Recent advances in recirculating aquaculture systems and role of microalgae to close system loop.

In the recirculating aquaculture systems (RAS), waste management of nutrient-rich byproducts accounts for 30-50% of the whole production costs. Integrating microalgae into RAS offers complementary solutions for transforming waste streams into valuable co-products. This review aims to provide an overview of recent advances in microalgae application to enhance RAS performance and derive value from all waste streams by using RAS effluents as microalgal nutrient sources. Aquaculture solid waste can be converted by hydrothermal liquefaction (HTL), then the resultant aqueous phase of HTL can be used for microalgae cultivation. In addition, microalgae generate the required oxygen while sequestering carbon dioxide. The review suggests a novel integrated system focusing on oxygenation and carbon dioxide capture along with recent technological developments concerning efficient microalgae cultivation and nutrient recovery techniques. In such system, microalgae-based biorefineries provide environmentally-conscious and economically-viable pathways for enhanced RAS performance and conversion of effluents into high-value products.

Integrated municipal wastewater treatment and lipid accumulation by a self-flocculating/floating microalga Limnothrix sp.

The present study evaluated the growth, self-flocculation, lipid content, and pollutants removal by Limnothrix sp. BASMWW-9 isolated from municipal wastewater treatment system and cultivated in municipal wastewater. The biomass yield and lipid content after 6 days of cultivation were 1.07 g dw/L and 27.34 %dw, respectively. In addition, its self-flocculating ability reached up to 90 % after harvesting time of 180 min. Moreover, COD,NH3-N, TN, and TP removalefficiencies were 71.65 %, 81.89 %, 74.64 %, and 80.16 %, respectively. The self-flocculation performance of Limnothrix sp. was greatly associated to its morphology and production of extracellular polymeric substances (EPS), with significant positive impact of the high calcium and magnesium content in municipal wastewater. Interestingly, blue light irradiation during harvest enhanced the aggregation and floc formation as a floating biomat, which was attributed to enhanced polysaccharides production. This study provides innovative harvest method for Limnothrix sp. BASMWW-9 cultivated in wastewater using blue light for enhanced lipid recovery.

Lipid-rich particles of processed food waste for microalgae harvest through lipid-enriched floating biomat formation.

Food waste was collected from the campus canteen and lipid-rich particles (LRP) phase was evaluated to harvest Tetradesmus obliquus. Box-Behnken design showed the highest harvest efficiency (HE) of 84.69 % in run#1 (LRP = 30 %; initial OD680 = 1.75; and harvest time = 6 h). Numerical optimization ramps suggested 24.15 % (v/v) LRP ratio, initial OD680 3.00, and harvest time 3.82 h for maximum HE. Two flocs were observed, a precipitate at the bottom (B-Floc) and a floating biomat (F-Floc). Experimental results showed HE of 88.3 %, with 67 % and 33 % of the harvested biomass forming F-Floc and B-Floc, respectively. Pre-heating of LRP in a boiling water bath for 10 min (HFB-T10) promoted F-Floc proportion up to 91.6 %. In addition, HFB-T10 showed the highest FAMEs yield of 11.17 g/L of the total used volume, which was significantly higher than that of the centrifuged cells and heat-untreated biomat. Moreover, HFB-T10 showed better iodine value and cetane number of the produced biodiesel.

Recent Advances in Micro-/Nanoplastic (MNPs) Removal by Microalgae and Possible Integrated Routes of Energy Recovery.

Reliance on plastic has resulted in the widespread occurrence of micro-/nanoplastics (MNPs) in aquatic ecosystems, threatening the food web and whole ecosystem functions. There is a tight interaction between MNPs and microalgae, as dominant living organisms and fundamental constituents at the base of the aquatic food web. Therefore, it is crucial to better understand the mechanisms underlying the interactions between plastic particles and microalgae, as well as the role of microalgae in removing MNPs from aquatic ecosystems. In addition, finding a suitable route for further utilization of MNP-contaminated algal biomass is of great importance. The present review article provides an interdisciplinary approach to elucidate microalgae-MNP interactions and subsequent impacts on microalgal physiology. The degradation of plastic in the environment and differences between micro- and nanoplastics are discussed. The possible toxic effects of MNPs on microalgal growth, photosynthetic activity, and morphology, due to physical or chemical interactions, are evaluated. In addition, the potential role of MNPs in microalgae cultivation and/or harvesting, together with further safe routes for biomass utilization in biofuel production, are suggested. Overall, the current article represents a state-of-the-art overview of MNP generation and the consequences of their accumulation in the environment, providing new insights into microalgae integrated routes of plastic removal and bioenergy production.

Microalgae and Bacteria Interaction-Evidence for Division of Diligence in the Alga Microbiota.

Microalgae are one of the most dominant forms of life on earth that is tightly associated with a distinct and specialized microbiota. We have previously shown that the microbiota of Scenedesmus quadricauda harbors less than 10 distinct microbial species. Here, we provide evidence that dominant species are affiliated with the genera of Variovorax, Porphyrobacter, and Dyadobacter. Experimental and transcriptome-based evidence implies that within this multispecies interaction, Dyadobacter is a key to alga growth and fitness and is highly adapted to live in the phycosphere. While presumably under light conditions the alga provides the energy source to the bacteria, Dyadobacter produces and releases mainly a large variety of polysaccharides modifying enzymes. This is coherent with high-level expression of the T9SS in alga cocultures. The transcriptome data further imply that quorum-quenching proteins (QQ) and biosynthesis of vitamins B1, B2, B5, B6, and B9 are expressed by Dyadobacter at high levels in comparison to Variovorax and Porphyrobacter. Notably, Dyadobacter produces a significant number of leucine-rich repeat (LRR) proteins and enzymes involved in bacterial reactive oxygen species (ROS) tolerance. Complementary to this, Variovorax expresses the genes of the biosynthesis of vitamins B2, B5, B6, B7, B9, and B12, and Porphyrobacter is specialized in the production of vitamins B2 and B6. Thus, the shared currency between partners are vitamins, microalgae growth-promoting substances, and dissolved carbon. This work significantly enlarges our knowledge on alga-bacteria interaction and demonstrates physiological investigations of microalgae and associated bacteria, using microscopy observations, photosynthetic activity measurements, and flow cytometry. IMPORTANCE The current study gives a detailed insight into mutualistic collaboration of microalgae and bacteria, including the involvement of competitive interplay between bacteria. We provide experimental evidence that Gram-negative bacteria belonging to the Dyadobacter, Porphyrobacter, and Variovorax are the key players in a Scenedesmus quadricauda alga-bacteria interaction. We impart strong evidence that Dyadobacter produces and releases polysaccharides degradation enzymes and leucine-rich repeat proteins; Variovorax supplies the consortium with auxins and vitamin B12, while Porphyrobacter produces a broad spectrum of B vitamins. We show not only that the microalgae collaborate with the bacteria and vice versa but also that the bacteria interact with each other via quorum-sensing and secretion system mechanisms. The shared currency between partners appears to be vitamins, microalgae growth-promoting substances, and dissolved carbon.

Draft Genome Sequence of the Green Alga Scenedesmus acuminatus SAG 38.81.

Scenedesmus acuminatus, also known as Tetradesmus acuminatus, is a promising green microalga for sustainable production of microalga products, including valuable compounds such as astaxanthin, ß-carotene, and lutein, polysaccharides such as ß-glucan, and polyunsaturated fatty acids. Here, we report the draft whole-genome sequence of Scenedesmus acuminatus SAG 38.81.

Efficient evaluation of a gene containment system for poplar through early flowering induction.

KEY MESSAGE: The early flowering system HSP::AtFT allowed a fast evaluation of a gene containment system based on the construct PsEND1::barnase-barstar for poplar. Transgenic lines showed disturbed pollen development and sterility. Vertical gene transfer through pollen flow from transgenic or non-native plant species into their crossable natural relatives is a major concern. Gene containment approaches have been proposed to reduce or even avoid gene flow among tree species. However, evaluation of genetic containment strategies for trees is very difficult due to the long-generation times. Early flowering induction would allow faster evaluation of genetic containment in this case. Although no reliable methods were available for the induction of fertile flowers in poplar, recently, a new early flowering approach was developed. In this study, early flowering poplar lines containing the gene construct PsEND1::barnase-barstar were obtained. The PsEND1 promoter was chosen due to its early expression pattern, its versality and efficiency for generation of male-sterile plants fused to the barnase gene. RT-PCRs confirmed barnase gene activity in flowers, and pollen development was disturbed, leading to sterile flowers. The system developed in this study represents a valuable tool for gene containment studies in forest tree species.

Darkness-induced effects on gene expression in Cosmarium crenatum (Zygnematophyceae) from a polar habitat.

Light is a key environmental regulator in all photosynthetic organisms. Many studies focused on the physiologic response to changes in light availability of species from the Zygnematophyceae, but the impact of the absence of light and the molecular acclimation process on the other side have been poorly understood. Here we present transcriptomic analyses of Cosmarium crenatum from a polar habitat exposed to darkness. The algae were cultured in dark for one week; cell number and quantum yield of photosystem II (Fv/Fm) were monitored. Cell number was stable, but the Fv/Fm decreased in both groups, darkness-treated and control. Gene expression analysis revealed a strong repression of transcripts associated with photosynthesis, photorespiration and cell wall development. General carbohydrate and lipid metabolism were differentially regulated, but starch is shown to be the primary energy source in these conditions. Additionally, C. crenatum induced mRNA responsible for epigenetic modifications which may be a specific response to an adaption and acclimation to polar conditions. Our study sheds light on the molecular acclimation process to darkness and provides ecological implications for new perspectives in this specialized group of green algae.

Effect of lipid-free microalgal biomass and waste glycerol on growth and lipid production of Scenedesmus obliquus: Innovative waste recycling for extraordinary lipid production.

In the present work, a novel approach of using growth medium with different substitutions of lipid-free algal hydrolysate (LFAH, 0, 5, 10 and 15%) and/or waste glycerol (WG, 0, 5, 10 and 20 g L-1) for enhanced biodiesel production from Scenedesmus obliquus was studied. Combination of different concentrations of WG with 15% LFAH showed the maximum significant biomass productivity, which represented 27.4, 30.5 and 28.9% over the control at combined 5, 10 and 20 g L-1 WG, respectively. The combinations of different LFAH with 20 g L-1 WG showed the maximum significant lipid accumulation, where lipid productivity showed its maximum significant value of 59.66 mg L-1 d-1 using LFAH15-WG10. In addition, LFAH15-WG10 significantly enhanced total FAMEs yield by 21.2% over the control. Moreover, it reduced polyunsaturated fatty acids (PUFAs) ratio from 52.1% to 47.8% of total FAMEs, and increased monounsaturated fatty acids (MUFAs) ratio from 26.6% to 31.3% of total FAMEs.

Responses of photosystems I and II of Acutodesmus obliquus to chemical stress caused by the use of recycled nutrients.

Nutrients derived from hydrothermal gasification of Acutodesmus obliquus were tested on its biological compatibility to support growth of the same microalgae. Photosynthetic parameters of photosystems I and II (PS I and PS II) were investigated to study physiological effects on the microalgal cell. The nutrients were collected as liquid residues. Dilutions of 1:500 showed no effect on both photosystems. Lower dilutions affected PS II initially and later also PS I. Cyclic electron flow around PS I compensated for loss of electrons due to partially inhibited PS II. The highest tested concentration of liquid residue erased any photosynthetic activity of PS II after 28 min and onwards. In contrast, PS I remained active. The results suggest that PS I is less susceptible than PS II and that the mixture of chemicals in the liquid residue did not directly affect PS I but PS II. The toxicants in the residues seemed to interfere with linear electron flow of PS II even though light-driven formation of radicals and subsequent damage to one of the photosystems can be excluded as demonstrated in darkness. Lowered photosynthetic activity of PS I during actinic irradiation was caused due to lack of supply of electrons from PS II. The cyclic electron flow might play a key role in delivering the energy needed to restore PS II activity and to biodegrade the toxicants when linear electron flow failed. These negative effects of liquid residue towards microalgal cells require a remediation step for direct application of the liquid residue to substitute commercial fertilizers in microalgal mass cultures.

Extracellular secretion of free fatty acids by the chrysophyte Ochromonas danica under photoautotrophic and mixotrophic growth.

Recently, microalgae have gained a lot of attention because of their ability to produce fatty acids in their surrounding environments. The present paper describes the influence of organic carbon on the different fatty acid pools including esterified fatty acids, intracellular free fatty acids and extracellular free fatty acids in Ochromonas danica. It also throws light on the ability of O. danica to secrete free fatty acids in the growth medium under photoautotrophic and mixotrophic conditions. Biomass production of photoautotrophically grown O. danica was higher than that of mixotrophically grown, where a cellular biomass formation of 1.8 g L(-1) was observed under photoautotrophic condition which was about five folds higher than that under mixotrophic conditions. Contrary, the esterified fatty acid content reached up to 99 mg g(-1) CDW under photoautotrophic conditions at the late exponential phase, while during mixotrophic conditions a maximum of 212 mg g(-1) CDW was observed at the stationary phase. Furthermore, O. danica cells grown under mixotrophic conditions showed higher intracellular free fatty acid and extracellular free fatty acid contents (up to 51 and 20 mg g(-1) CDW, respectively) than cells grown under photoautotrophic conditions (up to 26 and 4 mg g(-1) CDW, respectively). The intra- and extracellular free fatty acids consisted of a high proportion of polyunsaturated fatty acids, mainly C18:2n-6, C18:3n-3 and C20:4n-6.

Successful crossings with early flowering transgenic poplar: interspecific crossings, but not transgenesis, promoted aberrant phenotypes in offspring.

In forest tree species, the reproductive phase is reached only after many years or even decades of juvenile growth. Different early flowering systems based on the genetic transfer of heat-shock promoter driven flowering-time genes have been proposed for poplar; however, no fertile flowers were reported until now. Here, we studied flower and pollen development in both HSP::AtFT and wild-type male poplar in detail and developed an optimized heat treatment protocol to obtain fertile HSP::AtFT flowers. Anthers from HSP::AtFT poplar flowers containing fertile pollen grains showed arrested development in stage 12 instead of reaching phase 13 as do wild-type flowers. Pollen grains could be isolated under the binocular microscope and were used for intra- and interspecific crossings with wild-type poplar. F1-seedlings segregating the HSP::AtFT gene construct according to Mendelian laws were obtained. A comparison between intra- and interspecific crossings revealed that genetic transformation had no detrimental effects on F1-seedlings. However, interspecific crossings, a broadly accepted breeding method, produced 47% seedlings with an aberrant phenotype. The early flowering system presented in this study opens new possibilities for accelerating breeding of poplar and other forest tree species. Fast breeding and the selection of transgene-free plants, once the breeding process is concluded, can represent an attractive alternative even under very restrictive regulations.

Ultrastructure of Cosmarium strains (Zygnematophyceae, Streptophyta) collected from various geographic locations shows species-specific differences both at optimal and stress temperatures.

Plant species collected from various climatic zones and stressed in vitro at various temperatures reveal changes in cellular ultrastructure which are in accordance with the climate at their sampling sites. This observation initiated the investigation to establish if stress at different temperatures may cause diverse extents of changes in the ultrastructure of microalgal strains originating from different geographic zones. The study revealed that the six Cosmarium strains demonstrated ultrastructural characteristics that were consistent with their source location under optimal, low and high temperature conditions, pointing to their preference to specific climatic niches. Interestingly, chloroplasts of all of the Cosmarium strains correspond to a sun-adapted type, which is concomitant with earlier statements that these strains are rendered as high-light adapted algae. The Cosmarium strains developed multiple ultrastructural responses which enabled them to cope with excessive temperatures, occasionally occurring in desmid natural habitats. The appearance of cubic membranes and increased number of plastoglobules may represent the first line in protection against high-temperature stress, which is accompanied by the alteration of protein synthesis and the appearance of stress granules in order to preserve cell homeostasis. However, the prolonged warm- or cold-temperature stress obviously initiated the programmed cell death, as concluded from the appearance of several ultrastructural features observed in all of the Cosmarium strains. The fair acclimation possibilities and the ability to undergo programmed cell death in order to save the population, certainly favor the cosmopolitan distribution of the genus Cosmarium.

Xanthophyll cycle pool size and composition in several Cosmarium strains (Zygnematophyceae, Streptophyta) are related to their geographic distribution patterns.

The photosynthetic behaviour and composition of photosynthetic pigments of four Cosmarium strains collected from different geographic areas were examined under moderate and photoinhibitory white light by means of PAM fluorometry and high-performance liquid chromatography. Generally, all of the Cosmarium strains displayed the photosynthetic performance and the composition of xanthophyll cycle pigments corresponding to that of high-light adapted plants and algae, when grown under the standard laboratory conditions. However, photoinhibitory treatments provoked several strain- and species-specific characteristics despite the long-term cultivation in laboratory conditions. The typical arctic taxon, C. crenatum var. boldtianum, displayed an incomplete violaxanthin cycle yielding an accumulation of antheraxanthin during high light stress, which is considered as an adaptation to occasional high irradiances in the polar zone due to the albedo. So far, the violaxanthin/antheraxanthin turnover was known only in some prasinophycean algae. Antheraxanthin actively participated in the heat dissipation from PSII centres in C. crenatum, as concluded from a significant positive correlation between non-photochemical quenching (NPQ) and the quantity of antheraxanthin. In contrast, all the other Cosmarium strains displayed a complete violaxanthin de-epoxidase action during the high light treatments, as judged from the relatively high production of zeaxanthin which participated in thermal dissipation of excess energy.

Metagenome survey of a multispecies and alga-associated biofilm revealed key elements of bacterial-algal interactions in photobioreactors.

Photobioreactors (PBRs) are very attractive for sunlight-driven production of biofuels and capturing of anthropogenic CO2. One major problem associated with PBRs however, is that the bacteria usually associated with microalgae in nonaxenic cultures can lead to biofouling and thereby affect algal productivity. Here, we report on a phylogenetic, metagenome, and functional analysis of a mixed-species bacterial biofilm associated with the microalgae Chlorella vulgaris and Scenedesmus obliquus in a PBR. The biofilm diversity and population dynamics were examined through 16S rRNA phylogeny. Overall, the diversity was rather limited, with approximately 30 bacterial species associated with the algae. The majority of the observed microorganisms were affiliated with Alphaproteobacteria, Betaproteobacteria, and Bacteroidetes. A combined approach of sequencing via GS FLX Titanium from Roche and HiSeq 2000 from Illumina resulted in the overall production of 350 Mbp of sequenced DNA, 165 Mbp of which was assembled in larger contigs with a maximum size of 0.2 Mbp. A KEGG pathway analysis suggested high metabolic diversity with respect to the use of polymers and aromatic and nonaromatic compounds. Genes associated with the biosynthesis of essential B vitamins were highly redundant and functional. Moreover, a relatively high number of predicted and functional lipase and esterase genes indicated that the alga-associated bacteria are possibly a major sink for lipids and fatty acids produced by the microalgae. This is the first metagenome study of microalga- and PBR-associated biofilm bacteria, and it gives new clues for improved biofuel production in PBRs.

Protection strategies of Cosmarium strains (Zygnematophyceae, Streptophyta) isolated from various geographic regions against excessive photosynthetically active radiation.

Numerous in vitro investigations have suggested that macroalgae exhibit regular geographic and depth distribution patterns in accordance with the light and temperature predominance at their habitats; however, there have been only a few similar studies concerning microalgae. We examined the potential influence of irradiance on patterns of distribution of four Cosmarium strains isolated from various climatic zones and cultured long term (>15 years) under a constant temperature-light regime. All the Cosmarium strains demonstrated physiological responses that were consistent with the light intensity prevailing at their source location, confirming that these responses are genetically preserved, as concluded from chlorophyll fluorescence and oxygen evolution rates measurements. Addition of inhibitors of chloroplast-encoded protein synthesis (chloramphenicol and streptomycin) and violaxanthin de-epoxidase (dithiothreitol) indicated that the Cosmarium strains developed "sun- or shade-plant" protection strategies, in accordance with the climate at their sampling sites. The polar Cosmarium strains exhibited a "shade-plant strategy"-to suffer some photoinhibition, but acquire increasing protection from photoinhibited PSII centers, whereas the tropical strains displayed a "sun-plant strategy"-to counteract photoinhibition of PSII by a high rate of repair of photoinhibited PSII reaction centers and a high xanthophyll cycle turnover.

The effect of discontinuous airlift mixing in outdoor flat panel photobioreactors on growth of Scenedesmus obliquus.

Discontinuous airlift mixing was realized by injecting pressured air at time intervals with a frequency between 0.033 and 0.25 Hz (at 80 kPa; i.e., every 4-30 s; valve opening time 800 ms) into outdoor flat panel photobioreactors ([Formula: see text]). This caused a flow velocity between 2 and 20 cm s(-1) of the culture medium within the photobioreactor and the mixing time was between 38 and 103.5 s, requiring 0.175-1.340 L(gas volume) L(photobioreactor volume)(-1) min(-1) pressured air. In order to detect the effect on growth of Scenedesmus obliquus during outdoor experiments and to be able to compare obtained results, a batch run with an airlift frequency of 0.25 Hz was simultaneously used as control. Growth at different airlift frequencies was measured by the increase of cell dry weight (CDW) during 3-5 days and biomass yield on light energy was calculated. With increasing airlift frequencies, growth increased from 52 to 91 % compared to the control. When CDW was at around 1.0-1.5 g L(-1), airlift frequency had no effect on growth, indicating that mass transfer gradients of nutrients and gas were not the limiting factors of growth. Above 1.5 g CDW L(-1), growth increased with increasing airlift frequency and light limitation for a single cell occurred. This effect was observed during low and high irradiance and it is concluded that a higher mean flow causes a better light distribution, resulting in an enhanced growth. Biomass productivity and demand of pressured air are correlated logarithmically, which enables to save mixing energy during cultivation.

Optimization of biomass and fatty acid productivity of Scenedesmus obliquus as a promising microalga for biodiesel production.

Nowadays, microalgae are discussed as a promising feedstock for biodiesel production. The present study examines the possibility of enhancement of fatty acid productivity of Scenedesmus obliquus by modifications of the culture medium composition. The effect of different concentrations of sodium bicarbonate, salinity, potassium nitrate, glycerol and sugarcane molasses on the enhancement of biomass and esterified fatty acids production was studied. NaHCO3 caused an increase in the biomass productivity at low concentrations (0.5 g L(-1)), while negatively affected fatty acid productivity at all tested concentrations. Increase of salinity enhanced both biomass and fatty acid productivity. The optimum NaCl concentration and sea water ratio were 0.94 g L(-1) and 25 % which resulted in 56 and 39 % increase in fatty acid productivity, respectively. Nitrogen deficiency showed increase in fatty acid content by 54 % over control but fatty acid productivity was decreased as a result of growth inhibition. Nitrogen-free cultures and cultures treated with -50 % concentrations of KNO3 showed 96 and 42 % decrease in EFA productivity, respectively, as compared with the control. Addition of 0.05 and 0.1 M of glycerol increased the biomass productivity by 6 and 5 %, respectively but showed no significant effect on fatty acid productivity as a result of decrease in fatty acid content. Finally, usage of sugarcane molasses stimulated both biomass and fatty acid content. The increase in fatty acid productivity was 32, 65 and 73 % above the control level at 1, 3 and 5 g L(-1) of sugarcane molasses, respectively.

Irradiance optimization of outdoor microalgal cultures using solar tracked photobioreactors.

Photosynthetic activity and temperature regulation of microalgal cultures (Chlorella vulgaris and Scenedesmus obliquus) under different irradiances controlled by a solar tracker and different cell densities were studied in outdoor flat panel photobioreactors. An automated process control unit regulated light and temperature as well as pH value and nutrient concentration in the culture medium. CO2 was supplied using flue gas from an attached combined block heat and power station. Photosynthetic activity was determined by pulse amplitude modulation fluorometry. Compared to the horizontal irradiance of 55 mol photons m(-2) d(-1) on a clear day, the solar tracked photobioreactors enabled a decrease and increase in the overall light absorption from 19 mol photons m(-2) d(-1) (by rotation out of direct irradiance) to 79 mol photons m(-2) d(-1) (following the position of the sun). At biomass concentrations below 1.1 g cell dry weight (CDW) L(-1), photoinhibition of about 35 % occurred at irradiances of ≥1,000 µmol photons m(-2) s(-1) photosynthetic active radiation (PAR). Using solar tracked photobioreactors, photoinhibition can be reduced and at optimum biomass concentration (≥2.3 g CDW L(-1)), the culture was irradiated up to 2,000 µmol photons m(-2) s(-1) to overcome light limitation with biomass yields of 0.7 g CDW mol photons(-1) and high photosynthetic activities indicated by an effective quantum yield of 0.68 and a maximum quantum yield of 0.80 (F v/F m). Overheating due to high irradiance was avoided by turning the PBR out of the sun or using a cooling system, which maintained the temperature close to the species-specific temperature optima.

Prevailing sea surface temperatures inhibit summer reproduction of the kelp Laminaria digitata at Helgoland (North Sea).

The impact of abiotic factors on kelp sporophyte reproduction has rarely been investigated. Laminaria digitata (Hudson) J.V. Lamouroux is one of the few summer fertile Laminaria species worldwide and reproduction is subjected to relatively high water temperatures. We investigated the impact of prevailing summer temperatures (~18°C in August) on the induction of sporangia, meiospore release, and germination at the island of Helgoland (North Sea). At Helgoland, fertile sporophytes are found between April and December with a maximum in late summer. While released meiospore numbers were constant between June and October, germination rates decreased significantly in summer. Short-term exposure of mature sori to 17°C-22°C induced a significantly higher meiospore release indicating enhancement of sporulation by elevated temperatures. Induction of sporangia on vegetative blade disks was not possible at 20°C, and fertility was only 20% at 18°C-19°C, but it was 100% in cool temperatures of 1°C-10°C. It was shown for the first time in a kelp species that "sporogenesis" is the life-cycle process with the narrowest temperature window compared to growth or survival of the sporophyte or reproduction, growth, and survival of the gametophyte. We incorporated several parameters (induction time, fertile area, and relative fertility) into a "Reproductive efficiency index." This indicates that sporogenesis of L. digitata is a cold-adapted process with an optimum at (5)-10°C. The results show that the population at Helgoland is at its reproduction limit despite the existence of other geographically more southerly located populations.