RESUMO
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.
Assuntos
Peptídeos Cíclicos/metabolismo , Prochloron/metabolismo , Urocordados/metabolismo , Animais , Humanos , Concentração de Íons de Hidrogênio , Peptídeos Cíclicos/biossíntese , Peptídeos Cíclicos/farmacologia , Prochloron/genética , Simbiose , Urocordados/genéticaRESUMO
The data in this article describe the population growth of Prochloron cells outside the hosts at the different salinity levels. The cultivation was performed in enriched standard culture media with continuous photoperiod. The culture stock of Prochloron cells which was made as inoculum (starter) in the laboratory was isolated from tunics of an ascidian Lissoclinum patella. The ascidian was obtained from 20 m depth at Malalayang coastal water in Manado Bay, North Sulawesi, Indonesia. The initial stock was kept in 20 ppt liquid medium. Then the cells were transferred into culture chambers, each prepared with different salinities: 15, 20, 25, 30, 35, 40 and 45 ppt. After the cells reached exponential phase, some drops of cell suspension were transferred into agar media with the same salinity level until the green colony appeared. Each of the colonies was transferred again into liquid media with the same salinity. Population growth was observed until the death phase. The results of the study show that: (1) the growth of Prochloron cells at different salinity showed a different growth rate; (2) Prochloron cells grew well in salinity of 15 and 35 ppt; (3) the maximum population growth of Prochloron from each treatment varied. Prochloron cells grown in a medium with 35 ppt salinity had a rapid adaptability to the new culture environment. However, the maximum population growth was reached on the 75th day with a cell density of 31.00 × 106 cells/ml in a medium with 15 ppt salinity, much higher than those of the other treatments (20, 25, 30, 35 and 45 ppt). The data presented here are the success of the cultivation of Prochloron cells outside the host.
RESUMO
BACKGROUND: Marine 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. RESULTS: This 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. CONCLUSIONS: The 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.
Assuntos
Bactérias/genética , Bactérias/metabolismo , Metagenoma/genética , Microbiota/genética , Filogenia , Poríferos/classificação , Poríferos/microbiologia , Animais , Bactérias/classificação , Bactérias/isolamento & purificação , Biodiversidade , Genômica , Poríferos/metabolismo , RNA Ribossômico 16S/genética , SimbioseRESUMO
Noburu Sensui and Euichi Hirose (2018) In didemnid ascidians with cyanobacterial symbionts, the tunic has a specific peak absorbing ultraviolet radiation (UV-R) due to the presence of ultraviolet (UV)-absorbing compounds, which probably include mycosporine-like amino acids (MAAs). The UV-R absorbing tunic is supposed to protect the symbionts in the common cloacal cavity of the host colony. The histological distribution of UV-R absorption in the tunic was examined using a UV light microscope equipped with a digital camera, from which the low-pass filter of the UV-sensitive image sensor was removed. The cell peripheries of tunic bladder cells and cell-like objects were visualized with the trans-illumination of UV light, indicating UV-R absorption at that site. In contrast, tunic matrix and vacuolar content of tunic bladder cells appeared to lack of UV-R absorption, allowing damaging wavelengths to penetrate. Accordingly, UV-absorbing compounds are expected to be contained in the cytoplasmic matrix of tunic bladder cells and possibly other types of tunic cells.
RESUMO
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 ß-lactamase as well as α- and ß-glycosidase activity with kcat=(11.34±0.91)Ë10(-4)s(-1) for ß-lactamase, kcat=(1.55±0.13)Ë10(-4)s(-1) for α-glycosidase and kcat=(1.22±0.09)Ë10(-4)s(-1) for ß-glycosidase activity.
Assuntos
Proteínas de Bactérias/química , Cobre/química , Cianobactérias/enzimologia , Glicosídeo Hidrolases/química , Peptídeos Cíclicos/química , beta-Lactamases/química , AnimaisRESUMO
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 O(2) 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 minutes of darkness, while the same zone exhibited O(2) super-saturation and strongly alkaline pH after a few minutes 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.