced N uptake and translocation from N sources in the kelp canopy to sinks in the holdfast, 10 m below. This is the first evidence of long-distance (>1 m) transport of N in macroalgae. Patterns in physiological parameters suggest that M. pyrifera displays functional differentiation between canopy and basal tissues that may aid in nutrient-tolerance strategies, similar to those seen in higher plants and unlike those seen in more simple algae (i.e. non-kelps). This study highlights how little we know about N additions and N-use strategies within kelp compared to the wealth of literature available for higher plants.

Seasonal differences in the effects of oscillatory and uni-directional flow on the growth and nitrate-uptake rates of juvenile Laminaria digitata (Phaeophyceae).

The influence of oscillatory versus unidirectional flow on the growth and nitrate-uptake rates of juvenile kelp, Laminaria digitata, was determined seasonally in experimental treatments that simulated as closely as possible natural environmental conditions. In winter, regardless of flow condition (oscillatory and unidirectional) or water velocity, no influence of water motion was observed on the growth rate of L. digitata. In summer, when ambient nitrate concentrations were low, increased water motion enhanced macroalgal growth, which is assumed to be related to an increase in the rate of supply of nutrients to the blade surface. Nitrate-uptake rates were significantly influenced by water motion and season. Lowest nitrate-uptake rates were observed for velocities <5 cm · s(-1) and nitrate-uptake rates increased by 20%-50% under oscillatory motion compared to unidirectional flow at the same average speed. These data further suggested that the diffusion boundary layer played a significant role in influencing nitrate-uptake rates. However, while increased nitrate-uptake in oscillatory flow was clear, this was not reflected in growth rates and further work is required to understand the disconnection of nitrate-uptake and growth by L. digitata in oscillatory flow. The data obtained support those from related field-based studies, which suggest that in summer, when insufficient nitrogen is available in the water to saturate metabolic demand, the growth rate of kelps will be influenced by water motion restricting mass transfer of nitrogen.

Restricted use of nitrate and a strong preference for ammonium reflects the nitrogen ecophysiology of a light-limited red alga.

Ammonium and nitrate are important sources of inorganic nitrogen for coastal primary producers. Nitrate has higher energy requirement for uptake and assimilation, compared with ammonium, suggesting that it might be a more efficient nitrogen source for slow-growing, light-limited macroalgae. To address this hypothesis, we examined the nitrogen ecophysiology of Anotrichium crinitum, a rhodophyte macroalgae common in low-light habitats in New Zealand. We measured seasonal changes in seawater nitrate and ammonium concentrations and the concentration of nitrate and ammonium stored internally by A. crinitum. We determined the maximal uptake rates of nitrate and ammonium seasonally and grew A. crinitum in the laboratory with these nitrogen sources under two ecologically relevant saturating light levels. Our results show that field-harvested A. crinitum has a high affinity for ammonium and although it will grow when supplied exclusively with nitrate, internal nitrate pools are low and it is unable to take up nitrate without several days of acclimation to saturating light. Our data predict that A. crinitum would be able to sustain growth with ammonium as the sole source of nitrogen, a strategy that would help it survive under low-light conditions that prevail in the field.

Diurnal fluctuations in seawater pH influence the response of a calcifying macroalga to ocean acidification.

Coastal ecosystems that are characterized by kelp forests encounter daily pH fluctuations, driven by photosynthesis and respiration, which are larger than pH changes owing to ocean acidification (OA) projected for surface ocean waters by 2100. We investigated whether mimicry of biologically mediated diurnal shifts in pH-based for the first time on pH time-series measurements within a kelp forest-would offset or amplify the negative effects of OA on calcifiers. In a 40-day laboratory experiment, the calcifying coralline macroalga, Arthrocardia corymbosa, was exposed to two mean pH treatments (8.05 or 7.65). For each mean, two experimental pH manipulations were applied. In one treatment, pH was held constant. In the second treatment, pH was manipulated around the mean (as a step-function), 0.4 pH units higher during daylight and 0.4 units lower during darkness to approximate diurnal fluctuations in a kelp forest. In all cases, growth rates were lower at a reduced mean pH, and fluctuations in pH acted additively to further reduce growth. Photosynthesis, recruitment and elemental composition did not change with pH, but δ(13)C increased at lower mean pH. Including environmental heterogeneity in experimental design will assist with a more accurate assessment of the responses of calcifiers to OA.

Water motion and pH jointly impact the availability of dissolved inorganic carbon to macroalgae.

The supply of dissolved inorganic carbon to seaweeds is a key factor regulating photosynthesis. Thinner diffusive boundary layers at the seaweed surface or greater seawater carbon dioxide (CO2) concentrations increase CO2 supply to the seaweed surface. This may benefit seaweeds by alleviating carbon limitation either via an increased supply of CO2 that is taken up by passive diffusion, or via the down-regulation of active carbon concentrating mechanisms (CCMs) that enable the utilization of the abundant ion bicarbonate (HCO3-). Laboratory experiments showed that a 5 times increase in water motion increases DIC uptake efficiency in both a non-CCM (Hymenena palmata, Rhodophyta) and CCM (Xiphophora gladiata, Phaeophyceae) seaweed. In a field survey, brown and green seaweeds with active-CCMs maintained their CCM activity under diverse conditions of water motion. Whereas red seaweeds exhibited flexible photosynthetic rates depending on CO2 availability, and species switched from a non-CCM strategy in wave-exposed sites to an active-CCM strategy in sheltered sites where mass transfer of CO2 would be reduced. 97-99% of the seaweed assemblages at both wave-sheltered and exposed sites consisted of active-CCM species. Variable sensitivities to external CO2 would drive different responses to increasing CO2 availability, although dominance of the CCM-strategy suggests this will have minimal impact within shallow seaweed assemblages.

Effect of temperature on sporulation and spore development of giant kelp (Macrocystis pyrifera).

Rising ocean temperature is a major driver of kelp forest decline worldwide and one that threatens to intensify over the coming decades. What is not particularly well understood are the mechanisms that drive loss and how they operate at differing life stages. This study aimed to establish an understanding of the effects of increasing temperature on the early developmental stages of the giant kelp, Macrocystis pyrifera. Sporulation was carried out across 10 temperature treatments from 9.5 to 26.2°C ± 0.2°C at approximately 2°C intervals. Spores were then incubated at these temperatures under a 20.3±1.7 µmol photons m-2 s-1, 16L:8D photoperiod for 5 days. Results indicate that spore release was positively correlated with increasing temperature, whereas an inverse trend was observed between temperature and the growth of germ-tube. The thermal threshold for spore and germling development was determined to be between 21.7°C and 23.8°C. Spore settlement was the most drastically effected developmental phase by increasing temperature. This study highlights the vulnerability of early life stages of M. pyrifera development to rising ocean temperature and has implications for modelling future distribution of this valuable ecosystem engineer in a changing ocean.

Reducing the arbitrary: fuzzy detection of microbial ecotones and ecosystems - focus on the pelagic environment.

BACKGROUND: One of the central objectives of microbial ecology is to study the distribution of microbial communities and their association with their environments. Biogeographical studies have partitioned the oceans into provinces and regions, but the identification of their boundaries remains challenging, hindering our ability to study transition zones (i.e. ecotones) and microbial ecosystem heterogeneity. Fuzzy clustering is a promising method to do so, as it creates overlapping sets of clusters. The outputs of these analyses thus appear both structured (into clusters) and gradual (due to the overlaps), which aligns with the inherent continuity of the pelagic environment, and solves the issue of defining ecosystem boundaries. RESULTS: We show the suitability of applying fuzzy clustering to address the patchiness of microbial ecosystems, integrating environmental (Sea Surface Temperature, Salinity) and bacterioplankton data (Operational Taxonomic Units (OTUs) based on 16S rRNA gene) collected during six cruises over 1.5 years from the subtropical frontal zone off New Zealand. The technique was able to precisely identify ecological heterogeneity, distinguishing both the patches and the transitions between them. In particular we show that the subtropical front is a distinct, albeit transient, microbial ecosystem. Each water mass harboured a specific microbial community, and the characteristics of their ecotones matched the characteristics of the environmental transitions, highlighting that environmental mixing lead to community mixing. Further explorations into the OTU community compositions revealed that, although only a small proportion of the OTUs explained community variance, their associations with given water mass were consistent through time. CONCLUSION: We demonstrate recurrent associations between microbial communities and dynamic oceanic features. Fuzzy clusters can be applied to any ecosystem (terrestrial, human, marine, etc) to solve uncertainties regarding the position of microbial ecological boundaries and to refine the relation between the distribution of microorganisms and their environment.

Light Limitation within Southern New Zealand Kelp Forest Communities.

Light is the fundamental driver of primary productivity in the marine environment. Reduced light availability has the potential to alter the distribution, community composition, and productivity of key benthic primary producers, potentially reducing habitat and energy provision to coastal food webs. We compared the underwater light environment of macroalgal dominated shallow subtidal rocky reef habitats on a coastline modified by human activities with a coastline of forested catchments. Key metrics describing the availability of photosynthetically active radiation (PAR) were determined over 295 days and were related to macroalgal depth distribution, community composition, and standing biomass patterns, which were recorded seasonally. Light attenuation was more than twice as high in shallow subtidal zones along the modified coast. Macroalgal biomass was 2-5 times greater within forested sites, and even in shallow water (2m) a significant difference in biomass was observed. Long-term light dose provided the best explanation for differences in observed biomass between modified and forested coasts, with light availability over the study period differing by 60 and 90 mol photons m-2 at 2 and 10 metres, respectively. Higher biomass on the forested coast was driven by the presence of larger individuals rather than species diversity or density. This study suggests that commonly used metrics such as species diversity and density are not as sensitive as direct measures of biomass when detecting the effects of light limitation within macroalgal communities.

Diffusion boundary layers ameliorate the negative effects of ocean acidification on the temperate coralline macroalga Arthrocardia corymbosa.

Anthropogenically-modulated reductions in pH, termed ocean acidification, could pose a major threat to the physiological performance, stocks, and biodiversity of calcifiers and may devalue their ecosystem services. Recent debate has focussed on the need to develop approaches to arrest the potential negative impacts of ocean acidification on ecosystems dominated by calcareous organisms. In this study, we demonstrate the role of a discrete (i.e. diffusion) boundary layer (DBL), formed at the surface of some calcifying species under slow flows, in buffering them from the corrosive effects of low pH seawater. The coralline macroalga Arthrocardia corymbosa was grown in a multifactorial experiment with two mean pH levels (8.05 'ambient' and 7.65 a worst case 'ocean acidification' scenario projected for 2100), each with two levels of seawater flow (fast and slow, i.e. DBL thin or thick). Coralline algae grown under slow flows with thick DBLs (i.e., unstirred with regular replenishment of seawater to their surface) maintained net growth and calcification at pH 7.65 whereas those in higher flows with thin DBLs had net dissolution. Growth under ambient seawater pH (8.05) was not significantly different in thin and thick DBL treatments. No other measured diagnostic (recruit sizes and numbers, photosynthetic metrics, %C, %N, %MgCO3) responded to the effects of reduced seawater pH. Thus, flow conditions that promote the formation of thick DBLs, may enhance the subsistence of calcifiers by creating localised hydrodynamic conditions where metabolic activity ameliorates the negative impacts of ocean acidification.

Survival in low light: photosynthesis and growth of a red alga in relation to measured in situ irradiance.

Reduced light availability for benthic primary producers as a result of anthropogenic activities may be an important driver of change in coastal seas. However, our knowledge of the minimum light requirements for benthic macroalgae limits our understanding of how these changes may affect primary productivity and the functioning of coastal ecosystems. This knowledge gap is particularly acute in deeper water, where the impacts of increased light attenuation will be most severe. We examined the minimum light requirements of Anotrichium crinitum, which dominates near the maximum depth limit for macroalgae throughout New Zealand and Southern Australia, and is a functional analog of rhodophyte macroalgae in temperate low-light (deep-water) habitats throughout the world. These data show that A. crinitum is a shade-adapted seaweed with modest light requirements for the initiation of net photosynthesis (1.49-2.25 µmol photons · m(-2)  · s(-1) ) and growth (0.12-0.19 mol photons · m(-2)  · d(-1) ). A. crinitum maintains high photosynthetic efficiency and pigment content and a low C:N ratio throughout the year and can maintain biomass under sub-compensation (critical) light levels for at least 5 d. Nevertheless, in situ photon flux is less than the minimum light requirement for A. crinitum on at least 103 d per annum and is rarely sufficient to saturate growth. These findings reinforce the importance of understanding the physiological response of macroalgae at the extremes of environmental gradients and highlight the need to establish minimum thresholds that modification of the subtidal light environment should not cross.

CARBON-USE STRATEGIES IN MACROALGAE: DIFFERENTIAL RESPONSES TO LOWERED PH AND IMPLICATIONS FOR OCEAN ACIDIFICATION(1).

Ocean acidification (OA) is a reduction in oceanic pH due to increased absorption of anthropogenically produced CO2 . This change alters the seawater concentrations of inorganic carbon species that are utilized by macroalgae for photosynthesis and calcification: CO2 and HCO3 (-) increase; CO3 (2-) decreases. Two common methods of experimentally reducing seawater pH differentially alter other aspects of carbonate chemistry: the addition of CO2 gas mimics changes predicted due to OA, while the addition of HCl results in a comparatively lower [HCO3 (-) ]. We measured the short-term photosynthetic responses of five macroalgal species with various carbon-use strategies in one of three seawater pH treatments: pH 7.5 lowered by bubbling CO2 gas, pH 7.5 lowered by HCl, and ambient pH 7.9. There was no difference in photosynthetic rates between the CO2 , HCl, or pH 7.9 treatments for any of the species examined. However, the ability of macroalgae to raise the pH of the surrounding seawater through carbon uptake was greatest in the pH 7.5 treatments. Modeling of pH change due to carbon assimilation indicated that macroalgal species that could utilize HCO3 (-) increased their use of CO2 in the pH 7.5 treatments compared to pH 7.9 treatments. Species only capable of using CO2 did so exclusively in all treatments. Although CO2 is not likely to be limiting for photosynthesis for the macroalgal species examined, the diffusive uptake of CO2 is less energetically expensive than active HCO3 (-) uptake, and so HCO3 (-) -using macroalgae may benefit in future seawater with elevated CO2 .

TESTING THE EFFECTS OF OCEAN ACIDIFICATION ON ALGAL METABOLISM: CONSIDERATIONS FOR EXPERIMENTAL DESIGNS(1).

Ocean acidification describes changes in the carbonate chemistry of the ocean due to the increased absorption of anthropogenically released CO2 . Experiments to elucidate the biological effects of ocean acidification on algae are not straightforward because when pH is altered, the carbon speciation in seawater is altered, which has implications for photosynthesis and, for calcifying algae, calcification. Furthermore, photosynthesis, respiration, and calcification will themselves alter the pH of the seawater medium. In this review, algal physiologists and seawater carbonate chemists combine their knowledge to provide the fundamental information on carbon physiology and seawater carbonate chemistry required to comprehend the complexities of how ocean acidification might affect algae metabolism. A wide range in responses of algae to ocean acidification has been observed, which may be explained by differences in algal physiology, timescales of the responses measured, study duration, and the method employed to alter pH. Two methods have been widely used in a range of experimental systems: CO2 bubbling and HCl/NaOH additions. These methods affect the speciation of carbonate ions in the culture medium differently; we discuss how this could influence the biological responses of algae and suggest a third method based on HCl/NaHCO3 additions. We then discuss eight key points that should be considered prior to setting up experiments, including which method of manipulating pH to choose, monitoring during experiments, techniques for adding acidified seawater, biological side effects, and other environmental factors. Finally, we consider incubation timescales and prior conditioning of algae in terms of regulation, acclimation, and adaptation to ocean acidification.