The red imported fire ant, SolenopsisinvictaBuren, is a highly invasive eusocial insect pest that threatens native biodiversity, agriculture, and human health. The innate immune system and intricate social immune responses of S.invictapose challenges to the development of effective control strategies. MicroRNAs (miRNAs) play critical roles in the post-transcriptional regulation of gene expression, which influences various biological processes, including immunity and host-pathogen interactions. While the miRNA-mediated response of insects to pathogens has been extensively studied in solitary insects, little is known about the innate immune responses of individual members within a colony. To address this gap, we constructed small RNA libraries from Metarhiziumanisopliae-infected S.invictaworkers and investigated the temporal dynamics of miRNA-mediated immune responses to the entomopathogen. Several differentially expressed miRNAs were identified, and they were found to regulate genes involved in the Toll, IMD, and melanization immune pathways. Quantitative real-time PCR (qRT-PCR) was employed to analyze the spatiotemporal dynamics of key miRNAs/target genes, specifically miR-71/ModSP1-Relishand miR-7/Lysozyme2-Serineprotease7. A dual luciferase assay (invitro) was performed to validate the interactions between miRNAs and their target genes. Overexpression of miR-71 and miR-7 (viamiRNA mimics) efficiently suppressed their target genes, impaired the antifungal immune response of S.invictaand increased the susceptibility to M.anisopliaeinfection compared to controls. Furthermore, RNA interference-based gene silencing elucidated the roles of these immune genes in regulating fungal susceptibility, thus providing vital clues for developing virulent and effective mycoinsecticides using modern genetic engineering tools.

The development of skeletal muscle are complicated processes involving genes responsible for proper muscle morphology, contractility, cell proliferation, differentiation, interactions, migration, and death. The three-dimensional chromatin architecture of skeletal muscle development has not been studied intensively although dynamic transcriptional regulation during differentiation of muscle cells is one of the most deeply studied processes. The RNA-seq was used to analyze the transcriptome pattern during chicken muscle development across 12 stages. Hi-C was used to build chromatin architectures during four representative stages. ChIP-seq was conducted to identify enhancers and promoters in these four stages, which are occupied by histone H3K27ac and H3K4me3 peaks. Results show that large-scale genome architecture changes are mostly unidirectional, and coupled by complex on/off dynamic patterns of gene expression. Specifically, we observed 258.30Mb of the genome undergoing A/B compartment switching. Notable alterations (316.57Mb) of interaction frequencies within TADs were observed. Substantial aging-associated genes exhibited ascending connectivity with the compartment transition from repressive to active status during muscle development. Some muscle-related gene promoters that interacted with active enhancers during development, and some myopathy/aging-associated genes that were activated in aging muscle were founded. These results provide key insights into skeletal muscle development in vivo, and offer a valuable resource that allows in-depth functional characterization of candidate genes.

Primordial germ cells (PGCs) are the stem-cell population of adult animal gametes, which develop into sperm or eggs. It can be propagatedin vitroand injected into the host chicken for genome editing to obtain germline chimeric chicken. However, it has the limitation that the host embryo contains endogenous PGCs, which raises complications, resultantly donor PGCs fail to compete, and transmission efficiency reduced. Therefore, to increase the transmission efficiency, we generated a novel sterile chicken with the inducible elimination of endogenous PGCs in the host. This is the first study that applied the herpes simplex virus thymidine kinase (HSV-TK) cell ablation system in avian. CRISPR/Cas9-mediated homology-directed repair was performed to localize the HSV-TKsuicide gene to the last exon of the deleted in azoospermia-like (DAZL) gene, and ganciclovir (GCV) was added to induce the apoptosis in the germ cells of the host embryo. The sterilized host embryo introduced genome-edited PGCs to produce chimeric chicken carrying exogenous germ cells only. It was observed that the germline transmission efficiency was 100% achieved, and the obtained chicks were purely from donor breeds. The technologies established in the current study have important applications in germplasm conservation and gene editing in chicken.

NADC34-like porcine reproductive and respiratory syndrome virus (PRRSV), which first appeared in China in 2017, is currently one of the main epidemic strains in China. In this study, we found that a new variant of NADC34-like PRRSV evolved, named the L1A variant. The phylogenetics, epidemic status, and pathogenicity of the LA variants were subsequently comprehensively evaluated. Based on the results of the ORF5 phylogenetic analysis, the L1A variants were classified as NADC34-like PPRSV. All the strains had the same discontinuous 131-aa deletion in the NSP2 region (similar to that in the NADC30). Recombination analysis revealed that the L1A variants were recombinant viruses that contained an NADC30-like PRRSV skeleton, a nonstructural protein-encoding gene region obtained in part from JXA1-like PRRSV and a ORF2-ORF6 gene region partly obtained from NADC34-like PRRSV and that exhibited similar recombination patterns. We successfully isolated the L1A variant TZJ2756 from PAMs and Marc-145 cells. In animal experiments, TZJ2756 exhibited moderate pathogenicity in piglets, causing obvious clinical symptoms, namely, persistent fever, significantly reduced body weight, interstitial edema and severe interstitial pneumonia in the lungs, and prolonged high-load viremia. L1A variants have been detected in at least 12 provinces in China and share many similar epidemiological characteristics with the American L1C variant. This research will enhance our understanding of the prevalence of L1A variants and furnish valuable data for the ongoing monitoring of NADC34-like PRRSV in China.

A microbial strain designated BacilluslicheniformisQX928 was screened from hot springs in Sichuan Province, China, and a compound generated in the culture of this strain clearly inhibited PseudomonasaeruginosaATCC27853. The measured minimum inhibitory and lowest bactericidal concentrations were (13±0.17) and (22±0.72) mg L^–1, respectively. The compound was identified as 3-isopropylhexahydro-4H-pyrido[1,2-α]pyrazine-1,4(6H)-dione (IPHPPD). A SciFinder search revealed that IPHPPD could be the first compound synthesized by microorganisms that had both antibacterial and anti-quorum sensing properties. At low concentrations, IPHPPD interfered with the signaling factors and population effects of P.aeruginosa, thereby altering the biofilm morphology and structure. IPHPPD more strongly inhibited P.aeruginosaat high concentrations, primarily by reducing its virulence factors, cell membrane permeability and energy metabolism. A transcriptome analysis highlighted the role of IPHPPD in the transcriptional regulation of cellular metabolism and quorum sensing. Thus, the results of this study provide critical evidence that IPHPPD is a potential target for drug development to prevent and treat diseases in animals.

Long-term manure application has the potential to alleviate soil acidification, and increase carbon sequestration and nutrient availability, thus improving cropland fertility. However, the mechanisms behind greenhouse gas N₂O emissions from acidic soil mediated by long-term manure application remain poorly understood. Herein, we investigated N₂O emission and its linkage with gross N mineralization and nitrification rates, as well as nitrifying and denitrifying microbes in an acidic upland soil subjected to 36-year fertilization treatments, including an unfertilized control (CK), inorganic fertilizer (F), 2× rate of inorganic fertilizer (2F), manure (M), and the combination of inorganic fertilizer and manure (FM) treatments. Compared to the CK treatment (1.34 μg N kg^–1d^–1), fertilization strongly increased N₂O emissions by 34-fold on average, with more pronounced increases in the manure-amendment (10.6–169 μg N kg^–1d^–1) than those in the inorganic fertilizer treatments (3.26–5.51 μg N kg^–1d^–1). The manure amendment-stimulated N₂O emissions were highly associated with increased soil pH, mean weight diameter of soil aggregates, substrate availability (e.g., particulate organic carbon, NO₃⁻and available phosphorus), gross N mineralization rates, denitrifier abundances and the (nirK+nirS)/nosZratio. These findings suggest that the increased N₂O emissions primarily resulted from alleviated acidification, increased substrate availability and improved soil structure, thus enhancing microbial N mineralization and favoring N₂O-producing denitrifiers over N₂O consumers. Moreover, ammonia-oxidizing bacteria (AOB) rather than ammonia-oxidizing archaea (AOA) positively correlated with soil NO₃⁻concentration and N₂O emissions, indicating that nitrification indirectly contributed to N₂O production by supplying NO₃⁻for denitrification. Collectively, manure amendment potentially stimulates N₂O emissions, primarily resulting from alleviated soil acidification and increased substrate availability, thus enhancing N mineralization and denitrifier-mediated N₂O production. Our findings suggest that consideration should be given to the greenhouse gas budgets of agricultural ecosystems when applying manure for managing the pH and fertility of acidic soils.

As the global leader in rice production, China’s paddy fields contribute substantially to greenhouse gas emissions through methane (CH₄) and nitrous oxide (N₂O) releases. Aromatic rice cultivation practices have been optimized to enhance the aroma, so the relationship between its cultivation and greenhouse gas emissions from paddy fields is unclear. To investigate how aroma-enhancing cultivation practices drive microbial community dynamics in aromatic rice paddies and their implications for greenhouse gas emissions, a two-year experiment in five ecological locations (Xingning, Nanxiong, Conghua, Luoding, and Zengcheng) compared two farming practices: partial organic substitution for inorganic fertilizers combined with water-saving irrigation (IOF+W) and traditional cultivation (CK). The CH₄and N₂O emissions, soil microbial composition and function, global warming potential (GWP), nitrogen use efficiency, yield, and the content of 2-acetyl-1-pyrroline (2-AP) were measured and analyzed. The main purpose was to investigate the impact of IOF+W on CH₄and N₂O emissions and their relationship with soil microorganisms. The results showed that IOF+W significantly reduced CH₄emission fluxes and totals (36.95%) and GWP (31.29%), while significantly increasing N₂O emission fluxes and totals (14.82%). The soil microbial community structure was reshaped by the IOF+W treatment, which suppressed methanogens but enhanced the abundances of nitrifying and denitrifying bacteria. Key enzymatic activities involved in CH₄production, such as methyl-coenzyme M reductase, formylmethanofuran dehydrogenase, and methyltransferase, decreased. In contrast, the activity of the key CH₄-oxidizing enzyme methanol dehydrogenase increased.This shift led to an overall attenuation of the CH₄production metabolism while enhancing the CH₄oxidation metabolism. In addition, the activities of pivotal enzymes involved in denitrification and nitrification were improved, thus enhancing nitrogen nitrification and denitrification metabolism. Moreover, the IOF+W treatment significantly increased nitrogen use efficiency (47.83%), yield (14.77%), and 2-AP content (13.78%). Therefore, the IOF+W treatment demonstrated good efficacy as a sustainable strategy for achieving productive, green, resource-efficient, and premium-quality aromatic rice cultivation in South China.

The continuous supply of phosphorus (P) is indispensable in crop production. However, P resources are non-renewable, and environmental concerns like eutrophication associated with its loss from agroecosystems make the sustainable management of P resources essential for ensuring global food security. This study was designed to reduce mineral P inputs through management practices. A field experiment comprising a wheat–maize rotation system was conducted in the Guanzhong Plain of Shaanxi Province, China from 2018–2023. The eight treatments included CK (without P), FP (conventional P application); RP (recommended P); RP₈₀(20% reduction in RP); SRP₈₀(20% reduction in RP with straw wrapping); ARP₈₀(20% reduction in RP with ammonium sulfate instead of urea); SARP₈₀(20% reduction in RP with straw wrapping and ammonium sulfate instead of urea); and SARP₆₀(40% reduction in RP with straw wrapping and ammonium sulfate instead of urea). Crop yield, P uptake, and P fertilizer use efficiency were measured during harvest and throughout the entire period of the study. At the end of the experiment, P fractions were estimated using the Tiessen-Moir P classification method. The results revealed that the grain yields of all the treatments except for RP₈₀were significantly increased compared to CK, with increases of 14.9–28.8%. Furthermore, agronomic efficiency, apparent P use efficiency, P recovery rate, and partial factor productivity were significantly improved for the treatments that received 20% less P with straw wrapping. Moreover, the enhancement measures significantly increased labile and moderately labile P in the soil. Therefore, straw wrapping with ammonium sulfate instead of urea is one of the most effective ways to reduce mineral P inputs while increasing the efficiency of P in wheat–maize rotation systems.

Nitrogen use efficiency in rice is lower than in upland crops, likely due to differences in soil nitrogen dynamics and crop nitrogen preferences. However, the specific nitrogen dynamics in paddy and upland systems and their impact on crop nitrogen uptake remain poorly understood. The N dynamics and impact on crop N uptake determine the downstream environmental pollution from nitrogen fertilizer. To address this poor understanding, we analyzed 2,044 observations of gross nitrogen transformation rates in soils from 136 studies to examine nitrogen dynamics in both systems and their effects on nitrogen uptake in rice and upland crops. Our findings revealed that nitrogen mineralization and autotrophic nitrification rates are lower in paddies than in upland soil, while dissimilatory nitrate reduction to ammonium is higher in paddies, these differences being driven by flooding and lower total nitrogen content in paddies. Rice exhibited higher ammonium uptake, while upland crops had over twice the nitrate uptake. Autotrophic nitrification stimulated by pH reduced rice nitrogen uptake, while heterotrophic nitrification enhanced nitrogen uptake of upland crops. Autotrophic nitrification played a key role in regulating the ammonium-to-nitrate ratio in soils, which further affected the balance of plant nitrogen uptake. These results highlight the need to align soil nitrogen dynamics with crop nitrogen preferences to maximize plant maximize productivity and reduce reactive nitrogen pollution.

Green manuring is essential for improving soil quality and nutrient uptake. With the gradual depletion of phosphorus (P) resources, more attention is being paid to the role of green manures in cultivation systems, such as maize–green manure intercropping, to find possible pathways for enhancing soil P utilization. A maize–green manure intercropping experiment was started in 2009 to investigate the effects and mechanisms for enhancing P uptake and yield in maize. Three species of green manures (hairy vetch (HV), needleleafpea (NP), sweet pea (SP)) and a sole maize treatment (CK) were used, resulting in four treatments (CK, HVT, NPT, and SPT) in the experiment. During 2020–2023, the intercropping treatments enhanced maize yields in 2020 and 2021, particularly in HVT with increases of 13.7% (1.96t ha^–1) and 13.0% (2.13t ha^–1) compared with CK, respectively. Grain P accumulation of maize was significantly higher in the intercropping treatments than CK in 2020, 2021, and 2023, and with an average increase of 10.6% over the four years (5.2% for NPT, 10.8% for SPT and 15.9% for HVT) compared with CK. Intercropping promoted maize growth with a greater root length density and a higher organic acid release rate. HVT changed the soil properties more dramatically than the other treatments, with increases in the acid phosphatase and alkaline phosphatase activities of 29.8 and 38.5%, respectively, in the topsoil (0–15cm), while the soil pH was reduced by 0.37 units compared to CK (pH=8.44). Intercropping treatments facilitated the conversion of non-labile P to mod-labile P and stimulated the growth of soil bacteria in the topsoil. Compared with CK, the relative abundance of Gemmatimonadota,known for accumulating polyphosphate, and Actinobacteriota, a prominent source of bioactive compounds, increased significantly in the intercropping treatments, especially in HVT and SPT. A PLS-PM analysis showed that intercropping promoted soil P mobilization and the enrichment of beneficial bacteria by regulating maize root morphology and physiology. Our results highlight that maize–green manure intercropping optimizes root traits, soil properties and bacterial composition, which contribute to greater maize P uptake and yield, providing an effective strategy for sustainable crop production.