Porcine alveolar macrophages (PAMs) are important members of the body's first line of defense in innate immunity. Their strong plasticity provides a significant cell model for investigating the disease resistance mechanism of pigs (Sus scrofa). In order to establish different polarized isoforms of porcine alveolar macrophages and to compare the polarization heterogeneity and reversibility of PAMs and 3D4/21 cells, PAMs isolated from 30-day-old healthy Yorkshire piglets and commercialized 3D4/21 cell line perserved by laboratory were used as experimental materials in this study. A combination of lipopolysaccharide (LPS) with interferon gamma (IFN-γ), interleukin 4 (IL-4) were used to induce the polarization of PAMs and 3D4/21 cells to construct M1 and M2 cell models, respectively. PAMs and 3D4/21 cells without polarization were used as the control group (M0 type). PCR and qRT-PCR relative quantitative results showed that the M1-type marker genes IL-1β, CXC chemokine ligand 10 (CXCL10), tumour necrosis factor (TNF), IL-6, and IL-12B were significantly more highly expressed in PAMs and 3D4/21 cell M1 types than M0 and M2 types (P<0.05); M2-type marker genes arginase-1 (Arg1), peroxisome proliferators-activated receptor γ (PPARγ), IL-10 and chitinase 3-like 3 (Chi3L3 or Ym-1) were significantly more highly expressed in M2 type than in M0 and M1 types in the 3D4/21 cell (P<0.05). In PAMs, the expression levels of Arg1, PPARγ and Ym-1 in M0 and M2 were significantly higher than that of in M1 (P<0.05). Western blot detections of Arg1 protein in the 2 types of cells were consistent with the relative quantitative results of qRT-PCR. At the same time, the reversibility test results of PAMs and 3D4/21 cell polarization showed that the reversibility of PAMs polarization was stronger than that of 3D4/21 cell, and the ability of PAMs to switch from M2 to M1 was stronger than from M1 to M2 type conversion capability. This study provides selection materials and basic data for further exploration of porcine alveolar macrophage polarization, immune mechanisms, and pig disease-resistant breeding.

Nutrition and energy metabolism affect the reproductive ability of male animals, and AMP-activated protein kinase (AMPK) plays an important role in its reproductive function as an energy receptor molecule. This study aims to study the effect of AMPK on autophagy and apoptosis of sheep (Ovis aries) leydig cells (LCs) induced by starvation. Sheep LCs were used as experimental materials and cultivated to the fourth generation. After 0, 3, 6, 12, and 24 h in serum-free medium, used cell counting kit-8 to detect cell viability and selected appropriate treatment time. After starving for 12 h, the autophagy and apoptosis of LC were observed by transmission electron microscopy and flow cytometry, qRT-PCR and Western blot were used to detect the changes in gene and protein expression related to autophagy and apoptosis. The results showed that compared with the control group, there were more autophagosomes in starvation-induced LCs, and the apoptosis rate was significantly higher (P<0.05), programmed cell death 1 (Beclin1) and microtubule-associated protein 1 light chain 3 (LC3) and other genes expression level were significantly increased (P<0.05), Becline1 protein expression and LC3Ⅱ/LC3Ⅰ ratio were significantly increased (P<0.05), pro-apoptosis related genes Bcl-2 related X protein (BAX) and other genes expression level were significantly increased (P<0.05), anti-apoptosis related genes B-cell lymphoma 2 (Bcl-2) gene expression level was significantly reduced (P<0.01), and the ratio of BAX/Bcl-2 protein was increased; AMPK/mammalian target of rapamycin (mTOR)/UNC-51-like kinase 1 (ULK1) pathway key genes expression level were increased extremely significantly (P<0.01). In summary, starvation-induced sheep LCs could activate the AMPK/mTOR/ULK1 pathway and promote the occurrence of autophagy and apoptosis. This research provides a theoretical reference for further exploring the impact of AMPK on testicular development.

MYB transcription factor family is one of the largest families of transcription factors in plants, which are extensively involved in many physiological and biochemical processes. In order to explore the function of MYB transcription factor family of pepper (Capsicum annuum), this study conducted genome-wide identification and bioinformatics analysis, and detected the effects of Phytophthora capsici on the transcription level of the family members by transcriptome technique. The results showed that 102 R2R3-MYB transcription factors were identified from C. annuum genome database, which were distributed on each chromosome. Phylogenetic tree analysis found that R2R3-MYB proteins of pepper could be classified into 11 subgroups, and subfamily Ⅺ had a clear distinction from other subfamily in conserved domains, shared motifs and gene structure. Conserved domains included 2 classes with significant differences, but the conserved domains in each class were highly similar. The R2R3-MYB gene structure was significantly different among the subgroups. In the pepper genome, there were 5 pairs of homologous R2R3-MYB genes from tandem replication. The R2R3-MYB genes of Arabidopsis thaliana, Solanum lycopersicum and C. annuum could be classified into 22 subgroups by phylogenetic analysis. Above results indicate that R2R3-MYB proteins initially diversified through genome replication, intron insertion and domain shuffling, but remained relatively conserved throughout the subsequent plant evolution. Further analysis of R2R3-MYB gene expression pattern using pepper transcriptome data revealed that some of them could response to P. capsici. The expression of some R2R3-MYB transcription factors were up-regulated in disease-resistant varieties, suggesting that expressions of MYB transcription factor could be induced by P. capsici. The present study could provide a reference for in-depth exploration of the mechanism of pepper R2R3-MYB transcription factor family involved in P. capsic resistance.

Long peduncle varieties of melon (Cucumis melo) are beneficial to large-scale production and mechanized harvesting. In present study, long peduncle melon '1244' and short peduncle melon 'MS-5' were used to make crossing and constructed F₂ segregation population, using specific locus amplified fragment sequencing sequencing (SLAF-seq) technology to construct melon genetic map, combing with SSR molecular markers, to detect fruit peduncle length (fpl) QTL locus. The results showed, a genetic map of 12 linked melon populations, with 10 595 markers were constructed, the total distance was 1 383.88 cM and the average distance between the markers was 0.13 cM; melon peduncle length was controlled by 2 pairs of main genes, fpl1.1 was located on chromosome 1, the phenotypic contribution rate was 12.2%, QTL candidate interval was between 57 524 and 270 836 in melon scaffold00060, located between CMSSR05887 and CMSSR05892; fpl8.1 was located on chromosome 8, the phenotypic contribution rate was 13.2%, QTL candidate interval was between 94 748 and 105 498 in melon scaffold00058, located between CMSSR20549 and CMSSR20495. The results provide a theoretical basis for the study of melon molecular marker assisted selection breeding and melon mechanized harvesting.

Linoleic acid is main polyunsaturated fatty acids in plant lipids, which plays key roles in plant metabolism. Fatty desaturase 2 (FAD2) is a key enzyme for linoleic acid biosynthesis, CtFAD2-1 gene has the highest activity and was mainly responsible for the biosynthesis of linoleic acid in safflower (Carthamus tinctorius) seed. In this study, an 821 bp promoter region of CtFAD2-1 gene from safflower was cloned. Four 5'-deletion constructs of 821 bp (P1), 564 bp (P2), 230 bp (P3), 60 bp (P4) were constructed and functionally characterized in Arabidopsis thaliana by using transgenic approach. Computational analysis found that the abiotic stress responsive cis-elements like heat responsive element (HSE), low temperature response (LTR), MYB response (MBS) etc, as well as wound, fungus, methyljasmonate responsive motifs existed in the promoter. The histochemical staining of GUS revealed that the flower, mature seed and silique wall were stained. In addition, the transgenic plants treated by drought, salt, cold, high-temperature, wound and ABA hormone as well as control plants were also stained. The -230~-60 bp might be the key region for GUS activity in A. thaliana tissues. CtFAD2-1 gene was not seed-specific and it could be induced by an assortment of abiotic stresses. This research plays an indispensable role in furthering the research related to the regulation of CtFAD2-1 gene.

Anthrax is an important disease in litchi (Litchi chinensis) production, which severely limits the development of the litchi industry. In order to clarify the species composition, the dominant population of the pathogen of litchi anthracnose in Hainan, and the correlativity between the distribution of different populations in the spatial area of Hainan and the geographical source. This article conducted an investigation and sampling of the occurrence of anthracnose tissues in the litchi planting areas of Hainan from 2018 to 2019, and the isolation and purification of pathogenic microorganisms and the verification of Koch's law was proceeded; Using combined analysis of multiple genes (internal transcribed spacers (ITS), partial actin (ACT), glyceraldehyde-3-phosphate dehydrogenase (GAPDH), chitin synthetase (CHS-1), β-tubulin (TUB), giutamine synthetase (GS) and mating type (ApMat)) to research the species composition, distribution and geographical relationship of Hainan litchi anthracnose, as well as the genetic diversity of populations in different regions. A total of 79 strains of litchi anthracnose pathogens were isolated in this study. The results showed that the mainly species of pathogens causing litchi anthracnose were C. acutadum and the complex group C. gloeospoiorides species, including C. fructicola, C. siamense, C. asianum, C. musae, C. kahawas subsp. kahawae and C. horri, which had greater abundance in species composition in Hainan, as the dominant pathogenic group was C. gloeosporioides; Various strains do not have geographic regionality in origin. The geographical regionality and the genetic distance of each geographical strain were relatively close, and the distribution of pathogens had no obvious correlation with geography; the source of genetic variation was mainly among geographic populations. This study provides a theoretical basis for a more in-depth study of the occurrence rules, control measures of litchi anthracnose and further breeding for disease resistance.

Cumulus cells are important supporting cells of oocytes. Quality of cumulus cells influences in vitro maturation and developmental competence of oocytes. Low oxygen tension was beneficial for in vitro maturation and developmental competence of yak (Bos grunniens) oocytes, whereas whether the beneficial effects were exerted through cumulus cells has not been reported. In order to investigate the effects of low oxygen tension on transcriptome of yak cumulus cells, maturation of yak cumulus-oocyte complex (COC) was carried out at 5% and 20% O₂, respectively. Then cumulus cells were collected for RNA-seq, screening of differentially expressed genes (DEGs) and qRT-PCR verification. The results showed that 8 yak cumulus cell transcriptomic libraries were obtained and total data size was 56.17 Gb; gene expression analysis showed that a total of 13 584 genes were expressed of the two groups and 270 DEGs were identified, among which 229 DEGs were up-regulated in 5% O₂ and 41 DEGs were up-regulated in 20% O₂; Gene Ontology (GO) and KEGG pathway analysis showed that DEGs were mostly enriched in glycolysis, cumulus cell expansion and cumulus-oocyte signaling et al.; 6 711 new genes and 7 209 new transcripts, among which 829 new transcripts had protein coding potential, were found in unannotated region of the yak reference genome. qRT-PCR results were consistent with the sequencing data. Taken together, above results indicated that low oxygen tension might induce yak cumulus cells to have a similar transcriptomic profile to those matured in vivo and therefore promote the maturation and developmental competence of yak oocytes. The present study provides a reference for further study of the effect of oxygen tension on the mechanism of cumulus cell-oocyte interaction in the process of in vitro maturation of yak oocytes.

Circular RNA (circRNA) play important roles in the regulation of cell differentiation by acting as competing endogenous RNA (ceRNA). However, the regulatory mechanisms by which circRNAs and the circRNA-associated ceRNA network regulate chicken preadipocyte differentiation remain elusive. In this study, RNA-seq technology was used to analyze the expression of circRNAs, microRNAs (miRNAs) and mRNAs during the differentiation of chicken (Gallus gallus) preadipocytes. Bioinformatics methods were used to screen differentially expressed circRNAs, miRNAs and mRNAs, construct a circRNA-associated ceRNA regulatory network, and screen important circRNA-related ceRNA regulatory relationship pairs. A total of 4 904 circRNAs were identified in chicken preadipocytes. 143, 145 and 660 differentially expressed circRNAs, miRNAs and mRNAs were screened during the differentiation of chicken preadipocytes. Based on gene expression correlation analysis and target relationship prediction, a circRNA-associated ceRNA network was successfully constructed. Twelve candidate circRNA-miRNA-mRNA pairs were screened using qRT-PCR, which may play important regulatory roles in chicken preadipocyte differentiation. The present study lays a foundation for further analysis of the mechanism by which circRNAs regulating chicken fat deposition, and enriches the genetic regulation theory of chicken fat deposition.

Liancheng white duck (Anas platyrhynchos) is a precious and rare waterfowl resource in China, and has high meat quality and unique flavor, while the formation mechanism of meat quality is still unclear. Exploring the genes related to meat quality and flavor can not only be helpful for the development and utilization of Liancheng white duck, but also provide a reference for breeding and meat quality evaluation of meat ducks. In this study, RNA-seq was used for transcriptome sequencing to screen differentially expressed genes in breast muscle transcriptome of Liancheng white duck and Cherry Valley duck; and functional annotation, enrichment analysis and qRT-PCR verification were carried out to explore the potential regulatory genes of meat quality formation in Liancheng white duck. A total of 44.31 Gb clean data was obtained by sequencing, and the mapping rate with reference genome was over 77.54%. The edge R analysis identified 912 differentially expressed genes between the two breeds, among which 424 were highly expressed genes and 488 were low expressed genes in Liancheng white duck. In Gene Ontology (GO) annotation, 772 differentially expressed genes were annotated, including 12 biological processes related to the formation of meat flavor substances, such as lipid metabolism, fatty acid metabolism, threonine catabolism, and carboxylic acid metabolism. KEGG pathway analysis showed that the differentially expressed genes were enriched in 179 signaling pathways, among which 12 pathways were significantly enriched, including flavor-related pathways such as the interaction between cytokines and receptors, linoleic acid metabolism and histidine metabolism, glycerophosphate metabolism and transforming growth factor (TGFβ) signaling pathway. According to the annotation of pathway function, the genes such as acidic amino acid decarboxylase 1 (GADL1), coiled-coil domain-containing protein 57 (CCDC57), and pyruvate dehydrogenase kinase, isozyme 4 (PDK4) were screened out and might play an important role in regulating the formation of delicious amino acids and fatty acids in Liancheng white duck, and further affecting the formation of chest muscle quality. Furthermore, 2 high expressed genes and 3 low expressed genes were selected for qRT-PCR verification, and the results were consistent with that of transcriptome sequencing, which indicated that the sequencing results were reliable. The present study provides a reference for further studying on the formation mechanism of meat flavor of Liancheng white duck and selection of genetic markers.

Sodium channel nonvoltage-gated 1β (SCNN1B) plays an important role in cation transport, which can transport Na⁺ and promote the secretion of Ca²⁺, thus affect the quality of eggshell. In this study, the egg duck variety Sansui duck (Anas platyrhyncha domestica) was used as the research object. The tissue expression of SCNN1B gene was detected by qRT-PCR, the genetic variation of SCNN1B gene was detected by direct sequencing, and the single factor analysis of variance (ANOVA) of SPSS 18.0 was used to analyze the effect of the variation on eggshell quality. The results showed that SCNN1B gene was expressed in different degrees in 12 tissues of Sansui duck, and the relative expression level was in the following order: uterus>kidney>leg muscle>duodenum>glandular stomach>chest muscle>liver>lung>brain>heart>spleen>muscular stomach, and the expression abundance of uterus was significantly higher than that of other tissues (P<0.05). Three moderately polymorphic loci were detected in SCNN1B gene, among which g.794619 C>T and g.794635 G>T were located in intron 8, and g.794564 G>A was a synonymous mutation in exon 8, which led to a change in the secondary structure of mRNA. The distribution of g.794635 G>T and g.794564 G>A genotypes deviated significantly from Hardy-Weinberg balance (P<0.01), and there was strong linkage disequilibrium among the 3 SNP loci, and there were 4 haplotypes and 5 diplotypes. The eggshell weight of individuals with TC genotype at g.794619 C>T was significantly higher than that of TT genotype (P<0.05), and that of H1H3 and H1H4 haplotypes was significantly higher than that of H1H1 (P<0.05). The results suggest that SCNN1B gene expression has tissue specificity in laying duck, the TC genotype at the locus of g.794619 C>T is beneficial to increase eggshell weight, and diplotype H1H3 is favorable to increase eggshell weight. This study provides a reference for the improvement of eggshell quality and the selection of genetic markers.

Edwardsiella tarda is the main pathogen causing large-scale death of flounder (Paralichthys olivaceus) in factory farming. Its infectivity is extremely strong. The mortality rate of infected flounder with E. tarda is very high, which seriously impedes the development of flounder industry. In order to study the function of neurotrophin 3 (NTF3) in the innate immunity of P. olivaceus, the full-length cDNA sequence of P. olivaceus NTF3 gene was cloned, with the total length 1 411 bp, in which the 5' noncoding region is 89 bp and the 3' non-coding region is 428 bp.The open reading frame (ORF) of this gene was 894 bp.Which encoded 297 amino acids. The predicted molecular weight (Mw) was about 33.73 ku, and the theoretical isoelectric point (pI) was 9.56. The homology comparison found that the NTF3 gene had a high similarity with different species, and the highest homology was 90.91% with the NTF3 gene of amberjack (Seriola dumerili). The NTF3 gene was expressed in lots of healthy tissues of flounder (blood, liver, spleen, kidney, intestine, skin, gill, heart and brain). The highest expression level was tested in skin followed by blood. To further study the relative expression of NTF3 during E. tarda infection, a bacterial infection experiment was conducted on healthy flounder, and then the relative expression of NTF3 in 4 different tissues (intestine, kidney, spleen and liver) and different time points were compared. It was found that, in the intestine, spleen and liver, the expression levels of NTF3 gene were up-regulated after bacterial stimulation, but in the kidney, the expression levels showed a downward trend. The above results indicated that the NTF3 gene may be involved in the immune process of P. olivaceus against the E.tarda infection, and the relative expression of NTF3 gene is different in different tissues infected by the bacteria, which provides basic research data for further study of the role of NTF3 gene in the immune response process of flounder.

Follicle stimulating hormone receptor (FSHR) and luteinizing hormone receptor (LHR) belong to gonadotropic hormone receptors (GtHR) and play a regulatory role in the reproduction of fish. Nile tilapia (Oreochromis niloticus) has fast growth, short reproductive cycle and high economic value. In order to explore the relationship between GtHR and the reproductive activity of Nile tilapia, LHR and FSHR genes of Nile tilapia were cloned by using rapid amplification of cDNA ends (RACE) and homology-based cloning, and the expression changes of these two genes in ovary and testis, as well as the changes of estradiol (E₂) in female and testosterone (T) in male were measured after different oxytocin injection. The results showed that the full-length of FSHR cDNA was 2 577 bp, which encoded 701 amino acids; The full-length of LHR cDNA was 3 177 bp, which encoded 693 amino acids.The results of qRT-PCR showed that FSHR and LHR genes were mainly expressed in the gonads. After the injection of oxytocin, with the change of E₂ in female and T in male, FSHR showed an expression pattern of first increase then decrease and LHR showed an expression pattern of first increase, then decrease and finally increase in the gonads, and the expression level of FSHR and LHR in the injection group was significantly higher than that in the control group at multiple time points, in addition, the expression level of FSHR and LHR in the oxytocin combination injection group was higher than in the single injection group. These results indicated that FSHR and LHR might be involved in reproductive activities of Nile tilapia. Different oxytocin injections could affect the expression of FSHR and LHR genes in Nile tilapia, and the combined effect of oxytocin was better than single oxytocin. This study provides a basic information for further elucidating the physiological function and molecular mechanism of FSHR and LHR, and provids reference for reproductive regulation of Nile tilapia.

Microphtalmia-associated transcription factor a gene (mitfa) plays an important role in the formation of animal body colour. This study was conducted to explore the regulatory role of mitfa gene in the body colour variation between wild-type (WT) and yellow mutant (YM) rainbow trout (Oncorhynchus mykiss). The full-length sequence of mitfa gene of rainbow trout was obtained by rapid-amplification of cDNA ends (RACE) technique, and the bioinformatics analysis was performed. Furthermore, qRT-PCR was used to investigate the expression levels of mitfa in the different developmental stages and various tissues of WT and YM rainbow trout, respectively. The full-length cDNA of mitfa (GenBank No. MT813117) was 2 700 bp containing 114 bp 5'UTR, 1 374 bp 3'UTR, and 1 212 bp ORF encoding 403 amino acids. Sequence analysis showed that Mitfa protein sequence was more conservative in fishes than that in other vertebrates and the basic helix-loop-helix-leucine zipper (bHLHZip) structural domain was highly conserved among all vertebrates. These results were further confirmed by phylogenetic analysis. qRT-PCR analysis revealed that mitfa expressed at different levels during embryo and post hatch stages, and higher expression levels were detected at the fertilized-stage, 4-cell, 16-cell and multi-cell, which were significantly higher than that of other stages (P<0.05). Moreover, there were extremely significant differences (P<0.01) in the expression of mitfa gene at the same stages between WT and YM rainbow trout, e.g., at the fertilized-stage, 16-cell, multi-cell, blastula, somites, heartbeating, 3 dph (days post hatch), 5 dph, 7 dph, 10 dph, 1 M (month post hatch) and 12 M dorsal skin. mitfa gene expressed in all kinds of tissues of 12 M WT and YM rainbow trout, which expressed the highest amount in dorsal skin (P<0.05), it also highly expressed in ventral skin and muscle, dorasl muscle and eye, but relatively lower in other tissues. The above results indicated that the mitfa gene might be involved in the formation of pigment cells in rainbow trout and closely related to the body colour variation. The results of this study provide basic data for the molecular regulation mechanism of body colour variation and the genetic improvement of body colour in rainbow trout.

The genetic variation of Setosphaeria turcica was mainly caused by sexual reproduction. In order to explore the role of the S. turcica mating type (MAT) genes StMAT1-1 and StMAT1-2 in sexual reproduction, some strains with different mating type genes were sexually induced, and qRT-PCR was used to analyze the expression patterns of StMAT1-1 and StMAT1-2 in different mating type strains at 0, 15, 30 and 60 d after sexual induction. The transcriptional activity of StMAT1-1 and StMAT1-2 was determined by yeast (Saccharomyces cerevisiae) self-activation test. The results showed that StMAT1-1 and StMAT1-2 genes were expressed before sexual induction, 15, 30 and 60 d after sexual induction. With the extension of induction time, the expression of mating type genes in different induction combinations also changes significantly, indicating that MAT genes played an important role in the sexual reproduction process of S. turcica. In yeast self-activation test, it was found that the encoded proteins of StMAT1-1 and StMAT1-2 genes were transcription factors with high transcriptional activation activity. This study has reference value to the further research on the key role of MAT genes in the regulation of sexual reproduction of S. turcica.

A new type of Daphnis nerii cypovirus-23 (DnCPV-23) can kill a variety of Sphingidae insects and has the potential to be a new insect virus insecticide. At present, Daphnis nerii is the main model to study the virus, the replication of DnCPV-23 in cells is still unclear. In this study, DnCPV-23 virus particles were inoculated into Spodoptera frugiperda cell line Sf9, and the replication of the virus in the cells was analyzed. The virus-infected cells were cultured at 27 ℃, and the effect of the virus on the Sf9 cells were observed with the optical microscope; RNA of the infected cells on day 1, 3 and 7 was extracted and used as a template to obtain the cDNA, and the DnCPV-23 RNA level was analyzed by qRT-PCR; the supernatant of infected Sf9 cells and the serial passage of virus in Sf9 cells were studied by the qRT-PCR method. DnCPV-23 could adapt to Sf9 cells well, could successfully infect Sf9 cells, and produce cytopathic effect (CPE); virus genes could be detected in the cell culture supernatant; the virus could be passaged in Sf9 cells. DnCPV-23 could replicate and passage well on Sf9 cells and make the cells undergo CPE. The above-mentioned research provides a good cell model for in-depth study of DnCPV-23 invasion, transcription and other mechanisms and evaluation of virus virulence. At the same time, it provides a certain reference and basis for the development of new Sphingidae virus insecticides.

VP6 protein (viral structural protein 6) of Bluetongue virus (BTV) plays an important role in virus replication and genome assembly. Developing cell line stably expressing the BTV VP6 protein will provide foundation for the research of VP6 protein function and genetic engineering vaccine. In present study, VP6 gene of BTV was amplified by RT-PCR to construct prokaryotic expression plasmid of pET32-BTV-VP6 and eukaryotic expression plasmid of pCDH-BTV-VP6. The pET32-BTV-VP6 was transferred into Escherichia coli and recombinant VP6 protein was expressed in soluble form at 16 ℃, which accounted for 24.5% of total bacterial protein. The recombinant VP6 protein was purified using Ni²⁺-NTA affinity chromatography and was used to immunize rabbits (Oryctolagus cuniculu) for preparation of polyclonal antibodies against VP6 protein of BTV. The antibody titer of the immunized rabbits reached 1∶12 000 as confirmed by indirect ELISA. VP6 protein expressed by BTV-infected cells was detected by Western blot and indirect immunofluorescence assay (IFA) using the prepared serum against VP6. To construct BHK-BTV-VP6 cell line which stably expressed VP6 protein of BTV, pCDH-BTV-VP6 was transfected into BHK-21 cells and continuously screened by puromycin. The stable expression of VP6 protein in the BHK-BTV-VP6 cell line was confirmed by Western blot and IFA using rabbit polyclonal antibody against BTV VP6 as primary antibody. Conclusively, the present study successfully expressed and purified BTV VP6 recombinant protein in E. coli, and constructed a cell line stably expressing BTV VP6 protein, which provided material basis for the research on VP6 protein function and the development of new genetic engineering vaccine.

Soybean (Glycine max) is an important grain and oil crop in the world, which provides abundant protein and oil sources for human and animals. The successful commercial utilization of transgenic technology in soybean has become a classic case of biotechnology in promoting crop production. In recent years, the rapid development of new generation biotechnology, such as genome editing, has provided new opportunities for further improvement of soybean agronomic traits. Recently, genome editing technology represented by CRISPR/Cas technology begins to be widely used in genetic improvement of soybean, which shows powerful potentials on improving quality, regulating growth period and stress resistance. This review summarizes the latest research progress in this field from the development of genome editing technology, the application of genome editing technology in soybean genetic improvement and the main problems in current application of genome editing technology in soybean, which provides a reference for the work of using genome editing technology as a tool to accelerate soybean genetic improvement.

The small cysteine-rich proteins (SCRs) are one class of proteins secreted by fungi, which are usually less than 200 amino acids in length containing 2%~20% cysteine residues. Some members are avirulence proteins of plant pathogenic fungi, indicating that these effector proteins may play an important role in the life cycles of fungi. Molecular structure, subcellular localization and biological functions of SCRs in fungi were reviewed in this paper. Especially, the biological functions of these proteins were elucidated by focusing on their involvement in the pathogenesis of the pathogenic fungi and their influence on the growth and development of the fungi. This review provides a summary for illuminating the biological functions of SCRs, particularly lays a foundation for further elucidating the roles of these effector proteins in the interactions between fungi and their hosts, and for the management of plant fungal diseases.

Soil microbial community is an important part of ecosystem, which plays an irreplaceable role in regulating plant growth, promoting the formation of soil structure and maintaining the function and stability of ecosystem. In this study, using CiteSpace document visualization software, data mining was conducted on 6 909 articles about soil microbial community collected from the Web of Science core database from 1990 to 2020. The results showed a steady increase in the number of publications in the soil microbial community area, a higher number of publications in China and the United States, and a greater number of cooperative communications between China and the United States, and a higher number of publications in the field of soil microbial community in the Chinese Academy of Sciences (CAS). Despite its late launch, CAS occupied a very important position. Based on deep excavation of highly cited literature in this field and the cross-citation analysis, it was speculated that the future research directions include dynamic microbial community, soil, soil ecosystem characteristics and many kinds of unculturable bacteria. Finally, using co-occurrence network and emergent detection, it was found that current research focuses on "functional diversity", "litter decomposition" and "microbial diversity". This study will be helpful to the development, layout and innovation of soil microbial community research.