Latent virus‐like infections are present in a diverse range of Symbiodinium spp. (Dinophyta)

Coral reefs are increasingly threatened by disease outbreaks, which affect the coral animal and/or its algal symbionts (Symbiodinium spp.) and can cause mass mortalities. Currently around half of the recognized coral diseases have unknown causative agents. While many of the diseases are thought to be bacterial in origin, there is growing evidence that viruses may play a role. In particular, it appears that viruses may infect the algal symbionts, causing breakdown of the coral‐algal mutualism. In this study, we screened a wide range of Symbiodinium cultures in vitro for the presence of latent viral infections. Using flow cytometry and electron microscopy, we found that many types of Symbiodinium apparently harbor such infections, and that the type of putative virus varied within and among host types. Furthermore, the putative viral infections could be induced via abiotic stress and cause host cell lysis and population decline. If similar processes occur in Symbiodinium cells in hospite, they may provide an explanation for some of the diseases affecting corals and other organisms forming symbioses with these algae.     

Non‐specific immunity and disease resistance are enhanced by the polysaccharide fraction of a marine chlorophycean macroalga in Oreochromis niloticus (Linnaeus, 1758)

Polysaccharides (PF) from marine macroalgae, Caulerpa scalpelliformis were extracted and tested for its potential immunostimulatory and disease resistance properties in fish. Five groups of Nile tilapia (n = 6), Oreochromis niloticus (Linnaeus, 1758) were intraperitoneally administered with the different doses of PF (2, 20 or 200 mg/kg body weight) or with yeast‐derived commercial immunostimulant, Macrogard^? (20 mg/kg body weight), to compare the effectiveness. An untreated control group was also maintained. A total of fifteen fibre reinforced plastic tanks (150 L, ambient temperature and light conditions) were used, with triplicate tanks for each group. Only four fish per tank (totally 12 fish from a group) were taken at random and assayed. PF enhanced all the tested non‐specific serum immune responses namely lysozyme, myeloperoxidase, antiprotease, and bactericidal activities. There was an upregulation of the genes encoding IL‐1β, lysozyme and TNF‐α in the spleen of PF injected fish as compared to the control group. In order to study the overall functional immunity, disease resistance test was conducted. Another five groups of fish (n = 10) were treated by intraperitoneal injection with different doses of PF or Macrogard^? or untreated as mentioned earlier in triplicates (30 fish per group in three tanks, totally 150 fish in 15 tanks). Seven days post treatment, fish were challenged by intraperitoneal administration of live virulent Aeromonas hydrophila. PF treated fish were protected with significant reduction in the mortality and the consequent increased relative percent survival (RPS) of 92 in the least (2 mg/kg) and middle dose (20 mg/kg) groups. The disease resistance experiment was repeated again but this time, fish were challenged 21 days post treatment that resulted in RPS of 50 for the middle dose. The results clearly show that the intraperitoneal administration of the polysaccharide fraction had a stimulating effect on the non‐specific immune responses, immune gene expression and disease resistance.     

Chemical profiling of mycosporine‐like amino acids in twenty‐three red algal species

Rhodophyta produce a variety of chemically different mycosporine‐like amino acids (MAAs), compounds that are known as some of the strongest ultraviolet (UV) absorbing molecules in nature. Accordingly, they primarily act as photoprotectants against harmful levels of solar ultraviolet radiation in the UV‐A and UV‐B range. In order to get a deeper understanding of the chemical diversity of MAAs in red algae, pure standards of eleven mycosporine‐like amino acids were isolated from three different species (Agarophyton chilense, Pyropia plicata and Champia novae‐zelandiae) using various chromatographic methods. Their structures were confirmed by nuclear magnetic resonance and mass spectrometry. Four out of the eleven MAAs are reported for the first time in algae. In addition, a new high‐performance liquid chromatography method was developed for the separation of all isolated MAAs and successfully applied for the analysis of twenty‐three red algal species of marine origin. All of them contained MAAs, the most abundant compounds were shinorine, palythine, asterina‐330 and porphyra‐334. For some samples, the direct assignment of MAAs based on their UV spectra was not possible; therefore, the target analytes were enriched by a simple concentration step, followed by liquid chromatography‐mass spectrometry analysis of the extracts. This approach enabled a deeper insight into the MAA pattern of red algae, indicating that not only the four dominant ones are synthesized but also many others, which were often described as unknown compounds in previous studies.     

Differential contributions of plant Dicer‐like proteins to antiviral defences against potato virus X in leaves and roots

Members of the plant Dicer‐like (DCL) protein family are the critical components of the RNA‐silencing pathway that mediates innate antiviral defence. The distinct antiviral role of each individual DCL protein has been established with mostly based on observations of aerial parts of plants. Thus, although the roots are closely associated with the life cycle of many plant viruses, little is known about the antiviral activities of DCL proteins in roots. We observed that antiviral silencing strongly inhibits potato virus X (PVX) replication in roots of some susceptible Solanaceae species. Silencing of the DCL4 homolog in Nicotiana benthamiana partially elevated PVX replication levels in roots. In Arabidopsis thaliana, which was originally considered a non‐host plant of PVX, high levels of PVX accumulation in inoculated leaves were achieved by inactivation of DCL4, while in the upper leaves and roots, it required the additional inactivation of DCL2. In transgenic A. thaliana carrying the PVX amplicon with a green fluorescent protein (GFP) gene insertion in the chromosome (AMP243 line), absence of DCL4 enabled high levels of PVX‐GFP accumulation in various aerial organs but not in the roots, suggesting that DCL4 is critical for intracellular antiviral silencing in shoots but not in roots, where it can be functionally compensated by other DCL proteins. Together, the high level of functional redundancies among DCL proteins may contribute to the potent antiviral activities against PVX replication in roots.     

Zika virus induces massive cytoplasmic vacuolization and paraptosis‐like death in infected cells

The cytopathic effects of Zika virus (ZIKV) are poorly characterized. Innate immunity controls ZIKV infection and disease in most infected patients through mechanisms that remain to be understood. Here, we studied the morphological cellular changes induced by ZIKV and addressed the role of interferon‐induced transmembrane proteins (IFITM), a family of broad‐spectrum antiviral factors, during viral replication. We report that ZIKV induces massive vacuolization followed by “implosive” cell death in human epithelial cells, primary skin fibroblasts and astrocytes, a phenomenon which is exacerbated when IFITM3 levels are low. It is reminiscent of paraptosis, a caspase‐independent, non‐apoptotic form of cell death associated with the formation of large cytoplasmic vacuoles. We further show that ZIKV‐induced vacuoles are derived from the endoplasmic reticulum (ER) and dependent on the PI3K/Akt signaling axis. Inhibiting the Sec61 ER translocon in ZIKV‐infected cells blocked vacuole formation and viral production. Our results provide mechanistic insight behind the ZIKV‐induced cytopathic effect and indicate that IFITM3, by acting as a gatekeeper for incoming virus, restricts virus takeover of the ER and subsequent cell death.     

Sequence capture and next‐generation sequencing of ultraconserved elements in a large‐genome salamander

Amidst the rapid advancement in next‐generation sequencing (NGS) technology over the last few years, salamanders have been left behind. Salamanders have enormous genomes—up to 40 times the size of the human genome—and this poses challenges to generating NGS data sets of quality and quantity similar to those of other vertebrates. However, optimization of laboratory protocols is time‐consuming and often cost prohibitive, and continued omission of salamanders from novel phylogeographic research is detrimental to species facing decline. Here, we use a salamander endemic to the southeastern United States, Plethodon serratus, to test the utility of an established protocol for sequence capture of ultraconserved elements (UCEs) in resolving intraspecific phylogeographic relationships and delimiting cryptic species. Without modifying the standard laboratory protocol, we generated a data set consisting of over 600 million reads for 85 P. serratus samples. Species delimitation analyses support recognition of seven species within P. serratus sensu lato, and all phylogenetic relationships among the seven species are fully resolved under a coalescent model. Results also corroborate previous data suggesting nonmonophyly of the Ouachita and Louisiana regions. Our results demonstrate that established UCE protocols can successfully be used in phylogeographic studies of salamander species, providing a powerful tool for future research on evolutionary history of amphibians and other organisms with large genomes.     

Effects of global change factors on fatty acids and mycosporine‐like amino acid production in Chroothece richteriana (Rhodophyta)

Under natural conditions, Chroothece richteriana synthesizes a fairly high proportion of fatty acids. However, nothing is known about how environmental changes affect their production, or about the production of protective compounds, when colonies develop under full sunshine with high levels of UV radiation. In this study, wild colonies of C. richteriana were subjected to increasing temperature, conductivity, ammonium concentrations and photosynthetically active radiation (PAR), and UV radiations to assess the potential changes in lipid composition and mycosporine‐like amino acids (MAAs) concentration. The PERMANOVA analysis detected no differences for the whole fatty acid profile among treatments, but the percentages of α‐linolenic acid and total polyunsaturated fatty acids increased at the lowest assayed temperature. The percentages of linoleic and α‐linolenic acids increased with lowering temperature. γ‐linolenic and arachidonic acids decreased with increasing conductivity, and a high arachidonic acid concentration was related with increased conductivity. The samples exposed to UVB radiation showed higher percentages of eicosapentaenoic acid and total monounsaturated fatty acids, at the expense of saturated fatty acids. MAAs accumulation increased but not significantly at the lowest conductivity, and also with the highest PAR and UVR exposure, while ammonium and temperature had no effect. The observed changes are probably related with adaptations of both membrane fluidity to low temperature, and metabolism to protect cells against UV radiation damage. The results suggest the potential to change lipid composition and MAAs concentration in response to environmental stressful conditions due to climate change, and highlight the interest of the species in future research about the biotechnological production of both compound types.     

Invasive submerged freshwater macrophytes are more plastic in their response to light intensity than to the availability of free CO2 in air‐equilibrated water

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Relationships between serotonergic and cannabinoid system in depressive‐like behavior: a PET study with [11C]‐DASB

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