Description: Zostera marina (eelgrass) is a marine foundation species with key ecological roles in coastal habitats. Its bacterial microbiota has been well studied, but very little is known about its mycobiome. In this study, we have isolated and identified 13 fungal strains, dominated by Penicillium species (10 strains), from the leaf and the root rhizosphere of Baltic Z. marina. The organic extracts of the fungi that were cultured by an OSMAC (One-Strain–Many-Compounds) regime using five liquid culture media under both static and shaking conditions were investigated for their chemical and bioactivity profiles. All extracts showed strong anti-quorum sensing activity, and the majority of the Penicillium extracts displayed antimicrobial or anti-biofilm activity against Gram-negative environmental marine and human pathogens. HPLC-DAD-MS-based rapid metabolome analyses of the extracts indicated the high influence of culture conditions on the secondary metabolite (SM) profiles. Among 69 compounds detected in all Penicillium sp. extracts, 46 were successfully dereplicated. Analysis of SM relatedness in culture conditions by Hierarchical Cluster Analysis (HCA) revealed generally low similarity and showed a strong effect of medium selection on chemical profiles of Penicillium sp. This is the first study assessing both the metabolite and bioactivity profile of the fungi associated with Baltic eelgrass Z. marina.

Description: Microorganisms have been reported to induce settlement in various marine invertebrate larvae but their specificity of inductive capacities for the settlement of coral larvae remains poorly understood. In this study, we isolated 56 microbial strains from the crustose coralline alga (CCA) Hydrolithon reinboldii using five different media either with or without additional antibiotics and/or spiked CCA extract. We tested the isolates for their potential to induce settlement behavior in larvae of the brooding scleractinian coral Leptastrea purpurea. From these 56 CCA-associated microbial strains, we identified six bacterial classes and 18 families. The culturable bacterial community associated with H. reinboldii was dominated by Gammaproteobacteria, Actinobacteria, and Alphaproteobacteria while the Illumina MiSeq analysis showed that the culture-independent bacterial community was dominated by Gammaproteobacteria, Alphaproteobacteria, and Flavobacteria. Furthermore, we found no correlation between inductive settlement capacities and phylogenetic relationships. Instead, settlement behavior of L. purpurea larvae was induced by specific isolated species. Strains #1792 (Pseudovibrio denitrificans), #1678 (Acinetobacter pittii), #1633 (Pseudoalteromonas phenolica), #1772 (Marine bacterium LMG1), #1721 (Microbulbifer variabilis), and #1783 (Pseudoalteromonas rubra) induced settlement behavior in coral larvae at mostly high and significant levels (〉= 40%) but the remaining isolates strongly varied in their activity. Multispecies biofilms consisting of four strains (#1792, #1678, #1633, and #1721) were observed to synergistically increase settlement behavior levels (〉 90%); however, the addition of #1772 to the multispecies biofilms negatively affected coral larvae and resulted in a total loss of inducing activity. The findings provide new insights into the role of bacteria in the settlement process of scleractinian corals and may help to identify the true nature of bacteria-derived morphogenic cues.

Description: Marine natural products (MNPs) exhibit a wide range of pharmaceutically relevant bioactivities, including antibiotic, antiviral, anticancer, or anti-inflammatory properties. Besides marine macroorganisms such as sponges, algae, or corals, specifically marine bacteria and fungi have shown to produce novel secondary metabolites (SMs) with unique and diverse chemical structures that may hold the key for the development of novel drugs or drug leads. Apart from highlighting their potential benefit to humankind, this review is focusing on the manifold functions of SMs in the marine ecosystem. For example, potent MNPs have the ability to exile predators and competing organisms, act as attractants for mating purposes, or serve as dye for the expulsion or attraction of other organisms. A large compilation of literature on the role of MNPs in marine ecology is available, and several reviews evaluated the function of MNPs for the aforementioned topics. Therefore, we focused the second part of this review on the importance of bioactive compounds from crustose coralline algae (CCA) and their role during coral settlement, a topic that has received less attention. It has been shown that certain SMs derived from CCA and their associated bacteria are able to induce attachment and/or metamorphosis of many benthic invertebrate larvae, including globally threatened reef-building scleractinian corals. This review provides an overview on bioactivities of MNPs from marine microbes and their potential use in medicine as well as on the latest findings of the chemical ecology and settlement process of scleractinian corals and other invertebrate larvae.

Description: 〈jats:p〉Sexually produced juvenile scleractinian corals play a key role in the adaptation process of coral reefs, as they are considered to possess an innate plasticity and thus can adjust to changing environmental parameters within a certain range. In this study we investigated in detail the early life stages of the brooding species 〈jats:italic〉Leptastrea purpurea〈/jats:italic〉 to identify, categorize and visualize the critical steps of the complex transformation process from a swimming coral larva to a sessile coral recruit and later to a coral colony. For that, we performed settlement experiments using previously known cues: cycloprodigiosin (CYPRO) and crustose coralline algae (CCA) as well as novel cues: crude extracts of 〈jats:italic〉Pseudoalteromonas espejiana〈/jats:italic〉 and 〈jats:italic〉P. piscicida〈/jats:italic〉 to identify a general, cue-independent settlement pathway. We monitored the development of 〈jats:italic〉L. purpurea〈/jats:italic〉 over 12 months using bright field and fluorescence microscopy. Also we identified the fluorescence signals of 〈jats:italic〉L. purpurea〈/jats:italic〉 with confocal microscopy at four crucial development steps: (A) swimming larva, (B) metamorphosing larva, (C) coral recruit and (D) adult coral. Our methodological approach allowed us to observe an ontogenetic shift of fluorescence signals which provokes the hypothesis that certain fluorescence patterns might be connected to distinct sequential functions in the early life cycle of scleractinian corals. Our observations showed great similarities to the early development of other brooding and spawning corals, making 〈jats:italic〉L. purpurea〈/jats:italic〉 a prospective candidate to be used as a model organism for coral research. Furthermore, our in-depth picture series provides a robust monitoring reference for coral nurseries or field applications and demonstrates the potential of fluorescence as an indicator to instantly determine the growth stage of a developing coral recruit.〈/jats:p〉

Description: 〈jats:title〉Abstract〈/jats:title〉〈jats:p〉The global degradation of coral reefs is steadily increasing with ongoing climate change. Yet coral larvae settlement, a key mechanism of coral population rejuvenation and recovery, is largely understudied. Here, we show how the lipophilic, settlement-inducing bacterial pigment cycloprodigiosin (CYPRO) is actively harvested and subsequently enriched along the ectoderm of larvae of the scleractinian coral 〈jats:italic〉Leptastrea purpura〈/jats:italic〉. A light-dependent reaction transforms the CYPRO molecules through photolytic decomposition and provides a constant supply of hydrogen peroxide (H〈jats:sub〉2〈/jats:sub〉O〈jats:sub〉2〈/jats:sub〉), leading to attachment on the substrate and metamorphosis into a coral recruit. Micromolar concentrations of H〈jats:sub〉2〈/jats:sub〉O〈jats:sub〉2〈/jats:sub〉 in seawater also resulted in rapid metamorphosis, but without prior larval attachment. We propose that the morphogen CYPRO is responsible for initiating attachment while simultaneously acting as a molecular generator for the comprehensive metamorphosis of pelagic larvae. Ultimately, our approach opens a novel mechanistic dimension to the study of chemical signaling in coral settlement and provides unprecedented insights into the role of infochemicals in cross-kingdom interactions.〈/jats:p〉

Description: Melanin is a widely distributed and striking dark-colored pigment produced by countless living organisms. Although a wide range of bioactivities have been recognized, there are still major constraints in using melanin for biotechnological applications such as its fragmentary known chemical structure and its insolubility in inorganic and organic solvents. In this study, a bacterial culture of Streptomyces cavourensis SV 21 produced two distinct forms of melanin: (1) a particulate, insoluble form as well as (2) a rarely observed water-soluble form. The here presented novel, acid-free purification protocol of purified particulate melanin (PPM) and purified dissolved melanin (PDM) represents the basis for an in-depth comparison of their physicochemical and biological properties, which were compared to the traditional acid-based precipitation of melanin (AM) and to a synthetic melanin standard (SM). Our data show that the differences in solubility between PDM and PPM in aqueous solutions may be a result of different adjoining cation species, since the soluble PDM polymer is largely composed of Mg2+ ions and the insoluble PPM is dominated by Ca2+ ions. Furthermore, AM shared most properties with SM, which is likely attributed to a similar, acid-based production protocol. The here presented gentler approach of purifying melanin facilitates a new perspective of an intact form of soluble and insoluble melanin that is less chemical altered and thus closer to its original biological form.