Porcine deltacoronavirus (PDCoV) is a emerging porcine intestinal coronavirus in recent years, causing huge economic losses to the pig industry. To investigate the pathogenicity and the ultrastructural changes of the intestinal tract from PDCoV-infected newborn piglets (Sus scrofa domesticus), the clinical manifestations, autopsy and histopathological changes, tissue tropism and intestinal ultrastructural changes in 1-day-old neonatal piglets infected with PDCoV CHN-GD16-05 strain were observed and analysed. Severe vomiting, diarrhea, listless, and lethargy were observed in PDCoV-challenged piglets at day 3 post-infection. An autopsy revealed that PDCoV-challenged newborn piglets displayed intestinal bloating and dilation, exhibited thin and transparent intestinal walls with an accumulation of yellow liquid in the intestinal lumen. Additionally, PDCoV mainly infected duodenum, jejunum and ileum of newborn piglets determined by qPCR and Western blot, with extremely significantly higher levels of PDCoV-Nucleocapsid (PDCoV-N) gene and protein in the jejunum than in the duodenum and ileum (P<0.01). Further hematoxylin eosin staining of jejunal tissue sections showed that PDCoV infection disrupted the structure of the intestinal villi, and increased crypt cells proliferation and crypt depth, causing diffuse intestinal villi atrophy and even fusion. Electron microscope also showed that the intestinal villi of PDCoV-infected piglets were atrophied, the microvilli of intestinal epithelial cells were broken and shed, the tight junctions between cells were disrupted, the cytoplasmic vacuolated cells were increased, and the mitochondria were enlarged in size and reduced in number. Moreover, PDCoV infection stimulated the expression of antiviral genes in the host, such as interferon α (IFN-α), IFN-β, IFN-λ, and their downstream interferon-stimulated genes, such as myxovirus resistance 1 (MX-1), interferon-stimulating gene 15 (ISG-15), interferon induced protein with tetratricopeptide repeats 1 (IFIT1), 2'-5'-oligoadenylate synthetase 1 (OAS1), 2'-5'-oligoadenylate synthetase like (OASL), inflammatory factors such as tumor necrosis factor α (TNF-α), interleukin 1β (IL-1β), IL-6, and C-C motif chemokine ligand 20 (CCL20). Taken together, the results indicated that PDCoV infection could rapidly invade the small intestine of newborn piglets, and effectively replicate in the jejunal epithelial cells, resulting in degeneration and necrosis of infected epithelial cells, which in turn caused intestinal villi atrophy, and ultimately result in osmotic diarrhoea. This study provides a theoretical basis for elucidating the pathogenic mechanism of PDCoV-infected piglets.

Fructokinase (FRK) is the main catalytic enzyme in the phosphorylation of fructose and plays a key role in the regulation of plant growth and development. In order to further study the FRK gene function of Cucumis sativus, the FRK gene family and its molecular characteristics were systematically identified and analyzed, and the gene expression patterns under abiotic stress were analyzed. The results showed that there were 12 FRK genes distributed on 7 chromosomes. The number of encoded amino acids was 331~588, and the molecular weight was 35.63~65.90 kD. The FRK gene family of cucumber was divided into 3 subfamilies by phylogenetic analysis, and the gene structure and motif in each subfamily were basically the same. The promoter sequences mainly contained cis-acting elements such as light response, low temperature response, drought response, hormone response, defense stress and plant growth and development. Tissue-specific expression analysis showed that the expression levels of some FRK genes in cucumber were low or not expressed in roots, stems, leaves, female flowers, and male flowers, indicated that different cucumber FRK genes had tissue-specific expression patterns. Abiotic stress expression analysis showed that CsaFRK6, CsaFRK8 and CsaFRK12 was significantly up-regulated under low temperature stress (P<0.05). The expression of CsaFRK2, CsaFRK5 and CsaFRK7 was significantly up-regulated under salt stress (P<0.05). CsaFRK1~CsaFRK9 were significantly up-regulated under drought stress (P<0.05). The expression patterns of FRK genes in different tissues and organs were different in response to abiotic stress. This study provides a reference for elucidating the biological gene function of cucumber and its response to abiotic stress.

As an annual herbaceous plant, tomato (Solanum lycopersicum) is rich in nutrients, and with the change in market demand, breeding high-yield and high-quality tomato varieties has become one of the main goals of breeding. The number of seeds, soluble solids, transverse diameter and longitudinal diameter of tomato fruit are of great significance to improve the yield and quality of tomato fruit. This study was conducted using insertion/deletion (InDel) markers to locate QTLs for tomato fruit seed number, soluble solids, transverse diameter and longitudinal diameter, and candidate genes were identified based on RNA-seq. In this study, F₂ populations of 60 lines were obtained by crossbreeding 'Tomato microflora', which has low fruit seed number and soluble solids content and small transverse and longitudinal diameters, and 'M82' was used as the female parent, which has high fruit seed number and soluble solids content and large transverse and longitudinal diameters. Correlation and cluster analysis were carried out through the investigation of phenotypic data. Based on tomato genome information, candidate genes related to tomato fruit traits were explored. The results showed that the variation coefficients of F₂ tomato material seed number, soluble solid content, longitudinal diameter and transverse diameter were all greater than 10%, and skewness and kurtosis showed that these 4 characteristics were normally distributed. Seed number was positively correlated with soluble solid content, longitudinal diameter and transverse diameter, and soluble solid content was negatively correlated with longitudinal diameter and transverse diameter. Longitudinal and transverse diameters were positively correlated. Twenty-five parental polymorphic InDel markers were detected in 62 tomato samples, and a total of 149 sites were detected. Unweighted pair-group method with arithmetic means (UPGMA) cluster analysis showed that 62 tomato samples could be divided into 4 types when the genetic similarity coefficient was 0.70~0.75. Based on 25 polymorphic InDel markers, a total of 5 QTLs were obtained, 4 candidate genes were mined, and their expression patterns were provided by RNA-seq. Solyc12g027770 was located in both seed number and soluble solid. This study provides reference for genetic analysis, molecular mechanism research and the development of molecular markers of seed number, soluble solids and longitudinal diameter of tomato fruit.

The NAC is one of the plant specific transcription factor and plays a key role in plant growth, development and stress responses. In this study, 84 NAC genes were identified from Pisum sativum genome database by sequence alignment using the NAC family genes of Arabidopsis thaliana and Glycine max. They were distributed unevenly on 7 chromosomes and divided into 13 subfamilies. All subfamily members had highly homologous sequences at the N-terminal region and highly divergent sequences at the C-terminal region. Members of the same subfamily had similar types and quantities of conserved motifs, and contain specific conserved motifs. Most NAC genes contained 3 exons. There were 29 and 50 NAC genes in the pea genome, which were collinear with the NAC genes in the Arabidopsis and soybean genome, respectively. Seventy-nine genes contained at least one abiotic stress related element such as drought responsive element and low-temperature responsive element. The drought stress expression profile showed that 60 PsNAC genes had obvious response to drought stress. 75.0% and 81.7% of the 60 PsNAC showed significant response in the early stage and late stage of drought stress, respectively, indicating that PsNAC gene played an important role in different stages of drought in peas. Among them, five PsNAC genes had higher response level in the shoot or root within 48 h of drought stress. This study analyzed the information of PsNAC gene family members which could provide a reference for further research on identification of stress resistance related genes.

Hyperosmolarity gate calcium-permeable channels (OSCA) is a mechanosensitive porous ion channel, which can regulate Ca²⁺ concentration and water transport rate in plant cells under osmotic stress. It plays an important role in hypertonic stress in plants. In order to understand the structural characteristics and expression characteristics of OSCA gene family in ginger, this study identified and analyzed the OSCA gene family in ginger (Zingiber officinale). The results showed that 38 members of OSCA gene family were identified from ginger genome, which were located on 14 chromosomes respectively. The prediction of subcellular localization shows that the members of OSCA family in ginger were mainly located in plasma membrane. The analysis of cis-acting elements of promoters showed that 38 ZoOSCAs contained various response elements to adversity and hormone, such as methyl jasmonate, abscisic acid and drought. The transcriptome of salt stress and fluorescence quantitative results of drought treatment showed that the expression levels of 18, 23 family members in leaves and rhizomes were up-regulated under salt stress, and 11 out of 14 family members were up-regulated under drought treatment, and the expression levels of ZoOSCAs in different tissues were different. This study provides basic data for revealing the function and mechanism of ZoOSCAs.

TCP (teosinte branched1/cycloidea/proliferating cell factor) is a plant specific transcription factor that is widely involved in plant growth and development and response to environmental stress. TCP9 is a class Ⅰ member of TCP gene family and plays an important role in respose to plant drought stress. Pitaya (Hylocereus sp.) is an ideal material for drought resistance research because of the strong drought resistance. Hylocereus polyrhizus variety 'Da Hong' were used to isolated the HpTCP9 gene by RT-PCR technology in order to investigate the biological function of the TCP9 gene in the drought resistance of pitaya. Bioinformatics analysis showed that the ORF length of the HpTCP9 gene was 729 bp, encoding 242 amino acids, with a molecular weight of 24.86 kD, an isoelectric point of 5.04, and a hydrophilicity index of 1.021. Domain analysis showed that the HpTCP9 protein sequence had a typical TCP conserved domain. The protein structure prediction results indicated that α spiral was 12.40%, irregular curls were 66.12%, and extended chains were 12.81%, The β-turn was 8.68%. To detect the subcellular localization of HpTCP9, a pCAMBIA2300-HpTCP9-GFP expression vector carrying a GFP tag was constructed. HpTCP9 was detected as a nuclear localization protein through Arabidopsis protoplast transformation. Real time fluorescence quantitative PCR analysis showed that the HpTCP9 gene was significantly upregulated by 20% PEG6000 simulated drought treatment, and the expression level was positively correlated with the treatment duration. The above results indicated that the HpTCP9 belonged to the ClassⅠsubfamily of the TCP family, had a TCP conserved domain, was a hydrophobic protein located in the nucleus, and played a role in the drought stress defense mechanism of H. polyrhizus. This study provides a theoretical basis for further exploring the function of the HpTCP9 gene in H. polyrhizus.

Superoxide dismutase (SOD) is a key enzyme in the plant antioxidant system that protects plants from biotic and abiotic stresses by removing reactive oxygen species produced by living organisms. As a pioneer tree species with strong drought resistance, the study on the response mechanism of masson pine (Pinus massoniana) to high temperature and drought will help to understand the stress tolerance mechanism of plants. In this study, the transcriptome data of masson pine in drought, high temperature and high temperature-drought combined stress were mined, the PmSOD gene family was identified and bioinformatically analyzed, and the gene expression pattern of PmSOD gene family under high temperature, drought and high temperature-drought stress was detected. The results showed that 8 members of the PmSOD gene family were identified, including 4 Cu/Zn-SODs (CSDs), 2 Fe-SODs (FSDs) and 2 Mn-SODs (MSDs). Family members of the same type had the same protein domain, similar subcellular localization, and conserved motifs. The gene expression analysis showed that 3 types of stresses induced specific expression of PmSOD gene family. The expression mode of PmSOD gene family under combined stress was different from that of single high temperature or drought stress. The expression of PmCSD1 and PmCSD4 were significantly up-regulated under various stress treatments, indicating that PmCSD1 and PmCSD4 might be key genes in the abiotic stress response of masson pine. This study provides basic data for in-depth discussion of the function and application of SOD in the growth and stress adaptation of masson pine.

Mastitis has a significant impact on the health and milk production of dairy cows (Bos taurus), resulting in reduced quality of dairy products and significant economic losses to the entire dairy industry. Prunetin (Pru) had biological activities such as anti allergic and anti-inflammatory properties. The aim of this study was to investigate the effects and mechanisms of prunetin on the inflammatory response and lipid peroxidation induced by lipopolysaccharide in bovine mammary epithelial cells (BMECs). The experiment was divided into control (CK) group, lipopolysaccharide (LPS) group, and lipopolysaccharide plus prunetin (LPS+Pru) groups at concentrations of 10, 20, 40, and 80 μmol/L, as well as a lipopolysaccharide plus ferrostatin-1 (Fer-1)(LPS+Fer-1) group. The expression levels of inflammatory cytokines, nuclear factor κB (NF-κB), and nuclear factor erythroid 2-related factor 2 / Kelch-like ECH-associated protein 1 (Nrf2/Keap1) proteins were detected in each group. The results showed that LPS significantly upregulated the gene and protein levels of inflammatory cytokines such as tumor necrosis factor-α (TNF-α), interleukin-1 (IL-1β), IL-6, and IL-8. LPS also increased the content of Fe²⁺, reactive oxygen species (ROS), and malondialdehyde (MDA). Prunetin downregulated the genes of TNF-α, IL-1β, IL-6, IL-8, and their protein expression, upregulated the gene of Nrf2 and its protein expression, inhibited the NF-κB signaling pathway, and reduced cell death. Additionally, prunetin decreased the levels of Fe²⁺, ROS, and MDA while increasing the gene expression levels of glutathione peroxidase 4 (GPX4) and ferritin heavy polypeptide 1 (FTH1).These findings suggested that LPS induced inflammatory responses and lipid peroxidation in BMECs. Prunetin inhibited lipid peroxidation by activating Nrf2 and inhibiting NF-κB,and reducing the inflammatory response of BMECs induced by LPS, thereby exerting cytoprotective effects. This study provides theoretical support for the reduction of LPS induced inflammatory response and oxidative damage in BMECs by prunetin, and provides new ideas for protecting breast health in B. taurus.

Plemorphic adenoma gene 1 (PLAG1) plays an important role in regulating the growth and development of livestock and poultry. The purpose of this study was to investigate the relationship between SNP of PLAG1 gene and growth performance in Jiangquan black pig, Jiangquan black pigs weighing about 100 kg were used as research materials. PLAG1 gene mutations in 277 Jiangquan black pigs were detected by Mass ARRAY technology, and the association between SNP and growth traits was analyzed. The results showed that there were 3 SNP in the 5' regulatory region of PLAG1 genes, and 1 in the 3' regulatory region. The polymorphic information content (PIC) of g.75645278A>T and g.75693827A>G sites was 0.35, indicating moderate polymorphism (0.25<PIC<0.50). g.75686279G>A and g.75686320T>C sites showed low polymorphism (PIC<0.25). The genetic analysis showed that only g.75686279G>A locus was in Hardy-Weinberg equilibrium, and the other loci were significantly deviated from Hardy-Weinberg equilibrium (P<0.05). The results of linkage disequilibrium analysis and growth trait correlation showed that the g.75645278A>T and g.75693827A>G sites were completely linked, and had extremely significant effects on days to 100 kg (D100), chest circumference and abdominal circumference (P<0.01). In g.75645278A>T sites, AA individuals with D100 were significantly higher than TT and TA individuals (P<0.05). Chest and abdominal girth of TT and TA individuals were significantly higher than those of AA individuals (P<0.05). Four dominant haplotypes (GCA, GTG, GTA, ATG) and 7 diplotypes were constructed with the loci g.75686279G>A, g.75686320T>C and g.75693827A>G. The 7 combined genotypes had significant effects on D100 (P<0.05), and had extremely significant effects on chest and abdominal circumference (P<0.01). There was extremely significant interaction between haplotype GCA and ATG in chest and abdominal circumference (P<0.01), and significant interaction between haplotype GCA and ATG in body height (P<0.05). There was significant interaction between haplotype GTG and ATG on back height traits (P<0.05). In summary, SNP of PLAG1 gene locus are significantly correlated with growth performance of Jiangquan black pigs, and g.75645278A>T and g.75693827A>G sites could be used as candidate molecular marker sites for growth performance of Jiangquan black pigs. This study provides theoretical support for the breeding of Jiangquan black pigs.

Methyltransferase-like 23 (METTL23) plays an important role in the development of early porcine (Sus scrofa) embryos, tissues and organs. To analyze the genetic effect of SNP locus of METTL23 gene on reproductive traits, the correlation between SNP locus and reproductive trait of Kele pigs was analyzed by SPSS 22.0 software, including live piglet number, birth litter weight, birth average weight,weaning piglet number, weaning litter weight and weaning weight in this study. The results showed that 3 SNPs mutation sites of g.4804958G>T (intron 1), g.4805082C>T (exon 2) and g.4806821A>G (exon 4) were found in Kele pig METTL23 gene, and there were 3 genotypes. GT, CC and AA were the dominant genotypes of the corresponding mutation sites, and G, C and A were the dominant alleles of the corresponding mutation sites. The polymorphic information content (PIC) was moderately polymorphic (0.25<PIC<0.50). Linkage disequilibrium analysis showed that there was no strong linkage disequilibrium effect among the 3 SNPs. Correlation analysis showed that g.4804958G>T locus had significant effects on live piglets, birth litter weight and weaned piglets (P<0.05), and g.4806821A>G loci had significant effects on birth litter weight, average birth weight and weaning piglets (P<0.05). Diplotype H4H4 (GGCCGG) had significant effects on live piglets, birth litter weight, weaned piglets, weaning litter weight and average weaning weight (P<0.01). G.4805082C>T and g.4806821A>G had significant effects on the reproductive performance of Kele pigs, and the diplotype H4H4 (GGCCGG) had a very significant effect on the reproductive performance, so METTL23 could be used as an auxiliary marker for molecular breeding. This study provides a theoretical reference for pig genetics and breeding.

The identification of sheep (Ovis aries) molecular markers related to growth traits is of great significance to improve the economic benefits of sheep industry. Serine protease 12 (PRSS12), also known as neurotrypsin, is specifically expressed in the central nervous system and is associated with a variety of neurological functions. In order to explore the polymorphism of PRSS12 gene and its correlation with the growth traits of Hu sheep and Ujumqin sheep, this study used gene chip technology to genotype 1 008 Hu sheep and 336 Ujimqin sheep, and conducted association analysis with growth traits to identify molecular markers related to sheep growth and development. The results of genotyping showed that two SNP loci were detected in Ujimqin sheep population, in which rs402399470G>A presented low polymorphism, while rs418440631G>A showed moderate polymorphism. And only one SNP locus, namely rs418440631G>A, was detected in Hu sheep population, which showed low polymorphism. The results of association analysis with growth traits showed that rs402399470G>A locus was significantly correlated with 4-month-old body weight and 6-month-old chest width (P<0.05), while rs418440631G>A locus was significantly correlated with 4-month-old chest circumference, tube circumference and 6-month-old chest circumference of Ujimqin sheep (P<0.05). The rs418440631G>A locus was extremely significantly correlated with birth weight and 5-month chest circumference of Hu sheep (P<0.01), and significantly correlated with 3-month tube circumference, 5-month body length and chest width of Hu sheep (P<0.05). The correlation coefficient between body weight and body size of Ujimqin sheep and Hu sheep was calculated by R language corrplot package, showing a significant positive correlation between weight and body size (P<0.05), with the highest correlation coefficient between weight and chest circumference. There was a significant positive correlation between body weight and body size, and the correlation coefficient between weight and chest circumference was the highest. In conclusion, the rs402399470G>A and rs418440631G>A loci of PRSS12 gene could be used as potential molecular genetic markers for the growth traits of sheep. This study provides a reference basis for mutton sheep breeding and selection.

Thyroid hormone responsive spot 14 (THRSP) is a transcriptional regulator involved in the regulation of lipid metabolism in tissues. In order to understand the role of THRSP gene in lipid metabolism and its molecular mechanism, this study determined the expression of THRSP gene in 12 kinds of chicken (Gallus gallus) tissues, amplified the full-length CDS sequence of THRSP gene using the reverse transcription product of total RNA from chicken adipose tissue as a template, and constructed the eukaryotic expression vector of THRSP gene. After the THRSP gene was overexpressed and down-regulated in LMH (Leghorn male hepatoma) cells, the effect of lipid drop deposition was detected by oil red O staining, the content changes of total cholesterol (TC) and total triglyceride (TG) were determined using colorimetry method, and the expression changes of lipid metabolism-related genes were determined by qRT-PCR. The results showed that THRSP gene was highly expressed in adipose tissues and liver. Western blot analysis showed that THRSP-Myc fusion protein was successfully expressed in LMH. It was found that overexpression of THRSP gene could promote lipid drop deposition in LMH cells (P<0.001), TC (P<0.05) and TG (P<0.01) content significantly increased, the gene expression of peroxisome proliferator activated receptor γ (PPARγ)(P<0.01) and diacylglycerol acyltransferase 2 (DGAT2)(P<0.05) were up-regulated, and the gene expression of propionyl-CoA carboxylase subunit α (PCCA)(P<0.05) and phosphoenolpyruvate carboxykinase 1 (PCK1)(P<0.001) were down-regulated. When THRSP gene was knocked down in LMH cells, lipid drop deposition were inhibited (P<0.05), TC and TG contents were decreased (P=0.06), DGAT2 gene expression was down-regulated (P<0.05), while PCCA (P<0.01) and PCK1 (P<0.001) gene expression were up-regulated. The above results indicate that THRSP gene might play a role in promoting cellular lipid deposition by regulating the gene expression of PPARγ, DGAT2, PCCA, and PCK1. This study provides a new reference for further exploring the function of chicken THRSP gene.

Frankliniella occidentalis is an invasive pest that harms global agricultural production. Exploring the biological functions of immunophilins FK506-binding protein 6 (FKBP6) of F. occidentalis can provide a new target for the prevention and control of F. occidentalis. In this study, FKBP6 (GenBank No. XP_026292958.1) of F. occidentalis was analyzed through bioinformatics, the sequence of FKBP6 gene (GenBank No. XM_026437173.1) was cloned, expression of FKBP6 protein was induced through prokaryotic cells, and the polyclonal antibody was prepared. Then, the protein was expressed by the Bac-to-Bac baculovirus system, the polyclonal antibody and its subcellular localization were detected through Western blot and immunofluorescence (IF) labeling. The amino acid sequence alignment and phylogenetic tree analysis showed that FKBP6 proteins among 20 insect species had conserved regions, but they were only highly related among species. FKBP6 protein in Sf9 cells was expressed through baculovirus systems, and then IF and Western blot dection showed that polyclonal antibodies specifically bound to FKBP6 protein. Furthermore, subcellular localization results showed that FKBP6 was localized in the cytoplasm. This study provides molecular conditions for the study of the biological function of FKBP6.

Ustilago esculenta can induce the formation of the enlarged fleshy stem of Zizania latifolia after obligate infection, and its pathogenicity is closely related to the phenotype and yield of the swollen stem called Jiaobai. Pep1 (protein essential during penetration 1), as a fungal effector, plays an important role in the pathogenicity of Ustilago fungi. In this study, UePep1 (GenBank No. OR088111), the homologous protein coding gene of Pep1 in U. esculenta, was cloned by BLASTP mapping and sequence analysis, with an ORF of 528 bp, encoding 175 amino acids and no intronic sequences. The deletion of UePep1 gene did not affect the haploid morphology, growth rate, mycelial formation and filamentous growth of U. esculenta in vitro. However, it significantly inhibited the invaded mycelial proliferation of the U. esculenta in vivo, and could not induce the formation of the enlarged fleshy stem. Further supplementation of UePep1 into the SG200ΔPep1 strain of Ustilago maydis partially restored the infectivity of the SG200ΔPep1 strain, indicating that UePep1 is a functional complementary protein of the Pep1 in the smut fungi. The results provide a reference and theoretical support for further understanding of the interaction mechanism between U. esculenta and Z. latifolia and the formation mechanism of enlarged fleshy stem.

Low temperature will inhibit the growth and metabolism of denitrifying bacteria, resulting in poor nitrogen removal efficiency and substandard effluent quality in winter sewage treatment plants. The key measure to solve the problem is to screen the heterotrophic nitrifying and aerobic denitrification strains with high denitrification ability at low temperature. In this study, the low-temperature denitrifier strain was isolated from the activated sludge of a chemical plant in Inner Mongolia, and the strains was identified. Nitrogen removal characteristics were studied by using ammonia nitrogen, nitrate nitrogen and nitrite nitrogen as the only nitrogen sources. The effects of different single factors (carbon source, C/N, pH, rotational speed, temperature) on nitrogen removal performance were studied. The response surface method was used to optimize the key factors of nitrogen removal efficiency in order to determine the optimal nitrogen removal conditions. Finally, the isolated strain was applied to 2 different types of sewage in order to test its nitrogen removal effect. The results showed that the low-temperature resistant bacteria screened from sludge were identified as Enterobacter cloacae, and named N2 (GenBank No. OQ834747). The removal efficiency of strain N2 on ammonia nitrogen (304 mg/L), nitrate nitrogen (37 mg/L) and nitrite nitrogen (30 mg/L) reached 100% at 8 ℃. The strain N2 showed excellent removal ability of 300 mg/L ammonia nitrogen when carbon source of sucrose, carbon-to-nitrogen ratio (C/N) of 8~20, pH of 7~9, and rotation speed of 150~180 r/min, the optimal nitrogen removal conditions of strain N2 were as follows: carbon source of sucrose, C/N of 8.97, pH of 6.62, under these conditions, the removal rate of total nitrogen reached the highest (83.2%). Under the condition of low temperature of 8 ℃, the ammonia nitrogen removal rate of chemical plant sewage and coking plant sewage reached 100% and 91%, and the total nitrogen removal rate reached 99% and 80%, respectively. Therefore, N2 strain has excellent growth and nitrogen removal performance at low temperature and could be applied to wastewater nitrogen removal in cold areas.

The soil physicochemical properties, microbial community structure, and functional groups of artificially cultivated Pinellia ternata have undergone changes, resulting in low yield and unstable quality. However, the processing residue of Cinnamomum longepaniculatum is rich in organic matter and antibacterial components. Therefore, this study applied the waste residue of C. longepaniculatum to the cultivation of P. ternata in order to improve the yield and quality of P. ternata and realize the sustainable utilization of C. longepaniculatum. In this study, 3 treatment groups were set: The whole soil (CK), waste residue∶soil=3∶1 (V/V)(T6), the whole waste residue (T8), which began on March 15, 2022 and ended on November 15, 2022. The growth indicators of P. ternata were measured on April 15 and May 1, and the physiological indicators of P. ternata were measured on May 15. The disease situation of P. ternata was statistically analyzed throughout the process. After P. ternata fell, metagenome platform was used to conduct sequencing studies on the differences of rhizospheric microbial community and function, and determined the yield of P. ternata. The results showed that T6 and T8 both increased the number of seedlings, seedling height, leaf length and width of P. ternata, increased its protein content, malondialdehyde content, peroxidase activity, and proline content, but decreased its SPAD (soil and plant analyzer development) value and superoxide dismutase activity; T6 treatment could alleviate the disease of P. ternata, but T8 treatment was more serious; The yield of T6 treatment increased, but the yield of T8 treatment decreased; The proportion of bacteria in P. ternata rhizosphere soil (CK) was very high (96.32%~99.57%), while the proportion of fungi was very low (0.27%~0.82%). The number of bacteria in the T6 and T8 groups increased by 3.16% and 3.25% compared to CK, respectively, while the number of fungi decreased by 0.41% and 0.55%, respectively; After adding waste residue of C. longepaniculatum, the relative abundance of beneficial bacteria Proteobacteria, Steroidobacter, Povalibacter, Steroidobacter, Agromyces and Conexibacter increased. The relative abundance of harmful bacteria Acidobacteria was decreased; The abundance of COG (clusters of homologous groups of proteins) functional genes in the T6 and T8 groups increased, which was closely related to the genera Agromyces, Conexibacter, and Streptomyces. In summary, waste residue of C. longepaniculatum could improve the soil condition of P. ternata, and the T6 treatment group had a better effect. This study provides theoretical basis for the utilization of waste residue of C. longepaniculatum in P. ternata cultivation.

Epigenetics mainly studies the heritable changes in gene function that lead to phenotypic changes in the absence of alterations in DNA sequence, and is essentially a change in chromatin state. As an important part of epigenetic research, histone modifications play an important role in maintaining the genome stability, gene expression regulation and chromatin structure of eukaryotes. Their essence is the process of methylation, acetylation and ubiquitination under the action of related enzymes. Histone modifications are variety of various biological processes that regulate the development of tissues and organs. Mesenchymal stem cells (MSCs) are important members of the animal stem cell family and important sources of animal myogenic and adipogenic cells, and the lineage differentiation of MSCs is accompanied by reprogramming of gene expression, a process largely dependent on epigenetic regulation. Histone methylation and acetylation modification regulators act as a bridge between signal transduction and downstream gene reprogramming, playing a crucial role in the epigenetic remodeling of cell differentiation. This paper reviewed the types and mechanisms of histone modifications, and recent advances in the study of histone modifications on animal myogenesis and adipogenesis. This review provides a reference for exploring the mechanism of mesenchymal stem cell differentiation and provides a theoretical basis for further improving mammalian muscle quality.

Somatic cell nuclear transfer (SCNT) technology can be used to expand the production of top livestock and gene editing animals. So far, humans have successfully cloned more than 20 kinds of animals, but the efficiency of cloned animals obtained through somatic cell nuclear transfer technology is relatively low. Most somatic cell nuclear transfer embryos die before implantation. After nuclear transfer, the donor nucleus will undergo a series of reprogramming events to obtain the totipotency of embryonic development. The abnormal reprogramming of somatic cell nuclear transfer embryos is the main reason leading to the low efficiency of somatic cell nuclear transfer. In this paper, the abnormal reprogramming mechanisms of somatic cell nuclear transfer embryos were analyzed, such as chromatin remodeling, DNA methylation, variations and modifications of histones, sperm small RNA, etc., so as to provide reference for further research on the reprogramming of somatic cell nuclear transfer embryos.

Heavy metal pollution has become a major difficulty in soil environmental problems due to its refractory degradation and accumulation. Mycorrhizal fungi are able to form a reciprocal symbiotic relationship with the host plant, thereby improving the heavy metal resistance of the plant, which conducive to accelerating the vegetation cover of contaminated land, and can hold heavy metals to achieve the reuse of contaminated soil. The detoxification mechanism of heavy metals in host plants by fungi mainly includes structural fixation, nutrient dilution, gene regulation, chelation and auxiliary chelation, and antioxidant. In order to systematically understand the effects of different mycorrhizal fungi (endomycorrhizae and ectomycorrhizae) on hosts under heavy metal stress, the effect and mechanism of mycorrhizal fungi in enhancing the resistance of host plants to heavy metals were reviewed in this article, and future research directions were discussed. This review provides a theoretical reference for the application of mycorrhizal fungi in the remediation of heavy metal-contaminated soil.

At present, excellent yellow fruit tomato (Solanum lycopersicum) varieties are relatively scarce in the domestic market. In order to meet the market demand and improve the competitiveness of the seed industry, the tomato variety 'MoneyMaker' ('MM') was used to create yellow fruit by targeted gene editing of the tomato phytoene synthase 1 (SlPSY1) using CRISPR/Cas9 technology. Two sequences specific sgRNAs in the exon region of the SlPSY1 were selected as target sites to construct the CRISPR/Cas9 binary expression vector pKSE401. The 'MM' tomato material was genetically transformed using Agrobacterium tumefaciens mediated leaf disc transformation method. The results showed that 12 transgenic-positive plants were detected in 16 regenerated seedlings, with a positive transformation rate of 75%; Sequencing analysis of the target sites of the positive transgenic plants showed that 8 plants had a total of 21 editing events at 2 target sites, with a gene editing efficiency of 66.7%, and none of them was off-target; Five of the edited plants showed yellow fruit phenotype at maturity. In this study, CRISPR/Cas9 technology was used to successfully created SlPSY1 gene editing plants in 'MM' tomato, which will provide a technical and material basis for the rapid creation of tomato yellow fruit resources in the future.

The expression level of leucocyte cell-derived chemotaxin 2 (LECT2) reflects the health status of fish. In this study, the recombinant PaLECT2 (rPaLECT2) protein, expressed by Pichia pastoris, was used to prepare anti-PaLECT2 antisera in mice (Mus musculus) and New Zealand white rabbits (Oryctolagus cuniculus). Then the antibodies were purified from antisera by Protein G affinity chromatography. The mouse anti-PaLECT2 antibody was used as the capture antibody and the rabbit anti-PaLECT2 antibody conjugated with horseradish peroxidase (HRP) as the detecting antibody. A double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) method based on PaLECT2 protein was established after optimization. Finally, it was applied to detect the PaLECT2 in serum the Aeromonas hydrophila-infected ayu model and the prognosis model of antibiotic treatment. The results showed that the titer of PaLECT2 antibody in mice were 1∶12 800 and that of HRP-conjugated antibody in rabbits were 1∶25 600, respectively. The DAS-ELISA method for PaLECT2 used the optimal coating concentration of 8 μg/mL for the capture antibody and the best working dilution of 1∶1 600 for the detection antibody. The minimum detection limit of the method was 11.22 ng/mL with good specificity and stability. The protein concentration of PaLECT2 ((412±31.5) ng/mL) in the infected group was extremely significantly higher than that in the healthy group (54±7.2) ng/mL (P<0.001). The protein concentration of PaLECT2 in the treated group (225±56.1) ng/mL was extremely significantly lower than that in the infected group (P<0.01). In conclusion, the establishment of this method provides a more efficient and standard method for the quantitative measurement of PaLECT2 protein levels, which will be applied to the diagnosis and monitoring of ayu health status.