Bta-miR-365-3p-targeted FK506-binding protein 5 participates in the AMPK/mTOR signaling pathway in the regulation of preadipocyte differentiation in cattle.

OBJECTIVE: MicroRNAs (miRNAs) are endogenous non-coding RNAs that can play a role in the post-transcriptional regulation of mammalian preadipocyte differentiation. However, the precise functional mechanism of its regulation of fat metabolism is not fully understood. METHODS: We identified bta-miR-365-3p, which specifically targets the 3' untranslated region (3'UTR) of the FK506-binding protein 5 (FKBP5), and verified its mechanisms for regulating expression and involvement in adipogenesis. RESULTS: In this study, we found that the overexpression of bta-miR-365-3p significantly decreased the lipid accumulation and triglyceride content in the adipocytes. Compared to inhibiting bta-miR-36 5-3p group, overexpression of bta-miR-365-3p can inhibit the expression of adipocyte differentiation-related genes C/EBPα and PPARγ. The dualluciferase reporter system further validated the targeting relationship between bta-miR-365-3p and FKBP5. FKBP5 mRNA and protein expression were detected by quantitative real-time polymerase chain reaction and Western blot. Overexpression of bta-miR-365-3p significantly down-regulated FKBP5 expression, while inhibition of bta-miR-365-3p showed the opposite, indicating that bta-miR-365-3p negatively regulates FKBP5. Adenosine 5'-monophosphate (AMP)-activated protein kinase/mammalian target of rapamycin (AMPK/mTOR) signaling pathway is closely related to the regulation of cell growth and is involved in the development of bovine adipocytes. In this study, overexpression of bta-miR-365-3p significantly inhibited mRNA and protein expression of AMPK, mTOR, and SREBP1 genes, while the inhibition of bta-miR-365-3p expression was contrary to these results. Overexpression of FKBP5 significantly upregulated AMPK, mTOR, and SREBP1 gene expression, while inhibition of FKBP5 expression was contrary to the above experimental results. CONCLUSION: In conclusion, these results indicate that bta-miR-365-3p may be involved in the AMPK/mTOR signaling pathway in regulating Yanbian yellow cattle preadipocytes differentiation by targeting the FKBP5 gene.

Nitrogen removal performance and mechanisms of three aquatic plants for farmland tail water purification.

Constructed wetlands (CWs) are commonly used to control excessive nitrogen from farmlands; however, the interactions between vegetation and microorganisms, nitrogen removal performance, and the mechanisms involved remain unclear in subtropical areas. This study aimed to investigate the nitrogen removal performance and mechanism of CWs containing Canna indica, Acorus calamus, and Thalia dealbata. The results show that CWs with plants had significantly higher nitrogen removal efficiencies than those without, with those planted with T. dealbata having the highest efficiency. T. dealbata performed better than the other two plants due to its high biomass and excellent nitrogen uptake capacity; more importantly, CWs with it had the highest abundance of nitrogen functional genes. Microbial nitrification-denitrification, the primary process of nitrogen removal in CWs, contributed to 88 %, 91 %, and 84 % of the TN removal in the CWs with C. indica, A. calamus, and T. dealbata, respectively, 29 %-158 % higher than that in CWs without plants. Microorganisms played a crucial role in nitrogen removal in the CWs, while plants significantly stimulated microbial activity by enhancing microbial abundance and creating a suitable environment for growth and metabolism. These results can help in understanding the contribution of plants in cleaning farmland tailwater and further optimization of plant configuration and management strategies in wetland ecosystems to improve nitrogen removal efficiency.

Response strategies of stem/leaves endophyte communities to nano-plastics regulate growth performance of submerged macrophytes.

Research on the toxicity effects of nano-plastics on submerged macrophytes has been increasing over the past several years. However, how the endophytic bacteria of submerged macrophytes respond to nano-plastics remains unknown, although they have been widely shown to help terrestrial plants cope with various environmental stressors. Here, a microcosm experiment was performed to unravel the effects of high concentration of nano-plastics (20 mg/L) on three submerged macrophyte (Vallisneria natans, Potamogeton maackianus, Myriophyllum spicatum) and their endophytic bacterial communities. Results indicated that nano-plastics induced antioxidative stress in plants, but significantly reduction in relative growth rate (RGR) only occurred in V. natans (from 0.0034 to -0.0029 day-1), accompanied by change in the stem/leaves endophyte community composition. Further analysis suggested nano-plastics caused a reduction in environmental nutrient availability and the proportion of positive interactions between endophyte communities (43%), resulting in the lowest RGR of V. natans. In contrast, endophytes may help P. maackianus and M. spicatum cope with nano-plastic stress by increasing the proportion of positive correlations among communities (70% and 75%), leaving their RGR unaffected. Collectively, our study elucidates the species-specific response strategies of submerged macrophyte-endophyte to nano-plastics, which helps to reveal the different phytoremediation potential of submerged macrophytes against nano-plastic pollution.

Bacterial community are more susceptible to nanoplastics than algae community in aquatic ecosystems dominated by submerged macrophytes.

As a ubiquitous emerging pollutant, microplastics can interact with algal and bacterial communities in aquatic ecosystems. Currently, knowledge on how microplastics influence algae/bacteria is mostly limited to toxicity tests using either monocultures of algae/bacteria or specific algal-bacterial consortium. However, information on the effect of microplastics on algal and bacterial communities in natural habitats is not easily available. Here, we conducted a mesocosm experiment to test the effect of nanoplastics on algal and bacterial communities in aquatic ecosystems dominated by different submerged macrophytes. The community structure of algae and bacteria suspended in the water column (planktonic) and attached to the surface of submerged macrophytes (phyllospheric) were identified, respectively. Results showed that both planktonic and phyllospheric bacteria were more susceptible to nanoplastics, and these variations driven by decreased bacterial diversity and increased abundance of microplastic-degrading taxa, especially in aquatic systems dominated by V. natans. The community composition of both algae and bacteria were influenced to varying degrees by nanoplastics and/or plant types, but RDA results showed that only bacterial community composition was strongly correlated with environmental variables. Correlation network analysis showed that nanoplastics not only reduced the intensity of associations between planktonic algae and bacteria (average degree reduced from 4.88 to 3.24), but also reduced proportion of positive correlations (from 64% to 36%). Besides, nanoplastics also decreased the algal/bacterial connections between planktonic and phyllospheric habitats. Our study elucidates the potential interactions between nanoplastics and algal-bacterial community in natural aquatic ecosystems. These findings suggest that in aquatic ecosystems, bacterial community are more vulnerable to nanoplastics and may serve as a protective barrier for algae community. Further research is needed to reveal the protective mechanism of bacteria against algae at the community level.

miR-15a regulates the preadipocyte differentiation by targeting ABAT gene in Yanbian yellow cattle.

MicroRNAs (miRNAs) are small, single-stranded, noncoding RNAs of approximately 21 to 23 nucleotides in length. Owing to their regulation of gene expression and many physiological processes including fat metabolism, they have become a popular research topic in recent years; however, the exact functional mechanisms by which they regulate fat metabolism have not been fully elucidated. Here, we identified miR-15a, which specifically acquired the 3' untranslated region (UTR) containing 4-aminobutyrate aminotransferase (ABAT), and validated the regulation of its expression and involvement in adipogenesis mechanisms. We used a dual-luciferase reporter assay and transfection-mediated miR-15a overexpression and inhibition in Yanbian yellow cattle preadipocytes to investigate the role of miR-15a in adipogenesis. The results showed that miR-15a directly targets the 3'UTR of ABAT and downregulates its expression. Additionally, at the protein and mRNA levels, miR-15a overexpression using a miRNA mimic inhibited triglyceride accumulation and downregulated lipogenic peroxisome proliferator-activated receptor γ and CCAAT enhancer-binding protein α, whereas miR-15a inhibition had the opposite effect. The above results indicated that miR-15a regulated the differentiation of Yanbian yellow cattle preadipocytes by inhibiting the expression of ABAT. Furthermore, our findings suggested that miR-15a and its target gene(s) might represent new targets for investigating intramuscular fat deposits in cattle and treating human obesity.

Individual and combined toxicity of microplastics and diuron differs between freshwater and marine diatoms.

Microplastics are considered as the emerging pollutants, which not only directly affect aquatic organisms, but also causes combined pollution by adsorbing other pollutants. Diuron, as one of the most widely used herbicides, is frequently monitored in the aquatic environment for its adverse effects on aquatic organisms. However, little is known about the combined toxicity of microplastics and diuron to aquatic organisms, especially diatoms. In this study, freshwater diatom (Cyclotella meneghiniana) and marine diatom (Skeletonema costatum) were selected to study the individual and combined toxicity of microplastics (polystyrene, 0.6- 1.0 µm) and diuron. Experimental concentrations of microplastics and diuron were set at 50 mg/L and 100 µg/L, respectively, which have been shown to significantly inhibit the growth of aquatic organisms. Results suggested that both single microplastics and single diuron significantly inhibited the growth of the two diatoms, while significant SOD and MDA increase were only found in single diuron exposure. For diatoms exposed to individual microplastics, the microplastic particles adsorbed inside Cyclotella sp. and those aggregated around Skeletonema sp. were the major factor inhibiting the growth of diatom, respectively. According to the independent action model, the combined toxicity for both diatoms were all antagonistic. The adsorption behavior of microplastics to diuron alleviated the intracellular damage to diatoms caused by diuron, and the oxidative stress induced by diuron mitigated the physical damage to diatoms caused by microplastics. Collectively, our findings suggest that the co-existence of microplastics and diuron may affect their respective toxicity to diatoms. The mechanism of this "cross-phenomenon" between microplastics and diuron and their combined toxicity to different aquatic organisms need to be further studied.

Effects of Vitamin A on Yanbian Yellow Cattle and Their Preadipocytes by Activating AKT/mTOR Signaling Pathway and Intestinal Microflora.

In this study, the effects of vitamin A and its metabolite, all-trans retinoic acid (ATRA), on the proliferation and differentiation of preadipocytes and the intestinal microbiome in Yanbian yellow cattle were investigated. Preadipocytes collected from Yanbian yellow cattle treated with different concentrations of ATRA remained in the G1/G0 phase, as determined by flow cytometry. Quantitative reverse-transcription polymerase chain reaction and western blotting analyses showed that the mRNA and protein expression levels of key adipogenic factors, peroxisome proliferator- activated receptor gamma (PPARγ), CCAAT enhancer-binding protein α (C/EBPα), and extracellular signal-regulated kinase 2 (ERK2), decreased. ATRA was found to regulate the mTOR signaling pathway, which is involved in lipid metabolism, by inhibiting the expression of AKT2 and the adipogenic transcription factors SREBP1, ACC, and FAS; the protein and mRNA expression levels showed consistent trends. In addition, 16S rRNA sequencing results showed that a low concentration of vitamin A promoted the growth of intestinal microflora beneficial to lipid metabolism and maintained intestinal health. The results indicated that ATRA inhibited the adipogenic differentiation of preadipocytes from Yanbian yellow cattle through the AKT/mTOR signaling pathway, and that low concentrations of vitamin A may help maintain the intestinal microbes involved in lipid metabolism in cattle.

Seasonality and Species Specificity of Submerged Macrophyte Biomass in Shallow Lakes Under the Influence of Climate Warming and Eutrophication.

Climate warming and eutrophication caused by anthropogenic activities strongly affect aquatic ecosystems. Submerged macrophytes usually play a key role in shallow lakes and can maintain a stable clear state. It is extremely important to study the effects of climate warming and eutrophication on the growth of submerged macrophytes in shallow lakes. However, the responses of submerged macrophytes to climate warming and eutrophication are still controversial. Additionally, the understanding of the main pathways impacting submerged macrophytes remains to be clarified. In addition, the influence of seasonality on the growth responses of submerged macrophytes to climate warming and eutrophication requires further elucidation. In this study, we conducted a series of mesocosm experiments with four replicates across four seasons to study the effects of rising temperature and nutrient enrichment on the biomass of two submerged macrophytes, Potamogeton crispus and Elodea canadensis. Our results demonstrated the seasonality and species specificity of plant biomass under the influence of climate warming and eutrophication, as well as the main explanatory factors in each season. Consistent with the seasonal results, the overall results showed that E. canadensis biomass was directly increased by rising temperature rather than by nutrient enrichment. Conversely, the overall results showed that P. crispus biomass was indirectly reduced by phosphorus enrichment via the strengthening of competition among primary producers. Distinct physiological and morphological traits may induce species-specific responses of submerged macrophytes to climate warming and eutrophication, indicating that further research should take interspecies differences into account.

Algal migration and nutrient enrichment contribute to patterns in phytoplankton versus epiphyton communities.

Algal dominance between phytoplankton and epiphyton plays an essential role in predicting shallow lake shifts between clear-water and turbid-water states. However, compared to resources competition, studies on algal life-form shifts between phytoplankton and epiphyton have traditionally received less interest, as few studies have focused on algal communities in both habitats concurrently. We conducted a 4 × 3 factorial design microcosm experiment to explore the mutual feedback relationship between phytoplankton and epiphyton. The initial algal life-form (epiphytic algae and phytoplanktonic algae alone or together) and nutrients enrichment (ambient, enrichment with N and P alone or together) were manipulated. After 28 days of incubation, the results suggested that the nutrient effects on the phytoplankton and epiphyton communities differed among the three different initial algal life-forms. A significant competitive advantage of phytoplankton was found even in treatments containing only epiphytic algae as the initial algal community. The contribution of nutrient enrichment to phytoplankton abundance (13%) was similar to that of epiphyton abundance (11%). In the mutual influence between two algal communities, epiphyton was likely to be a beneficiary as the phytoplankton community contributed 15% of the variance in epiphyton abundance. In addition, significant algal life-form shifts between phytoplankton and epiphyton only occurred in treatments containing one algal life-form, but not in treatments containing both algal life-forms at the beginning of the experiment. Our results emphasized the competitive advantage of phytoplankton in utilizing nutrient resources in the water column of shallow lakes. Moreover, we demonstrated that algal life-form shift was an adaptive behavior closely correlated with environmental variation. These results will provide broader insights to explore algal succession between phytoplankton and epiphyton in shallow lakes. To better understand the mutual influence mechanism between two algal life-forms under different nutrient conditions, research on multiple short time-scales based on algal migration is needed in the future.

Effects of submerged vegetation on sediment nitrogen-cycling bacterial communities in Honghu Lake (China).

Sediment nitrogen (N) cycling is an important biological removal process for N permanently and driven by N-cycling microbial community. There is a growing interest in interactions between submerged vegetation (SV) and sediment N-cycling bacterial community, because of the close link between rooted aquatic plants and the sediment microbes. However, the effects of SV on the sediment N-cycling bacterial community are still controversial. Furthermore, the discrimination of direct and indirect effects of SV on the N-cycling bacterial community remains unclear. Here, we investigated the biomass and species richness of SV and determined the corresponding environment factors (water quality and sediment properties) in Honghu Lake (China). We also used functional genes as markers to unveil the bacterial diversity and community composition and abundance in lake sediments. Our results showed that biomass and species richness of SV affected the composition, diversity and abundance of sediment N-cycling bacterial communities through improving lake water quality and sediment properties. With the increasing richness and abundance of SV, the diversity of most N-cycling bacterial assemblages including nitrifying, denitrifying and DNRA bacteria decreased, while the abundance increased. However, the anammox bacterial assemblage in sediments showed inverse trends. Sediment carbon vs. nitrogen (C:N) ratio negatively affected the abundance of amoA and nirS + nirK + nosZ bacterial assemblages. Additionally, due to the presence of SV, positive interactions among N-cycling bacterial assemblages were found, such as amoA and nrfA bacterial assemblages. Overall, our findings confirmed the significant effects of SV on the N-cycling bacterial community structure and abundance. Moreover, the direct effects of SV on the N-cycling bacterial community and the indirect effects through altering the sediment C were clarified in our study. Our results casted a new light on the negative effects of high C:N ratio. From the study, we made a conclusion that the better SV develops, the greater nitrogen removal occurs in lake sediments.

Warming Effects on Periphyton Community and Abundance in Different Seasons Are Influenced by Nutrient State and Plant Type: A Shallow Lake Mesocosm Study.

Periphyton plays an important role in lake ecosystems processes, especially at low and intermediate nutrient levels where periphyton contribution to primary production can be similar to or exceed that of phytoplankton. Knowledge of how periphyton responds to key drivers such as climate change and nutrient enrichment is, therefore, crucial. We conducted a series of mesocosm experiments over four seasons to elucidate the responses of periphyton communities to nutrient (low and high, TN-0.33 mg L-1 TP-7.1 µg L-1 and TN-2.40 mg L-1 TP-165 µg L-1, respectively), temperature (ambient, IPCC A2 scenario and A2 + 50%) and plant type (two submerged macrophytes with different morphological structural complexity: Potamogeton crispus and Elodea canadensis, and their corresponding plastic imitations with similar size and structure). We found a noticeable seasonality in the abundance and composition of periphyton. In spring and summer, periphyton abundances were significantly higher in the turbid-high-nutrient state than in the clear-low-nutrient state, and in summer they were notably higher at ambient temperature than in climate scenario A2 and A2 + 50%. In contrast, periphyton abundances in autumn and winter were not influenced by nutrient and temperature, but they were notably higher on plants with a more complex morphological structure than simple ones. The genus composition of periphyton was significantly affected by nutrient-temperature interactions in all seasons and by plant type in winter. Moreover, periphyton functional composition exhibited noticeable seasonal change and responded strongly to nutrient enrichment and temperature rise in spring, summer, and autumn. Our results suggest that the effect of warming on periphyton abundance and composition in the different seasons varied with nutrient state and host plant type in these mesocosms, and similar results may likely be found under field conditions.

Citrate-based fluorophore-modified cellulose nanocrystals as a biocompatible fluorescent probe for detecting ferric ions and intracellular imaging.

In this contribution, citrate-based fluorophore (CF)-modified cellulose nanocrystals (CNCs) were prepared in a facile manner using sulfuric acid hydrolysis of citric acid/cysteine-treated microcrystalline celluloses. These rod-like CNCs have an average length of 156 nm and an average width of 7.9 nm. Because of conjugated CFs, these CNCs exhibit typical fluorescence characteristics, including a maximum excitation wavelength at 350 nm, maximum emission wavelength at 435 nm, high quantum yield of 83%, and good photostability. More importantly, these fluorescent CNCs exhibit a selective quenching effect toward Fe3+ ions; meanwhile, these CNCs exhibit negligible cytotoxicity and were internalized by cells. Therefore, these CNCs can be used as a fluorescence probe for detecting Fe3+ ions in living cells.

Multielement stoichiometry of submerged macrophytes across Yunnan plateau lakes (China).

Stoichiometric homeostasis of element composition is one of the central concepts of ecological stoichiometry. We analyzed concentrations of macroelements (C, N, P, Ca, K, Mg, S), microelements (Cu, Fe, Mn, Mo, Ni, Zn) and beneficial elements (Na, Se, Si) in submerged macrophytes, water and sediments across 20 Yunnan plateau lakes. We predicted that tissue element composition in submerged macrophytes is affected by lake trophic level and taxonomy, and submerged macrophytes have weak stoichiometric homeostasis for all above 16 elements. Canonical discriminant analyses successfully discriminated among trophic level groups and taxa groups. Of all the elements, C, N, P and S most effectively discriminated among trophic level groups across 20 lakes, revealing lake trophic level mostly affect tissue macroelement composition in submerged macrophytes; while Ca, K and Se most effectively discriminated among submerged macrophytes taxa groups, suggesting taxonomy mostly affect compositions of macroelements and beneficial elements in submerged macrophytes. In addition, the stoichiometric homeostatic coefficient of 1/HCa:C for all five taxa of submerged macrophytes were less than zero, suggesting submerged macrophytes in Yunnan plateau lakes have strong Ca stoichiometric homeostasis. Our findings, not only broaden the knowledge of multielement stoichiometric homeostasis, but also help to choose most appropriate lake management strategy.

Metal accumulation by submerged macrophytes in eutrophic lakes at the watershed scale.

Metal concentrations (Al, Ba, Ca, K, Li, Mg, Na, Se, Sr and Ti) in submerged macrophytes and corresponding water and sediments were studied in 24 eutrophic lakes along the middle and lower reaches of the Yangtze River (China). Results showed that these eutrophic lakes have high metal concentrations in both water and sediments because of human activities. Average concentrations of Al and Na in tissues of submerged macrophytes were very high in sampled eutrophic lakes. By comparison, Ceratophyllum demersum and Najas marina accumulated more metals (e.g. Ba, Ca, K, Mg, Na, Sr and Ti). Strong positive correlations were found between metal concentrations in tissues of submerged macrophytes, probably because of co-accumulation of metals. The concentrations of Li, Mg, Na and Sr in tissues of submerged macrophytes significantly correlated with their corresponding water values, but not sediment values.

Bioaccumulation of heavy metals by submerged macrophytes: looking for hyperaccumulators in eutrophic lakes.

To directly select submerged macrophytes with high accumulation capability from the field, 24 eutrophic lakes along the middle and lower reaches of the Yangtze River were investigated in the study. These eutrophic lakes have large amounts of heavy metals in both water and sediments because of human activities. The results showed that Najas marina is a hyperaccumulator of As and Cd, Ceratophyllum demersum is a hyperaccumulator of Co, Cr, and Fe, and Vallisneria natans is a hyperaccumulator of Pb. Strong positive correlations were found between concentrations of heavy metals in tissues of submerged macrophytes, probably because of coaccumulation of heavy metals. However, for most heavy metals, no significant correlations were found between submerged macrophytes and their surrounding environments. In conclusion, N. marina, C. demersum, and V. natans are good candidate species for removing heavy metals from eutrophic lakes.