Plant viruses pose significant threats to agriculture, with many vectored by insect pests. The entry of viruses and their encoded proteins into the host nucleus is a critical step for promoting some viral replication and enabling systemic infection. Laodelphaxstriatellus, also known as the small brown planthopper (SBPH), is an efficient vector for rice stripe virus (RSV), one of the most damaging viruses of rice. In this study, we demonstrate that RSV infection induces the expression of genes in both the classical and non-classical nuclear import pathways of SBPH. A gene belonging to the importin β family, importin5(LsIPO5), was upregulated by 84% in SBPH midguts infected with RSV. The nuclear localization signal (NLS, ¹⁶⁸YRSPSKKRHKYV¹⁷⁹) is located within the nonstructural protein NS3 directly bound to LsIPO5, thereby facilitating NS3 nuclear entry. Moreover, a RING-type E3 ligase (LsRING) in SBPH, which mediated the ubiquitination of NS3 in the insect vector, enhanced NS3 binding to LsIPO5 and facilitated NS3 perinuclear localization. Combined treatment of SBPH with both dsIPO5and dsRINGsignificantly reduced RSV loads, highlighting the importance of LsIPO5 and NS3 ubiquitination cooperation in facilitating viral replication. Our findings provide new insights into synergistic molecular mechanisms that govern RSV infection and suggest potential therapeutic targets to control viral transmission through their insect vectors.

The significance of purine base content as an important nutrition indicator in foods arises from its potential to trigger hyperuricemia or gout viahigh-purine diet. Livestock meats, including pork, generally contain moderate to high total purine content (TP). Recent research revealed substantial variations within and across pig breeds, implying genetic factors influencing this trait. Thus, this study aimed to unravel the genetic underpinnings governing purine base content in pork. The heritability estimates (h²) for the four purine traits ranged from 0.14 to 0.35. A total of 14, 36, 19 and 25 quantitative trait loci (QTLs) were identified for guanine, adenine, hypoxanthine, and TP, respectively. Our comprehensive gene set enrichment analysis and gene network analysis revealed 15 promising candidate genes intricately interwoven within diverse purine metabolism pathways, such as purine ribonucleotide metabolic process, purine nucleotide metabolism and transport, and purine salvage pathways, all contributing to TP. Strikingly, most genetic variants significantly associated with TP displayed analogous effects on multiple purine bases. Two distinct and highly significant QTLs (P<10^–12) emerged on Susscrofachromosome (SSC) 12: one impacting guanine content and the other concurrently influencing adenine and hypoxanthine levels. The peak of the guanine QTL on SSC12 resided 1.1 kb downstream of the transmembrane protein 238 like (TMEM238L) gene and is encapsulated within a genomic segment characterized by the histone modification H3K27me3. Focused fine-mapping for the SSC12 QTL associated with adenine and hypoxanthine levels narrowed its scope to around 172 kb, encompassing the growth arrest specific 7 (GAS7) and myosin heavy chain 13 (MYH13) genes. However, the observed QTL effect was not attributed to any missense mutations within the two genes. This pioneering study unveils the genetic variations and candidate genes associated with purine content in livestock, laying a robust foundation for the selective breeding of pig lines with reduced purine base content.

Genetic improvement of meat production traits has always been the primary goal of pig breeding. Geographical isolation, natural and artificial selection led to significant differences in the phenotypes of meat production traits between Chinese local pigs and Western commercial pigs. Comparative genomics and transcriptomics analysis provided powerful tools to identify genetic variants and genes associated with skeletal muscle growth. However, the number of available genetic variants and genes are still limited. In this study, a comprehensive comparison of transcriptomes showed that ribosomal protein S27-like (RPS27L)gene was highly expressed in skeletal muscle and up-regulated in Chinese local pigs when compared with Western commercial pigs. Functional analysis revealed that overexpression of RPS27Lpromoted myoblast proliferation and repressed differentiation in pig skeletal muscle cells. Conversely, the knockdown of RPS27Lled to the inhibition of myoblast proliferation and the promotion of differentiation. Notably, a 13-bp insertion-deletion (InDel) mutation was identified within the RPS27Lpromoter, inserted in Chinese local breeds and predominantly deleted in Western commercial breeds. Luciferase reporter assay suggested this InDel modulated RPS27Lexpression by influencing transcription factor 3 (TCF3) and myogenic differentiation antigen (MYOD) binding to the promoter. Furthermore, a positive correlation was observed between RPS27Lexpression and backfat thickness. Association studies demonstrated this InDel was significantly associated with the body weight of pigs at the age of 240 d. Together, our results suggested that RPS27Lwas a regulator of skeletal muscle development and growth, and was a candidate marker for improving meat production traits in pigs. This study not only provided a biomarker for animal breeding, but also was helpful for understanding skeletal muscle development and muscular disease in humans.

Myostatin (MSTN) is principally expressed in skeletal muscle and negatively regulates muscle growth and development. MSTN mutation can induce muscle overgrowth in cattle by activating cell proliferation, presenting a “double-muscle” phenotype. However, the specific regulatory mechanism is still unclear. Here, we found that Ca²⁺content in muscle tissue and muscle satellite cells of MSTN mutated (MSTN^–/–) cattle were significantly increased compared to wild-type (WT). Furthermore, transcriptome analysis of muscle satellite cells revealed that TRPC4 was significantly increased in MSTN^–/– cattle. And the expression of TRPC4 in muscle tissue of MSTN^–/–cattle was detected by RT-qPCR and Western blot, which was significantly higher than that of WT. These results suggested that MSTN mutation promoted muscle satellite cells proliferation through activation of TRPC4 channel. To further verify, ML204, a specific inhibitor of TRPC4, was used to treat MSTN^–/–muscle satellite cells. We found that cell proliferation was inhibited, calcineurin expression was downregulated, and the entry of NFATc3 into nuclei was reduced, which was similar to WT group. Thus, MSTN mutation leads to the activation of TRPC4 channel, which increases intracellular Ca²⁺ content, further activates calcineurin/NFATc3 pathway, and ultimately promotes the proliferation of muscle satellite cells.

Ketosis, a common metabolic disease during early lactation, is associated with high circulating levels of β-hydroxybutyrate (BHB). A portion of BHB that reaches the mammary gland is utilized as precursor for synthesis of fatty acids. Recent findings from nonruminant studies revealed that long chain fatty acyl-CoA ligase 4 (ACSL4) could play a role in the regulation of cellular fatty acid metabolism, but the mechanisms by which ACSL4 mediates cellular lipid metabolism in response to BHB remains unclear. To achieve the aims, we conducted invivoor invitroanalyses using bovine mammary gland biopsies and the immortalized mammary epithelial cell line (MAC-T). The invivostudy (n=6 cows per group) involved healthy cows (plasma BHB<0.60mmol L^–1) or ketotic cows (plasma BHB>2.0mmol L^–1) from which mammary gland tissue was biopsied. Invitro, MAC-T cells were challenged with 0, 0.3, 0.6, 1.2, or 2.4 mmol L^–1BHB for 24h to determine an optimal dose. Subsequently, MAC-T were incubated with 1.2mmol L^–1BHB for 0, 3, 6, 12, 24, or 48h. Furthermore, MAC-T cells were treated with small interfering ACSL4 (siACSL4) for 24 h or ACSL4 overexpression plasmid (pcACSL4) for 36h followed by a challenge with 1.2mmol L^–1BHB for 24h. Results showed that increased mRNA and protein abundance of lipogenic genes were linked to both mammary gland and invitrochallenge with BHB. BHB increased fatty acid content by activating ACSL4 expression, whereas inhibition of ACSL4 reduced BHB-induced reactive oxygen species (ROS) overproduction, enhancement of mitochondrial membrane potential, increase in fatty acid content, and lipid droplet accumulation. Furthermore, we also elevated ACSL4 expression with an overexpression plasmid to clarify its molecular role in response to BHB challenge. ACSL4 overexpression enhances BHB-induced lipid droplet accumulation by increased fatty acid content. Overall, the information showed that ACSL4 is crucial for the process of producing fatty acids from exogenous BHB. Reduced ACSL4 decreased fatty acid content and lipid droplet accumulation, improved mitochondrial function, directed more fatty acids towards oxidation. Thus, ACSL4 plays an important role in determining the fate of intracellular fatty acids and BHB in BMECs.

The bone morphogenetic protein (BMP) gene family comprises a group of multifunctional cytokines that play important roles in limb development, bone formation, fat deposition, and reproductive traits of vertebrates. However, no systematic and comprehensive investigations of the various traits of the whole family members have been conducted, particularly in chickens. Here, we performed genome-wide screening and identified 14 BMPgenes, which were classified into the BMP2/4, BMP5/6/7/8A, growth differentiation factor (GDF) 2/BMP10, GDF5/6/7, and GDF11/BMP3/15 subfamilies. Genetic variation pattern analysis showed that BMPgenes were responsible for the artificial selection of commercial broilers and layers, with BMP2, BMP6, and GDF7likely contributing significantly to the formation of both specialized meat- and egg-type lines, whereas BMP7likely contributed more to the formation of meat-type lines. Genetic association analysis showed that single nucleotide polymorphisms (SNPs) in the BMP7intron region were associated with body weight, breast muscle weight, leg weight, abdominal fat weights and contents of total cholesterol (T-CHO), triglyceride (TG), low-density lipoprotein (LDL), and high-density lipoprotein (HDL) in serum. Additionally, gain- and loss-of-function assays demonstrated that BMP7promoted the proliferation, myogenic differentiation, and lipid droplet accumulation in myoblasts; enhanced lipid synthesis in hepatocytes; promoted the proliferation and inhibited adipogenic differentiation of intramuscular preadipocytes; and induced the proliferation and adipogenic differentiation of abdominal preadipocytes. These results provide novel insights into the role of BMPgenes in chicken growth, reproductive regulation, and lipid deposition and could be used to develop genetic markers for breeding selection in chickens.

The objectives of this study were to evaluate the effect of hexanoic acid (HA) supplementations (0, as the control, CON; 0.05%, HA1; 0.1%, HA2; 0.2%, HA3) on β-carotene, and ascertain the way and key factors of HA influencing β-carotene content of alfalfa (Medicago sativaL.) after ensiled in an oxygen-free and dark conditions for 10, 40, and 80 d (from May to August, 2021). This was achieved by examining the dynamic change of β-carotene, activities of β-carotene-related enzymes, and bacterial community succession of ensiled alfalfa, using operoncrtNMidentification,crtEgene quantitation, and single-molecule real-time sequencing technology. The results revealed that when compared with the fresh material, terminal alfalfa silage treated with different level of HA supplementations (0, 0.05, 0.1, 0.2%; fresh weight basis) increased β-carotene content up to 2.86, 85.8, 159, and 133%, accordingly. Meanwhile, alfalfa silage treated with higher levels of HA (0.1 and 0.2%) showed superior effects compared to those treated with lower levels of supplementation (0 and 0.05%). HA supplementation specifically facilitated the increase abundance of Lactobacillus kullabergensisand the emergence of L.senioris.Multiple linear regression models inferred thatL.kullabergensis,L.apis, L.saniviri, L.senioris,peroxidase, phytoene desaturase, and lycopene β-cyclasepositively regulated β-carotene. Conversely,LactobacillusrenniniandL.brevisadjusted β-carotene, negatively. Positive regulations of the above bacterial species and enzymes had a stronger role in increasing β-carotene than L.renniniandL.brevis.In conclusion, the β-carotene increase of ensiled alfalfa may be regulated by HA supplementation viamultiple positive factors, including 4 specialLactobacillusspecies (L.kullabergensis, L.apis, L.saniviri, and L.senioris), and 3 vegetative β-carotene-related enzymes (peroxidase, phytoene desaturase, and lycopene β-cyclase).

Reducing aerobic spoilage and rumen greenhouse gas emissions from anaerobically fermented feeds remains critical challenges in energy saving and environmental protection of animal husbandry. This study investigated the effects of dandelions, both alone and combined with Lactobacillus plantarumand Lactobacillus buchneri(LAB) on fermentation quality, bacteriome and mycobiome after 180 d of anaerobic and 4 d of aerobic fermentation of whole-plant corn. In vitrodry matter digestion (IVDMD) and gas production from anaerobically fermented whole-plant corn were also assessed. The results demonstrated that dandelions, either alone or combined with LAB, effectively improved fermentation quality by reducing NH₃-N concentrations (22.72–25.99%) after anaerobic fermentation, decreasing the proliferation of yeast and molds to enhance the aerobic stability. Notably, the changes in the bacteriome were more pronounced than those in the mycobiome after aerobic exposure. The addition of dandelions or the combination reduced Acetobacter fabarumabundance, a member of the Acetobacterthat was spoilage-induced microbe indicated by correlation analysis. Besides, these treatments facilitated competition relations of microbiome which contributed to the enhanced aerobic stability. Furthermore, dandelions reduced CH₄and CO₂emissions by 14.88 and 13.73%, respectively, and also positively influencing IVDMD by 4.46%. Collectively, dandelion alone or combined with LAB are promising strategies to improve the aerobic stability of anaerobically fermented whole-plant corn, a process linked to the interactions between the bacteriome and mycobiome, and to contribute to clean production by reducing rumen CH₄and CO₂emissions.

Barren paddy fields characterized by poor soil structure, shallow tillage layers and low organic carbon content are a common limitation to rice production in subtropical China. As a novel approach to soil improvement, granulated organic amendments offer significant potential. Previous studies have shown that granulated straw can improve soil physicochemical properties and rapidly increase the soil organic carbon (SOC) content. However, their effects on barren paddies remain underexplored. This study evaluated four soil amendment strategies: no organic amendments (CK), 10 t ha^–1of composted manure (M10), 20 t ha^–1of granulated organic amendment (G20), and 40 t ha^–1of granulated organic amendment (G40). The objective was to assess the effects of these amendments on soil structure, the contents of aggregate-associated carbon (AAC), particulate organic carbon (POC) and mineral-associated organic carbon (MAOC), and the chemical stability of MAOC among various size aggregates in both topsoil (0–20 cm) and subsoil (20–40 cm). The results demonstrated that organic amendment inputs significantly increased the macroaggregate (>250 µm) proportion and improved soil structural stability. These amendments also elevated the carbon concentration within aggregates of various sizes and facilitated the redistribution of organic carbon from microaggregates (53–250 µm) and silt+clay fractions (<53 µm) to macroaggregates. The proportion of POC to AAC declined with decreasing aggregate size, whereas the proportion of MAOC increased. In the topsoil, macroaggregate formation enhanced the protection of POC, supported the accumulation of non-hydrolyzable carbon within MAOC, and accelerated the formation of intra-microaggregates. In the subsoil, mineral-bound organic carbon remained the dominant form of carbon sequestration. In conclusion, the application of 40 t ha^–1of granulated organic amendment proved to be a successful tactic for enhancing soil physicochemical structure, increasing SOC content, and improving carbon stability. This approach offers a promising and innovative solution for the sustainable management and restoration of barren paddy fields.

Legume-based intercropping enhances asymbiotic biological nitrogen fixation (BNF); however, the underlying mechanisms remain unclear, including the roles of soil keystone diazotroph taxa with varying niche breadths. A field experiment was conducted to evaluate soil BNF variations between rhizosphere and bulk soils in peanut/cotton intercropping systems and monocultures. BNF activities were measured by nitrogen fixation rates, nitrogenase activity, and nifHgene abundance. Phylogenetic null models, co-occurrence networks, and niche breadth analysis were applied to investigate the roles of diazotrophic keystone taxa and their ecological niches. Rhizosphere soils exhibited 7.8–125.5% higher BNF potentials than bulk soils, whereas intercropping systems showed 11.6–323.0% increases over monocultures for nitrogen fixation rate, nitrogenase activity, and nifHgene abundance (all P<0.05). Diazotrophic community composition and diversity differed significantly, with Proteobacteria (excluding Alphaproteobacteria) enriched in intercropping and rhizosphere soils, while Cyanobacteria and Firmicutes were less abundant. Deterministic processes, particularly heterogeneous selection, dominated community assembly in the rhizosphere (91.9%) and intercropping soils (86.3%). The co-occurrence networks consistently revealed more complex and interconnected communities in intercropping and rhizosphere soils that were dominated by opportunistic diazotrophs (78.8–85.9%), followed by specialists (10.2–18.5%) and generalists (1.38–3.80%). Keystone taxa, including opportunists such as Azoarcus, Azohydromonas, and Steroidobacter, and generalists like Pseudomonasand Azotobacter, correlated positively with microbial biomass carbon and nitrate nitrogen, contributing to enhanced BNF. Peanut/cotton intercropping enhances BNF by selectively enriching the keystone diazotrophic taxa with varying ecological roles, particularly opportunists and generalists. Such targeted intercropping strategies can optimize BNF, improve soil fertility, and promote sustainable agricultural production.