Circadian patterns and photoperiodic modulation of clock gene expression and neuroendocrine hormone secretion in the marine teleost Larimichthys crocea.

The light/dark cycle, known as the photoperiod, plays a crucial role in influencing various physiological activities in fish, such as growth, feeding and reproduction. However, the underlying mechanisms of this influence are not fully understood. This study focuses on exploring the impact of different light regimes (LD: 12 h of light and 12 h of darkness; LL: 24 h of light and 0 h of darkness; DD: 0 h of light and 24 h of darkness) on the expression of clock genes (LcClocka, LcClockb, LcBmal, LcPer1, LcPer2) and the secretion of hormones (melatonin, GnRH, NPY) in the large yellow croaker, Larimichthys crocea. Real-time quantitative PCR (RT-qPCR) and enzyme-linked immunosorbent assays were utilized to assess how photoperiod variations affect clock gene expression and hormone secretion. The results indicate that changes in photoperiod can disrupt the rhythmic patterns of clock genes, leading to phase shifts and decreased expression. Particularly under LL conditions, the pineal LcClocka, LcBmal and LcPer1 genes lose their rhythmicity, while LcClockb and LcPer2 genes exhibit phase shifts, highlighting the importance of dark phase entrainment for maintaining rhythmicity. Additionally, altered photoperiod affects the neuroendocrine system of L. crocea. In comparison to the LD condition, LL and DD treatments showed a phase delay of GnRH secretion and an acceleration of NPY synthesis. These findings provide valuable insights into the regulatory patterns of circadian rhythms in fish and may contribute to optimizing the light environment in the L. crocea farming industry.

Response of growth and Pb accumulation characteristics of plants with intercropping Arabis alpina-Zea mays to exogenous oxalic acid.

In order to study the effects of oxalic acids on plant growth and Pb accumulation in different parts of the plants of intercropping Arabis alpina and Zea mays, pot experiment was conducted to investigate the changes of oxalic acid contents of the plants and Pb accumulation through exogenous oxalic acid addition (0, 5, 25 and 50 mmol kg-1). The results showed the root biomass of intercropped A. alpina and total biomass of Z. mays increased by 3.22 folds and 2.97 folds with 5 mmol kg-1 oxalic acid treatment. The oxalic acid contents of shoots and root secretions decreased by 86.5% and 44.3%, respectively. The BCF (bio-accumulation factor) and TF (translocation factor) of intercropping A. alpina reduced under 25 - 50 mmol kg-1 oxalic acid treatments. There were relationships between exogenous oxalic acid treatment concentrations and oxalic acid contents of A. alpina shoots, Z. mays root secretions. The Pb contents of shoots of A. alpina and Z. mays were related to exogenous oxalic acid additions and oxalic acid contents of shoots. In general, 5 mmol kg-1 oxalic acid treatment, that can improve plant growth of intercropped A. alpina and Z. mays, which Pb translocation and accumulation of A. alpina were promoted, whereas Pb accumulation of A. alpina was inhibited with 25 - 50 mmol kg-1 concentrations addition. This study will provide a basis for promoting the application of phytoremediation techniques and efficient crop production in heavy metal contaminated areas.

Hyperaccumulators intercropped with crops will remediate heavy metal soils or mitigate the damage caused by heavy metals to plants through oxalic acid secretion by the root system. However, the effect of oxalic acid changes on plant growth and Pb accumulation is lacking. Our study investigated the changes in oxalic acid content at different concentrations and sites affected the ability of intercropped plants to grow and accumulate Pb. This work shown that under intercropping conditions, exogenous oxalic acid promotes intercropped plant growth as well as soil pH reduction, Pb content in shoots both Arabis alpina and Zea mays is influenced by exogenous oxalic acid content, while lower Z. mays roots Pb content is determined by a combination of exogenous addition and aboveground oxalic acid content. Low concentrations of oxalic acid promoted Pb enrichment in roots of A. alpina, while reducing the uptake of Pb content in Z. mays. This article gives us a better understanding for the response of intercropping plants to the use of organic acids under heavy metal stress and how to modify their environment so as to favor growth.

Circadian rhythm regulation in the sea cucumber Apostichopus japonicus: Insights into clock gene expression, photoperiod susceptibility, and neurohormone signaling.

Sea cucumber Apostichopus japonicus displays the typical circadian rhythms. This present study investigated the molecular regulation of clock genes, as well as monoamines and melatonin, in multiple tissues of A. japonicus, responding to the photoperiod. In order to determine their pivotal role in circadian rhythms, the crucial clock genes, namely AjClock, AjArnt1, AjCry1, and AjTimeless, were identified and a comprehensive analysis of their expressions across various tissues in adult A. japonicus was conducted, revealing the potential existence of central and peripheral oscillators. Results demonstrated that the tissues of polian vesicle and nerve ring exhibited significant clock gene expression associated with the orchestration of circadian regulation, and that environmental light fluctuations exerted influence on the expression of these clock genes. However, a number of genes, such as AjArnt1 and AjCry1, maintained their circadian rhythmicity even under continuous light conditions. Moreover, we further investigated the circadian patterns of melatonin (MT), serotonin (5-HT), and dopamine (DA) secretion in A. japonicus, data that underscored the tissue-specific regulatory differences and the inherent adaptability to dynamic light environments. Collectively, these findings will provide the molecular mechanisms controlling the circadian rhythm in echinoderms and the candidate tissues playing the role of central oscillators in sea cucumbers.

Comparative transcriptome analysis reveals key genes and pathways related to gonad development in the sea cucumber Apostichopus japonicus.

The sea cucumber Apostichopus japonicus is an economically important marine species in China, and understanding the mechanisms underlying its gonad development is crucial for successful reproduction and breeding. In this study, we performed transcriptome comparisons and analyses of A. japonicus gonadal and non-gonadal tissues to identify genes and molecular pathways associated with gonadal development. We also supplemented the annotation of the A. japonicus genome. Collectively, results revealed a total of 941 ovary-specific genes and 2499 testis-specific genes through different expression analysis and WGCNA analysis. The most enriched pathways in ovary and testis were "DNA replication" and "purine metabolism", respectively. Additionally, we identified key candidate gene modules that control gonad development and germ cell maturation, with CDT1 and DYNC2LI1 serving as hub genes. Our findings provide important insights into the gonadal development system of A. japonicus and offer valuable references for further research on reproductive biology in this marine invertebrate species.

Melatonin modulates the hypothalamic-pituitary neuroendocrine axis to regulate physiological color change in teleost fish.

Melatonin (MT) is a crucial neuroendocrine regulator of various physiological activities in vertebrates, especially in circadian or seasonal rhythm control. In the present study, the large yellow croaker (Larimichthys crocea), a marine bony fish with circadian body color change behavior, is chosen for functional investigation on teleost MT signaling systems that remain uncharacterized. All five melatonin receptors (LcMtnr1a1, LcMtnr1a2, LcMtnr1b1, LcMtnr1b2, and LcMtnr1c) were significantly activated by MT, triggering extracellular signal-regulated kinase 1/2 (ERK1/2) phosphorylation through different G protein coupling signaling pathways, with exclusive Gαi-dependency for LcMtnr1a2 and LcMtnr1c, and Gαq-dependency for two LcMtnr1b paralogs, whereas LcMtnr1a1 activated Gαi and Gαs dual-dependent signaling pathways. A comprehensive model of the MT signaling system in the hypothalamic-pituitary neuroendocrine axis was further constructed based on ligand-receptor interaction analysis using single-cell RNA-seq data, as well as spatial expression patterns of Mtnrs and related neuropeptides in central neuroendocrine tissues. A novel regulatory pathway of MT/melanin-concentrating hormone (MCH) and MT/(tachykinin precursor 1 (TAC1)+corticotropin-releasing hormone (CRH))/melanocyte-stimulating hormone (MSH) was discovered that functions in chromatophore mobilization and physiological color change and was further validated by pharmacological experiments. Together, our findings define multiple intracellular signaling pathways mediated by L. crocea melatonin receptors and provide the first in-depth evidence that uncover the upstream modulating roles of the MT signaling system in the hypothalamic-pituitary neuroendocrine axis of a marine teleost species, particularly in chromatophore mobilization and physiological color change.

Effects of Different Sulfur Compounds on the Distribution Characteristics of Subcellular Lead Content in Arabis alpina L. var. parviflora Franch under Lead Stress.

Sulfur plays a vital role in the phytoremediation of lead-contaminated soil. The effects of different sulfur forms (S Na2S, and Na2SO4) on lead (Pb) absorption in hyperaccumulator Arabis alpina L. var. parviflora Franch were studied in a soil pot experiment. The subcellular sulfur and lead enrichment characteristics in A. alpina were studied by adding sulfur in different forms and concentrations (0, 75, and 150 mg·kg-1) to Pb-contaminated soil. The results show that the root and shoot biomass increased by 1.94 times under Na2S and Na2SO4 treatment, and the root-shoot ratio of A. alpina increased 1.62 times under the three forms of sulfur treatments, compared with the control. Sulfur content in cell walls and soluble fractions of the root and shoot of A. alpina significantly increased 3.35~5.75 times and decreased 5.85 and 9.28 times in the organelles under 150 mg·kg-1 Na2SO4 treatment. Meanwhile, Pb content in the root and shoot cell walls of A. alpina significantly increased by 3.54 and 2.75 times, respectively. Pb content in the shoot soluble fraction increased by 3.46 times, while it significantly reduced by 3.78 times in the shoot organelle. Pb content in the root organelle and soluble fraction decreased by 2.72 and 2.46 times. Different forms and concentrations of sulfur had no regularity in the effect of Pb and sulfur content in the subcellular components of A. alpina, but the bioconcentration and translocation factors of A. alpina increased compared with the control. Under different concentrations of Na2SO4, there was a significant positive correlation between the contents of sulfur and Pb in the subcellular components of the root of A. alpina (p < 0.05). These results indicate that sulfur application can enhance the Pb resistance of A. alpina by strengthening the cell wall fixation and vacuolar compartmentalization.

Response of Cd, Zn Translocation and Distribution to Organic Acids Heterogeneity in Brassica juncea L.

In order to investigate the translocation, distribution, and organic acid heterogeneity characteristics in Brassica juncea L., a pot experiment with the exogenous application of Cd and Zn was conducted to analyze the effects of Cd, Zn, and organic acid contents and heterogeneity on the translocation and distribution of Cd and Zn. The results showed that the Cd and Zn contents of B. juncea were mainly accumulated in the roots. The Cd content in the symplast sap was 127.66-146.50% higher than that in the apoplast sap, while the opposite was true for Zn. The distribution of Cd in xylem sap occupied 64.60% under 20 mg kg-1 Cd treatment, and Zn in xylem sap occupied 60.14% under 100 mg kg-1 Zn treatment. The Cd was predominantly distributed in the vacuole, but the Zn was predominantly distributed in the cell walls. In addition, oxalic and malic acids were present in high concentrations in B. juncea. In the vacuole, correlation analysis showed that the contents of Cd were negatively correlated with the contents of oxalic acid and succinic acid, and the contents of Zn were positively correlated with the contents of malic acid and acetic acid. The contents of Cd and Zn were negatively related to the contents of oxalic acid and citric acid in xylem sap. Therefore, Cd in B. juncea was mainly absorbed through the symplast pathway, and Zn was mainly absorbed through the apoplast pathway, and then Cd and Zn were distributed in the vacuole and cell walls. The Cd and Zn in B. juncea are transferred upward through the xylem and promoted by oxalic acid, malic acid, and citric acid.

Janus kinase 1 in Megalobrama amblycephala: Identification, phylogenetic analysis and expression profiling after Aeromonas hydrophila infection.

Janus kinase 1 (JAK1), a member of the JAK family, plays an essential and non-redundant role in the mammalian immune system. However, the potential role of JAK1 in fish immune response remains largely unclear. In the present study, the JAK1 gene of Megalobrama amblycephala (MamJAK1) was identified and characterized. The open reading frame (ORF) of MamJAK1 was 3462 bp, encoding 1153 amino acids. MamJAK1 consists of four common domains of the JAK family, including B41, SH2, STyrKc (a pseudo kinase domain), and TyrKc (a kinase domain). Phylogenetic analysis showed that JAK1s are divided into two evolutionary clades, one containing fish JAK1s, and the other containing JAK1s from other vertebrates. The results of quantitative real-time PCR (qPCR) showed that in healthy M. amblycephala, MamJAK1 mRNA was highest expressed in blood, followed by spleen, intestine and mid-kidney, and lowly expressed in other tissues including gill, liver, head kidney, muscle, brain and heart. After Aeromonas hydrophila infection, the expression of MamJAK1 mRNA was significantly induced in four selected tissues including spleen, mid-kidney, liver and intestine, reaching a peak at 24 hpi (hour post infection) in spleen and mid-kidney, at 12 hpi in liver and at 4 hpi in intestine, and then the expression level was restricted to control levels at 72 or 120 hpi. In addition, the results of Western blot showed that the phosphorylation level of MamJAK1 protein in spleen and mid-kidney increased significantly after A. hydrophila infection, although MamJAK1 protein did not change obviously. Further, the JAK1 phosphorylation in Ctenopharyngodon idellus kidney (CIK) cells was found to be significantly induced by LPS stimulation and IL-6R over-expression. The results above suggest that MamJAK1 may play an essential role in the immune response against bacterial infection through the IL-6R mediated JAK1/STAT signaling pathway, which further deepen our understanding of JAK1 and provides a potential target for the treatment and prevention of bacterial diseases in teleost.

Adsorption properties and mechanism research of phosphorus with different molecular structures from aqueous solutions by La-modified biochar.

In order to explore the adsorption characteristics of phosphorus from molecules with different molecular structures and varying number of phosphate groups on metal-modified biochar, walnut shell biochar was modified with LaCl3 to prepare lanthanum-loaded biochar (BC-La). Adsorption of four polar components, namely phytic acid (IHP), adenosine-5'-disodium triphosphate (5-ATP), hydroxyethylidene diphosphonic acid (HEDP), and sodium pyrophosphate (PP), was studied. The adsorption properties and mechanism of phosphorus sorption by BC-La were analyzed by SEM-EDS and FTIR for the different structures. The results showed that the maximum adsorption capacity of BC-La for IHP, 5-ATP, HEDP, and PP was 85.85, 9.04, 15.80, and 14.45 mg/g, respectively. The adsorption capacity was positively correlated with the polarity of organic phosphorus. The adsorption behavior conformed to the quasi second-order kinetic fitting equation, and the increase of temperature was conducive to the removal of all four phosphorus pollutants. BC-La adsorbs IHP and HEDP mainly through electrostatic attraction. The adsorption of 5-ATP and PP is dominated by complexation. The La-modified biochar has broad prospects in water remediation, which can provide a theoretical basis for removal of different forms of phosphorus pollutants and prevention and control of water eutrophication.

Bio-accumulation effects of heavy metals Pb, Zn and Cd on Procecidochares utilis parasitism to Eupatorium adenophorum at Suzu metal mines, Yunnan.

Procecidochares utilis is an obligatory parasitic insect to Eupatorium adenophorum. Both organisms have been spread to some metal mines areas. The objective of this study is to comprehend the trend of heavy metals transfer and the process of their bio-accumulation in the soil-E. adenophorum-P. utilis system and particularly their impact on the parasitic effect of P. utilis to E. adenophorum to reflect the impact of heavy metals on obligate parasitic insect and its host. Therefore, a detailed investigation was carried out at the Suzu Lead-Zinc Mine in Yunnan Province using the concentric circle's method. The results showed that the parasitic rate of P. utilis to a single plant and branch is positively correlated with distance. The metals content of the soil in E. adenophorum and P. utilis, decreased dramatically with an increase in distance away from the center of the mining area. From which is cleared that these metals could enter to E. adenophorum and P. utilis through the soil-E. adenophorum-P. utilis system which likely to affect its parasitic activities. In addition, the parasitic rate is impacted by per Zn content greatly, and the parasitic rate per plant is affected by Cd content enormously. This work could provide important basis of data for further understanding and clarifying the effects of bioaccumulation and heavy metals pollution on various aspects of the food chain. Simultaneously, it could clarify and simplify whether heavy metal contamination affects the parasitic behaviour of some obligatory parasitic insects.

Identification of four STAT3 isoforms and functional investigation of IL-6/JAK2/STAT3 pathway in blunt snout bream (Megalobrama amblycephala).

Signal transducer and activator of transcription 3 (STAT3) is a major regulator of immune response and chronic inflammatory, which can be activated by interleukin-6 (IL-6). In mammals, STAT3 has multiple isoforms, and its function has been well studied. In teleost, a single stat3 has been cloned and identified in several species, but studies on its function are limited. In the present study, four stat3 isoforms including mastat3α1, mastat3α2, mastat3ß1 and mastat3ß2 were identified from blunt snout bream (Megalobrama amblycephala). The results of quantitative PCR (qPCR) showed that four mastat3 transcripts were ubiquitously expressed in all 10 tissues examined. After Aeromonas hydrophila challenge, the expression patterns of mastat3a1, mastat3a2 and mastat3ß2 were similar, but significantly different from that of mastat3ß1. In addition, western blot showed that rmaIL-6+rmasIL-6R (IL-6 trans-signaling) significantly up-regulated phosphorylation levels of the four maSTAT3 isoforms and mRNA levels of the il-10, il-11, tnf-a, socs3a and socs3b genes, while rmaIL-6 (IL-6 classical signaling) only significantly up-regulated phosphorylation levels of the two maSTAT3α isoforms and mRNA levels of the il-10, socs3a and socs3b genes. Meanwhile, overexpression or inhibition of JAK2 could significantly change the STAT3 phosphorylation. Finally, JAK2 and STAT3 inhibitors could significantly inhibit the up-regulation of il-10, il-11, tnf-a, socs3a and socs3b induced by rmaIL-6+rmasIL-6R or rmaIL-6. To sum up, this study reveals the functional distinctions and overlaps among the four maSTAT3 isoforms in blunt snout bream and reveals the differential regulation of IL-6 classical signaling and trans-signaling on downstream immune genes via the JAK2/STAT3 pathway, enriching our knowledge of fish's defense mechanisms against pathogens.

Classical Signaling and Trans-Signaling Pathways Stimulated by Megalobrama amblycephala IL-6 and IL-6R.

Interleukin-6 (IL-6) is a multipotent cytokine. IL-6 plays a dual role in inflammation through both classical signaling (IL-6 binds membrane IL-6 receptor/IL-6R) and trans-signaling (IL-6 binds soluble IL-6R). However, the regulation of IL-6 activity, especially the regulation of signaling pathways and downstream genes mediated by IL-6 trans-signaling, remains largely unclear in teleost. Grass carp (Ctenopharyngodon idellus) hepatic (L8824) cells, kidney (CIK) cells, and primary hepatocytes were used as test models in this study. First, the biological activity of recombinant blunt snout bream (Megalobrama amblycephala) IL-6 (rmaIL-6) and sIL-6R (rmasIL-6R) was verified by quantitative PCR (qPCR) and western blot. The western blot results showed that rmaIL-6 significantly upregulated signal transducer and activator of transcription 3 (STAT3) phosphorylation in L8824 cells and primary hepatocytes, while rmaIL-6 in combination with rmasIL-6R (rmaIL-6+rmasIL-6R) significantly upregulated STAT3 phosphorylation in all types of cells. Furthermore, maIL-6 and maIL-6+rmasIL-6R could only induce extracellular-signal-regulated kinase 1/2 (ERK1/2) phosphorylation in L8824 cells and CIK cells, respectively. Therefore, IL-6 mainly acts by activating the janus kinase (JAK)/STAT3 pathway rather than the mitogen-activated protein kinase (MEK)/ERK pathway. Finally, the activation of the JAK2/STAT3 pathway was shown to be essential for the generation of socs3a and socs3b induced by IL-6 trans-signaling after treatment by JAK2/STAT3 pathway inhibitors (c188-9 and TG101348). These findings provide functional insights into IL-6 classical signaling and trans-signaling regulatory mechanisms in teleost, enriching our knowledge of fish immunology.

The effects of blunt snout bream (Megalobrama amblycephala) IL-6 trans-signaling on immunity and iron metabolism via JAK/STAT3 pathway.

Interleukin-6 (IL-6) is a pleiotropic inflammatory cytokine, which plays a dual role in mammalian inflammation through both classical signaling (IL-6 binds to IL-6 receptor/IL-6R) and trans-signaling (IL-6 binds to soluble IL-6R). However, the function of IL-6, especially the regulatory mechanism of IL-6 trans-signaling in immunity and iron metabolism remains largely unclear in teleost. Here, L8824 cells (Ctenopharyngodon idella hepatic cells) were stimulated with blunt snout bream (Megalobrama amblycephala) IL-6 combination with sIL-6R protein (rmaIL-6+rmasIL-6R/maIL-6 trans-signaling) or STAT3 inhibitor (c188-9), and RNA-sequencing, global transcriptional analyses. The enrichment analysis of GO and KEGG showed that maIL-6 trans-signaling is mainly involved in stress and inflammation response, and the activation of STAT3 is mainly related to cell proliferation, apoptosis and immune regulation. Furthermore, after treated L8824 cells with JAK2 inhibitors, it was found that the induction of IL-6 trans-signaling on the selected immune-related genes could be inhibited. These results implied that in early stage after rmaIL-6+rmasIL-6R treatment, the maIL-6 trans-signaling played an important role in the immune regulation through the JAK2/STAT3 pathway. By extending the rmaIL-6+rmasIL-6R treatment time, it was found that maIL-6 trans-signaling could promote the expression of iron metabolism related genes (ft, tf, tfr1, hamp and fpn1) in L8824 cells, indicating that maIL-6 trans-signaling may be involved in iron metabolism in the non-acute immune phase. Finally, after treated L8824 cells with JAK2 and STAT3 inhibitors, it was found that only tf and fpn1 were regulated by maIL-6 trans-signaling through the JAK2/STAT3 pathway. These findings provided novel insights into IL-6 trans-signaling regulatory mechanism in teleost, enriching our knowledge of fish immunity and iron metabolism.

Evolution and expression analysis of STAT family members in blunt snout bream (Megalobrama amblycephala).

The Janus kinase/signal transducer and activator of transcription (JAK/STAT) pathway is involved in regulating the body's immunity, cell proliferation, differentiation, and apoptosis. Members of the STAT family have been extensively studied in different mammalian species. However, there are few studies on the STAT family genes in farmed economic fish. In this study, eight STAT genes including STAT1a, STAT1b, STAT2, STAT3, STAT4, STAT5a, STAT5b and STAT6, in blunt snout bream (Megalobrama amblycephala), an economically important fish in China, were identified and characterized. Analyses of gene location, phylogeny and conserved synteny were conducted to infer the evolutionary origin of these STAT family genes. Furthermore, the evolutionary origin model of STATs was constructed based on the 2R hypothesis and teleost genome duplication (TGD) hypothesis, which clarified the evolutionary origin of the eight STATs in blunt snout bream. Besides, expression of the eight STATs was detected in 10 tissues of healthy blunt snout bream, which showed different expression patterns, and all had the highest level in the blood. In addition, expression of the STATs was significantly induced in the spleen, liver, and kidney after infection of Aeromonas hydrophila, suggesting that they play an important role in protecting the host from pathogens. In general, the evolution of cytokine-related genes parallels that of the immune system, which has likely been a main evolutionary driver. Therefore, the evolutionary model of STAT genes, constructed in the non-model organism pioneeringly, may provide some enlightenment for the evolution of the fish STAT family genes and their involvement in the immune function.

Integrated analysis of lncRNA and mRNA in liver of Megalobrama amblycephala post Aeromonas hydrophila infection.

BACKGROUND: As non-coding RNA molecules of more than 200 bp in length, long non-coding RNAs (lncRNAs) play a variety of roles in biological processes, including regulating the immune responses to bacterial infections. In recent years, there have been many in-depth studies on mammalian lncRNAs, but the relevant studies in fish are very limited. Meanwhile, since lncRNAs are not conserved among species, it is difficult to apply the existing results directly to unstudied species. RESULTS: To obtain the information of lncRNAs in Megalobrama amblycephala, one of the most economically important freshwater fish in China, also to better understand the biological significance of lncRNAs in the immunity system, the fish liver at 0, 4, 12, 24, and 72 h post Aeromonas hydrophila infection (hpi) were obtained for lncRNA-sequencing (lncRNA-seq). A total of 14,849 lncRNAs were identified, and 2196 lncRNAs showed significant differences at different time points post A. hydrophila infection. Gene Ontology (GO) annotation and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed that the target genes of the differentially expressed lncRNAs were enriched in several pathways related to immune such as apoptosis, inflammation, and immune response. Time-specific modules were then identified, using weighted correlation network analysis (WGCNA), and 28 modules significantly correlated with different time point after infection were found. Furthermore, four immune-related genes and six lncRNAs in the time-specific modules were subsequently verified by RT-qPCR. CONCLUSIONS: The above findings reveal the discovery of widespread differentially expressed lncRNAs in the M. amblycephala liver post A. hydrophila infection, suggesting that lncRNAs might participate in the regulation of host response to bacterial infection, enriching the information of lncRNAs in teleost and providing a resources basis for further studies on the immune function of lncRNAs.

Organic Acid Excretion in Root Exudates as a Mechanism of Cadmium Uptake in a Sonchus Asper-Zea Mays Intercropping System.

This study assessed how the Sonchus asper L.-Zea mays intercropping influenced the plant Cd level, as well as the organic acid (low molecular mass) types and concentrations in root secretions, which was accomplished by conducting pot experiments with soil that was collected from a farmland surrounding a mining area. The results showed that the oxalic and citric acids were the prevailing organic acids in the plant root secretions, and that the intercropping led to prominently elevated root exudate concentrations of the two acids for S. asper by 43.8 and 75.4%, respectively, while decreased such concentrations by 18.5 and 18.7% for Z. mays, compared to monoculture. According to observations, the root exudate concentrations of citric and oxalic acids were significantly positively related to the available rhizosphere soil Cd and plant Cd. The results indicate that changes in root exudation of oxalic acid and citric acid result in different available Cd contents of the rhizosphere soil, thereby affecting the bioavailability of soil Cd, which increases Cd uptake and accumulation in S. asper but inhibits Cd accumulation in Z. mays.

Species diversity of Arabis alpina L. communities in two Pb/Zn mining areas with different smelting history in Yunnan Province, China.

In order to explore the species compositions and species diversity (Dominance (Do), Shannon-Wiener index (H'), Simpson diversity index (Dsi), Species richness (DMG), Pielou evenness index (Epi) and Frequency (Fr)) of twenty Arabis alpina L. var. parviflora Franch communities, field investigation was conducted in Huize county, Yunnan province, China. Some 20 A. alpina communities were distributed in two traditional smelting tailing piles of Minbingyingjiying (TST) and Shangduoduo village (SDD) with different Pb smelting history. The morphological characteristics (plant height, branch number, basal leaf length, basal leaf width and root morphology) of A. alpina, contents and accumulation characteristics of eight potentially toxic metals (As, Cd, Cr, Cu, Pb, Zn, Al, Fe) in soil and plants were analyzed. The results showed that the morphological characteristics of A. alpina were mean plant height 12.25 cm, the basal leaf length 17.69 cm, leaf width 5.40 cm, the total root length 117.86 cm plant-1, the root diameter 0.90 mm. The leaves of A. alpina in SDD were longer and narrower with higher plant height compared with in TST. The main companion plants of A. alpina were Sporobolus fertilis (Steud.) W. D. Clayt, Arenaria orbiculata Royle ex Edgew. et Hook. F. and Eulalia speciosa. (Debeaux). The H' and Dsi were 0.56-2.04 and 0.44-0.95, respectively. The plant species numbers, H', Dsi and Epi of A. alpina communities in TST were higher than those in SDD. Pb contents in shoot of two A. alpina samples were more than 1000 mg kg-1, Cd content in shoot of one A. alpina sample >100 mg kg-1 and Zn contents in shoot of seven A. alpina samples >10,000 mg kg-1 with the enrichment factors and transport coefficients greater than 1. The results indicate that A. alpina as a Zn, Pb and Cd hyperaccumulator, could be used for long-term phytoremediation of soils contaminated by Cd, Pb and Zn. Species diversity of A. alpina community during long-term natural vegetation restoration was high in the middle succession with single-peak model, and A. alpina would be dominant in the succession climax stage in potentially toxic metal contamination areas.

Novel insights into the immune regulatory effects of ferritins from blunt snout bream, Megalobrama amblycephala.

Ferritins play vital roles in maintenance of iron homeostasis as iron storage proteins. Recently, the immune function of ferritins have attracted increasing attention, especially their roles in defense against pathogenic infections. However, the immune regulatory mechanism of fish ferritins are not well known. In the present study, comparative digital gene expression (DGE) profiling was performed to explore the regulatory effects of the Megalobrama amblycephala ferritins (MamFers) using MamFers overexpressed and control L8824 cells (Ctenopharyngodon idella hepatic cell line). Clean reads were aligned to the C. idella genome and differential expression analysis was conducted with representative differentially expressed genes pointed out. On that basis, further studies were performed to verify two pivotal regulated pathways in L8824 and EPC (Epithelioma Papulosum Cyprini cell line) cells, respectively. The results showed that NLRC5 (NOD-like Receptor Family CARD Domain Containing 5) mediated the regulation of MamFers on expression of MHC I (Major Histocompatibility Complex Class I) and its chaperone ß2M (Beta-2-Microglobulin) in L8824 cells. Then, ß2M further mediated the regulation of MamFers on hepcidin expression, indicating that MamFers regulated the expression of hepcidin via NLRC5/MHC I/ß2M axis. In addition, MamFers regulated the adhesion of Aeromonas hydrophila to EPC cells by regulating the expression of two extracellular matrix proteins Intgß1 (integrin ß1) and FN (fibronectin). In a word, the present study provided novel insights into the immune regulatory functions of fish ferritins.

Effect of root exudates of intercropping Vicia faba and Arabis alpina on accumulation and sub-cellular distribution of lead and cadmium.

A field experiment in which a hyperaccumulator (Arabis alpina) was intercropped with winter crop (Vicia faba), was conducted to understand effect of the root exudates on the content and accumulated amounts, sub-cellular distribution of Cd and Pb of the intercropped plants during the ripening period of V. faba (120 d after sowing). The results showed that contents of soluble sugars exuded from the roots of intercropped A. alpina were 67.6% less than that of the monocropped plant, whereas contents of free amino acids was 57.9% greater. The total contents of organic acids exuded from roots of intercropped A. alpina and V. faba were 578.8% and 37.8% greater than that of monocropped plants, respectively. The contents of tartaric acid and malic acid exuded by roots of intercropped A. alpina were greater 31.9 times and 15.9 times than those of monocropped A. alpina, respectively. The contents and accumulated amounts of Cd and Pb in intercropped A. alpina were greater than those of monocropped A. alpina. The contents of Pb bound to organic matter in cell walls, cytoplasm and organelles of intercropped plants were greater than those of monocropped plants. These results demonstrate that increases in accumulated amounts of Pb and Cd caused by intercropping were closely related to migration of Cd and Pb in plants mediated by the composition and content of the root exudates.

Intelectin mediated phagocytosis and killing activity of macrophages in blunt snout bream (Megalobrama amblycephala).

Intelectin, a lectin discovered recently, has been identified in various vertebrate species, such as fish, amphibians, and mammals. In one of our previous studies, the efficient bacteria binding and agglutinating activity of the recombinant Megalobrama amblycephala intelectin protein (rMamINTL) and the enhanced immunopositive localization have been observed in the hepatic macrophage-like cells (kupffer cells) post Aeromonas hydrophila infection. Thus, the present study primarily focuses on the regulatory effects of rMamINTL on M. amblycephala macrophages. This study revealed a prominent LPS-binding activity of rMamINTL and a significantly increased phagocytosis of rMamINTL-treated A. hydrophila by M. amblycephala macrophages. However, the rMamINTL-treated M. amblycephala macrophages exhibited no evident regulatory effect on phagocytosis, whereas the enhanced killing activity of the rMamINTL-treated macrophages was observed, which may be attributed to the induced respiratory burst activity and the expression of inflammatory cytokines. In addition, the anti-proliferation effect of rMamINTL on two tumor cells was observed. However, its mechanism remains to be further studied. In short, these results show that MamINTL is a multifunctional immune protein with effective immunomodulatory activity.