Nuclear transcription factor Y (NF-Y) is composed of 3 subunits, NF-YA, NF-YB, and NF-YC, and can specifically recognize CCAAT-BOX. In order to identify the members of NF-YB family in maize (Zea mays) and analyze their expression patterns in different tissues and physiological conditions, the NF-YB coding genes in maize genome were systematically identified and analyzed by bioinformatics methods. The expression patterns of NF-YB family members under hormone treatment were determined by spraying inbred line B73 plants with salicylic acid, jasmonic acid, and abscisic acid. The results showed that there were 18 members of the maize NF-YB gene family, all of which had NFYB domains and upstream binding sites for salicylic acid, jasmonic acid and abscisic acid. The experiment proved that the expression levels of maize NF-YB family genes increased significantly within 24 h after salicylic acid, jasmonic acid and abscisic acid treatment. It was predicted that NF-YB was involved in salicylic acid, jasmonic acid and abscisic acid pathway. Under cold, heat and UV stresses, the overall expression levels of family genes were down regulated; under salt and drought stress, the overall expression levels of family genes were increased. Under the infection of Fusarium graminearum, the expression levels of Zm00001d042196, Zm00001d010574 and Zm00001d019101 were increased. In this study, the maize NF-YB family genes were systematically identified and analyzed, the members of the maize NF-YB family were clarified, and the expression rules of maize NF- YB family members in maize biotic and abiotic stresses were revealed. This study provides an important theoretical basis for further elucidating the function of this family.

Dwarf tomato (Solanum lycopersicum) has both ornamental and edible values, and its abundant germplasm resources are the basis for germplasm innovating and varieties breeding. In order to study the genetic diversity of germplasm resources and screen more diverse types of germplasm resource materials for parental selection and variety breeding, 25 traits of 45 dwarf tomato germplasm resources from domestic and foreign were investigated, and genetic diversity analysis as well as cluster analysis were carried out; 33 pairs of InDel primers were used to PCR amplification to analyze their genetic diversity. The results showed that the genetic diversity index of 25 traits ranged from 0.26 to 2.03, the diversity index of the 7 mass traits ranged from 0.26 to 1.31, with the average value of 0.92, and the coefficients of variation of the 18 quantitative traits were distributed from 8.51% to 32.51%, with the average value of 19.01%. The mean number of alleles effective for InDel marker detection was 1.64, and the polymorphic information content (PIC) ranged from 0.18 to 0.43, with the average value of 0.31. Both agronomic trait cluster analysis and InDel marker cluster analysis divided 45 materials into 3 categories, and the agronomic traits of most germplasm were consistent with the clustering results of InDel markers. The results showed that 45 dwarf tomato germplasm resources had rich genetic diversity, which could provide reference basis for new varieties breeding.

Zizania latifolia stem swelling is caused by infection of Ustilago esculenta, following the inhibition of apical dominance of the stem, which may be related to the regulation of polar transportation of auxin. In this research, 6 ZlPINs (Z. latifolia PIN formed) of indole-3-aceticacid (IAA) polar transport vector related genes were cloned from Jiaobai 'Zhejiao 7'. The Mem_trans structural domains were found among the 6 ZlPINs. Phylogenetic tree analysis showed that ZlPIN1a, ZlPIN1b, ZlPIN2 and ZlPIN3a were highly similar to wild rice (Oryza brachyantha) with the similarity of 96.80%, 89.27%, 90.52% and 82.88% respectively. The expression of different tissues of Z.latifolia showed that all of the 6 ZlPINs were significantly higher expressed in root than those in leaf and sheath (P＜0.05). The expression of ZlPIN1a, ZlPIN1b and ZlPIN4 could be detected in the stem, while the expression level of ZlPIN1a was significantly higher than other ZlPINs genes (P＜0.05) . It was speculated that ZlPIN1a would be the major regulatory gene for the polar transport of IAA in the stem of Z. latifolia. The cloned sequence of ZlPIN1a (GenBank No. om782294) had a total length of 1 770 bp and encoded 589 amino acids, including 6 exons and 5 introns. Based on the infection of U. esculenta in Z. latifolia in vitro, it was showed that the expression of ZlPIN1a in stem was inhibited by the infection of U. esculenta (P＜0.05). Compared to Male Jiaobai, the significantly higher expression of ZlPIN1a in stem were found in Normal Jiaobai and Grey Jiaobai (P＜0.05). During the development of swollen stem, the expression of ZlPIN1a in Normal Jiaobai increased firstly and then decreased, however, the expression of ZlPIN1a increased continuously in Grey Jiaobai. The expression change of ZlPIN1a would be related to the 2 stem swollen development phenotypes induced by the infection of U. esculenta. In this research, ZlPIN1a was screened as the IAA polar transport related gene expressed in the stem of Z. latifolia, which could be inhibited by the infection of U. esculenta, The expression mode of ZlPIN1a in the development of swollen stem was discussed, which could provide a new idea for the regulation mechanism of swollen stem caused by the infection of U. esculenta in Z. latifolia.

Nucleotide binding site-leucine rich repeats (NBS-LRR) class of resistant gene takes the most part of resistance genes (R) and plays an important role in peanut (Arachis hypogaea) disease resistance. Our group discovered a candidate gene Arachis hypogaea resistance rust and late leaf spot 1 (AhRRLLS1) conferring resistance to late leaf spot in peanut by the QTL-seq approach. In order to explore the gene expression regulation, the AhRRLLS1 gene and its promoter were cloned using cDNA and gDNA of 'Yueyou 92' as templates. Sequence analysis of the AhRRLLS1 gene and its promoter was conducted via bioinformatic techniques. The qPCR detected the gene expression of AhRRLLS1 in the peanut varieties induced by exogenous hormone, the resistant and susceptible peanut varieties for late leaf spot disease. The GUS gene driven by AhRRLLS1 promoter was genetically transformed into Arabidopsis thaliana for histochemical GUS staining. The results showed that the AhRRLLS1 gene encompassed the coding region of 3 129 bp, and encoded the protein of 1 042 amino acids, containing TIR, NB-ARC and LRR conserved domains. Phylogenetic analysis showed that the AhRRLLS1 gene belonged to a TIR-NBS-LRR (TNLs) class of R genes. The qPCR analysis showed that the AhRRLLS1 gene could respond to abscisic acid, methyl jasmonate, ethylene, salicylic acid and Phaeoisariopsis personata. The gene expression level of AhRRLLS1 revealed a significant difference between the resistant variety of 'Yueyou 92' and the susceptible variety of 'Baisha 1016' (P＜0.05). The expression of AhRRLLS1 gene in 'Yueyou 92' was lower than that in 'Baisha 1016' in the first 3 days. The GUS staining for transgenic A. thaliana showed that roots, stems, leaves, flowers and pods could be stained. The results indicated that AhRRLLS1 belonged to a TIR-NBS-LRR class of resistance gene, playing a regulatory role in resistance against infection with Phaeoisariopsis personata in peanut, and the AhRRLLS1 promoter might drive the expression of the gene in different tissue of peanut. This study provides a theoretical guidance for characterization of AhRRLLS1 and the mechanism of disease resistance in peanut.

Transcription factor families play important roles in plant response to nitrogen stress. In order to reveal the expression pattern of transcriptome factor genes in the sesame (Sesamum indicum) under low nitrogen stress, high-throughput sequencing technology was used to analyze the gene expression profile of transcription factors in the roots of sesame varieties 'Zhengzhi HL05' (ZZ, nitrogen-tolerance) and 'Myanmar high-yielding' (MD, nitrogen-sensitive) with different nitrogen efficiency under low nitrogen stress for 3 and 9 d. The results showed that a total of 55 transcription factor families were detected by transcriptome sequencing, with a total of 1 662 transcription factor genes, and 275 differentially expressed genes, accounting for 16.55% of the total. The families with a large number of differentially expressed transcription factor genes including MYB (avian myeloblastosis viral oncogenehomolog), bHLH (basic helix-loop-helix), ERF (ethylene responsive factor), NAC (NAM, ATAF1/2, CUC1/2) and WRKY families. Compared with the control, under low nitrogen stress for 3 d, 86 and 74 differentially expressed transcription factor genes were detected in 'Myanmar high-yielding' and 'Zhengzhi HL05' cultivars, respectively, and the number of down-regulated genes was more than that of up-regulated genes. Under low nitrogen stress for 9 d, 178 and 128 differentially expressed transcription factor genes were detected in each cultivar, and the number of up-regulated genes was more than or equal to that of down-regulated genes. The transcription factor genes in response to low nitrogen stress showed cultivar- specificity. There were 113 and 54 transcription factor genes specifically expressed in 'Myanmar high-yielding' and 'Zhengzhi HL05', respectively. The Venn diagram of differentially expressed transcription factor genes showed that a total of 15 transcription factor genes were differentially expressed in different treatment groups, of which 8 genes up-regulated and 7 genes down-regulated. The results of qRT-PCR validation showed that the change trend of 15 differentially expressed genes was consistent with the sequencing results, which proved the validity of the sequencing results. This study provides a reference basis for further exploring the molecular response mechanism of sesame transcription factor family to low nitrogen stress.

The flavonoid-3', 5'-hydroxylase (F3′5′H) gene is crucial for the development of blue flowers in nature. To provide more resources and reference for blue flower breeding by cloning and expression analysis of F3′5′H in Lupinus polyphyllus, the petals of L. polyphyllus 'Lupine' were used as the material to clone LpluF3′5′H and the sequence was used for bioinformatics analysis by software such as MEGA 10.0 and online websites. The subcellular localization of LpluF3'5'H protein was determined by tobacco injection method. qPCR was used to analyze the expression of LpluF3′5′H in petals with different colors and developmental stages. The LpluF3′5′H cDNA sequence had the encoding region of 1 620 bp encoding 539 amino acids, and was named LpluF3′5′H (GenBank No. OQ657278). Bioinformatics predicted that LpluF3′5′H belongs to the CYP75A subfamily, cytochrome P450 superfamily. Phylogenetic analysis revealed that the protein encoded by LpluF3′5′H was the highest similarity to the F3'5'H protein of L. angustifolius, and the monocotyledonous plants F3'5'H protein clustered into another family. Subcellular localization showed that LpluF3'5'H was localized in chloroplasts. qPCR analysis indicated that the expression of LpluF3′5′H could be detected in different development stages and petals. Besides, LpluF3′5′H showed the stronger expression in 'Lupine Blue' and lower in 'Lupine White'. In the different developmental stages of 'Lupine Blue', the expression levels of LpluF3′5′H peaked in the second developmental stage and then decreased. Among the different petals of 'Lupine Blue', LpluF3′5′H was the most highly expressed in the wings, followed by the vexils, and lowest expressed in the keels. LpluF3′5′H might play an important role in the flower color formation of 'Lupine Blue', and the study provides a basis for further research into the flower color mechanism of blue-flowered L. polyphyllus formation.

Muscle water retention is one of the important characteristics of edible meat quality, and greatly affects the sensory characteristics and economic value of meat. In order to screen the differentially expressed genes associated with muscle water retention of Gannan dzo (Bos taurus), 3 yaks (Bos grunniens) and 3 dzoes, each with healthy growth and development, equivalent slaughter time and even weight, were slaughtered in line with standardization. The longissimus dorsi muscle was collected as the experimental material for the determination of cooking loss, pressure loss, dripping loss and RNA-seq transcriptome sequencing. The differential expression multiplier value |log2 (Fold change)|≥1 and the significance level P<0.05 were used as the conditions for selecting the differential expression genes. The cooking loss and dripping loss of yak were significantly higher than that of dzo (P<0.05), and the pressure loss was significantly lower than that of dzo (P<0.05). A total of 748 differentially expressed genes were obtained from the sequencing results, including 526 down-regulated and 222 up-regulated genes. In order to verify the reliability of the sequencing data, 6 differentially expressed genes were randomly selected for qRT-PCR verification. The gene expression trend was consistent with the transcriptome sequencing results, indicating that the sequencing results were reliable. GO function annotation and KEGG pathway enrichment analysis found that the items related to muscle water retention included fibrinolysis, protein decomposition, glucose metabolism and regulation of actin cytoskeleton, and the pathways involved in water retention include glycolysis/gluconeogenesis, peroxisome proliferator activated receptor (PPAR) and AMP-activated protein kinase (AMPK). A total of 55 nonredundant differentially expressed genes were obtained, of which troponin T type 2 (TNNT2), bone morphogenetic protein 1 (BMP1), fatty acid-binding protein 1 (FABP1) and low-density lipoprotein receptor- related protein-1 (LRP1) were related to the water retention of dzo, which could be used as candidate genes for genetic and breeding improvement of meat quality traits. This study provides basic data for further research on the molecular mechanism of water retention of bovine muscle.

Daqingshan goat (Capra hircus) is one of the typical meat and cashmere goat groups in Inner Mongolia. There is a certain correlation between the accumulation of fat in muscle or metabolic of farmed animals and breeding time, way and so on. The fat content in the muscle has a direct impact on meat quality. In order to explore the transcriptome differences of meat quality and the data information of related genes in Daqingshan goats at different breeding stages, 3 Daqingshan goats of different ages (1.5, 2.5 and 3.5 years old) were used as research objects, the meat quality index of the longissimus dorsi was determined and the transcriptome was sequenced and analyzed. Results showed that the 3 age groups had no significant difference in drip loss and pH, but the crude fat content and the shear strength 3.5 years of age were significantly higher than the other 2 age groups (P＜0.05). The results of transcriptome data showed that there were 468 differentially expressed genes (transcripts) between 1.5 and 2.5 years old, 487 differentially expressed genes (transcripts) between 1.5 and 3.5 years old, and 376 differentially expressed genes (transcripts) between 2.5 years old and 3.5 years old. The differentially expressed genes were enriched in 44, 41 and 28 metabolic pathways, respectively, and the differentially expressed genes in 3 age groups were enriched in 85 functional items. According to GO functional annotation and KEGG pathway enrichment analysis, 9 differentially expressed genes related to the metabolism or deposition of lipids were obtained. These genes were regulator of G protein signaling protein 2 (RGS2), estrogen-related receptor γ (ESRRG), peroxisome proliferators- activated receptor- γ coactivator 1 A (PPARGC1A), myotubularin-related protein 6 (MTMR6), ankyrin repeat domain 9 (ANKRD9), transmembrane protein 160 (TMEM160), XVI phospholipase A2 (group XVI phospholipase A2, PLA2G16), fos-like antigen 1 (FOSL1) and a disintegrin and metalloproteinase with thrombospondin motifs 4 (ADAMTS4). Randomly selected RGS2, ANKRD9, FOSL1 and ADAMTS4 genes were verified by qPCR technology. Results showed that the trend of 4 genes expressed in 3 different age group were basically identical with the transcriptome sequencing results. The differentially expressed genes obtained in this study can provide basic data for the further study of meat quality characteristics of Daqingshan goat.

Feeding high-concentrate diets often causes excessive visceral fat deposition and high incidence of metabolic diseases in ruminants. In this study, high throughput sequencing technology was used to systematically analyze the lipid metabolism of femoral and omental adipose tissue in sheep fed with different concentrate to forage ratios diets. 15 healthy Dumont male lambs (Ovis aries) aged (90±15) days were selected and randomly divided into 3 groups. Diets concentrate ratio were 30% (G30), 50% (G50), and 70% (G70). After 15 d of adaptation, sheep were raised for 60 d and slaughtered at the end. Femoral and omental fat from 3 lamb that close to average body weight of every group were sampled for transcriptome sequencing. KEGG pathway enrichment analysis were conducted to study more than 10% differentially expressed gene (DEG) between different groups. Relative gene expression of lipid metabolism related key enzymes in femoral fat were also determined by qPCR. The results showed that the dry matter intake of sheep in G50 group were significantly (P<0.05) higher than those in G70 and G30 groups. The body weight of sheep in G50 and G70 groups after 60 days of feeding was significantly (P<0.05) higher than that in G30 group. KEGG pathway enrichment result and essential gene expression changes combined to indicate fatty acid synthesis, elongation, desaturation, triglyceride metabolism and fatty acid degradation in femoral and omental fat of G50 group were all higher than those in the G30 group. The fatty acid elongation, desaturation and fatty acid degradation of femoral and omental fat in G70 group were higher than those in G30 group. The fatty acid synthesis and desaturation of femoral and omental fat in G50 group were higher than those in G70 group. The differential impact of diets concentrate ratio on femoral or omental fat were as follow: The triglyceride metabolism of omental fat in G70 group was higher than that in G30 group (Padjust<0.05). The fatty acid elongation and triglyceride metabolism of femoral fat in G50 group were higher (Padjust<0.05) than those in G70 group. The fatty acid degradation of omental fat in G50 group was significantly (Padjust<0.05) higher than that in G70 group. qPCR results showed that the relative expression of fatty acid synthase and stearoyl CoA desaturase in femoral fat of G50 group were significantly higher than those in G30 and G70 groups (P<0.05), which was consistent with the transcriptome results. This study revealed the similarities and differences in lipid metabolism of femoral fat and omental fat of sheep fed with different concentrate ratio diets which may provide insights for reducing visceral fat deposit and increasing feed efficience.

Quercetin (QUE), an ingredient of traditional Chinese medicine, is a flavonoid compound with anti-inflammatory and antioxidant effects. This study aimed to investigate the protective mechanism of QUE against lipopolysaccharide (LPS)-induced inflammatory damage in mouse (Mus musculus) mammary epithelial cells (MMECs), and provide a theoretical basis for the treatment of clinical mastitis in dairy cows (Bos taurus). First, the optimal inflammatory concentration of LPS, the optimal dose of drug QUE and LPS+QUE were successfully screened by the Cell Counting Kit-8 (CCK-8) in an in vitro experiment. After that, LPS was used to induce inflammation, and MMECs were divided into control, model (10 μg/mL LPS), LPS+QUE high dose (125 μg/mL), LPS+QUE medium dose (62.5 μg/mL), and LPS+QUE low dose (31.25 μg/mL) groups. The growth status of MMECs was observed by morphology, MMECs were identified by immunofluorescence, and the relative expression of inflammatory-related factors were measured by qRT-PCR and Western blot. The results showed the existence of specific protein CK-18 in the cells which proved that the cells were MMECs; after 12 h of cell inflammation induced by 10 μg/mL LPS, the gene expression of inflammatory-related factors TNFα and IL6 were significantly higher than that of the blank control (P<0.01), proving successful induction of inflammation in MMECs; After QUE acted on MMECs, qRT-PCR and Western blot showed that inflammatory- related factors TNFα, IL6, NF-κB, and AKT gene and protein expression were down-regulated and appeared to be dosage dependent. Therefore, QUE might ameliorate LPS-induced mastitis in mice by inhibiting the PI3K-AKT/NF- κB signaling pathway, thereby reducing the expression of downstream inflammatory factors TNFα and IL6, and exerting anti-inflammatory effects. This study provides basic materials for the treatment of clinical mastitis in cows and the development of potential anti-inflammatory drugs.

The phosphatase and tensin homologue deleted on chromosome 10 (PTEN) is a tumor suppressor gene. It plays an important role in cell growth, organ development and metabolism. In this study, the PTEN gene (GenBank No. OQ359981) was cloned using cDNA of Muscovy duck (Cairna moschata) ovary tissue as template, and bioinformatics analysis was performed. It was found that the full length of PTEN gene CDS was 1 212 bp, and the coded amino acid sequence was highly conserved. The expression of PTEN gene and protein in the kidney, liver, spleen, lung, heart, ovary and follicles of different grades were detected by qRT-PCR and Western blot. The results showed that PTEN was expressed in all the tissues, especially in ovary, spleen and heart; PTEN was expressed in both pre-hierarchical follicles and pre-ovulatory follicles, with the highest expression in small yellow follicles (SYF); Immunohistochemical staining confirmed that PTEN located in the granulosa cell layer of follicles. The above results suggest that the PTEN gene might be involved in the growth and development of duck follicles. The present study provides basic materials for further exploration of the regulation mechanism of PTEN gene in follicle development, and also provides new ideas for the study of duck genetic breeding.

Potato blackleg disease (PBD) is a bacterial soil-borne disease caused by the pathogen Pectobacterium atroseptica (Pat), which seriously obstructs the development of the potato (Solanum tuberosum) industry. In this study, endophytic fungi were isolated from the stem and leaf tissues of Heritiera littoralis using the patato dextrose agar medium (PDA). Fungal metabolites were obtained by solid fermentation of corn medium and rice medium, and the highly active metabolites with significant resistance to Pat were screened using the filter paper method. High activity strain HU0460 was identified by combining morphological observation and ITS sequence comparison. The active components in the metabolites were obtained by ethanol extraction, concentration, and normal silica gel column chromatography. The antibacterial mechanism of the active components on Pat was explored using determination of bacteriostatic and growth curve, observation of cell morphology by scanning electron microscope, and detection of membrane potential by DiBAC4 (3) fluorescence probe. Furthermore, the inhibitory effect of the active metabolites on Pat was analyzed through tuber inoculation. The results showed that the corn fermentation product of the endophytic fungus HU0460 had good antibacterial activity against Pat. HU0460 was identified as Nigrospora sp. by morphology and ITS sequence.The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the bacteriostatic components isolated from the HU0460 corn fermentation medium by ethanol extraction and normal silica gel column chromatography on Pat were 2.44×10-3 and 3.91×10-2 μg/mL, respectively. Results of scanning electron microscope (SEM) showed that the degree of morphological deformation of Pat increased with the increase of the concentration of active metabolites. When the concentration reached the MBC, the Pat cells broke and the content of the cells was lost. The DiBAC4(3) fluorescence probe detection showed that the treatment of HU0460 active metabolites resulted in the up-regulation of the fluorescence intensity of the bacteria and depolarization of the membrane potential of Pat fluorescenceintensity which led to the infectivity to the tube was further confirmed by the tuber inoculation experiment. In this study, the endophytic fungus Nigrospora sp. HU0460 against Pat was isolated and analyzed. The isolated bacteriostatic metabolites have a good antibacterial effect on Pat, providing strain and reference data for the biological control of the PBD and the subsequent agricultural antibiotic research and development.

Apple (Malus domestica) continuous cropping obstacle hinders the development of apple industry significantly, which inhibits the survival rate of apple seedlings, and lowers the yield and quality of apple seriously. In order to explore the effect of engineering soil improvement combined with biological fertilizer technology improvement on rhizosphere microbial community of replanted apple and the controlling and the apple rhizosphere soil samples after 1 and 2 years of engineered soil improvement from loam orchards were collected in Guandao Town, Qixia, Shandong Province. The effects of engineering soil modification on apple continuous cropping obstacle soil were investigated from the perspectives of microbial community composition and network stability by high-throughput sequencing of 16S rRNA and intergenic transcribed spacer (ITS) genes. The results showed that the α diversity of bacteria community was increased and community structure was changed significantly after soil improvement. At phylum level, the relative abundance of Proteobacteria and Actinobacteria were increased, while the abundance of Acidobacteria was decreased. At genus level, the abundance of Pseudomonas and Massilia were increased continuously. Arthrobacter, Streptomyces, Acinetobacter, Sphingobium and Williamsia were enriched significantly during the process of soil improvement. The α diversity of fungi was not affected obviously, however, the community structure of fungi was changed. At phylum level, the relative abundance of Ascomycota, Basidiomycota and Mortierellomycota were increased. At genus level, the abundance of Fusarium, Penicillium and Cladosporium were decreased, while Mortierella and Paraphaeosphaeria were enriched significantly. Meanwhile, the co-

： Porcine parvovirus (PPV) can cause porcine (Sus scrofa domesticus) reproductive disorder. Vaccination is an effective method to prevent pigs from PPV disease and improve the immunity and reproductive rate of sows. In order to efficiently express PPV structural protein VP2 gene and prepare PPV genetic engineering subunit vaccine, the PPV-VP2 gene was optimized and synthesized according to silkworm (Bombyx mori) codon frequency in this study. The VP2 gene was cloned to transfer vector and recombined into the parental virus BmBacmid with orf1629 gene deficient by co-transfection. The recombinant baculovirus rBmNPV (PPV-VP2) was obtained and used to infect silkworm. The expressed antigen was used to prepare subunit vaccine, and guinea pigs (Cavia porcellus) were used for vaccine efficacy evaluation. The expressed recombinant protein was confirmed by Western blot and mass spectrometry. The observation of electron microscope indicated that the virus like particles (VLP) could be self-assembled into a similar size and morphology to PPV. The results of hemagglutination assay (HA) indicated that expressed antigen titer per gram of silkworm pupae was equivalent to more than 200 doses of commercial vaccine. Guinea pigs were immunized with 3 different doses of PPV-VP2 antigen contents, all inoculated animals had a seroconversion, and the antibody could neutralize PPV in vitro. If a guinea pig was immunized with 5 mg of silkworm pupae (parallel to 200 doses of antigen in commercial vaccine), the hemagglutination inhibition (HI) and neutralizing

Senecavirus A (SVA), a new RNA virus belonging to the Picornaviridae family, which seriously endangers the development of China's pig industry. At present, there is no effective SVA vaccine. In order to study the new SVA subunit vaccine, this study was based on the viral structural protein 2 (VP2) gene sequences of SVA, and the B cell epitope sequences predicted by bioinformatic methods and the T cell epitope genes in the reported Foot-and-mouth disease virus (FMDV) 3A protein. The recombinant expression plasmids pET28a-rSVP2-B and pET28a-rSVP2-BT were constructed respectively after optimization codon and connecting in series with the flexible linkers. The recombinant tandem epitope proteins rSVP2-B and rSVP2- BT, which were expressed in prokaryotic system and purified by Ni+ ion chromatographic method, were identified by SDS-PAGE and Western blot, and then immunized BALB/c mice (Mus musculus) to evaluate their immunogenicity. The results showed that the target proteins were expressed correctly after the recombined plasmid pET28a-rSVP2-B and pET28a-rSVP2-BT were transformed into Escherichia coli BL21 (DE3) and induced by 1 mmol/L isopropy- β -D-thiogalactoside (IPTG). SDS-PAGE analysis showed that the relative molecular weights were 34 and 38 kD, respectively. Western blot results showed that rSVP2-B and rSVP2-BT had good reactivity with SVA positive sera. There was no significant difference in humoral immune levels between rSVP2-B and rSVP2-BT immunized mice with purification protein. Both of them induced strong specific IgG and IgG1 antibodies, and mainly induced humoral immune response biased towards Th2 type. The results of virus neutralization test showed that the neutralizing antibody titers induced by the two recombinant proteins ranged from 1∶64 to 1∶128. Meanwhile, mice immunized rSVP2-B and rSVP2-BT also produced strong cellular immunity. The mice spleen lymphocytes proliferation levels and interleukin-2 (IL-2) and interferon- γ (IFN- γ) expression levels in the mice vaccinated with the rSVP2-BT protein group was significantly higher than those in the rSVP2-B group. In conclusion, the recombinant tandem multi-epitope proteins rSVP2-B and rSVP2-BT with good immunogenicity were successfully prepared, and the immunogenicity of rSVP2-BT was superior to that of rSVP2-B. This study provides a theoretical basis for the development of SVA multi-epitope vaccine.

Semen quality is an essential contributor to reproductive efficiency in livestock production, as well as an important indicator of breeding selection and genetic potential of male livestock, which is of great significance for livestock production. As the largest microecosystem in domestic animals, gut microbiota participates in multiple physiological and pathological processes, including male infertility, obesity, inflammation and cancer, involved in material and energy metabolism, has an important impact on body health. In this paper, the progress on the efffect of gut microbiota on semen quality and reproductive performance of male livestocks were reviewed, This review provides novel ideas for improving semen quality and promoting efficient delivery of excellent genes.

Somatic cloning of animals has the problem of low efficiency, and its pregnancy success rate is only 1%~5%. Abnormal placenta of cloned animals is the main cause of pregnancy failure, including placental oversize, placental angiogenesis defect, trophoblast abnormality and so on. Although the mechanism of these abnormalities is unclear, the existing evidence shows that the abnormal expression of proteins and imprinted genes in the cloned placenta and the failure of key regulatory genes to establish correct apparent modification in the normal development of the placenta may be the main reasons for the morphological abnormalities and functional defects of the cloned placenta. This review describes the abnormal phenomena and possible regulatory mechanisms in cloned animals placentas, which could help us to improve the efficiency of subsequent somatic cell nuclear transfer in animals.

Rare sugars are a class of sugar molecules and derivatives with low availability in nature. Rare sugars and their derivatives show unique physiological functions and have been extensively used in the food and medicine industry. Recently, the production cost of rare sugars has been significantly reduced with the continuous development of new preparation methods, and its application in agriculture is becoming a new research hotspot among academia and industry. Starting from different functions of rare sugars in agriculture, this paper summarizes its application in regulating the expression of defense genes, inhibiting the development of pests and diseases, promoting plant growth, and protecting the storage of agricultural products, and outlines the biological preparation of agricultural rare sugars. Rare sugars have the advantages of being safe for food and not polluting the environment, therefore the application of rare sugars in agriculture is safe, green, and highly efficient, and expected to provide technical assistance for the sustainable development of our country's agriculture.

Mining and utilizing the allelic variation Wxa of high amylose gene particle bound starch synthase (waxy, Wx) is one of the hot spots in rice (Oryza sativa) breeding, but the key is to screen the gene Wxa conveniently. In this study, the visual loop-mediated isothermal amplification (LAMP) of rice high amylose gene Wxa was designed to explore a convenient screening method. Firstly, the Ex10-115(T/C) variant site and upstream and downstream consistent sequences of Wxa gene were identified as target sequences by sequence alignment. Then, using Primer3 Input, NEBcutter V2.0 and Primerexplore V5 software, cleaved amplified polymorphic sequences (CAPs) and LAMP markers were designed for genotyping. Finally, the visibility of the functional markers were tested. The results showed that there were conserved sequences near the Ex10-115(T/ C) variant site, from which CAPs markers (Wxa-2F and Wxa-2R) and LAMP markers (F3, B3, FIP and BIP) were designed. Using the CAPs marker to detect the Ex10-115(T/C) variant site in 10 rice cultivars, the gene Wxa was found in 'D62B', 'G46B', 'II-32B', 'ZS97B' and 'GLA4' those cultivars with higher amylose contents, while the other 5 cultivars did not carry Wxa that there's concordance between genotype and phenotype. The Wxa gene in rice was detected by LAMP marker combined with hydroxynaphthol blue (HNB) at 63° C for 60 minutes that the Wxa gene may be detected with the naked eye in rice varieties. The result was consistent with that of CAPs marker. In this study, a convenient method was designed to screen Wxa genes in rice. The results can provide references for the development and application of functional markers for other important genes in rice.