Saliva plays a crucial role in mediating plant–insect interactions, yet the functional diversity of salivary proteins remains poorly understood. Here, we identify NlSP6935, a salivary gland-specific protein conserved among rice planthoppers but absent in bamboo-feeding relatives. Silencing NlSP6935 causes severe lethality, feeding impairment, and infertility in Nilaparvata lugens, independent of host plant resistance. Transient expression assays reveal that NlSP6935 suppresses H₂O₂ accumulation in plants, while overexpression in rice downregulates terpenoid biosynthesis and enhances host attractiveness. However, transgenic NlSP6935 plants only weakly rescue RNAi-induced lethality, demonstrating its dual role in insect physiology and plant defense suppression. Our findings reveal a novel effector essential for both planthopper survival and host adaptation, providing new insights into pest control strategies.

The bean bug, Riptortus pedestris, is a major pest of soybeans in East Asian countries. Male-released aggregation pheromones attract both adults and nymphs, offering potential for eco-friendly pest control. However, the molecular mechanisms underlying the detection of the aggregation pheromones remain unclear. In the present study, functional analysis using the Xenopus oocyte expression system demonstrated that two ORs (OR23h and OR109d) were responsible for sensing aggregation pheromones, with the primary component (E)-2-hexenyl (E)-2-hexenoate (E2HE2H) being shared by the two ORs. Further quantitative PCR (qPCR) profiling indicated that OR109d was expressed only in male antennae, while OR23h was expressed in both sexes at similar levels. RNA interference (RNAi) assays demonstrated that dsOR23h-treatment significantly reduced the electroantennographic (EAG) response of (E)-2-hexenyl (Z)-3-hexenoate (E2HZ3H) in both sexes. Furthermore, simultaneous RNAi knockdown of the two ORs significantly reduced the male EAG response to E2HE2H and abolished male attraction to this compound. These results were consistent with the sex expression profile, demonstrating the sex and functional differentiation between the two ORs. Taken together, this study characterizes the ORs responsible for chemical perception and the associated aggregation behaviors driven by these pheromones. Thus, this study enhances our understanding of olfactory signaling in a hemipteran insect and contributes to the knowledge required for improved pest management.

Although pigs lack classical brown adipose tissue, several studies have demonstrated that porcine adipocytes possess the capacity to undergo thermogenesis through UCP1-independent mechanisms. However, the developmental processes and regulatory mechanisms underlying these thermogenic adipocytes remain poorly characterized. Here, we found that dorsal subcutaneous adipose tissues (subWATs) in pigs exhibits significant thermogenic potential under cold stress. Notably, we observed substantial cold-induced structural remodeling in dorsal subWATs, characterized by increased fibrotic deposition. An integrated analysis of snRNA-seq and RNA-seq data on dorsal subWATs identified MFAP5, which encodes a microfibril-associated glycoprotein in the extracellular matrix, as a potential regulator of the cold-induced plasticity of dorsal subWATs. Both MFAP5 overexpression and MFAP5-conditioned medium (MFAP5-CM) not only inhibited preadipocyte differentiation into adipocytes but also promoted their commitment to non-adipogenic fibrogenic lineages. Furthermore, MFAP5 treatments significantly enhanced the mitochondrial biogenesis of these fibrogenic cells. Mechanistic investigations showed that these phenotypic alterations are predominantly mediated through the Hippo signaling pathway. In summary, our findings elucidate the pivotal role of MFAP5 in regulating adipocyte development following cold exposure, thus providing crucial insights into the molecular mechanisms underlying porcine adaptation to cold stress.

Ovulation is paramount for female animal fertility, necessitating a thorough understanding of its process and molecular underpinnings. This study aimed to delineate the temporal dynamics of ovulation in the goat ovary. Utilizing single-cell sequencing, we analyzed follicular fluid samples obtained at 0, 6, 12, 18, and 24 h post-hCG administration, identifying 4 cell types and 6 myeloid cell subtypes. We elucidated gene expression and functional changes in granulosa cells (GCs) over the time course of ovulation. Notably, our study detected and confirmed immune cell infiltration at 6 h post-luteinizing hormone (LH) peak. Additionally, cell–cell communication analysis revealed strong predicted interactions between GCs and macrophages, involving signaling programs associated with immune-cell recruitment, extracellular-matrix remodeling, and oocyte maturation. Collectively, our investigation has established a comprehensive single-cell transcriptome atlas of the ovulatory goat follicular microenvironment for advancing exploration into ovulation mechanisms and developing therapies for ovulatory disorders.

Sulforaphane (SFN) is a naturally occurring isothiocyanate found in cruciferous vegetables known for its anti-inflammatory and antioxidant effects in the body. However, whether its dietary addition impacts porcine liver health remains unclear, and if it does, the mechanisms are unknown. In this study, supplementing the diet of growing pigs with 1 g kg^–1 SFN was found to improve growth performance and hepatocellular proliferation. Further analyses revealed that SFN reduced hepatic and serum malondialdehyde levels, while increasing glutathione peroxidase (GSH-PX) activity in the liver. Transcriptomic and proteomic studies demonstrated that SFN down-regulated multiple pathways, including oxidative phosphorylation, inflammatory responses, IL-6-JAK-STAT3 signaling, and TNFα signaling via NFκB. Meanwhile, it upregulated NRF2/GPX4/HO-1 expression and reduced IL-6 and TNFα expression. Mechanistic studies identified potential NR1D1 and NRF2 binding elements in the promoters of the GPX4 and HO-1 genes in the liver. Furthermore, metabolomic profiling revealed a decline in serum β-hydroxybutyrate levels after the administration of SFN, while further analysis confirmed that SFN enhanced a type of epigenetic modification in the liver, lysine β-hydroxybutyrylation (Kbhb). These results highlight the protective roles of SFN against liver inflammation and oxidative damage, and a novel mechanism involving NRF2 and NR1D1 synergy is proposed, although the promotion of hepatic Kbhb by SFN still requires further exploration.

Porcine deltacoronavirus (PDCoV) is a newly identified pathogen that can potentially undergo cross-species transmission to threaten the safety of swine and humans. The mechanism by which PDCoV nonstructural protein 14 (nsp14) inhibits the expression of IFN-β is unknown. In this study, we showed that PDCoV nsp14 degrades the MAVS, MyD88 and TRAF3 proteins in host cells by proteasomal and autophagy pathways. PDCoV nsp14 recruits the E3 ubiquitin ligase MARCH8 for catalyzing MAVS, MyD88 and TRAF3 protein ubiquitination. These proteins were recognized and transported to lysosomes by the cargo receptor NDP52 for degradation to inhibit the expression of IFN-β. Furthermore, MAVS, MyD88 and TRAF3 were also found to degrade PDCoV nsp14 by selective autophagy. These results reveal the dual function of selective autophagy in the PDCoV nsp14 and host proteins, which can promote the ubiquitination of viral particles and host antiviral proteins to degrade them both for regulating the relationship between virus infection and host innate immunity.

Avian pathogenic Escherichia coli (APEC) can cause colibacillosis, which is economically devastating to poultry industries worldwide. The bacterial membrane is critical to its environment adaptability and virulence. The inner membrane protein TolA maintains membrane integrity, but its roles in the fitness and pathogenesis of APEC are not completely understood. Thus, a tolA gene mutant and complemented strains of APEC were constructed and characterized. Mutant strain ΔtolA showed damage in the inner and outer membranes, as well as altered morphology, impaired flagella production, reduced motility, increased outer membrane vesicle (OMV) production, and reduced resistance to antibiotics and environmental stress. Deletion of the tolA gene resulted in significant reductions in biofilm formation and interbacterial competition, due to the downregulated expression of biofilm-associated genes and type VI secretion system (T6SS) genes, respectively. In addition, the mutant strain exhibited reductions in serum bactericidal resistance, cell infection capacity, intracellular survival, consequently leading to attenuated bacterial survival and virulence in mice. Compared with the wild-type and complemented strains, the mutant strain induced less expression of inflammatory cytokine interleukin 1 beta (IL-1β) in HD-11 macrophages, consistent with the pathological damage in mice. In conclusion, inner membrane protein TolA contributes to the antibiotic resistance, environmental adaptability, biofilm formation and virulence of APEC.

Nitrogen (N) leaching is a major pathway of N loss in subtropical crop production systems, contributing to groundwater pollution and thus posing serious threats to human health. However, the characteristics of annual N leaching in subtropical open-field vegetable systems and the effectiveness of integrative N fertilization management practices in reducing N leaching remain poorly understood. In this study, two plot-based field experiments were conducted with open-field Chinese cabbage–pepper rotation system in subtropical Southwest China to quantify annual N leaching and evaluate the effectiveness of integrated N fertilization management practices. Experiment 1 compared five N fertilizer application rates using conventional urea, while Experiment 2 compared different N sources including conventional urea, organic fertilizer, nitrification inhibitor-based fertilizer, and controlled-release urea which were all applied at the optimized N rate. Results showed that the annual N leaching under farmers’ N practice (FNP) was 251 kg N ha⁻¹, with contributions of 55, 31, and 14% from the pepper season, Chinese cabbage season, and fallow period, respectively. Total N leaching increased exponentially with N rate. The seasonal N leaching factor was 32% for pepper and 17% for Chinese cabbage in the FNP treatment, respectively. Compared to FNP, optimizing N rate based on crop requirement and soil supply significantly reduced N leaching by 68% and gray water footprint by 66−75%, while improving N use efficiency (NUE) from 35 to 54%. In Experiment 2, mixing organic and inorganic fertilizers, applying nitrification inhibitor, and using controlled-release urea further reduced annual N leaching by 27, 54, and 25%, respectively, compared to conventional urea. These practices also improved crop yields by 2−11% and NUE by 10−13%, and lowered gray water footprint by 28−58%. In summary, integrative N stewardship practices, particularly use of nitrification inhibitors under optimized N rates, effectively reduced N leaching while achieving high NUE and vegetable yields, providing a promising strategy for sustainable subtropical vegetable production

While straw mulching has the potential to reduce fertilizer-nitrogen (N) losses in intensively managed cropland, how soil organic carbon (SOC) regulates this fate of fertilizer-N at soil aggregate or profile scales remains unresolved. Here, micro-plots were nested within a four-year field experiment to assess fertilizer-N fates and their linkages with SOC fractions and stabilization processes via ¹⁵N-tracing and ¹³C natural abundance analyses. Three treatments were included: (i) conventional N application (FN), (ii) reduced N application (RN), and (iii) reduced N with straw mulching (RS). While RN reduced crop yields compared to FN, RS achieved comparable yields and 7.71% higher N recovery efficiency (P<0.05). The δ¹³C fractionation between aggregates and bulk soil was significantly positively correlated with the fertilizer-N content in the >2 mm and <0.053 mm fractions, indicating that N retention was coupled with SOC stabilization processes. Compared with RN, RS resulted in a 2−3.4 times greater SOC conversion probability into the <0.053 mm fraction and a 1.4 times higher aggregate-associated fertilizer-N content. SOC fractions differentially regulated the profile distribution of fertilizer-N, with nonlabile organic carbon (C) correlated positively, while dissolved organic C correlated negatively but increased plant N recovery. Compared with RN, RS increased the SOC stock by 24%, reduced NO₃⁻-N accumulation by 37%, and immobilized 36% more N into the microbial biomass (P<0.05). Our findings demonstrate that straw mulching increases N recovery by mediating SOC fractionation, stabilization, and microbial N immobilization. These results provide new insights into SOC–N interactions that could aid in the development of optimal soil C and N management strategies.

Improving soil organic matter (SOM) maintenance is crucial for terrestrial carbon (C) sequestration and ecosystem functioning. Conservation tillage favors SOM pool buildup; however, it remains unclear how the decomposition of heterogeneous components is manipulated by microbial substrate utilization strategy from the view of SOM stability. Here, a one-year microcosm incubation was conducted using surface soils developed under 12 years of conservation tillage (high-C soil) and maize residue removal (low-C soil). Temporal changes in lignin phenols, neutral sugars, and amino sugars in the soil were monitored along with microbial phospholipid fatty acids (PLFAs) and enzyme activities. Throughout incubation, lignin phenols declined more (20.8–26.3%) than the SOM (12.3–14.5%) and amino sugars (10.6–12.3%), highlighting the key role of plant debris in SOM mineralization, and complementarily, the greater contribution of microbial necromass to SOM stabilization. Moreover, the decomposition dynamics of neutral sugars and lignin were strongly influenced by C availability. In the low-C soil, these two types of compounds decomposed with similar temporal patterns and extents, and such substrate co-metabolism was dominantly mediated by actinomycetes. In contrast, in the high-C soil, a lower oxidases-to-carbohydrolases ratio regulated the sequential decomposition of labile neutral sugars followed by recalcitrant lignin. Such microbial substrate selectivity was associated with a shift in microbial community from bacterial dominance toward increased fungal contribution. Overall, our findings underscore the significant interplay between soil C availability and flexible microbial substrate utilization strategy in regulating decomposition of heterogeneous SOM components, as well as their distinct contributions in SOM turnover and stabilization.