Heat shock transcription factor (Hsf) exists in most plants and plays an important role in the response to heat tolerance and other stresses. Family genes showed diverse characteristics and functions. In this study, 3 homoeologous genes TaHsfA1-1 (GenBank No. MW756130), TaHsfA1-2 (GenBank No. MW756131) and TaHsfA1-3 (GenBank No. MW756132) were obtained by homologous cloning technology from wheat (Triticum aestivum), their cDNA lengths were 1 590, 1 566 and 1 569 bp, respectively. Three protein sequences all contained the typical DNA binding domain (DBD), the same nuclear localization signal (NLS) sequence RRKP/KKRR and transcription activation domain sequence (aromaromatic, large hydrophobic and acidic amino acid residues, AHA) DSFWEQFLCA. The similarity of TaHsfA1-1, TaHsfA1-2 and TaHsfA1-3 to Aegilops tauschii AetHsfA1 was as high as 96%. Chromosome location analysis confirmed that TaHsfA1-1, TaHsfA1-2 and TaHsfA1-3 were located on chromosomes 4A, 5B and 5D, respectively. Through transient reporter assay with tobacco (Nicotiana tabacum) epidermal cells, it was found that the proteins was localized in the nucleus under the normal conditions. The qPCR analysis showed that the expression levels of the 3 genes were up-regulated by heat stress (HS) and salicylic acid (SA) treatment, respectively and down-regulated by abscisic acid (ABA) treatment, and TaHsfA1-3 expression was up-regulated by H₂O₂. Through assay in Saccharomyces cerevisiae AH109, TaHsfA1-1, TaHsfA1-2 and TaHsfA1-3 all had transcriptional activation activity. According to the expression level responsive to heat stress, TaHsfA1-1 was selected and transformed into Arabidopsis thaliana. Through observation the phenotype, it was found that TaHsfA1-1 could improve the basic thermotolerance and acquired thermotolerance of Arabidopsis thaliana seedlings and the chlorophyll contents of different transgenic lines under high temperature stress. The chlorophyll contents were consistent with the phenotype and survival rate. Hsp genes were induced to expression under the normal conditions and after heat stress in TaHsfA1-1 transgenic A. thaliana plants. The results indicated that TaHsfA1 could regulate thermotolerance, providing a theoretical basis and technical support for further exploring the characteristics and function of heat tolerance of wheat Hsf family.

NAC (NAM-ATAF1/2-CUC2) proteins are plant specific transcription factors, and play important roles in regulation of growth, development and stress response in plant. In the previous studies, TaNAC35 was cloned from wheat (Triticum aestivum) near iso-geneic line TcLr3ka based on the database of transcriptome RNA-seq from wheat and Puccinia triticina (Pt) interaction, in which the expression of TaNAC35 was significantly different between inoculated and uninoculated wheat. This study isolated the genome DNA (gDNA) sequence of TaNAC35 in TcLr3ka, and compared with CDS sequence by MEGA software. The results showed 4 exons and 3 introns in the gDNA sequence of TaNAC35. The promoter elements were predicted and analyzed, and the results showed that there were various of hormone responsive elements, light responsive elements and transcription factor binding sites. Recombinant plasmid pGR107-TaNAC35-GFP was constructed and transiently transformed into tobacco (Nicotiana tabacum), which revealed that TaNAC35 localized in the nucleus and the cell membrane. qPCR analysis showed TaNCA35 was highly expressed in leaves and might be involved in the response of wheat to Pt, abscisic acid and salicylic acid. Virus-induced gene silencing (VIGS) was employed to silence part of TaNAC35 sequence, the results showed that compared with TaNAC35 non-silenced wheat (control), the growth and development of Pt on TaNAC35 silenced wheat decreased significantly, which could inhibit the infection of the pathogen. The results in this study showed that TaNAC35 played a negative regulatory role in the resistance of wheat to Pt, which provides a reference for further exploration of the role of NAC transcription factors in the interaction between wheat and Pt.

VQ protein is a kind of non-specific plant protein with the conserved amino acid sequence of "FxxxVQxLTG". It is named as the VQ protein because it contains two highly conserved dipeptides (valine and glutamine). This family plays an important role in plant growth and development and stress response, but little research has been done in potatoes (Solanum tuberosum). In order to explore the functions of potato VQ gene family, the StVQ gene family members were analyzed by bioinformatics method, and the responses to various stresses were verified by qPCR. The results showed that 37 StVQs were identified in potato genome, named as StVQ1~StVQ37, which were distributed on 12 chromosomes in order, and 34 of them did not contain introns. According to the sequence characteristics, the family can be divided into 9 subfamilies. Sequence alignment results showed that there were 7 variation types in the StVQ family proteins. A couple of tandem repeat genes and 7 pairs of segmental duplication genes were founded with intraspecific collinearity analysis method. The protein network prediction analysis indicated that the StVQ family proteins interacted with the StWRKY. The RNA-seq data suggested that StVQ genes expressions varied at different tissue, and variations of expression level were also distinct under different conditions of stress. The further experiments of qPCR and promoter analysis demonstrated that those genes could respond to the abiotic stress of cold, salinity, drought, etc. The results of this study provide a reference for further functional verification and mechanism analysis of StVQ.

Polyphenol oxidase (PPO) widely exits in plants, animals and insects with multigene family members. It is the key enzyme causing browning for fruits and vegetables. It will be an effective means to reduce PPO gene expression and enzyme activity through grasp of its family members and their function. In order to further understand the function of PPO gene family members in lotus (Nelumbo nucifera), bioinformatics methods were used to identify members of the PPO gene family: PPO1, PPO4, aureusidin synthase 1 (AS-1) and AS-2. Its physicochemical properties, transmembrane region, signal peptide, subcellular localization, gene structure, conserved motif, secondary and tertiary structures were predicted, and PPO activity and gene response in different tissues, growth time and induction factors were detected by kit and qPCR, respectively. The results showed that the length of the CDS region of the 4 genes was between 1 740 and 1 833 bp, the number of encoded amino acids was 579 to 610, the molecular weight was 66.21 to 68.65 kD, and the isoelectric point was 5.87 to 8.59, all of which belonged to hydrophilic unstable proteins. PPO1 and PPO4 had no transmembrane region and no signal peptide, which were non-secretory proteins. AS-1 had no signal peptide but one transmembrane region. AS-2 was a secreted protein with one transmembrane region. PPO1 and PPO4 were located in the chloroplast, AS-1 was located in the endoplasmic reticulum, and AS-2 was located in the cytoplasm. PPO1 and PPO4 had no introns but both had 12 conserved motifs. AS-1 and AS-2 had one intron and 10 conserved motifs, respectively. The 4 proteins were very similar in structure, and the random coil was the main structural element of the proteins. The α-helix, β-turn and extension chain were dispersed throughout the polypeptide chain. The activity of PPO and expression of PPO1 and PPO4 were the highest in relatively young tissue buds, those of AS-1 was the highest in roots, and those of AS-2 was the highest in leaves. The activity of PPO and expression of PPO1, PPO4, and AS-1 were increased under different induction treatments, but AS-2 had no significant change; with the change of growth time, the PPO activity tended to increase first and then decrease. The expression of PPO1, PPO4 and AS-1 increased rapidly in around 30 d, while AS-2 did not change significantly. This study laid the foundation for further exploration of the characteristics and functions of the lotus PPO gene family.

Ground cover chrysanthemum (Chrysanthemum morifolium) is a perennial herb of the genus Chrysanthemum in the Compositae family, with neat flowers and diverse colors, which has high ornamental value. The low temperature in the north severely limits the cultivation range of ground cover chrysanthemum. The basic helix-loop-helix (bHLH) transcription factors are widely involved in plant growth and development and abiotic stress response. In this study, ground cover chrysanthemum 'Yingjie' was used as material, and 17 CmbHLHs were cloned based on transcriptome data, with ORF between 603 bp and 1 242 bp, and their physical and chemical properties were predicted. The physical and chemical properties of 17 CmbHLHs showed that 8 proteins were acidic and 9 proteins were basic, and these proteins were all located on the nucleus; The proportion of α-helix in the secondary structure of bHLH protein was 25.49%~51.50%, and the proportion of β-turn was 1.42%~5.88%; miRNA prediction analysis showed that 9 miRNAs might regulate the post-transcriptional expression of 8 CmbHLHs. Using MEME to predict the composition of CmbHLH motifs, a total of 6 conserved sequences were identified, except CmbHLH8 and CmbHLH15, all the other proteins contained motif 1 and motif 2. Phylogenetic analysis showed that 17 CmbHLHs were distributed in 12 bHLH subfamilies. Ground cover chrysanthemum seedlings were treated with low temperature, qPCR analysis found that the expression of 7 CmbHLHs increased under low temperature stress. This study provides references for further research on the function of ground cover chrysanthemum bHLH gene.

Tea plants (Camellia sinensis), as an important commercial crop, are one of the popular and economic beverages worldwide. Root formation of tea plants is essential for higher yields and stable quality. To elucidate the programmed root developmental processes, the root development associated genes (CsRDAs) were isolated and cloned by reverse transcription-PCR (RT-PCR), which was based on the root transcriptome library of Guizhou landrace tea cultivars, called C. sinensis cv. Qiancha 1 (QC1) and C. sinensis cv. QianMei601 (QM601). They were named CsRDA1 (GenBank No. MW451597), CsRDA2 (GenBank No. MZ516824), CsRDA3 (GenBank No. MZ516825) and CsRDA4 (GenBank No. MZ516826), respectively. Phylogenetic and the domain analysis showed that the CsRDAs were evolutionally conservative and contained similar motifs. qPCR results showed that the 4 members of CsRDAs could specifically express in different tissues and developmental stages, which showed that their functions may be tissue specific and developmentally controlled. Meanwhile, transient expression of CsRDAs fusion protein in Nicotiana benthamiana leaves revealed that CsRDA1 and CsRDA2 mainly localized in the nucleus and cytoplasm, while CsRDA3 localized in the chloroplast and cytoplasm. Furthermore, the transgenic rice (Oryza sativa) plants which overexpressed CsRDA1 were successfully obtained by Agrobacterium tumefaciens-mediated callus transformation. Preliminary results showed that constitutive expression of CsRDA1 increased the number of roots in rice plants. These data suggested that CsRDA1 may participate in the regulation of the development of tea plant root system. It provides a theoretical reference for further elucidating the regulation network of tea plant radicle development.

Seasonal estrus limits reproduction efficiency of sheep (Ovis aries), therefore, it is of great significance to explore key genes that affect the seasonal estrus of sheep. In order to explore the expression level and polymorphism of the F-box and leucine-rich repeat protein 3 (FBXL3) gene which was found by whole genome resequencing (WGRS) and its association with seasonal estrus in sheep (Ovis aries), the present study selected Sunite sheep as research object, qPCR was used to investigate the expression of FBXL3 gene in different tissues and the influence of SNP on its expression. SequenomMassARRAY^® SNP was applied to detect polymorphism of g.52551820G>A locus of FBXL3 gene in 2 different estrus patterns (year-round estrus group and seasonal estrus group). Then correlation analysis was carried out with seasonal estrus traits of sheep. Meanwhile, bioinformatics methods were used to analyze the effects of mutation on mRNA secondary structure and protein properties. The results showed that FBXL3 gene was widely expressed in the tissues of Sunite sheep, and 3 genetypes of GG, GA, and AA existed in the locus of FBXL3 g.52551820G>A. The expression of GG genotype was significantly higher than AA genotype in liver (P<0.01), kidney (P<0.01), and adrenal gland (P<0.05). The G was the dominant allele in seasonal estrus breed, while A was the dominant allele in year-round estrus breed. The genotype frequency and allele frequency of this locus were extremely significant different between seasonal estrous and year-round estrous breeds (P<0.01). Population genetic analysis showed that the g.52551820G>A locus of FBXL3 gene was at low polymorphism (PIC<0.25) in Prairie Tibetan sheep and Dorper sheep, and was at moderate polymorphism (0.25<PIC<0.50) in Tan sheep, Sunite sheep, Small Tail Han sheep and Suffolk sheep. Moreover, bioinformatics analysis showed that the hydrophilicity of FBXL3 amino acid residues near the mutation site decreased slightly, and the phosphorylation of the protein also changed. Altogether, the mutation at g.52551820G>A locus of FBXL3 gene might be a potential molecular marker of seasonal estrus in sheep, which provides a basic reference for studying the molecular mechanism of seasonal estrus.

Intramuscular fat (IMF) content was one of the important indicators to measure meat quality. However, the deposition of intramuscular fat content was affected by heredity, environment and nutrition level, and the deposition of intramuscular fat content was different among different varieties and strains, even among different individuals of the same strain. The aim of this study was to explore the differences in IMF content among different individuals of Guizhou yellow chickens (Gallus gallus domesticus) and to analyze the molecular regulation mechanism of IMF deposition in muscle of Guizhou yellow chickens by RNA-seq technique. 30 Guizhou yellow chickens hens were selected and raised in the same conditions to 120 d of age, and the IMF content in breast muscle was determined one by one after slaughter. There were 2 groups according to the level of IMF content: The high-IMF group (H group, n=4) and the low-IMF group (L group, n=4). In this study, RNA-seq technique was used to screen differentially expressed genes affecting IMF deposition in breast muscle of Guizhou yellow chicken; It also analyzed the differentially expressed genes for GO functional annotation and KEGG pathway enrichment. The results showed that a total of 259 differentially expressed genes were screened out from breast muscle tissues of individuals with high and low IMF content, including 64 up-regulated genes and 195 down-regulated genes. 6 genes were randomly selected for qPCR validation, and the qPCR results were consistent with the sequencing results, proved that the sequencing results were reliable. KEGG signaling pathway analysis showed that 5 signaling pathways, including focal adhesion, extracellular matrix (ECM) -receptor interaction, regulation of actin cytoskeleton, cell adhesion molecules, and TGF (transforming growth factor)-β signaling pathway, these pathways were significantly enriched. Calproteinase 2 (CAPN2), collagen type Ⅰ alpha 1 chain (COL1A1), COL1A2, collagen type Ⅵ alpha 1 chain (COL6A1), COL6A2, COL6A3, lipin1 (LPIN1) and phospholipid transfer protein (PLTP) were found in adhesion spots, ECM-receptor interaction, TGF-beta signaling pathway and PPAR signaling pathway. These genes may be key genes affecting IMF deposition in Guizhou yellow chickens. The differential expression genes identified in this study can be used as key genes for IMF deposition and provide a reference to explore the molecular regulation mechanism of IMF deposition in poultry in future.

Interleukin-1 receptor accessory protein (IL-1RAcP) is an important molecule involved in the pro-inflammatory cytokine interleukin-1 (IL-1) signal transduction, which plays critical roles in the inflammation, immune response and antimicrobial defense. To explore the mechanism of IL-1RAcP in the fish immune defense response to pathogen infection, the IL-1RAcP gene named shIL-1RAcP of snakehead (Channa argus) was cloned, and its immune response to the infection with 2 pathogenic bacteria and the stimulation with 3 pathogen analogues or recombinant IL-1β protein was preliminarily analyzed. The results showed that the full-length ORF of shIL-1RAcP was 1 848 bp (GenBank No. MW928496), which encoded a putative protein of 615 amino acids. Sequences analysis showed that shIL-1RAcP possessed a typical structural characterization composed of three extracellular Ig-like domains, one transmembrane domain, and one intracellular Toll/IL-1 receptor (TIR) domain. The phylogenetic tree analysis revealed that shIL-1RAcP clustered within a subgroup of other teleost species and shared the closest evolutionary relationship with the IL-1RAcP proteins of Micropterus salmoides and Seriola lalandi. Moreover, qPCR analysis showed that shIL-1RAcP was widely expressed with different expression levels in all tested tissues of healthy snakehead. The highest expression of shIL-1RAcP was in the liver, followed by several immune tissues, including the head kidney, spleen, and truck kidney, whereas the lowest expression was detected in the heart. After infection with Nocardia seriolae or Aeromonas schubertii, the relative expressions of shIL-1RAcP were mainly up-regulated in the head kidney and spleen in vivo at different time points, respectively. Similarly, after stimulation with three pathogen analogues and recombinant shIL-1β protein, the expressions of shIL-1RAcP in head kidney leukocytes in vitro could be induced by lipoteichoic acid (LTA), lipopolysaccharides (LPS), polyinosinic-polycytidylic acid [Poly (I:C)], and recombinant shIL-1β protein. In conclusion, these results illustrated that shIL-1RAcP was involved in the immune defense against the two pathogenic bacteria. Furthermore, this research provides several basic data for further revealing the mechanism of IL-1RAcP in fish species against bacterial infection.

Northern corn leaf blight (NCLB) occurrs on maize (Zea mays) leaf and is a foliar disease caused by Setosphaeria turcica. The physical properties of maize leaf, such as hardness, hydrophilicity, and hydrophobicity, can be perceived by S. turcica firstly when it infects maize leaves. The conidia germination rate, appressorium formation rate, lipid droplets, and glycogen distribution were observed. The results showed that the rate of conidia germination and appressorium formation on the hydrophobic substrate were higher than those on the hydrophilic substrate. The diameter of lipid droplets in germ tubes cultured on the hydrophilic substrate was larger than those cultured on the hydrophobic substrate. The distribution of glycogen in germ tubes was not significantly different. The surface morphology of the susceptible ('Xianyu 335') and resistant ('Zhengdan 958') variety to S. turcica was significantly different. The epicuticular wax layer of 'Zhangdan 958' leaf was rougher than that of 'Xianyu 335' leaf. These results indicated that the hydrophilicity of glass slide affects the development of the S. turcica conidia and provided data support for the breeding of maize.

Phosphorus solibilizating fungus can convert the potentially ineffective phosphorus into available phosphorus in the soil, which is an effective way to alleviate the phenomenon of soil phosphorus deficiency and increase the utilization rate of phosphorus fertilizer. The strain WR1-4 (Talaromyces sp.) was screened from the rhizosphere soil of soybean (Glycine max) in preliminary work. In order to clarify its phosphorus solubility and acid production, the phosphate solubilizing ability of strain WR1-4 was measured by plating method and broth method in present study; the molybdenum antimony anti colorimetric method was used to detect the phosphorus solubilizing characteristics of strain WR1-4 under different conditions of insoluble phosphorus, carbon and nitrogen sources; HPLC was used to detect the organic acids produced by WR1-4. The results showed that WR1-4 had different ability to dissolve 5 insoluble phosphates (calcium phosphate, zinc phosphate, aluminum phosphate, phosphate rock powder, iron phosphate), and the effective phosphorus increments were 693.07, 613.47, 260.14, 137.84 and 386.33 mg/L, respectively. Strain WR1-4 used 6 carbon sources in the order of glucose, sucrose, maltose, starch, lactose and cellulose. As to different nitrogen sources, strain WR1-4 had strongest phosphate solubilizing capacity under the condition of ammonium sulfate, and least phosphate solubilizing ability under the condition of potassium nitrate. Strain WR1-4 could produce 8 kinds of organic acids, including oxalic acid, lactic acid, citric acid, malic acid, acetic acid, fumaric acid, tartaric acid, and maleic acid. According to Pearson analysis, the correlation between the increase of available phosphorus and organic acid was positively correlated. At the same time, while strain WR1-4 was in the stage of phosphate solubilization, the pH of the environment decreased. Above results indicate that strain WR1-4 had a strong ability to dissolve phosphorus and a wide range of growth and metabolism conditions, which makes it has potential application prospects in microbial fertilizer development and agricultural production.

Biogas slurry irrigation in farmland can solve the problem of subsequent disposal and the reduction of chemical fertilizers, which has become one of the important ways of resource utilization of livestock wastewater. In this study, a 6-year field experiment was conducted around a pig farm in Jian'ou City, Fujian Province to investigate the effects of biogas slurry application on soil bacterial community of areca taro (Colocasia esculenta) in different seasons and depths. Soil samples were collected in summer and winter at two different depths (0~20 cm and 20~40 cm). The soil with no application of biogas slurry was also collected as the control group. Bacterial community composition and diversity were investigated using Illumina MiSeq sequencing technology, and its relationship with soil properties was discussed. The results showed that the contents of soil organic matter, alkali-hydrolyzable nitrogen and available potassium increased in the biogas slurry treatment over control treatment. Also the contents of them were generally higher at the 0~20 cm than that at the 20~40 cm soil layers. Compared with the control, the long-term application of biogas slurry had a positive effect on improving the bacterial richness. For bacterial diversity, it varied according to seasons. The richness and diversity of soil bacteria in different seasons basically decreased with increasing soil depth, and the application of biogas slurry changed their seasonal variation. Bacterial community structure in the soil varied with biogas slurry irrigation. Proteobacteria, Actinobacteria, Firmicutes were the dominant plyumn of all soil samples. Long-term application of biogas slurry generally increased the relative abundance of Proteobacteria, Planctomycetes and Nitrospirae, but it decreased Actinobacteria. Moreover, Massilia and Geobacter belonging to Proteobacteria increased significantly in the long-term biogas slurry applied soil (P<0.05), while the proportion of Mizugakiibacter showed an opposite trend. Through redundancy analysis, it was found that soil bacterial community structure was greatly affected by soil available potassium. Long-term application of biogas slurry caused changes in soil microenvironment in different seasons and depths, and then led to changes in community composition and diversity of soil bacteria. These results can serve as a theoretical basis for evaluating the effects of biogas slurry application on the soil micro-ecological environment.

Channa are economically important freshwater fish, high-density and intensive culture have led to the deterioration of its breeding environment and frequent diseases, which greatly hinders the development of snakehead breeding industry. To study the effects of lactic acid bacteria (LAB) on growth performance and intestinal microflora of hybrid snakehead (Channa argus (♀) × Channa maculata (♂)), the fish in experimental group (QXSY group) were fed in the basal diet supplemented with LAB, while those in control group (DZ group) were fed in the basal diet. The intestines and their contents were collected from DZ and QXSY groups on 0, 26, 60 and 73 d, respectively, for high-throughput sequencing to detect the intestinal microbial structure. At the same time, 90 fish were randomly sampled from each group to measure body length and weight at each sampling, and the feed consumption, morbidity and the rate of surviving fish were recorded. The results showed that the diversity of intestinal microflora in hybrid snakehead could be changed by supplementing LAB The diversity of intestinal microflora in QXSY3 group (the experimental group was sampled on the 60^th day) was higher than that in DZ3 group (the control group was sampled on the 60^th day), while there was no significant difference in Lactobacillus content between the two groups. Lactobacillus content in QXSY4 group the experimental group was sampled on the 73^rd day) was much higher than that in DZ4 group (the control group was sampled on the 73^rd day), at the same time, the diversity of intestinal microflora in QXSY4 group was lower than that in DZ4 group, indicating that LAB 'settled' and became the dominant microflora. Some bacterial communities also changed with supplement of LAB, Cetobacterium and Lactobacillus in DZ and QXSY group showed an opposite trend. At the end of the experiment, the growth rate of QXSY group was higher than that of DZ group by (12.0±2.3)%, and the feed coefficient of QXSY group (1.02) was lower than that of DZ group (1.13). The results showed that LAB in the feed of hybrid snakehead could improve the community structure of intestinal bacterial, increase the number of potential probiotics, reduce the number of potential harmful bacteria, optimize the intestinal environment of hybrid snakehead. This research provides a theoretical basis for the application of LAB in the healthy breeding of hybrid snakehead.

Chitinases (CHI) are a series of glycoside hydrolases (GHs) that break chitin into chitooligosaccharides ((GlcNAc)_n, 10≥n≥2) or N-acetylglucosamine (GlcNAc). In this study, a chitinase gene derived from Bacillus thuringensis, BtCHI1, was cloned and heterologously expressed in Escherichia coli BL21(DE3)plysS. The enzymatic properties, kinetic parameters, and substrate hydrolytic patterns recombinant enzyme BtCHI1 were characterized. The results showed that the molecular mass of BtCHI1 was approximate 75 kD. Its optimum temperature and pH were 40 ℃ and 7.0, respectively. The enzyme was unstable after heat-challenge at 50 ℃ for 1 h, retaining only 20.11% residual activity. However, the enzyme was considerably stable in broad pH buffers ranging from 3.0 to 10.0, retained over 80% residual activities after incubation for 1 h, respectively. Substrate kinetics assays indicated the recombinant BtCHI1 was active against chitin, colloidal chitin and chitosan, with V_max values of (4.62±0.46), (0.52±0.03) and (0.22±0.02) μmol/(min·mg), respectively. Nevertheless, the enzyme was inactive towards hydroxypropyl chitosan or glycol chitosan. Meanwhile, the enzyme was considerably resilient to 0.5~10 mmol/L Na⁺, Al³⁺ or EDTA (P>0.05). However, after treatment with 0.5~10 mmol/L Ca²⁺ or 5~10 mmol/L SDS for 1 h, the activities of BtCHI1 decline significantly (P<0.01). Thin-layer chromatography and high-performance liquid chromatography analysis revealed that BtCHI1 releases chitobiose from chitin and colloidal chitin, and further converted it into GlcNAc. Additionally, BtCHI1 (0.28 U) was observed to liberate (1.00±0.04) mg/mL and (1.71±0.11) mg/mL from natural shrimp and crab shell substrates. These results provide new insights into chitooligosaccharides/GlcNAc development and natural chitin substrate utilization.

Wnt is a secreted glycoprotein, which can bind to receptors in the form of paracrine or autocrine, activate intracellular signaling pathways, regulate target gene transcription, and then participate in the regulation of various cytological processes such as embryonic development and cell proliferation, especially playing an important role in skeletal muscle development. In this review, Wnt protein receptors and Wnt signaling pathways were reviewed, and the regulatory roles of Wnt pathways in skeletal muscle development and regeneration were expounded from the perspectives of ligands, regulatory factors and interactions with other pathways. This review not only provides ideas for the study of research on the regulation mechanism of skeletal muscle growth and development, but also lays a theoretical foundation for animal husbandry production.

The fate of SSC is divided into two kinds in mammalian spermatogenesis, one is self-renewal through mitosis, and the other is differentiation into sperm through meiosis. The balance and orderly transition between SSC's mitosis and meiosis is the key to spermatogenesis in male animals. The review focuses on the key regulatory and influencing factors of SSC's mitosis and meiosis, and key point is the regulation of retinoic acid, methylation of mRNA, and hormones during mitosis to meiosis , it would help us to understand the reproductive mechanism of male mammals, and provide references for the treatment of male infertility or the breeding of excellent sires.

In order to establish a rapid, efficient and simple method for detecting African swine fever virus (ASFV) antibodies, this study used a prokaryotic expression system to express ASFV capsid protein (p72) as a capture antigen, and anti-p72 polyclonal antibody and rabbit anti-pig IgG were coated on nitrocellulose membrane, as the quality control line (C line) and detection line (T line), respectively. The reaction conditions were optimized to prepare colloidal gold immunochromatographic test strip (ICTS), used for clinical sample detection. The results showed that the pCold-TF-p72 recombinant protein was successfully constructed in this study. Western blot and agar expansion experiments showed that the prepared anti-p72 polyclonal antibody had good reactogenicity and immunogenicity. The test strips could specifically detect anti-ASFV antibody and had no cross-reaction with positive sera against Pseudorabies virus, Porcine circovirus 2, Porcine reproductive and respiratory syndrome virus and Classical swine fever virus; In the detection of clinical samples, the positive detection rate of ICTS was 55.17% (48/87), the positive detection rate of a commercial ELISA kit was 59.77% (52/87), respectively. The agreement between ICTS and a commercial ELISA kit (IDVET) was 92.31% (48/52). The ICTS prepared in this study provides a reference for clinical ASFV antibody detection and disease prevention.

The yield of distiller's grains is huge, but it is not suitable for storage due to its high acidity and high-water content. Besides, the high proportion of rice husk in distiller's grains makes the fiber content of distiller's grains higher, which increases the difficulty of the utilization of distiller's grains. The resource utilization of distiller's grains is wide, for example, protein feed, fertilizer, chemical products and so on. The distiller's grains feed is the most common, such as silage feed, fly maggot feed, microbial fermentation feed, etc, and microbial fermentation feed is the research hotspot. However, most of distiller's grain feed has a single function, only pursuing high protein content, and other nutrients mainly rely on external addition. This increases the cost of distiller's grains feed, making it difficult to promote, and most of them can only stay at the laboratory level. In this study, the preparation of multifunctional microbial fermentation feed from distiller's grains by steam explosion pretreatment technology combined with microorganisms including Candida utilis, Saccharomyces cerevisiae, Bacillus subtilis, Lactobacillus, and Molds was studied. The distiller's grains treated under different steam explosion conditions were detected and analyzed. The results showed that the steam explosion pretreatment could effectively degrade the hemicellulose and lignin in the distiller's grains, which was beneficial to the reuse of the distiller's grains. The 1.7 MPa pressure treatment for 15 minutes was most conducive to the degradation of hemicellulose and lignin. Compared with untreated distiller's grains, the lignin and hemicellulose contents of distiller's grains treated by the optimum steam explosion conditions decreased by 14.65% and 51.72%, respectively, and the content of reducing sugar increased by 8.66 times. Then according to the ability of producing short chain organic acids and bacteriostasis, the feeding lactic acid bacteria were determined by using 3 strains of lactic acid bacteria combined with S. cerevisiae, C. utilis, B. subtilis and Trichoderma koningii to ferment, respectively. Finally, the optimal combination of S. cerevisiae, C. utilis, L. plantarum, B. subtilis, T. koningii, Aspergillus niger H7, A. niger H1496 and Myceliophthora thermophila was selected from different fermentation combinations of four kinds of molds, with the content of crude protein, crude fiber, nicotinic acid and acetoin as indexes. Compared with the distiller's grains pretreated under the optimized conditions of steam explosion, the content of crude protein and acetone increased by 22.55% and 76.38%, respectively, the content of crude fiber decreased by 17.85%, and the content of niacin reached 0.31 mg/g dry distiller's grains. In addition, there was also an appropriate amount of short chain organic acids, which had a pleasant sour fragrance. The results revealed that the study of multi-functional distiller's grains could not only make up for the lack of protein feed, but also solve the problem of distiller's grains with single function and high cellulose ratio. Therefore, functional distiller's grains feed will become the main research direction of distiller's grains feed in the future, which will help to realize the rational utilization of resources, alleviate the current situation of human and animal food competition, and promote the green development of traditional industries such as Baijiu industry.