Maize (Zea mays) is an important model plant to decipher the molecular mechanism of sex determination. The research on sex determination of tassel and ear is of great significance to improve the inflorescence architecture and increase yield. In this study, a maize sex determination mutant tasselseed10 (ts10) that could produce kernel on tassel was identified by treating B73 pollen with ethyl methane sulfonate (EMS). In fields, the vegetative growth of the ts10 was normal. When tassels emerged, the spikelets on middle and upper of main spikes and branches showed sex reversal, which could produce a large quantity of silks. While the apical spikelets could shed pollens normally. Phenotype observation under scanning electron microscope indicated when the male inflorescence meristem of ts10 was about 12 mm long, the florets produced feminized structure. Using map-based cloning strategy, the ts10 gene was narrowed down to a region of about 600 kb between the SSR markers PL4 and PR5 on Chr.1, which harbored 7 annotated genes. Sequencing analysis revealed that only the myelocytomatosis 7 (ZmMyc7) produced a mutation in which the 1 594^th base G was substituted by base a resulting in the 532^nd amino acid glutamic acid substituted by lysine. Further analysis suggested the mutation was located at the conserved domain HLH of ZmMyc7. Expression analysis showed that ts10 was expressed in root, stem, leaf, immature tassel, immature ear and kernel, among which the expression level in ear was the highest, followed by tassel; in addition, except for leaf, there was no significant difference in expression level of ZmMyc7 between ts10 and normal plants in root, stem, immature tassel, immature ear, embryo and endosperm. The subcellular localization results showed ZmMyc7 functions in nucleus. This study provides further theoretical support and genetic resources for enriching the molecular mechanism of sex determination in maize.

Class Ⅲ peroxidases (PRXs) are a family of genes unique to higher plants that are widely involved in catalyzing hydrogen peroxide, phenolic compounds and reactive oxygen species. To date, the study of the PRX gene family at the genome-wide level in Zea mays has not been reported. This study utilized bioinformatic methods to conduct a comprehensive analysis of the PRX gene family at the whole-genome level in maize, and examining their expression patterns in different tissues and under various stress conditions. The results indicated that a total of 90 ZmPRX genes were identified, unevenly distributed across the 10 chromosomes of maize. Specifically, 12 of these genes were found to be involved in 6 segmental duplication events, while 15 genes participated in 6 tandem duplication events. Phylogenetic analysis classified them into 8 branches, with genes in the same branch having similar gene structures and conserved motifs. Cis-element analysis revealed that there were 5 categories of elements distributed in the promoter region of ZmPRX genes, including hormonal responsiveness, stress responsiveness, growth and development, light responsiveness, and universal elements. The expression pattern analysis revealed that the expression of ZmPRX genes exhibited tissue-specific characteristics and were significantly induced under different abiotic stress conditions. Furthermore, under drought stress conditions, the expression level of the ZmPRX gene in the drought-tolerant inbred line 54358 significantly increases, suggesting that this variety possesses strong capabilities in resisting drought stress. This study provides a theoretical basis for further investigating the functional characteristics of PRX genes in maize.

Allitride is an important secondary metabolite in garlic (Allium sativium), which has an important effect on its flavor quality. Sulfur, as one of the essential elements for plant growth, is involved in the synthesis of allitride. In order to determine the effect of leaf surface sulfur application on allitride accumulation and expression of related genes of allitride synthesis, garlic varieties 'Zhengyuezao' with low allitride content and 'Siliuban' with high allitride content were used as materials, the leaves were sprayed with 6 g/L potassium sulfate solution, the content of allitride was determined and the expression of allitride synthesis-related genes was analyzed. The results showed that sulfur application promoted bulb enlargement, induced the expression of allitride synthesis-related genes, and increased the allitride content in bulbs. After sulfur application, the fresh weight of 2 varieties of garlic bulbs significantly increased, indicating that sulfur application promoted the expansion of bulbs. Allitride content in bulbs continuously increased during bulbous expansion stage after sulfur application. The accumulation of allitride in bulbs of both varieties was promoted by sulfur application, and the expression levels of most allitride synthesis-related genes in bulbs and leaves were up-regulated. Allitride content in 2 varieties of garlic bulbs was positively correlated with the expression levels of allitride synthesis-related glutathione synthase 1 (GSH1) genes AsGSH1a, AsGSH1b, AsGSH1c, AsGSH1d and AsAly, among which AsGSH1d was significantly positively correlated, indicating that AsGSH1d might play an important role in the synthesis of allicin. This study provides a theoretical basis for increasing the content of allitride.

Plant cyclic nucleotide-gated ion channel (CNGC) plays an important role in plant growth, development and stress response. In order to investigate the function of CNGC gene family in pepper (Capsicum annuum), the CNGC family members of pepper were identified, and the physical and chemical properties, subcellular localization, phylogenetic development, structural characteristics, promoter cis-acting elements and gene expression patterns were analyzed. The results showed that a total of 14 CaCNGC gene family members were identified in pepper, which were not uniformly distributed on each chromosome. The isoelectric point of CaCNGC was 8.64~9.61, the molecular weight was 72.87~91.49 kD. The instability index was greater than 40, indicating that 14 members of CaCNGC were unstable proteins. All CaCNGC proteins were located in the plasma membrane, had transmembrane structures, and were hydrophilic proteins. The analysis of cis-element in promoter showed that, most of the CaCNGC contained cis-elements related to growth and development, hormone regulation, and abiotic stresses response. Transcriptome data analysis showed that most members of CaCNGC had low or no expression in leaves, flowers and fruits of pepper. Most members of CaCNGC were expressed under low temperature, high temperature, abscisic acid, gibberellin, hydrogen peroxide, indoleacetic acid, jasmonic acid and salt stress. qRT-PCR analysis showed that the expression of CaCNGC2, CaCNGC7 and CaCNGC10, was up-regulated under low and high temperature stress, which might be involved in the response of pepper to temperature stress. These results provide a theoretical reference for further investigating the function of CaCNGC gene family, and new insights into the molecular mechanisms of pepper tolerance to temperature stress.

The plant hormone abscisic acid (ABA) was the key hormone in the regulation of plant growth and stress response, and played a key role in the regulation of seed dormancy and germination and growth and development, and the ABA receptor pyrabactin resistance (PYR) / PYR1-like (pyrabactin resistance-like, PYL) proteins (PYLs) / regulatory components of ABA receptor (RCAR), as one of the core components of the ABA signaling pathway, plays an important function in the ABA signaling pathway. In this study, bioinformatics methods and transcriptome sequencing were used for genome-wide identification of PYL family members in Malus sieversii, the chromosomal locations, evolutionary relationships, gene structures, protein motifs, covariate and cis regulatory elements were analyzed by bioinformatics methods were used for genome-wide identification of PYL family members in M. sieversii, as well as the expression patterns in different organs, and expression profiles characterized by the low-temperature stratification process of seeds of PYL members by transcriptome sequencing. The results showed that 14 PYL genes were identified in the whole genome of M.sieversii, which were distributed on 7 chromosomes with clustered gene distribution. Phylogenetic analysis classified them into 5 subgroups: groups A~E. The family members contained 0~4 introns, and the encoded proteins of the 14 PYL genes were identified to have PYR_PYL_RCAR_like conserved structures. Cis-acting element analysis showed that a variable number and variety of cis-acting elements involved in hormone response, growth and developmental response, abiotic and biotic stress response were prevalent in the 2 000 bp upstream sequence of the PYL gene in M. sieversii. Gene ontology enrichment showed that the MsiPYL gene mainly played a role in abscisic acid activation signaling pathway, regulation of phosphatase activity and phosphoprotein phosphatase activity, etc. KEGG pathway analysis showed that it was mainly involved in mitogen-activated protein kinase (MAPK) signaling pathway-plant and phytohormone signaling. The MsiPYL genes were constitutively expressed in 6 different tissues, with levels of expression in reproductive organs (flowers and fruit) significantly higher than those in nutrient organs (roots and stems). The results of RNA-seq and qRT-PCR showed that MsiPYL8 (MsiPYR1) had the lowest expression in seeds, which was down-regulated under the low-temperature stratification of M. sieversii seeds, and it might be an important candidate gene in response to seed germination in M. sieversii, which provides a basis for revealing the roles of MsiPYL genes in the release of seed dormancy in M. sieversii.

MicroRNA (miRNA) mediated post transcriptional regulation plays a crucial role in plant responses to pathogens. In order to screen miRNAs in grapes (Vitis vinifera) that respond to Botrytis cinerea infection, this study infected grape leaves with B. cinerea spore suspension and constructed a small RNA library for high-throughput sequencing. A total of 171 known miRNAs and 251 new miRNAs were obtained in response to B. cinerea, of which 53 miRNAs showed differential expression after inoculation, especially miRNA399a, miRNA399b, and miRNA399h belonging to the miRNA399 family, which showed significant downregulation. The susceptibility of grapes to B. cinerea was determined by analyzing the phenotype, lesion area, and pathogenicity of leaves. 8 key target genes were obtained by predicting the downstream target genes of the 3 key miRNAs mentioned above. qPCR analysis showed that the target genes of miRNA399b and miRNA399h were significantly upregulated when infected with B. cinerea. This study provides a theoretical basis for further understanding the resistance response of fruit trees to B. cinerea during domestication and for breeding resistant grape varieties.

Atropa belladonna is a commercial medicinal plant of tropane alkaloids (TAs) listed in the 《Chinese Pharmacopoeia》, and WRKY transcription factors may regulate the biosynthesis of TAs. In this study, tissue expression profile, phylogenetic tree and inducible expression analyses of the WRKY transcription factor family in A. belladonna were performed to screen out WRKY gene that positively regulated TAs biosynthesis. AbWRKY1 was identified and its function was verified at the biochemical and molecular genetic levels. AbWRKY1 and TAs synthetic pathway genes shared the same tissue and inducible expression characteristics that they were specifically expressed in the adventitious roots and were induced by the calcium channel inhibitor verapamil. AbWRKY1 clustered together with WRKYs from other species that had functions in regulating secondary metabolism. AbWRKY1 protein contained 326 amino acid residues, and localized in nuclear. Yeast one-hybrid result showed that AbWRKY1 could interact with the W-box rich region of hyoscyamine 6β-hydroxylase (H6H) gene promoter. Knockdown of AbWRKY1 expression by virus-induced gene silencing (VIGS) in A. belladonna plants reduced scopolamine and hyoscyamine levels by 44.91% and 79.33%, respectively. AbWRKY1 overexpression raised the amount of scopolamine, anisodamine, and hyoscyamine in A. belladonna's hairy roots, while the expression of the 4 TAs synthesis pathway genes, namely putrescine N-methyltransferase (PMT), tropinone reductaseⅠ(TRⅠ), hyoscyamine dehydrogenase (HDH) and H6H, were increased concomitantly. This study provides a theoretical basis for the elucidation of the regulatory mechanism of TAs biosynthesis and the molecular breeding of A. belladonna.

Structural variation (SV) can affect gene phenotype by affecting the stability and expression regulation of related genes. To investigate the polymorphic distribution of the 282 bp SV in intron 1 of the CD2-associated protein (CD2AP) gene in the Xiang pig (Sus scrofa) population and its effect on the expression of the CD2AP gene, Xiang pig and Large White were taken as research subjects, and the SV locus was genotyped by PCR sequencing, and the distribution pattern and the association between this SV and pulmonary diseases in Xiang pig and Large White populations were studied. Bioinformatics methods were used to detect the repetitive elements and transcription factor binding sites in the SV region of intron 1 of the CD2AP gene. qPCR and Western blot were used to detect the effect of SV on CD2AP gene and protein expression. The effect of short interspersed nuclear element (SINE) on the transcriptional activity of CD2AP gene was examined by constructing pEGFP fluorescent expression vector. The results showed that the CD2AP-I1-sv282 structural variant fragment was 282 bp long and contained a 260 bp SINE element in the interval. It belonged to the tRNA family and was located in pig Chr7:42249739~42249998 bp, containing an RNA polymeraseⅢ promoter, 8 transcription factor binding sites, and 2 AluⅠsites. This SV exhibited polymorphism in both Xiang pig and Large White genomes, with II genotype (homozygous insertion), DD genotype (homozygous deletion), and ID genotype (heterozygous). χ² test showed that the SV was in Hardy Weinberg equilibrium between the Xiang pig and Large White populations (P>0.05), and there was extremely significant difference in genotype frequencies between the Xiang pig and Large White populations (P<0.01). The frequency of the D allele was significantly higher than that of the I allele (P<0.001). The D allele was the dominant allele in the diseased lung samples, while the I allele was the dominant allele in the healthy lung samples. The Kendall's tau-b correlation analyses indicated that the presence of the pathological features was positively correlated with genotype, with the DD genotype group being more susceptible to structural mutations. As detected by using qPCR and Western blot assays, it was found that the mRNA and protein expression of the CD2AP gene in Xiang pig spleens was highly significantly higher in the DD genotype than in the ID genotype (P<0.01). The pEGFP fluorescent reporter vector carrying the SINE element showed a decrease in fluorescence intensity and EGFP protein in cells. In summary, the 282 bp structural variation in intron 1 of the CD2AP gene exhibited polymorphism in Xiang pig population, and was predominantly deletion type, leading to upregulation of gene expression. The results indicated that this SV affected gene transcriptional activity, and was associated with susceptibility to lung disease in Xiang pigs. The results are of great significance to analyze the molecular mechanism of disease susceptibility in Xiang pigs.

Peroxisome proliferator activated receptor γ coactivator-1 alpha (PGC-1α) is a transcriptional coactivator that influences various energy metabolism processes and plays a crucial role in fat deposition. To investigate the function and underlying mechanisms of the PGC-1α gene in lipid metabolism, a series of experiments were performed. The expression of PGC-1α in 12 different tissues was examined in chickens (Gallus gallus). Using RT-PCR, the CDS of the chicken PGC-1α gene was amplified and obtained, and a eukaryotic expression vector was constructed. The vector was then transfected into the leghorn male hepatoma (LMH) cell line, and Western blot analysis was used to confirm the expression of the PGC-1α protein. After overexpressing and knocking down PGC-1α gene in LMH cells, lipid droplet deposition was assessed using oil red O staining, total triglyceride (TG) content was measured by colorimetric assay, and the expression of lipid metabolism-related genes was analyzed through qPCR. The results showed that PGC-1α had the highest expression in cardiac tissue, with relatively high levels in leg muscle, kidney, and brain tissues. Western blot analysis confirmed that the overexpression vector successfully expressed the fusion protein in LMH cells. Overexpression of PGC-1α led to a extremely significant reduction in lipid deposition and TG content (P<0.01), while knockdown of PGC-1α resulted in a trend of increased lipid deposition and a significant rise in TG content (P<0.05). Moreover, overexpression of PGC-1α significantly downregulated the expression of fatty acid synthetase (FASN), peroxisome proliferator activated receptor γ (PPARγ) and acetyl-CoA carboxylase alpha (ACACA) genes (P<0.05 or P<0.01), while extremely significantly upregulated phosphoenolpyruvate carboxykinase 1 (PCK1) (P<0.01). Knockdown of PGC-1α extremely significantly decreased PCK1 expression (P<0.01). These findings suggested that the chicken PGC-1α gene likely exerts its effects by regulating the expression of these functional genes. By constructing overexpression and knockdown models of the PGC-1α gene, this study reveals its key regulatory role in chicken lipid metabolism, particularly through modulating the expression of FASN, PPARγ, ACACA, and PCK1, which affect lipid deposition and triglyceride levels. This study provides a valuable reference for further exploration of the functional mechanisms of PGC-1α in chicken lipid metabolism.

Poultry primordial germ cells (PGCs) migrate from the blood and colonize the reproductive ridge. Therefore, PGCs can not only replace fertilized eggs for genetic modification, but also have important application prospects in germplasm resource conservation and the recovery of endangered species. In vitro culture systems for PGCs exhibit differences in supporting the growth of male and female PGCs. So it is difficult to meet the requirements of PGCs in germplasm resource conservation. In response to this problem, this study aims to explore the function of fatty acid oxidation (FAO) involved in the formation of PGCs, and provide theoretical guidance for the subsequent establishment of a perfect in vitro culture system of PGCs.The role of FAO in the differentiation of embryonic stem cells (ESCs) into PGCs was analyzed based on transcriptomics, and the optimal addition concentrations of FAO activator BMS-687453 (BMS) and inhibitor perhexiline maleate (Per) were investigated by CCK8 and EdU tests. Finally, the role of FAO in the differentiation process from ESCs to PGCs was detected by cell morphology, qRT-PCR, indirect immunofluorescence and flow cytometry.The results showed that the genes involved in FAO were differentially expressed in ESCs and PGCs by transcriptomic analysis. Moreover, it was enriched in germ cell differentiation related pathways (MAPK signaling pathway, GnRH signaling pathway, PI3K-Akt signaling pathway). qRT-PCR results showed that acetyl-Coacyltransferase 2 (ACAA2) and enoyl-CoA hydratase 1 (ECH1) were highly expressed in PGCs.CCK8 and EdU experiments determined that the optimal addition concentrations of BMS and Per were 80 and 160 nmol/L, respectively. The results of cell morphology showed that the number of blastoid bodies in BMS group was significantly higher than that in Per group at 6 d (P<0.05). In BMS group, the expression levels of chicken VASA homolog (CVH) and c-Kit proto-oncogene receptor tyrosine kinase (C-KIT) were significantly higher than those in Per group (P<0.05). Indirect immunofluorescence and flow cytometry results showed that the number of DDX4 positive cells in BMS group was significantly higher than that in Per group (P<0.05). The above results indicated that promoting FAO could promote the formation of PGCs. This study provides a theoretical basis for establishing a perfect PGCs system in vitro culture of poultry, and also helps to systematically clarify the genesis mechanism of PGCs from the perspective of cell energy metabolism.

Adapter protein myeloid differentiation factor 88 (MyD88) is involved in the interleukin-1 receptor (IL-1R) and Toll-like receptor (TLR)-mediated activation of nuclear factor-kappaB (NF-κB), which plays important role in innate immunity. In this study, the full-length cDNA of largemouth bass (Micropterus salmoides) MyD88 was isolated. Its ORF was 867 bp in length which encoded 288 amino acid residues. Protein secondary structure analysis showed that MyD88 protein had a N-terminal death domain and a TIR (Toll-like/IL-1) domain, which are known as important functional structural domains in mammalian MyD88. The largemouth bass MyD88 protein had high identity (58.5%~99.3%) with other vertebrates. Phylogenetic analysis showed that largemouth bass MyD88 gathered together with MyD88 of other fish species. In healthy largemouth bass, MyD88 mRNA was detected in all sampled tissues, and MyD88 mRNA had the highest expression levels in the liver. To study the role of MyD88 in innate immunity, its mRNA expression profile after stimulation with polyinosinic-polycytidylic acid (PolyI:C) and Nocardia seriolae was studied in the intestine, gill, spleen and kidney. The results showed that in all 4 detected issues, 2 kinds of immune stimuli both upregulated the transcription level of largemouth bass MyD88, and the induction by N. seriolae in the kidney was the strongest. In kidney tissue, MyD88 showed the maximum upregulation at 9 d after infection, and the expression level in the infection group was 8.5 times of that in the control group (P<0.05). The induction by PolyI:C in the gill was initiated earliest. In gill tissue, after 8 h of stimulation, the relative expression level of the experimental group was 5.6 times of that in the control group. Largemouth bass MyD88 were distributed in the HeLa cytoplasm. Overexpression of MsMyD88 could activate NF-κB. These results provide basic data for elucidating the role of MyD88 in the innate immunity of largemouth bass.

The hybridoma cell technique is a crucial method for monoclonal antibody production. Monoclonal antibody-producing hybridoma cells against Porcine epidemic diarrhea virus (PEDV) have been prepared in this research group. In this study, the hybridoma cell culture system was optimized to improve the efficiency of antibody secretion from hybridoma cells by adding different concentrations of additive factors, including glutamine (0~10 mmol/L), antimicrobial peptides (0~20 μg/mL), peptides (0~20 ng/mL), human epidermal growth factor (hEGF) (0~20 ng/mL), basic fibroblast growth factor (bFGF)(0~20 ng/mL), and vitamin C (VitC)(0~20 ng/mL). These factors were introduced into the hybridoma cell culture system to investigate their impact on the growth and antibody secretion of hybridoma cells by plotting growth curves and using enzyme-linked immunosorbent assay (ELISA), respectively. The results showed that the peak of cell density was reached on the 5th day when glutamine and antimicrobial peptides were added, and the maximum cell density was achieved at the addition concentration of 6 mmol/L glutamine and 10 μg/mL antimicrobial peptides. The peak of cell density was reached on the 4th day when hEGF, bFGF and VitC were added, and the maximum cell density was achieved at the addition concentration of 10 ng/mL hEGF, 20 ng/mL bFGF and 20 ng/mL VitC. The results of antibody titer detection showed that the highest antibody titer (1∶8000) was obtained in the glutamine addition group at a concentration of 6 mmol/L, the highest antibody titer (1∶16000) was obtained in the antimicrobial peptide addition group at a concentration of 10 μg/mL, the highest antibody titer (1∶16000) was obtained in the hEGF and bFGF addition groups at concentrations of 5 and 10 ng/mL, respectively, and the highest antibody titer (1∶8000) was obtained in the VitC addition group at a concentration of 10 ng/mL. In conclusion, the growth and antibody production capabilities of hybridoma cells have been effectively enhanced through the modification of medium components. This study establishes a solid groundwork for the formulation of specialized hybridoma cell culture media aimed at production purposes.

Fusarium proliferatum is a phytopathogenic fungus with a wide host range and serious damage, but its pathogenic mechanism is not well understood. Effectors play an important role in the infection process of phytopathogenic fungi. The prediction, screening, and identification of effectors are of great importance in exploring the mechanism of host-pathogen interaction. In this study, based on the transcriptome sequencing results of F. proliferatum, 43 putative effectors were screened by signal peptide identification, transmembrane domain screening, cysteine number analysis, and Pathogen Host Interactions database (PHI-base) analysis. Functional studies were carried out on one of the candidate effectors, feruloyl esterase FpESD. Yeast system validation revealed that the signal peptide encoded by the N-terminal 20 amino acids of FpESD possesses secretion activity. The homologous recombination strategy was used to generate the FpESD knockout mutant (ΔFpESD), and the complementary strain (ΔFpESD-C) was obtained through complementation experiments. Growth rate measurements, spore yield assessments, and spore length analysis demonstrated that the deletion of the FpESD gene significantly reduced fungal growth rate and conidia production (P<0.05), with shorter conidia length observed for the knockout mutant. Additionally, environmental stress tests revealed that ΔFpESD exhibited significantly reduced sensitivity to CaCl₂, MgCl₂, and Congo red (P<0.05), indicating that FpESD was involved in regulating the response to environmental stress. Pathogenicity assays using plate inoculation showed that the disease index of alfalfa (Medicago sativa) seedlings infected by ΔFpESD was significantly lower than that of the wild-type strain (P<0.05), suggesting that FpESD positively regulated the pathogenicity of F.proliferatum. This study helps to elucidate the pathogenic molecular mechanism of F. proliferatum, and also provides ideas for the in-depth study of F. proliferatum effectors.

With the promulgation of the ban on antibiotics in feed, there is an urgent need for a feed additive capable of inhibiting Gram-negative bacteria in the process of livestock and poultry breeding, so as to reduce the occurrence of diseases caused by such bacteria. The aim of this study was to design an antimicrobial peptide PC that can specifically inhibit the growth of Gram-negative bacteria, but has no significant inhibitory effect on Gram-positive bacteria. Using (RP)₃WW (RP)₃ as the basic unit (where P is proline, R is arginine, and W is tryptophan), the trypsin inhibitor (SFTI-1) binding loop (CTKSIPPIC) derived from sunflower (Helianthus annuus) was added to the amino terminus of the antimicrobial peptides. The secondary structure of the antimicrobial peptide PC was detected by circular dichroism (CD). And further to detect the antimicrobial activity, hemolysis, cytotoxicity and stability of the antimicrobial peptide PC. The antimicrobial peptide PC exhibited an α-helical structure under the simulated membrane hydrophobic environment. The results showed that the antimicrobial peptide PC had high inhibitory activity against Gram-negative bacteria such as Escherichia coli, Salmonella pullorum and Pseudomonas aeruginosa. In contrast, for Gram-positive bacteria such as Staphylococcus aureus, Staphylococcus epidermidis and Enterococcus faecalis, antimicrobial peptide PC did not show inhibitory activity, but could promote bacteria to produce reactive oxygen species (ROS), and had low hemolytic activity and cytotoxicity. The antimicrobial peptide PC obtained in this study had the characteristics of anti-Gram-negative bacteria, good biological safety and stability, and had the development potential to be a substitute for forage antibiotics. This study provides a basis for the design and development of antimicrobial peptides against gram bacteria..

Disease-resistant breeding is a crucial strategy to enhance the resilience of pig (Sus scrofa) populations against pathogen infections. The success of this approach relies on the comprehensive identification and investigation of candidate genes associated with disease resistance in the host. Therefore, identifying high-quality candidate genes is essential for disease-resistant breeding. This review summarized recent progress in research on candidate genes related to resistance against several significant viral infectious diseases in pigs, including Porcine reproductive and respiratory syndrome virus (PRRSV), African swine fever virus (ASFV), Classical swine fever virus (CSFV), Porcine epidemic diarrhea virus (PEDV), Pseudorabies virus (PRV), and Japanese encephalitis virus (JEV). Furthermore, the review discussed the challenges associated with the study of candidate genes for disease resistance and their application in breeding practices, and provides valuable insights for swine disease-resistant breeding research and practice in China.

Phosphorus in soil exists in complex forms, such as phytic acid, which is difficult to be absorbed and utilized by plants. This problem can be effectively solved through degrading soil phytic acid by phytase and releasing phosphorus. In this study, high-yielding phytase strains were selected from the soil, and the composition of the fermentation medium and culture conditions were optimized by the Response Surface Methodology. Phytase was prepared into microcapsule enzyme preparation with sodium alginate (SA) and CaCl₂ as embedding and crosslinking agents, and its storage stability and growth-promoting effect on corn (Zea mays) were investigated. The results showed that a strain of phytase-producing bacteria was identified as Pseudomonas corrugata by 16S rRNA. The optimal medium composition and fermentation conditions were determined through fermentation optimization, and the phytase activity after optimization was (435.70±1.87) U/mL, which was increased by 82.16% compared with that before optimization. The microcapsule enzyme preparation prepared with 3% SA and 3% CaCl₂ had the characteristics of good pellet formation and high enzyme activity. After 30 d of storage, the enzyme activity of (85.63±1.73)% remained. The experiment showed that microcapsule enzyme preparation significantly promoted corn growth, increasing corn seedling dry weight, plant height, root dry weight, root length and fresh seedling weight by 48.59%~304.46%. This study provided a method of strain screening, fermentation optimization and preparation of microcapsule enzyme preparation, which is of great significance for the extensive application of microcapsule enzyme preparations in agricultural production and new bacterial fertilizers.

Pepper mottle virus (PepMoV) is a member of the genus Potyvirus, capable of infecting important Solanaceous crops such as pepper (Capsicum annuum) and tomato (Solanum lycopersicum). In this study, PepMoV was successfully detected in pepper samples from Hangzhou, Zhejiang province, using small RNA sequencing technology. The complete genome, 9 640 nucleotides in length, was amplified using RT-PCR. To overcome difficulties in constructing infectious clones of PepMoV, such as large fragment assembly challenges and lethal genes for Escherichia coli, yeast (Saccharomyces cerevisiae) homologous recombination technology was employed, which enabled the assembly 5 DNA fragments into the full-length infectious clone of PepMoV in a single step in yeast. Importantly, the yeast plasmid can be directly transformed into (Agrobacterium tumefaciens) for infiltration, thereby avoiding the E. coli cloning step and its associated toxicity issues. Results indicated that, compared with traditional enzyme ligation methods, this construction strategy reduced the required time from several months to just one week. The obtained infectious clone of PepMoV-HZ effectively infected Nicotiana benthamiana, exhibiting clear symptoms. The successful construction of the PepMoV infectious clone not only provides a positive sample control for the rapid detection method of the virus but also offers crucial experimental tools for in-depth studies of the virus's gene functions and its interaction mechanisms with the host.

High mobility group nucleosomal binding domain 2 (HMGN2) is highly expressed in mammalian embryonic development, which can bind to nucleosomes, change chromosome structure, and participate in the regulation of cell differentiation and embryonic development. In order to explore the mechanism of HMGN2 protein in the uterus of yak (Bos grunniens), in this study, yak HMGN2 gene was cloned and HMGN2 polyclonal antibody was prepared, and the relative expression of HMGN2 gene in yak uterus at different stages was detected by qPCR. Oryctolagus cuniculus was immunized with purified yak HMGN2 recombinant protein, and yak HMGN2 polyclonal antibody was purified by A/G-Sepharose column chromatography. Western blot was used to detect the expression of HMGN2 protein, and immunohistochemistry (IHC) was used to detect the localization of HMGN2 protein. The results showed that the recombinant protein was induced by the successful cloning of yak HMGN2 gene (GenBank No. OR713742) and the construction of recombinant plasmid pKMD-SUMO-HMGN2 in this study were in line with the expected results. The results of qPCR and Western blot showed that HMGN2 gene and protein were the highest in the first trimester of yak uterus. The results of IHC showed that HMGN2 was mainly expressed in the uterine gland and endometrium suggesting that HMGN2 might play a regulatory role in embryo implantation and fetal growth and development in the early pregnancy of yak uterus. These results indicated that the yak HMGN2 antibody prepared in this study could be used for molecular experiments using yak as a model, which provides basic data for further study of the mechanism of action of HMGN2 protein.

Human papillomavirus (HPV) is a common genital tract virus HPV subtypes 16 and 18 have a close relationship with cervical cancer. Currently, there are 6 types of preventive HPV vaccines available worldwide. However, these vaccines are only for preventive and cannot be used for the treatment of infected cases. Therefore, this study constructed the TC-1-HPV18 E7 cell model co-expression HPV16 E7 and HPV18 E7 using mouse (Mus musculus) cervical cancer cell line TC-1 as a base model. The attenuated Listeria monocytogenes (LM) was used as a vector for the recombinant strain vaccine LM-HPV18 E7 to deliver tumor antigens, and the therapeutic efficacy of the recombinant strains in the TC-1-HPV18 E7 tumor-bearing mouse model was evaluated. The results showed that the TC-1-HPV18 E7 cell model and mouse model were successfully constructed. There was no difference in tumor formation between TC-1-HPV18 E7 tumor-bearing mice and TC-1 tumor-bearing mice, indicating that stable expression of exogenous genes had no impact on the cell and mouse models. LM-HPV18 E7 vaccine exhibited significant therapeutic effects on tumors in TC-1-HPV18 E7 tumor-bearing mice compared with TC-1 tumor-bearing mice, which proving that LM was a potent live vaccine vector platform. This study expands the use of TC-1-based cell models and provides a reference for the research of LM as a therapeutic vaccine vector.