Floridean Starch and Floridoside Metabolic Pathways of Neoporphyra haitanensis and Their Regulatory Mechanism under Continuous Darkness

Floridean starch and floridoside are the main storage carbohydrates of red algae. However, their complete metabolic pathways and the origin, function, and regulatory mechanism of their pathway genes have not been fully elucidated. In this study, we identified their metabolic pathway genes and analyzed the changes in related gene expression and metabolite content in Neoporphyra haitanensis under continuous dark conditions. Our results showed that genes from different sources, including eukaryotic hosts, cyanobacteria, and bacteria, were combined to construct floridean starch and floridoside metabolic pathways in N. haitanensis. Moreover, compared with those in the control, under continuous dark conditions, floridean starch biosynthesis genes and some degradation genes were significantly upregulated with no significant change in floridean starch content, whereas floridoside degradation genes were significantly upregulated with a significant decrease in floridoside content. This implies that floridean starch content is maintained but floridoside is consumed in N. haitanensis under dark conditions. This study elucidates the “floridean starch–floridoside” metabolic network and its gene origins in N. haitanensis for the first time.

Chronobiological Effect on the Reproductive Behavior of Chrysomya megacephala (Diptera: Calliphoridae)

The most widely used entomological method of determining the time since death (minimum postmortem interval, mPMI) has been calculating the developmental time of blow flies (Diptera: Calliphoridae) on the deceased body. However, because blow flies are known to be diurnal, nocturnal oviposition has been excluded from standard mPMI calculations. This has been challenged by recent studies demonstrating nocturnal oviposition due to an unknown reason. Therefore, this study investigated the role of chronobiology. We recorded the locomotion amount and pattern of Chrysomya megacephala (Fabricius, 1794) (Diptera: Calliphoridae) under different chronobiological conditions and examined whether Ch. megacephala can oviposit under nighttime conditions in field and laboratory settings. Subjects were found to have a daily activity pattern under normal darkness conditions (12:12 L:D) and under continuous darkness (DD), but they exhibited no pattern under continuous light (LL). Free-running period was approximately 1,341 min/d (22.35 h/d). In the field, no flies were observed during nighttime. Oviposition occurred in the laboratory setting during daytime with no lights and during nighttime with artificial lights. Free-running subjects oviposited in both active and resting periods, with more eggs laid during active than resting periods. The result of this study indicates it is possible to induce oviposition behavior during evening hours on Ch. megacephala. However, this was only observed in the laboratory setting and could only happen during the flies’ subjective day.

Earthworm Depuration: Analysis of Coprophagy and Light Impacts

Earthworms are used as biomarkers to determine the bioavailability of contaminants. As such, their uptake of contaminants has been studied extensively. Protocols have been established to ensure that laboratory-obtained data are valid and comparable. However, the method of removing the organism’s gut content (depuration) before assessing the contaminant in the tissue is not standardized. The aim of this research project is to investigate some parameters for earthworm depuration: light conditions and coprophagy prevention. Eisenia fetida were depurated for 48 hours in two separate studies according to guidelines ASTM-E1676 and OECD Test No. 317. In one study, 2 frequencies of egesta removal were employed during depuration to prevent coprophagy and compared to the control (egesta and worms removed after 48 hours). In another study, the subjects and material egested were assessed under conditions of continuous darkness and the control (continuous light). The depuration methods that included egesta removal every 12 and 24 hours resulted in 62% and 10% more egested material per mg of earthworm than the control (filter paper disc change after 48 hours), respectively. The earthworms depurated in continuous darkness egested 94% more material per mg of earthworm than the control. The results indicate that depuration would be more total under continuous darkness and employing a coprophagy prevention method. These findings could lead to more efficient depuration methods.

Circadian regulation of vertebrate cone photoreceptor function

Eukaryotes generally display a circadian rhythm as an adaption to the reoccurring day/night cycle. This is particularly true for visual physiology that is directly affected by changing light conditions. Here we investigate the influence of the circadian rhythm on the expression and function of visual transduction cascade regulators in diurnal zebrafish and nocturnal mice. We focused on regulators of shut-off kinetics such as Recoverins, Arrestins, Opsin kinases, and Regulator of G-protein signaling that have direct effects on temporal vision. Transcript as well as protein levels of most analyzed genes show a robust circadian rhythm-dependent regulation, which correlates with changes in photoresponse kinetics. Electroretinography demonstrates that photoresponse recovery in zebrafish is delayed in the evening and accelerated in the morning. Functional rhythmicity persists in continuous darkness, and it is reversed by an inverted light cycle and disrupted by constant light. This is in line with our finding that orthologous gene transcripts from diurnal zebrafish and nocturnal mice are often expressed in an anti-phasic daily rhythm.

Asymbiotic seed germination and in vitro seedling development of Orchis militaris, an endangered orchid in Siberia

Background Terrestrial orchids belonging to the Orchis genus are difficult to propagate and are under great pressure in their natural habitats. Studies regarding the influence of photoperiod and temperature regimes on Orchis militaris germination and morphological changes during immature seed development in vitro are scarce. Our aim was to identify photoperiod, temperature, and different nutrient media requirements for optimization of O. militaris seed germination and vigorous seedling production. Results Post-germination morphological changes were recorded with O. militaris seeds collected from 32-day-old fruits, where the percentage of O. militaris seeds without embryo was 38.4%. The highest rate of O. militaris seed germination (82.6%) was obtained on Malmgren modified terrestrial orchid medium (mM), enriched by 5% coconut water, 5% birch sap, and 0.1% AC. Nine percent of seedlings were able to reach the advanced seedling stage (stage 6) after 12 months of maintenance on this medium. In all 3 modified media (Harvais, Knudson С and Malmgren), regeneration was via the production of protocorms and seedlings without callus formation. It was proved that more abundantly vigorous protocorms were formed on the modified Harvais 2 under continuous darkness, while the subculture in Knudson C medium with AC addition could be necessary to stimulate their further development. The regeneration success of the species for in vitro conditions was increased by following its natural seasonal cycle. Conclusion This study demonstrated an efficient micropropagation system for O. militaris using immature seeds and thus widely opened the perspectives for its conservation in nature. The favorable conditions of seed germination periods for in vitro culture, identified as the definite shift of temperatures and photoperiod regimes intrinsic to the species in nature, could improve seedling survival of this medicinally important orchid.

Dusky Grouper Epinephelus marginatus Growth and Survival When Exposed to Different Photoperiods

Photoperiod is considered an environmental factor that influences reproduction and the growth processes of fish throughout the year. In the present study, we subjected dusky grouper Epinephelus marginatus to different photoperiods in order to evaluate growth and survival. Juvenile dusky grouper were randomly distributed in twelve 100 L tanks containing four fish per aquarium. The fish were exposed to continuous light, normal photoperiod or continuous darkness for 50 days. Fish were fed 3% of their total biomass, twice a day, and the diet consisted of ground commercial dry pellets (42% crude protein, 12% humidity, 9% ether extract, 15% mineral matter, 4% crude fiber, 3.5% calcium and 3% vitamin C, in accordance with the manufacturer’s instructions). There was no mortality during the experiment. After 50 days, the best performance was found for exposure to continuous light (24 h artificial light). The final weight of the fish reared under continuous light was significantly higher than that of the fish exposed to continuous darkness. The specific growth rate of the fish exposed to continuous light was significantly higher than that of the fish exposed to the natural photoperiod and to continuous darkness. Modifications to the photoperiod can be a key factor in increasing the efficacy of current production and the improvement of current aquaculture protocols.

Gut Microbiota Mediates Oolong Tea Polyphenols Alleviation of Circadian Rhythm Disturbance

BackgroundTea polyphenols can be digested in the intestinal tract so as to promote the growth of helpful gut microbiota, and through the production of catechin, tryptophan, acetic acid and other active substances which involved in the modulation of circadian rhythms mechanism to improve the circadian rhythm disorders and neurological weakness, while its specific mechanism is still unclear. The interaction between host circadian rhythm and gut microbes through the gut-brain axis (GBA) provides new clues for tea polyphenols to improve host health. Our present research mainly investigated the metabolites of the gut microbiota and the heterogeneous expression of circadian rhythm genes in the liver and hypothalamic, and then revealed the modulatory role of oolong tea polyphenols (OTP) of circadian rhythm disorder via the GBA. We used 16S rDNA and untargeted metabolomics sequencing techniques to analyze the effects of OTP on intestinal flora diversity and abundance of specific flora in mice with diurnal disorders, and to screen out marker metabolites which may be involved in circadian rhythm regulation. Transcriptomics and 10X single-cell sequencing were used to evaluate the effects of OTP on circadian rhythm genes in liver and hypothalamus and hypothalamus cell types in mice with diurnal disorders. The Y maze and Morris water maze experiments were investigated the effects of OTP on long-term and short-term memory impairment.ResultsBy establishing a circadian rhythm disorder mouse model, our experimental results showed that OTP improved the structural disorder of the intestinal microbiota caused by continuous darkness, especially significantly decreased the ratio of Firmicutes/Bacteroidetes (F/B), thereby modulating the production of metabolites related to pyruvate metabolism, glycolysis/gluconeogenesis, and tryptophan metabolism to alleviate the steady-state imbalance caused by circadian rhythm disorders. In addition, OTP can significantly ameliorate the rhythm oscillation disorder of specific gut microbes and liver clock genes induced by continuous darkness, and made dysrhythmic mice perform well in cognitive behavior tests. Simultaneously, OTP intervention increased the number of rhythmic expression genes in the liver which in the CD group has 156, while in the OTP group has 208. Transplanting fecal microbiota from the OTP group into germ-free mice exhibited that OTP significantly increased the number of hypothalamus cell clusters, up-regulated the number of astrocytes and fibroblasts, and enhanced the expression of circadian rhythm genes Cry2, Per3, Bhlhe41, Nr1d1, Nr1d2, Dbp, and Rorb in hypothalamic cells. ConclusionsOur results confirmed that OTP reduced the F/B ratio, made the daily oscillation of the intestinal microbiota tended to be regular, actively improve the intestinal microecological status, the content of important metabolite closely associated with circadian rhythm such as Tryptophan, Glutamine, 2-indolecarboxylic acid and some others has been significantly increased, the poor expression of clock genes (such as Cry2, Per3, Bhlhe41, Nr1d1, Nr1d2, Dbp and Rorb) in liver and hypothalamus cells has been improved. These results indicated that OTP can significantly improve the metabolic imbalance and cognitive impairment caused by the circadian rhythm disorder, maintain the host's homeostasis, which with potential prebiotic functional characteristics to positively contribute to host health.

Effect of relative humidity, temperature, light and plant age on infection of lowbush blueberry by Sphaerulina vaccinii

Leaf spot and stem canker caused by Sphaerulina vaccinii is associated with premature defoliation in lowbush blueberry resulting in reduced yields. In this study, we investigated the impact of free water, relative humidity (RH), temperature, light, and plant age on leaf infection under controlled conditions. On potato dextrose agar, germination of conidia was usually polar. Growth was minimal at 5 and 10°C, increased at 15 and 20°C, was maximal at 25°C and decreased at 30°C. Percentage of germinated conidia on inoculated blueberry leaves incubated in dark controlled-humidity chambers for 3 days (25°C) was 86.0, 90.5, 81.3 and 28.3% in free water, 100, 97.5 or 95% relative humidity (RH), respectively. Germination did not occur at 90 or 85% RH. Infection of inoculated plants, however, was not favored by free water, but rather by high RH (>95%) and a 14-h photoperiod (180 µmol/m2/s). Infection failed in continuous darkness, continuous light or continuous darkness followed by 4, 8 or 12 h of light. Light and scanning electron microscopy showed that hyphal penetration into stomata on abaxial leaf surfaces was strongly tropic. When germ tubes grew in close proximity to a stomate, a penetration hypha formed at about 90° angles to the germ tube and took the closest path to the stomate. Stomatal penetration was usually direct, but occasionally appressorium-like hyphal swellings formed over stomatal openings. When inoculated plants were exposed to high RH (>95%) at various temperatures, infection occurred after 4 days at 10°C, after 3 days at 15°C and after 1 day at 20 and 25°C. Infection failed to occur at 30°C. Disease severity also increased with duration of the humid period. When leaves were examined microscopically, those that had been incubated for 6 days showed a substantially greater network of epiphytic growth with more stomatal penetrations compared to those incubated for 3 days. Infection was substantially reduced when the humid period was interrupted by alternating days of low RH (60%). Two-week-old leaves were 2.7 times more susceptible than 8-week-old leaves.

GR24, A Synthetic Strigolactone Analog, and Light Affect the Organization of Cortical Microtubules in Arabidopsis Hypocotyl Cells

Strigolactones are plant hormones regulating cytoskeleton-mediated developmental events in roots, such as lateral root formation and elongation of root hairs and hypocotyls. The latter process was addressed herein by the exogenous application of a synthetic strigolactone, GR24, and an inhibitor of strigolactone biosynthesis, TIS108, on hypocotyls of wild-type Arabidopsis and a strigolactone signaling mutant max2-1 (more axillary growth 2-1). Owing to the interdependence between light and strigolactone signaling, the present work was extended to seedlings grown under a standard light/dark regime, or under continuous darkness. Given the essential role of the cortical microtubules in cell elongation, their organization and dynamics were characterized under the conditions of altered strigolactone signaling using fluorescence microscopy methods with different spatiotemporal capacities, such as confocal laser scanning microscopy (CLSM) and structured illumination microscopy (SIM). It was found that GR24-dependent inhibition of hypocotyl elongation correlated with changes in cortical microtubule organization and dynamics, observed in living wild-type and max2-1 seedlings stably expressing genetically encoded fluorescent molecular markers for microtubules. Quantitative assessment of microscopic datasets revealed that chemical and/or genetic manipulation of strigolactone signaling affected microtubule remodeling, especially under light conditions. The application of GR24 in dark conditions partially alleviated cytoskeletal rearrangement, suggesting a new mechanistic connection between cytoskeletal behavior and the light-dependence of strigolactone signaling.