Regulatory roles of miRNA-530 in the post-transcriptional regulation of NF-κB signaling pathway through targeted modulation of IκBα in Sebastesschlegelii.

MicroRNAs (miRNAs) are a crucial type of non-coding RNAs involved in post-transcriptional regulation. The playing essential regulatory roles in the NF-κB signaling pathway and modulate the host immune response to diverse pathogens by targeting IκBα. However, the regulatory mechanism of miRNAs in relation with IκBα in Sebastes schlegelii remains unclear. In our study, we identified two copies of IkBα gene in black rockfish (Sebastes schlegelii), namely IkBα1 and IkBα2. Moreover, we have discovered that miRNA-530 can activate the NF-κB signaling pathway by inhibiting the expression of IκBα, thereby inducing the inflammatory response. This project comprehensively investigated the interactive regulatory roles of miRNA-530 in the NF-κB signaling pathway at both cellular and in vivo levels, while also elucidating the regulatory relationships between miRNA-530 and IκBα. In conclusion, our research confirmed that miRNA-530 can target the 3'UTR region of IκBα, resulting in a decrease in the expression of IκBα at the post-transcriptional level and inhibiting its translation. The findings contribute to the understanding of the regulatory network of non-coding RNA in teleosts and its subsequent regulation of the NF-κB signaling pathway by miRNAs.

Maize/peanut rotation intercropping improves ecosystem carbon budget and economic benefits in the dry farming regions of China.

Monoculture is widely practiced to increase crop productivity, but long-term adaptation has drawbacks as it increases the depletion of soil nutrients and reduces soil quality, especially in dryland areas. Conversion from traditional maize monoculture to intercropping improves sustainable production. However, maize/peanut intercropping, especially rotation of planting strips impacts of maize/peanut intercropping in dryland on carbon (C) budgets and economic benefits remain unclear. In this study, a 5-year field experiment was conducted to evaluate the influence of maize/peanut intercropping with rotation of planting strips on soil health, indirect CO2-eq greenhouse gas emissions, and ecosystem C inputs. Four intercropping treatments viz. maize monoculture, peanut monoculture, maize/peanut intercropping, and maize/peanut rotation-intercropping were tested from 2018 to 2022. Maize/peanut rotation intercropping significantly improved the land equivalent ratio followed by intercropping and monoculture. Rotation-intercropping also improved economic benefits over intercropping and monoculture which were mainly associated with increased peanut yield where the border rows contributed the maximum, followed by the middle rows. Moreover, rotation-intercropping significantly increased the soil organic C and nitrogen (N) content. Rotation-intercropping decreased indirect CO2-eq greenhouse gas emissions and ecosystem C inputs by 3.11% and 18.04%, whereas increased ecosystem C outputs and net ecosystem C budget by 10.38% and 29.14%, respectively, over the average of monoculture. On average for intercropping and monoculture, rotation-intercropping increased ecosystem C emission efficiency for economic benefits by 51.94% and 227.27% in 2021 and 2022, respectively, showing the highest C utilization efficiency than other treatments. In the long run, maize/peanut rotation-intercropping can be practiced in dryland agriculture to achieve sustainable agriculture goals.

Comprehensive analysis of circRNA-miRNA-mRNA networks in the kidney of snakehead (Channa argus) response to Nocardia seriolae challenge.

Non-coding RNAs (ncRNAs) play vital roles in regulating the expression levels of genes that control essential biological functions, including immune response to bacterial infections in teleost. To dissect the roles of ncRNAs in the Channa argus (snakehead), a systematic analysis of the expression profiles of circRNA, miRNA and mRNA, as well as competing endogenous RNAs (ceRNA) regulatory networks in the kidney of snakehead following Nocardia seriolae infection were performed in the present study. A total of 111 differentially expressed circRNAs, 706 differentially expressed miRNAs, and 2548 differentially expressed mRNAs were identified in the N. seriolae infected snakehead. Based on these differently expressed RNAs, we identified 55 circRNA-mRNA pairs, 124 miRNA-mRNA pairs, and 35 circRNA-miRNA-mRNA regulatory networks, including dre-miR-103-CD302, dre-miR-27e-IGSF3, novel_circ_0005462/novel_403-IGKC, novel_circ_0001750/novel_circ_0002162-novel_477-OCLN, and novel_circ_0003847-novel_4-KCNAB3. In addition, luciferase reporter assay was employed to detect the target relationships of several circRNA-miRNA-mRNA pairs. Taken together, this study demonstrates that the genes associated with immunity and structures in the kidney of snakehead can be regulated by circRNAs and miRNAs at post-transcription levels, and provided theoretical guidance for ncRNAs studies for other teleost. However, further studies are still in great need to validate the regulatory mechanisms of ncRNAs in snakehead.

Characterization of caspase gene family in Sebastes schlegelii and their expression profiles under Aeromonas salmonicida and Vibrio anguillarum infection.

The caspase, functioning as a proteinase, plays a crucial role in eukaryotic cell apoptosis, regulation of apoptosis, cellular growth, differentiation, and immunity. The identification of caspase gene family in Sebastes schlegelii is of great help to understand its antimicrobial research. In S. schlegelii, we totally identified nine caspase genes, including four apoptosis initiator caspases (caspase 2, caspase 8, caspase 9 and caspase 10), four apoptosis executioners (caspase 3a, caspase 3b, caspase 6, and caspase 7) and one inflammatory executioner (caspase 1). The duplication of caspase 3 genes on chr3 and chr8 may have been facilitated by whole genome duplication (WGD) events or other complex evolutionary processes. In general, the number of caspase genes relatively conserved in high vertebrates, while exhibiting variation in teleosts. Furthermore, syntenic analysis and phylogenetic relationships analysis supported the correct classification of these caspase gene family in S. schlegelii, especially for genes with duplicated copies. Additionally, the expression patterns of these caspase genes in different tissues of S. schlegelii under healthy conditions were assessed. The results revealed that the expression levels of most caspase genes were significantly elevated in the intestine, spleen, and liver. To further investigate the potential immune functions of these caspase genes in S. schlegelii, we challenged individuals with A. salmonicida and V. anguillarum, respectively. After infection with A. salmonicida, the expression levels of caspase 1 in the liver and spleen of S. schlegelii remained consistently elevated throughout the infection time points. The expression levels of most caspase family members in the intestine exhibited significant divergence following V. anguillarum infection. This study provides a comprehensive understanding of the caspase gene families in S. schlegelii, thereby establishing a solid foundation for further investigations into the functional roles of these caspase genes.

Optimization of fertilization scheme based on sustainable wheat productivity and minor nitrate residue in organic dry farming: An empirical study.

The substitution of chemical nitrogen fertilizer with manure holds the potential for a synergistic rise in wheat grain yield and protein concentration, while minimizing residual nitrate in soil. We conducted a 6-year field fertilization experiment including two manure treatments (with or without) and five nitrogen applications rates (0, 60, 120, 180 and 240 kg ha-1). The study investigated the impact of single chemical nitrogen (CN) and manure substitution for nitrogen fertilizer (MN) on the grain yield (GY), grain protein concentration (GPC), plant nitrogen uptake (PNupt) and plant nitrogen requirement (PNR) of wheat, and the dynamic change of soil nitrate-N. The findings revealed that: (1) the MN demonstrated a greater advantage over CN, as evidenced by a 13.4-16.0 % increase in GY, a 2.6-3.8 % increase in GPC, a 7.2-15.7 % increase in PNupt and a 1.5-4.7 % reduction in PNR. (2) Soil nitrate accumulation (SNA) significantly increased when fertilizer rates ≥180 kg ha-1 and the peak annually shifted to deeper layer. The MN increased the SNA0-100 by 20.9-21.8 %, but significantly reduced SNA0-200 by 11.8-13.5 % compared with the CN. Topsoil nitrate content (SNC0-20) can be adopted as a substitute for SNA0-100 to make the fertilization schedule convenient. (3) Regression analysis revealed (taking the MN for example) that the optimum N rates for the maximum GY (5417 kg ha-1) and GPC (15.3 %) were 164 and 211 kg N ha-1, respectively. The nitrate-N safety threshold was 62 kg ha-1 at the fertilizer rate of 89 kg N ha-1. Based on this, nitrogen fertilizer input reduced by 44.8-57.2 % and SNA0-200 by 17.9-33.6 %, with achieving 91.8-95.0 % of maximum GY and 89.7-92.9 % of maximum GPC. Substituting manure for nitrogen fertilizer achieved the potential of maintaining the grain yield and protein concentration while the minimization in soil nitrate residue. This study offers a feasible way for fertilization recommendation and nitrate residue controlling in dry farming.

Molecular characterization and expression analysis of immunoglobulins (IgM and IgT) heavy chains in black rockfish (Sebastes schlegelii) that response to bacterial challenge.

Sebastes schlegelii is a kind of fish with great economic values. Recently, with the continuous expansion of aquaculture scale and the continuous improvement of aquaculture density, outbreak of various diseases has caused huge economic losses to its aquaculture industry. Study of fish immune system can help to understand the mechanism of immune response to external pathogens and can promote the development of immune prevention and control methods. Immunoglobulins (Igs) are complex glycoproteins that appear to be unique to the vertebrates that can recognize a wide variety of pathogens and recruit immune cells and molecules to destroy pathogens, which are generated by a series of rearrangement and somatic mutations. We therefore studied the immunoglobulin genes of S. schlegelii in view of their important roles in resisting to external pathogen infections. In this study, the immunoglobulin heavy chain genes (sIgM, mIgM, sIgT, and mIgT) of S. schlegelii were successfully identified and cloned. Phylogenetic analysis showed that the IgM and IgT genes of S. schlegelii were clustered together with homologous genes of other species, indicating that they were highly conserved during the evolutionary process. Collinearity analysis showed that the immunoglobulin genes and their adjacent genes were aligned with zebrafish, Atlantic salmon and tilapia, which further confirmed the conserved immunoglobulin gene of teleost. Expression analysis of healthy tissues showed that the expression levels of sIgM, sIgT and mIgT were the highest in the skin, while mIgM was the highest in spleen. After different bacterial infection, IgM and IgT were significantly expressed in skin and gill, which may be because skin and gill are the first line of defense against the infection pathogens. Subcellular localization showed that the mIgT protein was expressed in both the cell membrane and cytoplasm. Meanwhile, recombinant protein of mIgT was obtained in vitro, which laid a foundation for subsequent protein function studies. These results provide a theoretical basis for understanding the immunity role of immunoglobulin in S. schlegelii.

[Effect of Electroacupuncture Stimulation of Different Tissues at "Huantiao" (GB 30) Acupoint on Expression of Phosphorylated JNK and c-jun in Spinal Cord of Rats with Sciatic Nerve Injury].

OBJECTIVE: To observe the effects of electroacupuncture (EA) stimulation of different tissues (nerve stem, muscular layer) at "Huantiao" (GB 30) acupoint on expression of hosphorylated c-jun N-terminal kinase (p-JNK) and c-jun (p-c-jun) proteins in the lumbar spinal cord in rats with sciatic nerve injury, so as to explore its mechanism underlying improvement of peripheral neuropathic damage. METHODS: Forty-eight SD rats were randomly divided into normal, model (the left sciatic nerve severed), GB 30 deep needling (the acupuncture needle tip was inserted to the sciatic nerve trunk to elicit an instantaneous jerk of the hind limb) and GB 30 shallow needling (the needle tip was inserted to the muscle layer to evoke a local muscular contraction) groups (n = 12 rats in each group). EA stimuli were delivered at 2 Hz/100 Hz, 1 mA, 20 min in duration per treatment for 10 consecutive days. Histopathological changes were observed by Hematoxylin-eosin (HE) staining and immunohistochemical assay was carried out to examine the pathological change of spinal segments (L4-L5) and the expression of p-JNK and p-c-jun proteins, respectively. RESULTS: For rats with the sciatic nerve severed, the spinal neurons became swelling, degeneration or even apoptosis. Acupuncture intervention reduced the number of apoptosic neurons and improved the pathological change, which was relatively better in the.deep needling group than in the shallow needling group. Likewise, the elevated spinal p-JNK and p-c-jun expression levels of the model group were significantly reduced by EA intervention (deep needling vs shallow needling, P < 0.01. CONCLUSION: Acupuncture can improve the spinal pathological changes in rats with sciatic nerve injury, which is probably achieved by decreasing the p-JNK and p-c-jun expression and inhibiting the JNK signaling pathway, and thereby, reducing the apoptosis of the spinal neurons. Deep needling results in greater benefits than shallow needling.