Glyphosate can kill most herbaceous plants by inhibiting the 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) activity. Glyphosate-resistant (GR) crops can be developed by transforming the EPSPS gene encoding the glyphosate-resistant enzyme into crops. The current commercial application of GR EPSPS is mainly derived from microorganisms, and such "cross-kingdom" gene transfer is more likely to cause people's concerns about the safety of transgenic food. Therefore, improving the EPSPS of crops and then developing GR crops using crop endogenous genes is a desirable strategy to solve this problem. In this study, the Hotspots mutant library of EPSPS of Brassica napus was constructed, and the mutants resistant to high glyphosate levels were screened. 5 mutants with improved herbicide resistance were obtained, among which mBnEPSPS showed the highest glyphosate resistance. Compared with the wild type (BnEPSPS), the kcat of the mutant decreased from 515/s to 257/s, with a decrease of 50%. In the presence of glyphosate, the turnover numbers kcat (gly) of mBnEPSPS and BnEPSPS were 116/s and 13/s, respectively, meaning an 8.9-folds increase in glyphosate resistance. The mutation did not change the adaptation of enzymes to the plant cell microenvironment. After being transformed to tobacco (Nicotiana benthamiana), mBnEPSPS could confer tobacco resistance to roundup at 5 times higher than the commercial recommended dose of Roundup (41% glyphosate-isopropylammonium), suggesting that mBnEPSPS had a high breeding application value. This study provides genetic resources and a theoretical basis for developing herbicide-resistant rape seed through transgenic and gene editing.

Castration and long term fattening of bulls (Bos taurus) could promote visceral fat deposition, excessive fat deposition was easy to cause systemic chronic inflammation. However, there was very little known molecular mechanism of omental adipose tissue inflammation in cattle. The present study selected omental adipose tissue of cattle as research object, the aim of the present study was to evaluate the effects of sex and age on inflammation-related genes expression in omental adipose tissue of Holstein fattening cattle. Holstein bulls at 16 months of age (BF16 group, n=3) and Holstein steers at 16 and 26 months of age (SF16 and SF26 group, n=3) were selected. The growth performance and blood biochemical indexes were measured. After slaughtering, omental adipose tissue samples were collected for hematoxylin-eosin staining (HE) staining, transcriptome analysis was carried out for omental adipose tissue from three groups cattle by RNA- seq. Differentially expressed genes (DEGs) between groups was screened, GO function annotation, KEGG enrichment analysis, qPCR validation, candidate gene screening and protein-protein interaction network of differentially expressed genes were conducted. The results showed that a total 1 173 differentially expressed genes were identified, and enriched in 8 gene expression trend profiles. Functional analysis showed that the differentially genes were involved in multiple biological processes and signal pathways related to inflammation. Six differentially expressed genes were randomly selected for qPCR validation, and the results were consistent with transcriptome sequencing, which indicated that the sequencing results were reliable. Fifty non-redundant genes in 5 biological process items and 5 signaling pathways related to inflammation were screened. Cluster analysis showed that castration could upregulate the expression of anti-inflammatory and adipocyte differentiation genes (such as TNF alpha induced protein 3 (TNFAIP3), bone morphogenetic protein 2 (BMP2), etc) and downregulate the expression of proinflammatory genes (such as interleukin-1beta (IL1B)) in omental adipose tissue of SF16 group in comparison with BF16 group, long term fattening could upregulate the expression of genes related to aging and chronic inflammation (such as galectin 3 (LGALS3), C-C motif chemokine receptor 7 (CCR7), etc)) in omental adipose tissue of SF26 group in comparison with BF16 and SF16 groups. Protein-protein interaction network analysis showed that IL1B was the key central node. This research can serve as a theoretical basis for optimizing marbled beef production technology for Holstein steer.

Maize (Zea mays) is an important food crop, feed source and industrial raw material. With the development of bio-technology, transgenic and other new technologies are widely used in maize breeding. Cauliflower mosaic virus 35S (CaMV 35S) promoter and its enhancer are widely used in transgenic operation of maize and other crops. Therefore, studying the mechanism of CaMV 35S enhancer has important theoretical value for future application. Two trait mutants were screened from a maize activation-tagging mutant library previously constructed by the research group on genetic improvement of maize. Based on the identification of artificial transposon insertion sites in the chromosomes, the spatio-temporal variation of gene RNA expression in the range of about 100 kb upstream and downstream of the transposon insertion sites was analyzed using qPCR. The results showed that the enhancer of CaMV 35S significantly up-regulated the gene expression of adjacent genes (P<0.05), and the transcriptionally increased genes were distributed upstream and downstream of the transposon insertion site, with activation distance of up to 63.7 kb. Of the 4 genes up-regulated expression, the enhancement was dose-dependent, and the expression of the homozygous mutant was significantly higher than that of the heterozygous mutant (P<0.05). Although the CaMV 35S enhancer increased the transcription level of genes in the maize mutants, it did not significantly change the expression pattern of genes in different tissues. The scope, dosage effect, and tissue specificity of CaMV 35S enhancer in maize provide an experimental basis for subsequent use in maize molecular biology, and genetic engineering, and can also provide reference for the application of the promoter in other plants.

The NAC (NAM, ATAF1/2 and CUC2) transcription factor family is a collective name for genes with conserved NAC structural domains that play key regulatory roles in plant growth, development, and stress response and other life activities. In this study, 90 CcNAC candidate genes were initially identified from the Cajanus cajan genome by sequence alignment using the NAC family genes of Arabidopsis thaliana. Phylogenetic analysis showed that the 90 CcNAC genes could be divided into 13 subfamilies, and a total of 24 CcNAC family genes were screened as potential biotic stress response genes in subfamilies Group1~4 based on the study of Arabidopsis gene functions. Further physicochemical characterization and cis-acting element analysis of the 24 CcNAC family genes showed that the promoter region contains several phytohormone response elements, which might be closely related to hormone signaling and have potential responses to biotic and abiotic stresses function. Transcriptome analysis showed that 13 NAC genes were significantly up- regulated and 5 NAC genes were significantly down-regulated at 12 h of treatment under the effect of MeJA, among which CcNAC55 was significantly up-regulated at 3, 6 and 12 h of treatment, and the highest expression levels were found at 6 and 12 h. Further, using semi-quantitative assays, CcNAC55 was found that it was sustaining significantly up-regulated under fungal Cc1-1 stress, while there was no significant difference under high temperature stress, suggested that CcNAC55 might play an important role in the response of Cajanus cajan to biotic stress. To verify the function of CcNAC55 in response to biotic stress, in this study, the lines of Cajanus cajan overexpressed CcNAC55 were established by vacuum osmotic transformation and inoculated the leaves with the pathogenic fungus Cc1-1. The results showed that CcNAC55 overexpression could significantly reduce the infestation rate and proportion of infected area of Cajanus cajan leaves. This study provides a reference for the study of NAC gene function in Cajanus cajan and will help to identify and select candidate genes related to biotic stress tolerance.

Aquaporin (AQP) is a small molecule membrane intrinsic protein, which mediates the rapid and passive water transport and participates in transmembrane transport of plants. In this study, 41 genes of AQP gene family were identified according to non-redundant protein sequence database (Nr) annotation from the whole genome of Osmanthus fragrans. According to the analysis of amino acid sequence and phylogenetic tree, they were classified into 4 subgroups: 16 members of plasma membrane intrinsic proteins (PIP), 14 members of tonoplast intrinsic proteins (TIP), 8 members of nodulin-26-like intrinsic proteins (NIP), 3 members of small intrinsic proteins (SIP). The gene structure and conserved motif of amino acids in the same subgroup were highly conserved. The length of the AQP proteins ranged from 145 to 432 aa with molecular mass of 14.81 to 45.90 kD and the isoelectric points were from 4.84 to 9.81. The analysis of the expression patterns of OfPIPs and OfTIPs in different tissues of O. fragrans using qPCR showed that OfPIP1, OfPIP5, OfPIP12, OfPIP13, OfPIP14, OfPIP15, OfPIP16, OfTIP1, OfTIP3, OfTIP4, OfTIP6, OfTIP9 and OfTIP13 were specific expressed in flowers. Further study on their expression patterns during flower opening period of O. fragrans showed that, OfTIP6 was highly transcribed in the early flower opening period (S1~S2) of O. fragrans. It is speculated that OfTIP6 plays an important role in vacuole expansion at the early flower opening stage. However, the relative expression levels of OfPIP1, OfPIP13, OfPIP14, OfPIP15, OfPIP16 and OfTIP13 were significantly up-regulated at the late flower opening period (S4~S5) of O. fragrans (P<0.05). It is speculated that they are the key genes to exercise function at the late flower opening stage. This research provides a reference for further analyzing the biological function of OfAQPs protein.

The unusual floral organs (UFO) gene and its orthologous genes have an important role in the regulating the identity of floral meristem and the development of floral organs, and its functions have been revealed in many species. AmFimbriata (AmFIM) is the earliest cloned UFO like gene, compared with other UFO like genes, it has certain differences in function and gene interaction. In addition, the study of AmFIM in Antirrhinum majus was limited to transposon-mediated mutants. Therefore, the function of AmFIM gene needs further verification and improvement. In this experiment, the pBI35S:: AmFIM overexpression vector was heterologously transformed in Solanum nigrum, the morphological observation found that the transgenic S. nigrum plants showed obvious dwarf, early flowering phenotype, and the difference in AmFIM gene expression had different effects on S. nigrum cymes and floral organs. In weakly phenotypic lines, the number of flowers in each inflorescence was reduced, the apical inflorescence architecture was complicated, and the floral organs were not significantly different compared with the wild type; in strongly phenotypic lines, the cymes were transformed into single flowers, the floral organs were homozygously transformed, the sepals became larger, petaloid tissue appeared in sepals and stamens, staminoid tissue appeared in carpels, and sepals fused with petals, petals fused with stamens. The internal structure of stamenloid carpel of transgenic S. nigrum was further observed by paraffin section, and it was found that there were tissues similar to anthers and pollen. The results showed that the overexpression of AmFIM gene in S. nigrum affects the inflorescence architecture and the development of floral organs, reflecting the function of AmFIM gene in regulating the determinacy of floral meristems, floral organ differentiation and maintaining floral organ boundaries. This study further improves the research content of AmFIM gene and provides a reference for the molecular regulation mechanism of UFO like genes.

Tobacco (Nicotiana tabacum) is photophile crop, and its growth can be affected by shading. In order to deeply understand the response mechanism of tobacco under shading stress, explore the effect of shading on photosynthetic physiology of tobacco leaves and the changes of tobacco leaf transcriptome, this study took 'Yunyan 87' as the test material, measured the chlorophyll content and photosynthetic parameters of tobacco leaves under natural light and 40% shading rate, and sequenced the transcriptome of tobacco leaves. The results showed that compared with the control, the contents of photosynthetic pigments chlorophyll a (Chla), Chlb, carotenoids and Chla+Chlb increased (P<0.05) in the upper tobacco leaves of the shading treatment group, and Chla/Chlb did not change significantly. The photosynthetic parameters, net photosynthetic rate (Pn), decreased significantly (P<0.05), and there was no significant change in stomatal conductance (Gs), transpiration rate (Tr) and intercellular CO2 concentration (Ci). Transcriptome data analysis showed that there were a total of 10 110 differentially expressed genes (DEGs) between the control group and the shading group, of which 4 438 DEGs were up-regulated and 5 672 DEGs were down-regulated. GO analysis showed that DEGs were closely related to binding, catalytic activity, cellular fraction, membrane fraction, cellular process, and other items. KEGG analysis found that DEGs were mainly enriched in photosynthesis-antenna protein, plant hormone signal transduction, phagosome, glutathione metabolism, starch and sucrose metabolism, glycerolipid metabolism, porphyrin and chlorophyll metabolism. For photosynthesis-related genes, shading significantly increased the expression of some genes of photosystem Ⅰ (PSA) and photosystemⅡ (PSB), while PsaB, PsbS, PsbR and other genes were down-regulated, and the genes related to photosynthetic electron transport pathway and F-type ATPase were down-regulated. For porphyrin and chlorophyll metabolism-related genes, the genes related to chlorophyll synthesis were up-regulated, and the genes related to chlorophyll degradation were down-regulated. The results of qRT-PCR validation showed that the expression trend of DEGs was highly consistent with that of RNA-seq. This study provides experimental basis for the application of stress-resistant molecules in tobacco planting and new variety breeding.

CIDEA is a lipid droplet (LD) -associated protein that is highly expressed in the lactating mammary gland, mediates the transformation of small lipid droplets into large lipid droplets and promotes triglyceride (TAG) accumulation. In order to detect the effect of CIDEA gene on lipid metabolism-related gene expression and TAG synthesis in mammary epithelial cells of dairy cows (Bos taurus), the expression of CIDEA gene in mammary tissues of dairy cows during dry milk and lactation was detected by qPCR technique; the synthesis of siRNA targeting CIDEA gene was designed to detect the expression of genes related to lipid metabolism; and the intracellular TAG content was detected by TAG kit. Results showed that the sequence of the CDS region of the full-length 660 bp cow CIDEA gene (GenBank No. MW960011) was obtained by cloning. Bioinformatics analysis revealed that the nucleotide sequences of B. indicus (GenBank No. XM_019986952.1), B. mutus (GenBank No. XM_005892367.1) and Bubalus bubalis (GenBank No. XM_006043025.3) were 99.73%, 99.46% and 98.52%, respectively, and the amino acid sequence homology was more than 95%. The qPCR technique revealed that the expression of CIDEA gene was 1.5 times higher in dairy cows during peak lactation than dry period; siRNA was transfected in dairy cows mammary epithelial cells and the interference efficiency reached over 95%. Interfering with the CIDEA gene significantly downregulated the expression of sterol regulatory element binding protein 1 (SREBF1) and acetyl coenzyme A carboxylase alpha (ACACA), and inhibited the expression of diacylglycerol acyltransferase 1 (DGAT1), DGAT2, lipin 1 (LPIN1), and heart fatty acid binding protein 3 (FABP3), cluster of differentiation 36 (CD36)(P<0.05), and promoted TAG hydrolase (ATGL) expression (P<0.05). After CIDEA interference, a significant decrease in intracellular TAG content was detected (P<0.05). In conclusion, CIDEA plays an important role in regulating genes related to lipid metabolism and TAG synthesis in BMEC. This study further clarifies the regulation mechanism of lipid metabolism in dairy cows, which has important theoretical and practical implications for the healthy development of the dairy industry.

A certain concentration of lysobisphosphatidic acid (LPA) can promote oocyte maturation, while cumulus cells and oocyte-secreted factors (OSFs) can guarantee the normal development and function of oocytes. The purpose of this study was to investigate the expression of oocyte-secreted factors (OSFs) regulated by exogenous lysophosphatidic acid (LPA) in cumulus cells. The cumulus cells of Bos grunniens cultured in vitro were used as the research object, and exogenous LPA (0, 5, 15, 30 and 50 µmol/L) was supplemented into the cultured cumulus cells. PCR, qPCR, immunofluorescence tech and Western blot (WB) were used to evaluate the expression status of OSFs after exogenous LPA regulation. qPCR results showed that when the concentration of exogenous LPA was 15 µmol/L, the relative expression levels of growth differentiation factor 9 (GDF9) and fibroblast growth factors (FGF10) mRNA were the highest. And when the concentration was greater than 15 µmol/L, the expression levels of GDF9 and FGF10 mRNA decreased. The relative expression levels of bone morphogenetic proteins 1 (BMP1) and BMP15 mRNA were the highest when the concentration of LPA was 30 µmol/L. However, when the concentration was greater than 30 µmol/L, the expression of BMP15 mRNA decreased, and the relative expression level of BMP1 mRNA did not change significantly. Western blot results showed that when the concentration of LPA was 30 µmol/L, the relative expression level of GDF9, BMP1 and BMP15 protein was the highest. However, when the concentration of LPA was more than 30 µmol/L, the expression levels of BMP1 and BMP15 proteins were decreased, while the expression of GDF9 protein was inhibited. Compared with the response of other OSFs proteins to LPA, the relative expression level of FGF10 protein and the concentration of added LPA were dose-dependent within a certain range of LPA concentrations. Immunofluorescence results showed that the fluorescence labeling of OSFs in the cumulus cells of 0 µmol/L treatment group was mainly concentrated in the cytoplasm and nucleus, while the fluorescence labeling of OSFs in the cytoplasm of cumulus cells was significantly (P<0.05) enhanced after exogenous LPA treatment. This study confirmed that exogenous LPA can regulate the expression of OSFs in yak cumulus cells at the gene and protein levels during in vitro culture, and the addition of appropriate concentration of LPA can increase the secretion of related cytokines in yak cumulus cells. In turn, the development of oocytes is regulated. This study can be used as a reference for yak and other mammalian oocyte maturation in vitro, and is helpful to the development of related cell research.

Long-chain non-coding RNA (lncRNA) has become an important participant in almost all gene functions and regulatory levels. In order to reveal the potential effect of lncRNA on the milk yield of horses (Equus caballus) in early lactation, 8 healthy Kazakh horses of 3~4 years old with similar body shape and early lactation were selected in this study, and they were divided into high milk yield group (H) and low milk yield group (L) according to their milk yield. The non-coding RNA in the milk of 2 groups of horses was sequenced by transcriptome sequencing technology. Through the significant enrichment analysis of GO and Pathway functions, the main biological functions of differentially expressed genes, biochemical metabolic pathways and signal pathways involved were determined. According to P<0.05 and FC≥1, the differentially expressed lncRNA was screened out, and the function of the differentially expressed lncRNA was further studied. The results showed that a total of 30 differentially expressed lncRNA (up-regulated 16, down-regulated 14) were found in Kazakh horse milk of high-yield group and low-yield group in early lactation. Genes related to breast development and lactation, such as vascular endothlial growth factor, (VEGFD), phosphatidylinositol glycan, class A (PIGA), and aconitase closing odor (ACO1) were screened out. The results showed that lncRNA may regulate the process of lactation or mammary gland development by targeting genes related to mammary gland development and lactation. This study provides reference for the application of lncRNA in horse breeding.

Pregnant mare serum gonadotropin (PMSG) combined with gonadotropin-releasing hormone (GnRH) can improve reproductive efficiency by stimulating the ovaries of mice (Mus musculus), but its effect on embryo implantation remains unclear. This study aimed to explore the effect of PMSG combined with GnRH on embryo implantation in mice. In the experiment group, mice were used to inject 3.5 µg of GnRH agonist leuprorelin 48 h after 7.5 IU of PMSG injection (referred to as P+G group), and the control group was injected twice with saline. Normal blastocysts were transferred into the uterus of pseudo-pregnant mice in the P+G and control groups, then embryo implantation rates were analyzed after 5 d embryo transfer, and the expression of endometrial receptivity marker gene - leukemia inhibitory factor (Lif) was detected. Transcriptome sequencing data were analyzed to reveal the effect of P+G on endometrial receptivity, and qRT-PCR and Western blot were used to verify the data accuracy. The results showed that the embryo implantation rate of P+G group was significantly lower than that of the control group (P<0.05), and the expression of endometrial receptivity marker gene Lif was significantly reduced (P<0.05) in the P+G group. Transcription group sequencing analysis found 751 differentially expressed genes (DEGs), of which 587 up-regulated and 164 down-regulated. GO and KEGG analysis found that DEGs mainly enriched in the immune response, immune system process, cell adhesion molecules (CAMs), and cytokine-cytokine receptor interaction pathways. The results of qRT-PCR and Western blot showed that the expression of Lif gene and protein were down-regulated and the expression of Lcn2 and Prap1 gene and protein were up-regulated in the P+G group, which was consistent with the results of transcriptome sequencing. This study provides a certain theoretical basis for in- depth exploration of the regulatory mechanism of PMSG and GnRH involved in embryo implantation.

E2F transcription factor 1 (E2F1) and TATA-binding protein (TBP) jointly participate in the gene transcription regulation. To investigate the interaction between chicken (Gallus gallus) E2F1 and TBP proteins and their gene expression characteristics in tissues, in this study, the recombinant eukaryotic expression vectors of chicken E2F1 and TBP genes were constructed and then transfected into human embryonic kidney cells (HEK-293T). Indirect immunofluorescence and co-immunoprecipitation (Co-IP) assays were performed to verify their interaction. In addition, qPCR assay was used to examine the expression levels of E2F1 and TBP genes in different tissues at 14 embryonic days (E14 d) in chick embryos and 1-day-old (H1 d), 7-day-old (H7 d), 14-day-old (H14 d) after shelling. The results showed that the recombinant vectors pCMV-HA-E2F1 and pCMV-Myc-TBP of chicken were successfully constructed, and the recombinant proteins HA-E2F1 and Myc-TBP were correctly expressed and mainly localized in the nucleus in plasmid-transfected cells. The results of tissue characteristics analysis showed that chicken TBP and E2F1 genes were expressed in different chicken tissues at 4 stages, and the expression levels were significant different (P<0.05). At E14 d, the expression of TBP and E2F1 genes was higher in lung, but lower in brain; at H1 d, the 2 gens had high expression level in liver, but low in brain; at H7 d, they showed high expression in brain, but low in leg muscles; at H14 d, there were high expression in lung, liver and heart, but low in chest and leg muscles. Chicken E2F1 protein could interact with TBP protein in the nucleus, and the expression levels of E2F1 and TBP genes were relatively high in lung and liver at different developmental stages. The results provide references for further studying the mechanism of E2F1 and TBP protein interaction regulating the development of chicken lung and liver tissues.

Tilapia (Oreochromis niloticus) is one of the most important commercial freshwater fish in China. Understanding the effect of different initial feeding on the intestine microbiota of tilapia can provide a basis for the scientific cultivation of tilapia fry. This study aimed to explore the influence of different initial feeding on the morphology and the structure of microbial community of tilapia gut. All female fry of 11 days post-fertilization (dpf) were fed with 3 diets for 20 d, including Artemia salina (FA group), high protein artificial diet (protein level=25.5%) (FH group), and low protein artificial diet (protein level=16%) (FL group). The samples were collected on 11 (before feeding), 20 and 30 dpf, respectively. The results showed the survival rate of tilapia fry in FL group was significantly different from that in FA and FH group (P<0.05), with the highest value in group FA ((98.00±0.54)%) and the lowest value in group FL ((92.00±1.50)%). The results of intestinal histology showed that the folds were damaged and sparsely arranged in the fish intestine of the group FL. High-throughput sequencing analysis showed that the abundance and the diversity of intestinal microbiota in the group FA at 20 dpf were the lowest, the abundance of intestinal microbiota in the group FL at 30 dpf and the diversity of intestinal microbiota in the group FA at 30 dpf were the highest. At the phylum level, Proteobacteria was the dominant microbiota in the intestine of tilapia fry in FA, FH, and FL group, whereas Rhodobacter and Gemmobacter were dominated at the genus level. Compared with groups FH and FL, the relative abundance of Rhodobacter and Cetobacterium in group FA was significantly increased (P<0.05). Compared with group FA and FL, the relative abundance of Aeromonas and Pseudomonas in group FH was significantly increased (P<0.05). Compared with groups FA and FH, the relative abundance of Shinella and Brevifollis in group FL was significantly increased (P<0.05). The above results showed that, compared with the formulated feed, A. salina feeding could improve the survival rate of tilapia fry, improve the intestinal tissue structure, and have a beneficial effect on the intestinal microflora structure; The survival rate of tilapia in the high-protein diet group was significantly higher than that in the low-protein diet group, and there was no significant difference from the A. salina group, which could be used as initial feeding for tilapia. This study provides basic data for the selection of tilapia initial feeding.

Infectious hematopoietic necrosis virus (IHNV) infection causes much higher morbidity and mortality in small-sized rainbow trout (Oncorhynchus mykiss) than in large-sized fish. In order to explore the possible mechanism of this difference from the perspective of intestinal and hepatic immune response and microecological health of fish, 2 sizes of rainbow trout fingerling were selected from the same cage culture area where IHNV infection was detected. Using qPCR and 16S ribosomal DNA identification (16S rDNA) comparative analysis the Toll-like receptor 2/7 (TLR2/7), interleukin-8/18 (IL-8/18), interferon (IFN), interferon regulatory factor 9 (IRF9), Myxovirus resistant 1 (MX1) and vipern (VIG-1) genes were involved in the immune response mechanism and intestinal microflora differences. The results showed that among the 8 genes examined, TLR7 and MX1 were significantly higher expressed in the intestine of small-sized rainbow trout than large-sized fish (P<0.05), IL-8 was significantly higher expressed in the liver of small-sized rainbow trout than the large-sized (P<0.05), and the intestinal microbial diversity of large-sized rainbow trout was higher than that of small-sized fish. The abundance of Actinobacteria and Bacillus in the intestine of large- size rainbow trout (Oncorhynchus mykiss) fingerling was significantly higher than that of small-size fish (P<0.05), and the abundance of Serratia in the intestine of small-size rainbow trout was significantly higher than that of large-size fish (P<0.05). In conclusion, small-sized fish may have higher susceptibility to pathogenic stressors such as viruses that may be present in the environment, the intestinal micro-ecosystem was less stable compared to large-sized, and the abundance of beneficial bacteria present in the intestine of large-sized rainbow trout that may be related to a higher disease resistance. The results of this study elucidate the differences in immune genes and intestinal microorganisms of different sizes of rainbow trout, and provide theoretical support for the prevention and treatment of IHNV from immune genes and intestinal microbial regulation.

Soil nutrient cycling and plant growth are significantly affected by rainfall leaching in different forest stands in subtropical regions. This study aimed to investigate how the litter under Moso bamboo (Phyllostachys edulis) invasion into broadleaved forest affected the soil nitrogen mineralization through the release of water-soluble carbon and nitrogen from litter. The incubation experiment in broadleaved forest soil for 42 d was conducted by adding different concentrations of the litter extracts from invaded Moso bamboo and uninvaded broadleaved forests. This study investigated the effects of litter extracts from invasive and non- invasive forests on the soil net nitrogen mineralization (ammonification rate, nitrification rate, and nitrogen mineralization rate), hydrolase enzyme (β -glucosidase enzyme, β -1, 4-N-acetylglucosidase, and leucine aminopeptidase) activities, and the abundance of bacteria and fungi, and soil pH. The results showed that the net ammonification rate (9.2%~52.3%), net nitrification rate (8.4%~138.9%), net nitrogen mineralization rate (8.7%~37.9%) were significantly increased by adding the litter extract of Moso bamboo forest compared with the control (P<0.05). The net ammonification rate (6.1%~92.1%), net nitrification rate (13.8%~279.7%), and net mineralization rate (15.1%~65.4%) were significantly decreased by adding a litter-eluting extract of the broadleaved forest (P<0.05). The addition of the litter extracts from Moso bamboo were significantly decreased soil pH (P<0.05), but significantly increased the fungal-to-bacterial ratio (P<0.05), and significantly promoted the enzyme activities related to nitrogen mineralizations, e. g. β -1, 4-N- acetaminoglycosidase and leucine aminopeptidase (P<0.05). The correlation analysis showed that the nitrogen mineralization rate was affected by the soil pH, β -glucosidase, β -1, 4-N-acetylglucosidase, leucine aminopeptidase, C/N ratio of litter extracts and soil fungal-to-bacterial ratio. Thus, the litter extracts from invasive plant bamboo significantly increased the soil nitrogen mineralization rate, which will result in a conducive environment for bamboo growth and assured the availability of soil nutrients to promote its invasion into the native broadleaved forest. This study provides an important theoretical basis for understanding the plant-soil interaction during bamboo invasion into natural forest.

Histone monoubiquitination (H2Bub1) is conserved among eukaryotes, and is one of known histone modifications. H2Bub1 is essential for the growth and development of organisms. In recent years, a great number of studies have shown that H2Bub1 is widely involved in plant growth, development and stress response. In this review, diverse roles of H2Bub1 in regulating developmental processes were introduced, including seed dormancy, photomorphogenesis, and flowering time, and the biological function of H2Bub1 in response to biotic and abiotic stress were discussed. This review provides reference for the crop genetic improvement.

Single-cell sequencing is a new research area, which mainly includes transcriptome sequencing, whole genome sequencing, and epigenomic sequencing. Single-cell RNA-sequencing (scRNA-seq) refers to a new technique for high-throughput sequencing and analysis of RNA at the single cell level. Recently, single- cell related experimental technology has developed rapidly and has been widely used in the field of biology and medicine, mainly for the study of biological evolution process, tissue and organ development, tumor disease occurrence and precision medical diagnosis. However, few studies were reported in the field of animal science research and its application, currently, most of them focus on the field of animal physiology and evolutionary development. This article summarized the technical methods of single-cell transcriptome sequencing and its application in the field of animal science. This review provides a theoretical basis for the development of single-cell transcriptome sequencing technology and its application in the field of animal science.

For analyzing the expression of genes accurately by quantitative real-time PCR (qRT-PCR), screening of stabilized reference genes is of great important. Based on transcriptome data, 9 candidate genes including 18S ribosome RNA (18S rRNA), initiation factor 2 (IF2), elongation factor-1α 1 (EF-1-α1), EF-1-α2, adenine phosphoribosyltransferase (APT), cycloidea1 (cyc1), glyceraldehyde-3-phosphatedehydrogenase (GAPDH), GAPDH2 and β -tubulin (TUB) were amplified by qRT-PCR in leaves and roots of Cornus hongkongensis subsp. tonkinensis suffering from different salt stress time. The expression stability of 9 candidate genes were measured by GeNorm, NormFinder and BestKeeper. Suitable reference gene was finally screened based on comprehensive evaluation of expression stability with RefFinder. Then the selected reference genes were verified by target genes superoxide dismutase (SOD) and catalase (CAT). The results showed that 18S rRNA and GAPDH were the optimal candidate reference genes in various organs under salt stress, whereas EF-1- α2 was the least stable gene. However, the Ct value of 18S rRNA was higher, indicated that the expression of 18S rRNA was lower than that of GAPDH in various organs. The results also showed that the relative expression levels of target genes were consistent with the transcriptome sequencing results when using GAPDH as reference gene. This study determined that the most suitable reference gene was GAPDH for qRT-PCR, which could provide the reference for further study on the expression of related gene in Cornus hongkongensis subsp. tonkinensis under salt stress.

African swine fever (ASF), a devastating disease for pigs (Sus scrofa), usually causes hemorrhagic fever, which bring great economic losses to the pig industry. Multigene families (MGF) 505-5R was in the right variable region (RVR) of the African swine fever virus (ASFV) genome and could influence the virulence of the virus. The work aimed to analyze the molecular characteristics of the MGF505-5R gene and protein and construct a eukaryotic expression vector for the ASFV MGF505-5R gene. The MGF505-5R gene was obtained from the ASFV reference genome in GenBank (GenBank No. MK128995.1) for bioinformatics analysis, and the MGF505-5R gene was synthesized. The MGF505-5R gene was ligated into the pRK5M-C-2×Strep vector by homologous recombination. The recombinant plasmid pRK5M-C-2×Strep-MGF505-5R was transfected into IPEC-J2 cells after being identified by PCR and sequencing, and the expression of the MGF505-5R gene was confirmed by immunofluorescence and Western blot. Transcript levels of exosome 8 subunits, Mtr4 exosome RNA helicase (MTREX), terminal nucleotidyltransferase 4A (TENT4A) and zinc finger DHHC-type palmitoyltransferase 16 (ZDHHC16) in IPEC-J2 cells overexpressing pRK5M-C-2×Strep-MGF505-5R were detected by qPCR. Bioinformatics analysis showed that the MGF505-5R gene was divided into 3 branches in the phylogenetic tree and there was a positive selection at amino acids 20, 105 and 485. The secondary structure of MGF505-5R protein matched the tertiary structure, and the positive selection sites were all located on the irregularly coiling. PCR results showed that the MGF505-5R gene fragment was successfully ligated into the pRK5M-C-2×Strep vector. Immunofluorescence and Western blot assays showed that the pRK5M-C-2×Strep-MGF505-5R vector was successfully expressed in porcine intestinal columnar epithelial cells (IPEC-J2) cells. The qPCR results showed that the expression of exocytotic complex subunits exosome component (EXOSC) 1, EXOSC2, EXOSC3, EXOSC4, EXOSC5, EXOSC7, EXOSC8, EXOSC10, TENT4A and ZDHHC16 was extremely significantly decreased in IPEC-J2 cells overexpressing pRK5M-C-2×Strep-MGF505-5R (P<0.01); no significant change in MTREX expression. The analysis of the molecular characteristics of MGF505-5R in this study provided a basis for the evolutionary study of this gene; the construction of the MGF505-5R expression vector in pig IPEC-J2 cells provides the effective tool and experimental material for the study of protein interactions between ASFV and the host.