Reproductive containment in farmed fish holds significant application potential in improving economic traits, ensuring the industrialization of biological breeding, and protecting intellectual property (IP). Currently, interspecific hybridization and triploid sterility methods have achieved success in the production of sterile broodstock for a limited number of farmed fish species. However, due to technical limitations, these approaches have not yet been widely adopted on a large scale. With the identification of key genes regulating reproduction, the elucidation of gene functions, and advances in genetic engineering techniques, sterile fish can be generated by knocking down or knocking out genes associated with reproduction. However, sterile fish produced directly through gene loss cannot pass on desirable economic traits to the next generation, making it difficult to establish stable families. Therefore, further research is needed on methods to rescue the parental sterility phenotype, such as through exogenous hormone supplementation. Conditional induced sterility and reproductive on-off strategies based on two-line hybridization represent novel approaches to reproductive containment. These techniques enable reproductive containment in fish while ensuring the stable inheritance of parental desirable economic traits. The technical approaches have been preliminarily validated in model fish species, demonstrating promising development prospects. This review summarizes advances in fish reproductive containment technologies, aiming to provide references for breeding economically valuable, ecologically friendly, and IP-protected sterile farmed fish, such as grass carp (Ctenopharyngodon idellus).

The high temperature and small temperature difference between day and night in horticultural facilities are easy to occur in summer, which causes tomato (Solanum lycopersicum) seedlings to overgrow, especially the epicotyl, resulting in lower yields and economic losses. As an inhibitor of gibberellin synthesis, uniconazole can delay cell elongation and inhibit the seedling to overgrow. In order to clarify the effect of uniconazole on epicotyl dwarfing in tomato, 0, 15, 45 and 75 mg/L uniconazole solutions were sprayed on long internode 'DH' tomato seedlings, and the morphological indexes of tomato seedlings were measured at 7 d intervals after spraying, such as epicotyl length, epicotyl thickness, plant height, leaf width and leaf length, and measured the quality indicators of tomato fruits after planting. The results showed that 45 mg/L of uniconazole significantly reduced epicotyl length and plant height in tomato seedlings, and significantly increased soluble protein and soluble solids contents in tomato fruits. The expression of genes in the gibberellic acid (GA) and indole-3-acetic acid (IAA) regulatory pathway in the tomato epicotyl at 7 d after uniconazole treatment were analyzed. The results showed that the majority of genes in the GA regulatory pathway genes showed up-regulated expression patterns. Auxin response factor 13 (SlARF13) gene in the IAA regulatory pathway also showed up-regulated expression patterns, whereas SlIAA33 gene showed down-regulated expression patterns. Therefore, GA2ox3 and GA2-β-dioxygenase8 gene in the GA pathway and the SlARF13 and SlIAA33 genes could be used as candidate genes for tomato epicotyl dwarfing. This study provides a theoretical basis for the subsequent application of uniconazole in tomato seedling.

NAC transcription factors play an important role in regulating the response to both biotic and abiotic stresses during the plant growth and development. They are actively involved in the regulation of secondary cell wall biosynthesis and deposition. In this study, the ZlNAC105 gene (GenBank No. PP437561) was cloned from Zizania latifolia. The full-length cDNA sequence was 972 bp, encoding a 323 amino acid protein containing a conserved NAM domain. Phylogenetic analysis revealed that ZlNAC105 shared the highest sequence similarity with its ortholog in Oryza brachyantha. The qRT-PCR analysis revealed that ZlNAC105 exhibited the highest expression level in the stems of 'Wujiang wild jiaobai', and could be induced by Ustilago esculenta infection, showing significant upregulation (P<0.05) at 10 d post-inoculation, which was consistent with the previously reported results of ZlNAC29. Furthermore, expression analysis of ZlNAC105 during different developmental stages of stem swelling in Z. latifolia demonstrated that the expression level in the stems of 'Longjiao No.2' at the 8-leaf stage was significantly higher than that in 'Wujiang wild jiaobai' (P<0.05). During the swelling of stem development, ZlNAC105 expression was significantly downregulated in the upper, middle, and lower internodes of aged stems of Z. latifolia with higher lignification degrees, as well as in the floral tubes (P<0.05). In the middle-lower internodes of swollen stems, ZlNAC105 expression was significantly higher in 10 cm swelling stems compared with late swelling stages and aged Z. latifolia (P<0.05). Combined with scanning electron microscopy observations, ZlNAC105 exhibited lower expression levels in highly lignified swollen fleshy stems, demonstrating a generally negative correlation with the degree of stem lignification. Notably, this expression pattern showed significant dif-ferences compared with the previously reported expression profile of ZlNAC29. These results demonstrated that ZlNAC105 responded to the infection of U. esculenta and participated in regulating the stem lignification of Z. latifolia, potentially playing a crucial role in secondary cell wall biosynthesis and thickening. This study provides theoretical foundations for understanding the mechanism by which U. esculenta infection regulates stem swelling development in Z. latifolia.

Phytosulfokine (PSK), a class of essential plant peptide hormones, plays critical roles in plant disease resistance and defense responses. To systematically dissect the functions of the PSK gene family in watermelon (Citrullus lanatus) defense responses, a genome-wide identification and bioinformatics analysis of ClPSKs were performed, followed by expression profile analysis during fruit development and upon Fusarium oxysporum f. sp. niveum infection. The results showed that 4 ClPSKs (Cla97C01G016930, Cla97C08G160340, Cla97C10G193065 and Cla97C10G202430) were identified in the watermelon genome, distributed across 3 watermelon chromosomes. The molecular weight of ClPSKs-encoded proteins ranged from 9.276 to 11.491 kD. All ClPSKs were acidic and unstable proteins, and they all contained PSK family conserved motifs. The phylogenetic tree analysis showed that watermelon ClPSKs clustered with AtPSKs from Arabidopsis thaliana, indicating high evolutionary conservation. Promoter cis-acting element analysis revealed that the stress responsive cis-elements and the plant hormone responsive cis-elements were enriched in the promoter sequence of watermelon PSKs gene, which providing a molecular basis for ClPSKs involvement in stress responses. Expression analysis showed that ClPSKs exhibited tissue-differential expression, with the highest levels in leaf tissues, followed by roots. Cla97C01G016930, Cla97C08G160340 and Cla97C10G193065 exhibited the highest expression levels at 10 d after pollination of watermelon fruit, indicating that these genes played a role in the early stage of watermelon fruit growth and development. Fusarium oxysporum infection significantly induced the expression of ClPSKs genes. Cla97C01G016930, Cla97C08G160340, and Cla97C10G193065 had the highest expression levels at 12 h after F. oxysporum f. sp. niveum infection, while Cla97C10G202430 showed continuous upregulation, indicating distinct spatiotemporal roles of ClPSKs in disease defense. This study provides scientific references for further understanding the role of ClPSKs in watermelon disease resistance and defense responses.

Kiwifruit is rich in ascorbic acid (AsA), but the AsA content significantly decreases during postharvest ripening. AsA metabolism is regulated by transcription factors. The basic helix-loop-helix (bHLH) gene family has been proven to be involved in various metabolic pathways. To explore the role of the bHLH gene family in AsA metabolism in kiwifruit, this study conducted bioinformatics identification of the kiwifruit (Actinidia chinensis) bHLH gene family. The physicochemical properties, chromosomal localization, conserved motifs, evolutionary relationships, synonymous codon usage frequency, and protein interaction networks of these members were analyzed to determine gene nomenclature and classify the gene family, predicting their interaction relationships. Finally, by analyzing the expression patterns of bHLH transcription factors (TFs) and AsA metabolic genes during postharvest storage, combined with the prediction of cis-acting elements in the promoters of functional genes, the potential links between bHLH TFs and AsA metabolism were inferred. The results showed that the 144 members were distributed across 29 chromosomes, with most members possessing typical bHLH conserved motifs. The cis-acting elements included 7 major categories: Light response, stress response, MYB binding, hormone response, defense and stress response, endosperm and meristem expression and circadian rhythm. According to the phylogenetic tree analysis, 144 bHLH members were divided into 25 subfamilies, of which 7 members having a relative synonymous codon usage frequency greater than 2.0, indicating a strong codon usage preference in the AcbHLH gene family. Protein interaction network analysis revealed interactions among 60 members. qRT-PCR analysis indicated that AcbHLH072, AcbHLH120, AcbHLH134, and AcbHLH067 were significantly correlated with AsA content. Additionally, 8 AsA metabolism-related genes all contained cis-acting elements for bHLH binding, suggesting that AcbHLH072, AcbHLH120, AcbHLH134, and AcbHLH067 might regulate AsA synthesis and degradation by influencing the expression of related genes such as ascorbate peroxidase 2 (AcAPX2), AcAPX8, L-galactose dehydrogenase 1 (AcGalDH1), galacturonic acid uridylyltransferase 2 (AcGalUR2), and AcGalUR3, thereby modulating the AsA metabolic pathway in the fruit. This study provides insights into the mechanisms by which the kiwifruit bHLH gene family participates in ascorbic acid metabolism.

With the increasing research on the molecular biology of Camellia spp., it is crucial to screen the reference genes suitable for qPCR analysis of Camellia spp.. 3 Camellia varieties with different ornamental values were selected as experimental materials for this study. 7 common reference genes, including elongation factor-α (EF-1α), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), Actin 7 (ACT), α- tubulin (TUA), β-tubulin (TUB), protein phosphatase 2C (PP2C), and phosphatase activator TIP41 (TIP41), were detected using reverse transcription polymerase chain reaction (RT-PCR) and qPCR in various Camellia spp. flower tissues and flower developmental periods. The stability of the candidate genes was analyzed using ΔCt, geNorm, NormFinder and BestKeeper programs, and RefFinder was used for comprehensive evaluation to screen out the candidate reference genes suitable for qPCR in Camellia spp., and to verify the stability of the reference genes by the key genes of flower color and floral aroma biosynthesis. The results showed that the average expression of EF-1α was found to be higher than that of other candidate reference genes. The analysis results from ΔCt, NormFinder and BestKeeper all showed that EF-1α was the most stable in different tissues and stages of Camellia spp. flower development, while the results from geNorm showed that GAPDH and TIP41 were more stable. Additionally, the results from all programs showed that ACT was the least stable reference gene. Based on the comprehensive evaluation of RefFinder, EF-1α and ACT were validated by the expression pattern analysis of the floral color genes, chalcone synthase (CrCHS) and chalcone isomerase (CrCHI), and the floral scent genes, 1-deoxy-D-xylulose 5-phosphate synthase (ChDXS1) and 1-deoxy-D-xylulose 5-phosphate reductoisomerase (ChDXR). The findings demonstrated that EF-1α with superior stability could be chosen as an internal reference gene in the study of Camellia spp. floral color and floral scent. The results of the study provide a theoretical basis and technical support for further research on the functional genes related to the formation process of Camellia spp. floral color and floral scent.

InDel markers have been widely used in research fields such as genetic diversity analysis of crop germplasm resources and molecular marker-assisted breeding, but they are scarcely reported in flax (Linum usitatissimum) related studies. This study used resequencing data of 2 flax materials to identify InDel loci, develop polymorphic InDel markers and genotyped 93 flax cultivars to investigate the genetic diversity, phylogenetic relationships, and population genetic structure characteristics of flax cultivars. The results showed that, identified 66 851 InDel loci and designed 120 InDel markers based on the whole genome re-sequencing data, and 26 pairs of primers with good polymorphism were screened to analyze genetic diversity and population structure of 93 flax varieties. A total of 80 alleles were detected with an average of 3.08 alleles per locus, the gene diversity index and polymorphism information content ranged from 0.169 6 to 0.694 2 and 0.161 5 to 0.636 9, respectively, with an average of 0.485 2 and 0.415 0 respectively. All varieties were divided into 2 groups with cluster analysis, the fiber flax were clustered into one groups, the oil flax were divided into one groups; population structure analysis divided all varieties into 4 groups; principal component analysis separated fiber flax from oil flax; The results of analysis of molecular variance (AMOVA) showed that the variation mainly occurred within individuals and among individuals, there was also a certain amount of variation among populations. The results of cluster analysis and principal component analysis were consistent, indicated that genetic diversity of fiber flax populations was higher than that of oil flax populations, the genetic variation of oil flax was independent of geographical region. The results can reveal the relationship of 93 cultivated flax varieties, which has certain theoretical significance and application value of identification flax varieties, analysis flax resources' relationship and molecularly assisted breeding.

Walnut (Juglans regia) is highly valued by consumers for its distinctive aroma, yet the molecular mechanisms underlying its aroma formation remain unclear. In this study, fresh 'Qingxiang' walnut kernels were analyzed using headspace solid-phase microextraction (HS-SPME)-gas chromatography-mass spectrometry (GC-MS) and transcriptome sequencing (RNA-seq) to systematically investigate the dynamic changes in aroma-related genes and volatile compounds during kernel development from 85 to 141 d after flowering. GC-MS analysis identified 70 volatile organic compounds (VOCs), with aldehydes (e.g., nonanal), alcohols (e.g., 1-octen-3-ol), and ketones (e.g., 3-octanone) as key aroma contributors. Based on odor activity value (OAV>1), 11 core aroma-active compounds were screened, including (E)-2-butenal and hexanal. Transcriptome analysis revealed that differentially expressed genes (DEGs) during fruit maturation were significantly enriched in starch and sucrose metabolism (94), pyruvate metabolism (66), fatty acid biosynthesis (39), terpenoid backbone biosynthesis (35), α-linolenic acid metabolism (36), and fatty acid degradation (29). Further integration of omics data and aroma dynamics enabled the construction of a regulatory network for aldehyde and ketone biosynthesis, It was found that hydroperoxide lyase (HPL) coding gene JrHPL, alcohol dehydrogenase (ADH) coding genes JrADH_7 and JrADH_8, and aldehyde dehydrogenase (ALDH) coding gene JrALDH_7 play key roles in aroma metabolism. This study reveals the multi-omics regulatory mechanism of walnut aroma formation, offering a theoretical reference for molecular breeding strategies to enhance nut flavor.

Mitochondrial antiviral signaling protein (MAVS) is a key molecule involved in innate antiviral response through the RIG-1 like receptor pathway. As an essential adaptor in signal transduction, MAVS plays crucial role in innate immunity against viruses in mammals and fish. To investigate the role of MAVS gene in the innate immunity of largemouth bass (Micropterus salmoides), the cDNA sequence of MAVS gene (GenBank No. PP740757) was obtained by reverse transcription PCR (RT-PCR) and cloning, and bioinformatics was also analyzed. qRT-PCR was used to detect the expression pattern of MAVS in healthy individuals and in individuals intraperitoneal injection with Nocardia seriolae and polyinosinic polycytidylic acid (Poly I:C). Subcellular localization of the MAVS protein was determined using immunofluorescence assay, and the effects of the MAVS gene on the promoter activities of nuclear factor kappa-B (NF-κB) and β-type interferon (IFN-β) were analyzed by dual-luciferase reporter assay. The results showed that the MAVS cDNA was 1 921 bp in length and encoded a peptide of 586 amino acid residues. The deduced protein of MAVS in largemouth bass had a N-terminal CARD domain and a proline rich region, which were known to be important functional domains of mammalian MAVS. The MAVS protein of largemouth bass had high homology with other fish species (48.7%~96.1%, and 29.6% homology with grass carp (Ctenopharyngodon idella)). Phylogenetic analysis showed that largemouth bass MAVS was grouped with other fish MAVS. The MAVS mRNA was expressed in all tested tissues of healthy largemouth bass, with the highest expression level in the gills. In all 4 tested tissues, both immunostimulants could up-regulate the transcription level of MAVS in largemouth bass. Among them, on the 9 d after Poly I:C stimulation, MAVS mRNA expression level in the gills was as high as 53.17 folds that of the control group (P<0.05). In kidney, MAVS showed the highest increase at 6 d after N. seriolae infection, and the expression level in the infected group was 14.88 folds that of the control group (P<0.05). Subcellular localization showed that MAVS was presented in the cytoplasm of HeLa cells. Overexpression of MAVS in HeLa cells could increase the NF-κB and IFN-β promoter activity. These results indicated that MAVS played important roles in the innate immunity of largemouth bass. This study lays the foundation for further elucidating the role of MAVS gene in the innate immunity of largemouth bass.

The largemouth bronze gudgeon (Coreius guichenoti), a rare endemic fish species in the upper Yangtze River, faces critical survival challenges due to ichthyophthiriasis. In order to investigate the therapeutic efficacy of plant-derived gallic acid (GA) against Ichthyophthirius multifiliis infection and elucidate its molecular mechanisms, this study systematically evaluated the therapeutic efficacy, safety profile, and underlying mechanisms of action of GA through in vitro and in vivo antiparasitic assays, acute toxicity tests, and hepatic non-targeted metabolomics analysis. In vitro assays demonstrated that GA exhibited concentration-dependent antiparasitic effects on I. multifiliis. After 4 h of treatment with 50 mg/L GA, the mortality of trophonts and theronts reached 100.0%, while 20 mg/L GA resulted in 63.3% and 80.3% mortality, respectively. Microscopic observations revealed pathological alterations in parasites, including cell membrane disruption, cytoplasmic leakage, and mitotic arrest in the parasites following GA treatment. In vivo trials showed that 50 and 20 mg/L GA reduced fish mortality from 100% (control) to 2.3% and 20.0% within 48 h, respectively. Concurrently, parasite mortality reached 54.3% and 21.3%. Moreover, GA treatment alleviated clinical symptoms, including white spots, excessive mucus secretion, and respiratory distress. Acute toxicity tests determined that the safe concentration (SC) of GA for C. guichenoti was 64.6 mg/L, with 96 h half-lethal concentration (LC₅₀) values of 245.8 mg/L. Ultra performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) analysis revealed that 50 mg/L GA significantly reshaped the hepatic metabolic network in I. multifiliis-infected fish: (1) downregulating arachidonic acid metabolism, VEGF signaling, and Th17 cell differentiation pathways to suppress pro-inflammatory mediators; (2) upregulating endogenous antioxidants (e.g., glutathione and taurine) while activating FoxO signaling and lysosomal pathways to enhance reactive oxygen species scavenging; (3) optimizing mitochondrial energy metabolism and membrane homeostasis by enriching pathways such as oxidative phosphorylation, glycerophospholipid metabolism, and the malate-aspartate shuttle. These findings demonstrated that 50 mg/L GA effectively controled ichthyophthiriasis via a synergistic mechanism involving anti-inflammatory regulation, antioxidant defense activation, and metabolic reprogramming, thereby restoring hepatic metabolic homeostasis. This study provides novel strategies and a candidate agent for the environmentally friendly control of ichthyophthiriasis in fish.

Wheat common bunt is one of the major wheat diseases which caused by Tilletia foetida, not only reducing wheat yield, but also deteriorating the quality of the wheat. To investigate the interaction mechanism between the common bunt fungus T. foetida and Triticum aestivum, a candidate effector protein g5965 (GenBank No. XXN76190.1) belonging to the protein kinase superfamily was cloned via RT-PCR.The results of bioinformatics analysis revealed that the g5965 gene had a full length of 1 722 bp, encoded 573 amino acids, and contained a typical phosphotransferase domain. Its signal peptide secretion function had been verified through signal peptide secretion assays. Constructed the PGR107-g5965 fusion expression vector and transiently expressed it in the leaves of Nicotiana benthamiana via Agrobacterium-mediated transformation. The g5965 protein did not suppress the programmed cell death (PCD) response in tobacco cells induced by the pro-apoptotic protein BAX (Bcl-2 associated X protein). The green fluorescent protein (GFP) expression vector pBIN-g5965 was constructed and used to infect N. benthamiana leaves. The green fluorescence was observed at the cell membrane of tobacco cells under a laser scanning confocal microscope after 2~3 d.This study provides a certain theoretical basis for further elucidating the pathogenic mechanism of T. foetida against the wheat.

Gossypol (GOS) is a polyphenolic antinutritional factor naturally present in cottonseed, which severely limits the widespread application of cottonseed by-products in the food and feed industries. Our research group screened Bacillus subtilis M-15 for its ability to degrade GOS efficiently and found that catechol 2,3-dioxygenase (C23O) may be a key enzyme in the GOS degradation process. This study aimed to verify the degradation effect of BsC23O on GOS in Bacillus subtilis M-15. The changes in the expression level of BsC23O under GOS stress were verified by qPCR. The physicochemical properties of BsC23O were predicted using bioinformatics methods. Subsequently, its expression was induced in an Escherichia coli expression system, and the effect of the expressed product on the degradation of gossypol (GOS) was tested. Additionally, the potential interaction mechanism between BsC23O and GOS was explored using structural confirmation and molecular docking. The results showed that the expression of BsC23O was up-regulated significantly under GOS stress (P<0.05). Bioinformatics analysis revealed that the protein encoded by BsC23O consisted of 285 amino acids, with a relative molecular mass of 31.56465 kD and an isoelectric point of 5.48. This protein lacked a signal peptide, had a relatively stable structure, was a hydrophilic protein, and was mainly located in the cytoplasm. Its structure contained 2 main domains: the VOC_BsCatE_like_N domain located at the N-terminus and the VOC_BsCatE_like_C domain located at the C-terminus. Additionally, the protein contained 23 potential phosphorylation sites and was primarily composed of α-helices and random coils. In the prokaryotic expression system, the activity of BsC23O reached 97.79 U/L. The expression product degraded 29.42% of GOS within 1 h. Molecular docking showed that the multiple benzene rings of GOS formed hydrophobic and conjugated interactions with residues around the pocket, promoting stable binding. This study successfully expressed highly active BsC23O and validated its pivotal role in GOS degradation. These findings provide critical theoretical support for the development of efficient GOS degradation technology and practicing enzyme engineering for safe utilization of cottonseed byproducts.

The rice-crayfish integrated farming system is an ecological agricultural model characterized by resource recycling and mutualistic symbiosis between crop cultivation and aquaculture. This study investigated the effects of straw return on soil bacterial community composition and diversity in a rice-crayfish integrated farming system. Three experimental treatments were implemented: winter flooding alone, winter flooding with straw return, and winter flooding combined with crayfish (Procambarus clarkii) culture and straw return. Bacterial community structure was analyzed using Illumina MiSeq high-throughput sequencing. The results demonstrated that: (1) Winter flooding combined with crayfish culture and straw return significantly increased soil total organic carbon and alkali-hydrolyzable nitrogen content but reduced soil pH; (2) Compared to winter flooding with straw return, winter flooding combined with crayfish culture and straw return decreased microbial species richness, elevated the relative abundances of phylum Myxococcota and candidate phylum MBNT15 as well as genus unclassified_MBNT15, but suppressed genera Intrasporangium and MND1; (3) Alpha and beta-diversity analyses revealed that integrating crayfish culture with straw return under winter flooding significantly reduced bacterial richness and diversity, while altering community structure; (4) Redundancy analysis identified soil pH and available phosphorus as the dominant drivers of bacterial community variation. This study demonstrates that straw return under the rice-crayfish integrated farming system alters soil physicochemical properties, reduces bacterial community richness and diversity, and modifies microbial community structure, thus providing a theoretical basis for nutrient cycling mechanisms in paddy soils of the rice-crayfish integrated farming system.

Carbon nanomaterials demonstrate significant potential in plant gene transfection due to their superior physicochemical properties and biocompatibility. They can effectively penetrate plant cell walls, markedly enhancing gene delivery efficiency while protecting exogenous genes from degradation and inactivation. Surface functionalization further optimizes their targeting and gene expression capabilities, rendering them crucial tools for plant genetic engineering. However, their biosafety and environmental impact require further investigation. This review provides an in-depth analysis of their application progress in plant gene transfection, covering carbon dots, carbon nanotubes, graphene, fullerenes, and carbon nanofibers, and explores their potential in improving gene delivery efficiency and precise expression, and discusses their biosafety and environmental impact. This review offers theoretical and experimental guidance for plant genetic engineering applications.

As the world's largest producer and consumer of pork, the high-quality development of the pig (Sus scrofa) industry in China is directly related to the country's food safety and economic security. In recent years, outbreaks of African swine fever, Porcine reproductive and respiratory syndrome and Porcine epidemic diarrhea have caused significant economic losses to the pig farming industry. Traditional prevention and control methods face challenges such as lagging vaccine development and drug residues. Traditional disease resistant breeding has a long cycle and slow progress, and there is an urgent need to develop safer and more effective breeding technologies. The emerging gene editing technology, with advantages such as high efficiency, precision, and convenience, has shown great potential in agricultural animal breeding, including pigs, in recent years, providing strong technical support for the cultivation of new disease resistant pig breeds. This article systematically reviews the development history of gene editing technology, with a focus on analyzing the research progress of CRISPR/Cas9 and its derivative technologies in pig disease resistant breeding, aiming to provide theoretical basis and technical guidance for gene editing disease resistant pig breeding research.

Mitochondria play a pivotal role in the energy metabolism of eukaryotic cells and are closely associated with lipid metabolism in adipose tissue. Lipid droplets maintain intimate connections with mitochondria and could form unique structural architectures; multiple proteins localized at the lipid droplet-mitochondria membrane contact sites are involved in regulating the biogenesis and expansion of lipid droplets. Mitophagy is categorized into 2 subtypes, namely ubiquitin-dependent mitophagy and ubiquitin-independent mitophagy, which could selectively eliminate damaged mitochondria to preserve cellular homeostasis. In adipocytes, mitophagy modulates mitochondrial quantity and adipocyte-specific properties, exerting profound impacts on lipid metabolism and systemic health. Although mitochondria and lipid droplets are of great significance in regulating lipid metabolism, preventing lipotoxicity, and maintaining lipid homeostasis, the current understanding of their interplay remains limited. This paper aims to broaden the insights into the mechanisms underlying the regulation of lipid metabolism by mitochondria-lipid droplet interactions, thereby providing theoretical foundations and research frameworks for elucidating the pathological mechanisms of metabolic diseases and developing targeted therapeutic strategies.

The widespread application of CRISPR/Cas9 gene editing technology in crop breeding has demonstrated significant potential for improving crop resistance, yield, and quality. However, this progress has also heightened the need for effective detection of gene-edited components. A significant number of crop samples containing non-excised exogenous CRISPR/Cas9 elements remain prevalent during early breeding stages and under specific conditions. However, existing detection methods are associated with limitations such as low sensitivity, time-consuming procedures, or dependence on sophisticated instrumentation. To meet the growing demand for rapid detection of CRISPR/Cas9-edited crops, a highly sensitive method integrating PCR with lateral flow strips (PCR-LFS) was developed in this study. Cas9-specific primers were designed and the amplification system was optimized, allowing rapid visual detection using colloidal gold-labeled test strips. A detection limit of 0.012% (by mass fraction) and sensitivity of 17.5 copies/μL were achieved, outperforming conventional PCR-capillary gel electrophoresis (0.1%) and quantitative PCR (1.75×10⁴ copies/μL). Specificity testing confirmed that Cas9-positive tomato (Solanum lycopersicum) samples were accurately distinguished from non-transgenic crops without cross-reactivity. Practical validation demonstrated that reliable detection was achieved in mixed samples at a ratio of 1∶2¹³, with high repeatability and stability. Compared with traditional techniques, this method eliminated the need for complex instruments, reduced detection time to 3 h, and was suitable for on-site screening and port inspections. It was suggested that future optimization of primer design and LFS procedures might extend the application to other gene-edited components. This study provides strong technical support for precise monitoring of genetically modified crops.

Kelch-like protein 21 (KLHL21), a member of the Kelch gene family, plays critical roles in various physiological processes in mammals, including mediating protein-protein interactions, protein degradation, and signal transduction. In order to study the effect of KLHL21 on the replication of Vesicular stomatitis virus (VSV), in this study, a KLHL21-overexpressing HEK-293T cell line (HEK-293T/KLHL21) was established using a lentiviral packaging system, and the replication efficiency of VSV in the cells was compared with that in wild-type HEK-293T cells. Firstly, in HEK-293T cells, the auxiliary plasmids psPAX2, pMD2.G and the newly constructed recombinant plasmid pLV-puro-3×Flag -KLHL21 were co-transfected to produce Lentivirus. Then, it was used to infect HEK-293T cells, and the positive cells HEK-293T/KLHL21 were obtained through puromycin selection. Western blot was used to assess VSV protein levels, while indirect immunofluorescence was applied to visualize VSV-GFP expression. Additionally, viral titers were determined to evaluate viral replication. The results showed that the proliferation ability of VSV in the HEK-293T/KLHL21 cell line was significantly weakened compared with HEK-293T cells. The result of qPCR revealed that, compared with wild-type cells, the established cell line activated the typeⅠ interferon response after VSV infection, which consequently led to a significant upregulation of the expression levels of its downstream genes. These findings indicated that KLHL21 activated the typeⅠinterferon signaling pathway and significantly inhibited the proliferation of VSV. This study provides theoretical basis and material support for further research on the antiviral mechanism of KLHL21.

Histone H2B is an important part of the nucleosome, which affects gene transcription and biological processes, and is expected to become a molecular target for disease treatment. In this study, lentiviral transfection was applied to construct an HEK-293T cell line with stable expression of histone H2B. The recombinant plasmid pLVX-mCherry-puro-H2B was constructed using the H2B gene as the target and then co-transfected with auxiliary plasmids pMD2.G and psPAX2 into HEK-293T cells for lentiviral packaging. The Lentivirus was transduced to HEK-293T cells, and a stable histone H2B-expressing cell line was obtained through purinomycin pressure screening, which was named HEK-293T-H2B cell line. The replication capacity of Herpes simplex virus (HSV) in the HEK-293T-H2B cell line was determined by qPCR and Western blot assays. The results showed that the replication of HSV was significantly weakened in the HEK-293T-H2B cell line. qPCR was used to detect the expression of downstream interferon genes in HEK-293T-H2B and wild-type HEK-293T cells infected with HSV. The results showed that the HEK-293T-H2B cell line promoted the expression of genes induced by HSV downstream typeⅠinterferon. In conclusion, the HEK-293T-H2B cell line constructed in this study can significantly prompte the typeⅠinterferon signaling pathway and inhibit the proliferation of HSV, which provides a basis for further research on the antiviral function of histone H2B.