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BackgroundMarine sponges and their microbiomes contribute significantly to carbon and nutrient cycling in global reefs, processing and remineralizing dissolved and particulate organic matter. Lamellodysidea herbacea sponges obtain additional energy from abundant photosynthetic Hormoscilla cyanobacterial symbionts, which also produce polybrominated diphenyl ethers (PBDEs) chemically similar to anthropogenic pollutants of environmental concern. Potential contributions of non-Hormoscilla bacteria to Lamellodysidea microbiome metabolism and the synthesis and degradation of additional secondary metabolites are currently unknown.ResultsThis study has determined relative abundance, taxonomic novelty, metabolic capacities, and secondary metabolite potential in 21 previously uncharacterized, uncultured Lamellodysidea-associated microbial populations by reconstructing near-complete metagenome-assembled genomes (MAGs) to complement 16S rRNA gene amplicon studies. Microbial community compositions aligned with sponge host subgroup phylogeny in 16 samples from four host clades collected from multiple sites in Guam over a 3-year period, including representatives of Alphaproteobacteria, Gammaproteobacteria, Oligoflexia, and Bacteroidetes as well as Cyanobacteria (Hormoscilla). Unexpectedly, microbiomes from one host clade also included Cyanobacteria from the prolific secondary metabolite-producer genus Prochloron, a common tunicate symbiont. Two novel Alphaproteobacteria MAGs encoded pathways diagnostic for methylotrophic metabolism as well as type III secretion systems, and have been provisionally assigned to a new order, designated Candidatus Methylospongiales. MAGs from other taxonomic groups encoded light-driven energy production pathways using not only chlorophyll, but also bacteriochlorophyll and proteorhodopsin. Diverse heterotrophic capabilities favoring aerobic versus anaerobic conditions included pathways for degrading chitin, eukaryotic extracellular matrix polymers, phosphonates, dimethylsulfoniopropionate, trimethylamine, and benzoate. Genetic evidence identified an aerobic catabolic pathway for halogenated aromatics that may enable endogenous PBDEs to be used as a carbon and energy source.ConclusionsThe reconstruction of high-quality MAGs from all microbial taxa comprising greater than 0.1% of the sponge microbiome enabled species-specific assignment of unique metabolic features that could not have been predicted from taxonomic data alone. This information will promote more representative models of marine invertebrate microbiome contributions to host bioenergetics, the identification of potential new sponge parasites and pathogens based on conserved metabolic and physiological markers, and a better understanding of biosynthetic and degradative pathways for secondary metabolites and halogenated compounds in sponge-associated microbiota. Video Abstract.

Patellamides are highly bioactive compounds found along with other cyanobactins in the symbiosis between didemnid ascidians and the enigmatic cyanobacterium Prochloron. The biosynthetic pathway of patellamide synthesis is well understood, the relevant operons have been identified in the Prochloron genome and genes involved in patellamide synthesis are among the most highly transcribed cyanobacterial genes in hospite. However, a more detailed study of the in vivo dynamics of patellamides and their function in the ascidian-Prochloron symbiosis is complicated by the fact that Prochloron remains uncultivated despite numerous attempts since its discovery in 1975. A major challenge is to account for the highly dynamic microenvironmental conditions experienced by Prochloron in hospite, where light-dark cycles drive rapid shifts between hyperoxia and anoxia as well as pH variations from pH ~6 to ~10. Recently, work on patellamide analogues has pointed out a range of different catalytic functions of patellamide that could prove essential for the ascidian-Prochloron symbiosis and could be modulated by the strong microenvironmental dynamics. Here, we review fundamental properties of patellamides and their occurrence and dynamics in vitro and in vivo. We discuss possible functions of patellamides in the ascidian-Prochloron symbiosis and identify important knowledge gaps and needs for further experimental studies.

In chordates, obligate photosynthetic symbiosis has been reported exclusively in some colonial ascidians of the family Didemnidae. The vertical transmission of the symbionts is crucial in establishing the obligate symbiosis between the cyanobacteria and the host ascidians. The results of comparative surveys on the morphological processes of cyanobacterial transmission suggest the occurrence of convergent evolution of the vertical transmission in the host species harboring symbionts in the cloacal cavity. In Trididemnum species harboring cyanobacterial cells in the tunic, the symbiont cells are transported by the tunic cells to the tunic of embryos brooded in the tunic of the parent colony. The present study examined whether the mode of symbiont transmission is the same in host species harboring the symbionts in the tunic, regardless of host genera, or whether non‐Trididemnum hosts have a different vertical transmission mode. Our results showed that the vertical transmission process in Lissoclinum midui was almost the same as in the Trididemnum species, supporting the occurrence of convergent evolution in the two distinct didemnid genera, that is, Trididemnum and Lissoclinum. High plasticity of the embryogenic process in didemnid ascidians may be important in developing the mechanism of vertical transmission; this assumption may also explain why the obligate cyanobacterial symbiosis has been exclusively established in didemnid ascidians among chordates. [ABSTRACT FROM AUTHOR]

Ascidians are known to harbour bacterial symbionts in their tunics. In particular, the ascidian genus Lissoclinum can host abundant and diverse cyanobacterial associates. Here, we determined the diversity and host-specificity of cyanobacteria inhabiting 28 ascidian samples corresponding to eight Lissoclinum species: L. bistratum, L. midui, L. patella, L. punctatum, and L. timorense from Japan, L. aff. fragile and L. verrilli from the Bahamas, and L. perforatum from Spain and Chile. Cyanobacterial symbionts were characterized using both partial 16S rRNA gene sequences and sequences obtained for the entire 16S-23S rRNA internal transcribed spacer region (ITS). We found that both host species and geographic location played a role in structuring ascidian-cyanobacterial symbioses. Broad biogeographic trends included the dominance of Prochloron symbionts in Japanese ascidians and the presence of a novel cyanobacterial lineage in L. aff. fragile hosts from the Bahamas. Within each geographic region, a high degree of host-specificity was observed, where similar symbionts were recovered from ascidian hosts across multiple collection locations. Further, our analysis revealed the existence of nine distinct Prochloron clades in Japan, some of which corresponded to particular host species and sampling sites. For L. aff. fragile, further differences were observed between cyanobacterial symbionts in ascidians from reef and mangrove habitats. Our results showed high host-specificity in ascidian-cyanobacterial symbioses characterized by cryptic diversity and structured by host identity, location and habitat. [ABSTRACT FROM AUTHOR]

Prochloron spp. are obligate cyanobacterial symbionts of many didemnid family ascidians. It has been proposed that the cyclic peptides of the patellamide class found in didemnid extracts are synthesized by Prochloron spp., but studies in which host and symbiont cells are separated and chemically analyzed to identify the biosynthetic source have yielded inconclusive results. As part of the Prochloron didemni sequencing project, we identified patellamide biosynthetic genes and confirmed their function by heterologous expression of the whole pathway in Escherichia coli. The primary sequence of patellamides A and C is encoded on a single ORF that resembles a precursor peptide. We propose that this prepatellamide is heterocyclized to form thiazole and oxazoline rings, and the peptide is cleaved to yield the two cyclic patellamides, A and C. This work represents the full sequencing and functional expression of a marine natural-product pathway from an obligate symbiont. In addition, a related cluster was identified in Trichodesmium erythraeum IMS101, an important bloom-forming cyanobacterium.

Dinuclear CuII-patellamide complexes (patellamides are naturally occurring cyclic pseudo-octapeptides) are known to be efficient catalysts for hydrolysis reactions of biological importance, for example, those of phosphatase, carbonic anhydrase, and glycosidase. However, the biological role of patellamides is still unknown. Patellamides were originally extracted from the sea squirt Lissoclinum patella, but are now known to be ribosomally expressed by the blue-green algae Prochloron that live in symbiosis with L. patella. In a further step to unravel the metabolic significance of the patellamide complexes, the question as to whether these are also formed inside Prochloron cells is addressed. In this study, a biocompatible patellamide-fluorescent dye conjugate has been introduced into living Prochloron cells and, by means of flow cytometry and confocal microscopy, it is shown that CuII ions are coordinated to patellamides in vivo. [ABSTRACT FROM AUTHOR]

The patellamides (cyclic pseudo-octapeptides) are produced by Prochloron, a symbiont of the ascidians, marine invertebrate filter feeders. These pseudo-octapeptides are present in the cytoplasm and a possible natural function of putative metal complexes of these compounds is hydrolase activity, however the true biological role is still unknown. The dinuclear CuII complexes of synthetic patellamide derivatives have been shown in in vitro experiments to be efficient hydrolase model catalysts. Many hydrolase enzymes, specifically phosphatases and carboanhydrases, are ZnII-based enzymes and therefore, we have studied the ZnII and mixed ZnII/CuII solution chemistry of a series of synthetic patellamide derivatives, including solution structural and computational work, with the special focus on model phosphatase chemistry with bis-(2,4-dinitrophenyl)phosphate (BDNPP) as the substrate. The ZnII complexes of a series of ligands are shown to form complexes of similar structure and stability compared to the well-studied CuII analogues and the phosphatase reactivities are also similar. Since the complex stabilities and phosphatase activities are generally a little lower compared to those of CuII and since the concentration of ZnII in Prochloron cells is slightly smaller, we conclude that the CuII complexes of the patellamides are more likely to be of biological importance. [ABSTRACT FROM AUTHOR]

Diversity-generating metabolism leads to the evolution of many different chemicals in living organisms. Here, by examining a marine symbiosis, we provide a precise evolutionary model of how nature generates a family of novel chemicals, the cyanobactins. We show that tunicates and their symbiotic Prochloron cyanobacteria share congruent phylogenies, indicating that Prochloron phylogeny is related to host phylogeny and not to external habitat or geography. We observe that Prochloron exchanges discrete functional genetic modules for cyanobactin secondary metabolite biosynthesis in an otherwise conserved genetic background. The module exchange leads to gain or loss of discrete chemical functional groups. Because the underlying enzymes exhibit broad substrate tolerance, discrete exchange of substrates and enzymes between Prochloron strains leads to the rapid generation of chemical novelty. These results have implications in choosing biochemical pathways and enzymes for engineered or combinatorial biosynthesis. [ABSTRACT FROM AUTHOR]

Prochloron , a blue-green algae belonging to ancient prokaryotes, produces, like other cyanobacteria, cyclic pseudo -peptides, which are also found in its obligate symbiont ascidiae ( Lissoclinum patellum ). Although research has focused for some time on the putative metabolic function of these cyclic peptides, to date it is still not understood. Their role might be connected to the increased concentrations of divalent metal ions, especially Cu II , found in ascidiae. Dinuclear copper(II) complexes of cyclic pseudo -peptides revealed a broad hydrolytic capacity, including carboanhydrase and phosphatase activity. This study reports their β - lactamase as well as α- and β - glycosidase activity with k cat = (11.34 ± 0.91)ˑ10 − 4 s − 1 for β - lactamase, k cat = (1.55 ± 0.13)ˑ10 − 4 s − 1 for α-glycosidase and k cat = (1.22 ± 0.09)ˑ10 − 4 s − 1 for β - glycosidase activity. [ABSTRACT FROM AUTHOR]

The cyanobacterium P rochloron didemni is primarily found in symbiotic relationships with various marine hosts such as ascidians and sponges. P rochloron remains to be successfully cultivated outside of its host, which reflects a lack of knowledge of its unique ecophysiological requirements. We investigated the microenvironment and diversity of P rochloron inhabiting the upper, exposed surface of didemnid ascidians, providing the first insights into this microhabitat. The pH and O2 concentration in this P rochloron biofilm changes dynamically with irradiance, where photosynthetic activity measurements showed low light adaptation (Ek ∼80 ± 7 μmol photons m−2 s−1) but high light tolerance. Surface P rochloron cells exhibited a different fine structure to P rochloron cells from cloacal cavities in other ascidians, the principle difference being a central area of many vacuoles dissected by single thylakoids in the surface P rochloron. Cyanobacterial 16S rDNA pyro-sequencing of the biofilm community on four ascidians resulted in 433 operational taxonomic units ( OTUs) where on average −85% (65-99%) of all sequence reads, represented by 136 OTUs, were identified as P rochloron via blast search. All of the major P rochloron- OTUs clustered into independent, highly supported phylotypes separate from sequences reported for internal P rochloron, suggesting a hitherto unexplored genetic variability among P rochloron colonizing the outer surface of didemnids. [ABSTRACT FROM AUTHOR]

The photosymbiotic ascidianDiplosoma simileis characterized by a wide global distribution. The current study provides the first record ofD. similein the Red Sea, with remarks on its spatial distribution and phylogeny. Partial COI and 18S rDNA sequences ofD. similefrom the Red Sea revealed high similarity to those from the West Pacific, indicating a much broader range than previously recorded. In the Red SeaD. simileis common on natural coral reefs at depths ranging from 1–35 m.Diplosoma similecolonies were found to overgrow and would appear to suffocate several species of live stony corals, though this does not appear to cause mass coral mortality in the Red Sea, unlike that recorded in other locations. [ABSTRACT FROM AUTHOR]

The discovery of the cyanobacterium Prochloron was the first finding of a bacterial oxyphototroph with chlorophyll (Chl) b, in addition to Chl a. It was first described as Prochloron didemni but a number of clades have since been described. Prochloron is a conspicuously large (7-25 µm) unicellular cyanobacterium living in a symbiotic relationship, primarily with (sub-) tropical didemnid ascidians; it has resisted numerous cultivation attempts and appears truly obligatory symbiotic. Recently, a Prochloron draft genome was published, revealing no lack of metabolic genes that could explain the apparent inability to reproduce and sustain photosynthesis in a free-living stage. Possibly, the unsuccessful cultivation is partly due to a lack of knowledge about the microenvironmental conditions and ecophysiology of Prochloron in its natural habitat. We used microsensors, variable chlorophyll fluorescence imaging and imaging of O2 and pH to obtain a detailed insight to the microenvironmental ecology and photobiology of Prochloron in hospite in the didemnid ascidian Lissoclinum patella. The microenvironment within ascidians is characterized by steep gradients of light and chemical parameters that change rapidly with varying irradiances. The interior zone of the ascidians harboring Prochloron thus became anoxic and acidic within a few min of darkness, while the same zone exhibited O2 super-saturation and strongly alkaline pH after a few min of illumination. Photosynthesis showed lack of photoinhibition even at high irradiances equivalent to full sunlight, and photosynthesis recovered rapidly after periods of anoxia. We discuss these new insights on the ecological niche of Prochloron and possible interactions with its host and other microbes in light of its recently published genome and a recent study of the overall microbial diversity and metagenome of L. patella.

Two didemnid ascidians associating with cyanobacterial symbionts (Prochloronspp.) were firstly recorded from Caribbean Panama:Lissoclinum verrilli, which facultatively harbouredProchloroncells on the colony surface, andDiplosoma simile, which obligately harboured algal cells in the peribranchial and common cloacal cavities within the colonies. WhileL. verrilli sensu strictohas been exclusively recorded from the Bermudas and Caribbean islands,D. simileis widely distributed in tropical Indo-Pacific regions including oceanic islands such as Hawaii. Partial COI sequences ofD. similefrom the Caribbean were identical to those from the West Pacific, suggesting a high larval-dispersal ability and broad range of environmental tolerance. Molecular phylogenetics of the symbionts, based on 16S rRNA gene sequences, revealed that both ascidian species were associated withProchloron, while aSynechocystissp. sequence was also obtained forL. verrilli.In addition,L. verrilliandD. simileharboured different phylotypes within theProchloronlineage that included symbionts from various hosts and various Pacific sites. Our results indicate that multiple phylotypes ofProchloronexist in Caribbean Panama and that considering the abundance and the number of host species in the Pacific,ProchloronandD. similemay have come from tropical Indo-West Pacific. [ABSTRACT FROM AUTHOR]

Abstract: The marine cyanobacterium Prochloron is a unique photosynthetic organism that lives in obligate symbiosis with colonial ascidians. We compared Prochloron harbored in four different host species and cultured Prochlorothrix by means of spectroscopic measurements, including time-resolved fluorescence, to investigate host-induced differences in light-harvesting strategies between the cyanobacteria. The light-harvesting efficiency of photosystems including antenna Pcb, PS II–PS I connection, and pigment status, especially that of PS I Red Chls, were different among the four samples. We also discuss relationships between these observed characteristics and the light conditions, to which Prochloron cells are exposed, influenced by distribution pattern in the host colonies, presence or absence of tunic spicules, and microenvironments within the ascidians'' habitat. [Copyright &y& Elsevier]

Vertical transmission of cyanobacterial symbionts occurs in didemnid ascidians harboring Prochloron as an obligate symbiont; the photosymbionts are transferred from the parental ascidian colony to the offspring in various ways depending on host species. Although several didemnids harbor non-Prochloron cyanobacteria in their tunics, few studies have reported the processes of vertical transmission in these didemnids. Here we describe the histological processes of the transmission of cyanobacteria in two didemnids, Trididemnum nubilum harboring Synechocystis and T. clinides harboring three cyanobacterial species. In both species, the photosymbionts in the tunic of the parent colony were apparently captured by the tunic cells of the host and transferred to the embryos brooded in the tunic. The symbiont cells were then incorporated into the inner tunic of the embryo. This mode of transmission is essentially the same as that of T. miniatum harboring Proehloron in the tunic, although there are some differences among species in the timing of the release of the symbionts from the tunic cells. We suggest that the similar modes of vertical transmission are an example of convergent evolution caused by constraints in the distribution patterns of symbiont cells in the host colony. [ABSTRACT FROM AUTHOR]

Terpios hoshinota, a dark encrusting sponge, is known to be a competitor for space in coral reef environments, and facilitates the death of corals. Although numerous cyanobacteria have been detected in the sponge, little is known of the sponge-associated bacterial community. This study examined the sponge-associated bacterial community and the difference between the bacterial communities in the sponge and the coral partially covered by the sponge by analysis of 16S rRNA gene sequences of samples isolated from the sponge covering the corals Favia complanata, Isopora palifera, Millepora sp. , Montipora efflorescens and Porites lutea. The sponge-associated bacterial community was mainly (61-98%) composed of cyanobacteria, with approximately 15% of these alphaproteobacteria and gammaproteobacteria, although the proportions varied in different sponge samples. The dominant cyanobacteria group was an isolated group closely related to Prochloron sp. The comparison of the bacterial communities isolated from sponge-free and the sponge-covered P. lutea showed that covering by the sponge caused changes in the coral-associated bacterial communities, with the presence of bacteria similar to those detected in black-band disease, suggesting the sponge might benefit from the presence of bacteria associated with unhealthy coral, particularly in the parts of the coral closest to the margin of the sponge. [ABSTRACT FROM AUTHOR]

The complex evolution of secondary metabolism is important in biology, drug development, and synthetic biology. To examine this problem at a fine scale, we compared the genomes and chemistry of 24 strains of uncultivated cyanobacteria, Prochloron didemni, that live symbiotically with tropical ascidians and that produce natural products isolated from the animals. Although several animal species were obtained along a >5500 km transect of the Pacific Ocean, P. didemni strains are >97% identical across much of their genomes, with only a few exceptions concentrated in secondary metabolism. Secondary metabolic gene clusters were sporadically present or absent in identical genomic locations with no consistent pattern of co-occurrence. Discrete mutations were observed, leading to new chemicals that we isolated from animals. Functional cassettes encoding diverse chemicals are exchanged among a single population of symbiotic P. didemni that spans the tropical Pacific, providing the host animals with a varying arsenal of secondary metabolites. [ABSTRACT FROM AUTHOR]

The article focuses on a research study which aims at developing lantipeptides in Escherichia coli. It describes lantipeptides as polycyclic peptides which contain ribosimally synthesized and modified peptides containing thioether links and explains the methodology which was conducted on prochlorosins, a form of lantipeptides which are product of ubiquitous marine cynobacterium Prochlorococcus.

Prochlorococcus describes a diverse and abundant genus of marine photosynthetic microbes. It is primarily found in oligotrophic waters across the globe and plays a crucial role in energy and nutrient cycling in the ocean ecosystem. The abundance, global distribution, and availability of isolates make Prochlorococcus a model system for understanding marine microbial diversity and biogeochemical cycling. Analysis of 73 metagenomic samples from the Global Ocean Sampling expedition acquired in the Atlantic, Pacific, and Indian Oceans revealed the presence of two uncharacterized Prochlorococcus clades. A phylogenetic analysis using six different genetic markers places the clades close to known lineages adapted to high-light environments. The two uncharacterized clades consistently cooccur and dominate the surface waters of high-temperature, macronutrient-replete, and low-iron regions of the Eastern Equatorial Pacific upwelling and the tropical Indian Ocean. They are genetically distinct from each other and other high-light Prochlorococcus isolates and likely define a previously unrecognized ecotype. Our detailed genomic analysis indicates that these clades comprise organisms that are adapted to iron-depleted environments by reducing their iron quota through the loss of several iron-containing proteins that likely function as electron sinks in the photosynthetic pathway in other Prochlorococcus clades from high-light environments. The presence and inferred physiology of these clades may explain why Prochlorococcus populations from iron-depleted regions do not respond to iron fertilization experiments and further expand our understanding of how phytoplankton adapt to variations in nutrient availability in the ocean. [ABSTRACT FROM AUTHOR]

Some invertebrates have enlisted autotrophic unicellular algae to provide a competitive metabolic advantage in nutritionally demanding habitats. These symbioses exist primarily but not exclusively in shallow tropical oceanic waters where clear water and low nutrient levels provide maximal advantage to the association. Mostly, the endosymbiotic algae are localized in host cells surrounded by a host-derived membrane (symbiosome). This anatomy has required adaptation of the host biochemistry to allow transport of the normally excreted inorganic nutrients (CO2, NH3 and PO43−) to the alga. In return, the symbiont supplies photosynthetic products to the host to meet its energy demands. Most attention has focused on the metabolism of CO2 and nitrogen sources. Carbon-concentrating mechanisms are a feature of all algae, but the products exported to the host following photosynthetic CO2 fixation vary. Identification of the stimulus for release of algal photosynthate in hospite remains elusive. Nitrogen assimilation within the symbiosis is an essential element in the host's control over the alga. Recent studies have concentrated on cnidarians because of the impact of global climate change resulting in coral bleaching. The loss of the algal symbiont and its metabolic contribution to the host has the potential to result in the transition from a coral-dominated to an algal-dominated ecosystem. [ABSTRACT FROM AUTHOR]

Prochloron is an oxygenic photosynthetic bacterium that lives in obligate symbiosis with didemnid ascidians, such as Diplosoma spp., Lissoclinum spp. and Trididemnum spp. This study investigated the genetic diversity of the genus Prochloron by constructing a phylogenetic tree based on the 16S rRNA gene sequences of 27 isolates from 11 species of didemnid ascidians collected from Japan, Australia and the USA. The 27 isolates formed three phylogenetic groups: 22 of the samples were identified to be closely related members of Prochloron. Two samples, isolated from Trididemnum nubilum and Trididemnum clinides, were found to belong to the species Synechocystis trididemni, the closest relative of Prochloron. Three isolates formed a separate group from both Prochloron sp. and S. trididemni, potentially indicating a new symbiotic phylotype. Genomic polymorphism analysis, employing cyanobacterium-specific highly iterative palindrome 1 repeats, could not delineate the isolates further. For the Prochloron sp. isolates, the phylogenetic outcome was independent of host species and geographic origin of the sample indicating a low level of host specificity, low genetic variation within the taxon and possibly a lack of a host–symbiont relationship during reproductive dispersal. This study contributes significantly to the understanding of Prochloron diversity and phylogeny, and implications for the evolutionary relationship of prochlorophytes, cyanobacteria and chloroplasts are also discussed. [ABSTRACT FROM AUTHOR]

In obligate symbioses between didemnid ascidians and prokaryotic algae (cyanobacteria), the larvae of the host ascidian usually possess Prochloron cells that are acquired from their mother colony. The process of vertical transmission of the photosymbionts has been histologically described in several species harboring Prochloron in the common cloacal cavity, where Prochloron cells are attached to the hairy surface of the tunic of the larvae during or just before larval spawning. Since the process has never been described in species harboring the photosymbionts in the tunic of the colony, here we describe the histological and ultrastructural process of vertical transmission in a photosymbiotic didemnid, Trididemnum miniatum. No photosymbionts were associated with gametes of the ascidian. Prochloron cells appeared in the epithelial pouch enclosing the embryo and were then incorporated in the tunic of embryos in late embryonic stages. Although Prochloron cells in the tunic of the colony were rarely associated with the host cells, some amoeboid-shaped tunic cells containing Prochloron cells were occasionally found around the epithelial pouch. Therefore, the host cell is thought to be a vehicle transporting the photosymbionts in the tunic of the colony to the tunic of the embryos. The photosymbiont cells were directly embedded in the tunic of the larval trunk. They were not contained in the host cells, as are those in the tunic of the colony. These observations revealed that the mechanism of vertical transmission in T. miniatum is very different from that in didemnids harboring Prochloron in the common cloacal cavity of the colonies, such as Trididemnum cyclops. The mechanisms of symbiont transmission are diverse even within the genus Trididemnum. [ABSTRACT FROM AUTHOR]

The contents of mycosporine-like amino acids (MAAs) were compared in the two color morphs (dark-gray and brown colonies) of the tropical ascidian Didemnum molle (Herdman, 1886), which harbors the photosymbiotic prokaryote Prochloron. The colonies of each color morph were exclusively distributed in shallow reef lagoons at the different sites. Spectroscopic and chromatographic analyses showed that the Prochloron cell density and MAA concentration in the dark-gray colonies were an estimated 1.4 and 2.4 times higher, respectively, than in the brown colonies. The significant difference in MAA contents between the color morphs was primarily due to the difference in shinorine contents ( p < 0.01, Mann–Whitney U-test). The high concentration of MAAs in the dark-gray colonies may provide better conditions for Prochloron cells, compared to the brown colonies with lower MAA concentrations. [ABSTRACT FROM AUTHOR]

Abstract: In the tropics, certain didemnid ascidians harbor the prokaryotic photosymbiont Prochloron. To date, this photosymbiosis has been found in four didemnid genera that include non-symbiotic species. Here, we report the molecular phylogeny of symbiotic and non-symbiotic didemnids based on their 18S rDNA sequences. The data cover all four genera containing symbiotic species and one other genus comprised of only non-symbiotic species. Near-complete nucleotide sequences of 18S rDNAs were determined for four non-didemnid species and 52 didemnid samples (five genera), including 48 photosymbiotic samples collected from the Ryukyu Archipelago, the Great Barrier Reef, Hawaii, and Bali. Our phylogenetic trees indicated a monophyletic origin of the family Didemnidae, as well as each of the didemnid genera. The results strongly support the hypothesis that establishment of the ascidian-Prochloron symbiosis occurred independently in the Didemnidae lineage at least once in each of the genera that possess symbiotic species. [Copyright &y& Elsevier]

This is a mini-review demonstrating that investigation of the genomics of marine microbes from all three domains has the potential to revolutionize the search for secondary metabolites originally thought to be the product of marine invertebrates. The basis for the review was a symposium at the 2005 Annual Meeting of the SIM covering some aspects of the potential for marine microbes to be the primary producers of such metabolites. The work reported at that symposium has been integrated into a fuller discussion of current published literature on the subject with examples drawn from bacteria, cyanophytes and fungi. [ABSTRACT FROM AUTHOR]

Focuses on a study which localized the ultraviolet-absorbing substances in the tissues of two colonial didemnid ascidians that contain the symbiotic photo-oxygenic prokaryote Prochloron sp. Risks posed by ultraviolet radiation for organisms in marine environments; Information on mycosporine-like amino acids; Reception of light for photosynthesis.

The effects of light intensity, pH, temperature, and UV irradiation on the photosynthetic rate of Prochloron isolated from the ascidian host Lissoclinum patella, collected from Palau, were examined. Photosynthesis increased with light intensity with saturation at 500 µmol/m² per second. It was maximum at pH 8 to 9 but almost completely suppressed below pH 7. The optimum temperature was 35° to 40°C, but the photosynthesis was absent at ≤20°C and at 45°C. It was recovered when the symbiont was transferred from 1 hour of incubation at ≤20°C to 35°C but not when transferred from incubation at 45°C. Ultraviolet irradiation severely inhibited the photosynthesis of Prochloron in isolation but not in vivo. This protection was brought about by the tunic covering the ascidian colony, which contains UV-absorbing mycosporine-like amino acids. These results indicate that the characteristic condition of the tropical marine environment largely determines the ecological distribution of Prochloron, and the ascidian tunic protects the organism from UV radiation. [ABSTRACT FROM AUTHOR]

Patellamides are highly bioactive compounds found along with other cyanobactins in the symbiosis between didemnid ascidians and the enigmatic cyanobacterium Prochloron . The biosynthetic pathway of patellamide synthesis is well understood, the relevant operons have been identified in the Prochloron genome and genes involved in patellamide synthesis are among the most highly transcribed cyanobacterial genes in hospite. However, a more detailed study of the in vivo dynamics of patellamides and their function in the ascidian- Prochloron symbiosis is complicated by the fact that Prochloron remains uncultivated despite numerous attempts since its discovery in 1975. A major challenge is to account for the highly dynamic microenvironmental conditions experienced by Prochloron &nbsp;in hospite, where light-dark cycles drive rapid shifts between hyperoxia and anoxia as well as pH variations from pH ~6 to ~10. Recently, work on patellamide analogues has pointed out a range of different catalytic functions of patellamide that could prove essential for the ascidian- Prochloron symbiosis and could be modulated by the strong microenvironmental dynamics. Here, we review fundamental properties of patellamides and their occurrence and dynamics in vitro and in vivo. We discuss possible functions of patellamides in the ascidian- Prochloron symbiosis and identify important knowledge gaps and needs for further experimental studies.

Nucleotide sequence data from DNA-dependent RNA polymerase (rpoC) genes were used to examine the phylogenetic relationships among the phycobiliprotein- and three known chlorophyll b-containing (prochlorophyte) cyanobactera. The phylogenetic tree obtained confirm the polyphyletic nature of the prochlorophytes. Data from Prochloron cells obtained from six different tunicate host species suggest that at least two closely related groups of Prochloron exist in the same area in Polau, West Caroline Island. Overall, however, the genetic diversity within the analyzed samples was much smaller than within the nonsymbiotic Prochlorococcus. [ABSTRACT FROM AUTHOR]

Seven Indo-West Pacific Prochloron Lewin samples one Caribbean cyanobacterial sample were compared by DNA-DNA reassociation experiments. The quality and quantity of extracted DNA were affected byseveral cell preservatin methods and DNA extraction and purification procedures. We conclude that Prochloron samples from six different hosts and three different geographical locations spanning ca. 9000 km bilong to a single species but are not closely related to the strain of cyanobacteria used. The intraspecific genotypic variation with in prochloron is discussed with respect to host specificity and geographical distribution. [ABSTRACT FROM AUTHOR]

We studied the distribution of the DNA-containing region and the ribulose 1, 5-bisphosphate carboxylase-oxygenase (RuBisCo) content of polyhedral bodies in three different prochlorophyte cell types in a search for broad evolutionary affinities of these chlorophyll b-containing prokaryotes. DNA was localized by DAPI staining and electron microscopy utilizing monoclonal anti-DNA antibody 2C-10 plus a secondary antibody labeled with tolloidal gold. Antibodies against the large RuB isCo subunit from a higher plant raised in rabbits were used to localize RuBisCo in polyhedral bodies. We studied Prochloron Lewin cells from two different didemnid ascidian hosts (Lissoclinum patella and Didemnum molle) collected in Palau, West Caroline Islands, and cells of Prochlorothrix hollandica Burger-Wiersma, Stal, and Mur grown in laboratory culture. Cells of the blue-green alga Anabaena 7120 were studied for comparison. The DNA distribution was markedly difference in the two Prochloron cell types. The thylakoids in cells from L. patella were concentrically arranged around a large central vacuole; the DNA-containing stromal areas appeared in thin sections as concentric arcs between the thylakoid stacks. The central vacuole was lacking in cells from D. molle, and the thylakoid stacks and strands of DNA-containing stroma showed a more haphazard arrangement. In the filamentous Prochlorothrix the DNA-containing stroma was largely limited to a central nucleoid structure running the length of the cell. Although the DNA arrangements in Prochloron might be considered "chloroplast-like" since DNA-containing storma is distributed, as in chloroplasts, in scattered sites among photosynthetic membranes, this is not so in Prochlorothrix, where there is an axial nucleoid, as in many filamentous cyanobacteria. Our anti-RuBisCo antibodies were selectively bound to the polyhedral bodies of all three cell types, indicating that Pro- chloron and Prochlorothrix, Iike many other auto-trophic prokaryotes, possess typical carboxysomes. [ABSTRACT FROM AUTHOR]

Two new PAM fluorometers (pulse amplitude modulated) were used in an investigation of photosynthetic performance of Prochloron resident as a symbiont in the ascidian Lissoclinum patella, growing in a coral reef of Heron Island on the Great Barrier Reef. With a new DIVING-PAM in situ measurements of effective PSII quantum yield (δF/Fm′) as a function of quantum flux density (rapid light curves) were carried out in 2.5 m depth in the reef and in a seawater tank. Photosynthetic electron transport rates were measured on in hospite Prochloron both in situ and in collected material. Both light-limited and light-saturated yields were exceptionally high. Maximal yields (Fv/Fm) were ˜0.83. A new TEACHING-PAM was employed for analysing dark-light induction and light-dark relaxation kinetics in collected samples with Prochloron in hospite. Considerable variability in kinetic responses was observed which was found to be at least in part due to differences in O2 concentration. It is suggested that endogenous reductants feed electrons into the intersystem transport chain, which normally is reoxidized by O2 (chlororespiration), and that in the dark, the reduction level of PSII acceptors is increased due to a decline in O2 concentration. The pattern of fluorescence responses differed markedly from those found in cyanobacteria and provides new insights into light-harvesting responses of a photosynthetic prokaryote with a membrane bound light-harvesting system, as contrasted with an extrinsic light-harvesting system. [ABSTRACT FROM AUTHOR]