Wheat (Triticum aestivum) awn,an important agronomic character of wheat controlled by many genes,plays an important role in resisting insect pests,seed reproduction and increasing wheat yield. CASL4AL is a chromosome arm substitution line (CASL) of wild emmer (T. dicoccoides,TTD) accession TTD140 in the genetic background of common wheat variety Chinese Spring (T. aestivum cv. Chinese Spring,CS),the long arm of chromosome 4A of CASL4AL is replaced by TTD140,showing long awn. In this study,a F2 population with 134 lines was constructed by using CASL4AL with long awn and the line of 442 with hook awn (derived from the cross of CASL4AL×CS,hooded awn as CS). The SSR molecular markers were used for QTL mapping for the awn length of wheat by ICIM (inclusive composite interval mapping) method. A QTL with LOD value of 30.71 was detected on the chromosome 4AS related to awn length,explaining 82.63% of the phenotypic variation. In this mapping interval,the annotation of the variants (SNP and InDel) for the CASL4AL resequencing data were conducted according to reference wheat genome,combined with the differential analysis of transcriptome sequencing,five differentially expressed genes with sense variation were obtained,among which,TraesCS4A02G078700 translates MADS-box transcription factor 1,which involved in the differentiation and proliferation of the meristem of lemma and palea,and probably involved in regulating the growth of wheat awn. This study provide reference for the screening of fine mapping materials and fine mapping of awn gene Hd (Hooded).

Homeobox transcription factor (HTF) is a type of transcriptional regulatory factors with conservative structure and function. These genes can affect the growth and development and morphogenesis of eukaryotic organisms by participating in regulating the expression of functional genes. StHTF2 is a member of the HTF transcription factor family of Setosphaeria turcica playing an important role during the infection process. In this study,StHTF2 gene (GenBank No. XP_008029609.1) was cloned. The analysis results of gene sequence and its encoded protein characteristics showed that StHTF2 gene has 1 747 bp in total length and contained an open reading frame of 1 641 bp,which encoded 546 amino acids. Moreover,StHtf2 contained a highly conserved homologous domain HOX,which was conserved in eukaryotic organisms. The spatial structure formed from this domain including 3 helix regions could bind to a specific DNA motif to perform the function of a transcription factor. The expression and identification results of the StHtf2-His fusion protein showed that the fusion protein can be expressed in the Escherichia coli system,and the expression efficiency was higher at an induction temperature of 42 ℃,an induction pH of 7,and an induction time of 6 h. Furthermore,analysis of StHTF2 expression level during infection showed that compared with the conidium stage of the strain,the expression level of StHTF2 gene in susceptible maize (Zea mays) 'B73' and resistant maize 'B73Ht' were increased by 4.5 and 2 times,respectively,when the pathogens were infected for 24 h on the leaves of maize,which predicted that StHTF2 gene was involved in the infection process of pathogens and its expression was affected by the host. The above research results will not only provide a prerequisite for preparing StHtf2 protein antibodies and further studying the interaction of StHtf2 protein with other proteins but also create the prerequisite for revealing the molecular mechanism when the pathogen infect the hosts.

Sucrose non-fermented related protein kinase 2 (SnRK2) family members are plant-specific serine/threonine kinases involved in response to abiotic stresses. In this study,to explore the biological roles of SnRK2 gene in growth,development,and abiotic stress tolerance in oat (Avena sativa),one of the SnRK2 family members,designated AsSnRK2.10 (GenBank No. MN729578),was isolated from oat variety 'Monida' by reverse transcription-PCR (RT-PCR). The molecular characteristics of AsSnRK2.10 encoded protein were predicted by bioinformatics analysis. Then subcellular localization of target protein was performed using a transient expression system in tobacco (Nicotiana benthamiana). And lastly,the expression profiles of AsSnRK2.10 in different tissues and under abiotic stress conditions were detected by qRT-PCR,respectively. The results showed that AsSnRK2.10 gene contained a 1 086 bp of open reading frame (ORF),which encoded a polypeptide of 361 amino acid polypeptide with a predicted molecular weight of 40.66 kD and a theoretical isoelectric point of 4.75. The AsSnRK2.10 protein contained the SnRK2-specific functional domains,including a protein kinase adenosine triphosphate (ATP)-binding signaling region,a N-myristoylation site,a serine/threonine protein kinase active site,an activation loop,a transmembrane helix site,a domain 1 required for the osmotic stress response,and a domain 2 required for the abscisic acid (ABA) response. The smallest secondary structure element of the AsSnRK2.10 protein was beta turn,and the largest was random coil. Phylogenetic analysis showed that the AsSnRK2.10 protein had the closest evolutionary relationship with SnRK2 members from seven species,and was clustered into one branch. The subcellular localization analysis revealed that the AsSnRK2.7 protein was localized primarily in the nucleus. Tissue-specific expression showed that AsSnRK2.10 was expressed in the roots,stems,leaves,and panicles of oat during different development stages. However,the highest transcript levels of the AsSnRK2.10 gene in the roots,stems,leaves,and panicles occurred at the heading stage,filling stage,tillering stage,and heading stage,respectively. Furthermore,AsSnRK2.10 gene was involved in ABA signal transduction and positively responded to dehydration,high salinity,and low-temperature stresses. The results provide a theoretical basis for further study on the biological function of AsSnRK2.10 gene and its application in oat stress resistance breeding.

Jasmonic acid substances,as endogenous regulatory hormones in plants,play important roles in plant growth and responding to stress. Jasmonic acid carboxyl methyltransferase (JMT) is a key enzyme that catalyzes synthesizing methyl jasmonate (MeJA) from jasmonate. It controls the transformation of jasmonate in plants and affects plants resistance to biological and abiotic stress. Based on sugarcane (Saccharum spp. hybrids) transcriptome database,a full-length of sugarcane jasmonate methyltransferase ScJMT (GenBank No.MK784565) was cloned and analyzed. The expression levels of ScJMT in different organs and expression profiles under different stresses were detected by qRT-PCR. Bioinformatics results showed that the open reading frame (ORF) of ScJMT gene was 1 110 bp encoding 369 aa. The isoelectric point,instability coefficient,and average hydrophobicity value of the ScJMT protein were 5.28,56.76,-0.135,respectively. The ScJMT was an unstable acid hydrophilic protein by prediction. The α-helix structure and random curl constituted its secondary structure and tertiary structure,and it also contained the methylation_7 conserved domain. Through phylogenetic tree analysis showed ScJMT was on the same branch as SvJMT from Sorghum vulgare and ZmJMT from Zea mays. In addition,the spatial expression level of ScJMT was detected and expression profiles under different stress were also determined. There were no significant differences among 3 sugarcane organs,and ScJMT was increased in abscisic acid (ABA),drought (PEG6000),high salinity treatments and oriental armyworm Mythirnna separata feeding stress,while treatments of MeJA and Acidovorax avenae subsp. avenae showed no effect on ScJMT expression. The results make foundations for further research on the role of this gene,and also provide gene resources for the cultivation of new resistant sugarcane varieties.

Plant basic helix-loop-helix (bHLH) transcription factors are the second largest class of transcription factors in eukaryotes,and play important roles in plant growth,secondary metabolism and stress response. In this study,bHLH family genes were screened by transcriptome sequencing of root samples of Pinus massoniana seedlings. The subcellular location,conservative motifs,expression pattern and phylogenetic analyses of these genes were performed by bioinformatics methods. The results showed that 64 bHLH family genes were identified from the transcriptome of Pinus massoniana,and these genes were predicted to be mainly located in the nucleus. Motif1 and motif2 appeared most frequently in conserved motifs. Under the condition of low-phosphorus stress,PmbHLH01/09/27/29/32/39/41 showed significantly differential expression,suggesting that they might be involved in phosphorus stress. These results provide a reference for further revealing the biological function of bHLH transcription factors in response to phosphorus change of P. massoniana.

The absence of horns or polled phenotype is desirable in the modern cattle industry for their beneficial to animal welfare and reduction of breeding costs. In this study,Illumina-based whole-genome resequencing of seven Mongolia cattle (Bos Taurus) (3 horn,4 polled) was reported. The whole genomic DNA was used to prepare 350 bp pair-end libraries and the high-throughput sequencing yielded 380.88 Gb. Comparative genomic analyses identified 11 362 382 genome-wide single nucleotide polymorphisms (SNPs) and 1 150 664 insertions/deletions (InDels),Most of the variations (96% of SNPs and 94% of InDels) were located in intergenic,transcript and intron regions,noted 3 768 genes. After screening,20 differential genes were related to the horn growth and development. Meanwhile,the population stratification analysis of SNPs and InDels showed that hornless traits did not cause genetic differentiation of Mongolian cattle. Further analysis by sequence,polymerase chain reaction (PCR),agarose gel electrophoresis and Sanger sequencing showed that the Mongolia cattle POLLED genotype belongs to P_219ID mutation. This study provides baseline data for further research to investigate the molecular mechanisms of polled trait,and provides a valuable resource for genetic characteristics of Mongolia cattle.

The myostatin gene (MSTN) is a negative regulator of skeletal muscle development,and its function lose in mammals leads to overdevelopment of muscle,significant improvement of production performance,and certain effects on meat quality traits. The present study utilized the muscle tissues from MSTN gene edited Luxi beef cattle (Bos taurus) (Exp for short) and wild type (Ctr for short) to perform label free quantitative comparative proteomics,and investigate the effect of MSTN artificial mutation on meat quality traits. A total of 1 595 quantified proteins containing at least one unique peptide per protein were identified. After scatter plotting analysis used for determining the internal error of the biological replicates and student t-test analysis of the data set,the proteins with fold change of Exp/Ctr in relative abundance more than 1.5 or less than 0.67 and a P-value less than 0.05 from t-test were identified as differentially expressed proteins (DEPs). As a result,160 DEPs were identified which contained 28 proteins in increased abundance and 132 in decreased abundance. Gene Ontology (GO) enrichment and KEGG pathway analysis were performed to explore molecular mechanism of DEPs in the determination of meat quality. There were 5 GO terms of biological process,7 of cellular component,and 5 of molecular function enriched significantly. The biological process were related to oxidation reduction process,skeletal muscle contraction,transition between fast and slow fiber,mitochondrial electron transport nicotinamide adenine dinucleotide (NADH) to ubiquinone,cardiac muscle hypertrophy in response to stress. The cellular component were extracellular exosome,mitochondrial respiratory chain complexⅠ,mitochondrion,Z disc,focal adhesion,extracellular matrix,mitochondrial inner membrane. The molecular function were actin binding,NADH dehydrogenase (ubiquinone) activity,protein C-terminus binding,structural constituent of muscle,thioredoxin peroxidase activity. KEGG pathway analysis results included non-alcoholic fatty liver disease (NAFLD),oxidative phosphorylation,Alzheimer's disease,Parkinson's disease,Huntington's disease. Among the DEPs there were 27 proteins related to meat quality traits,which had been reported. The proteins related with muscle fiber were MYH (myosin heavy chain) 1,MYH6,and MYL6 (myosin light polypeptide 6),MYBPH (myosin binding protein H) and MYOM2 (myomesin (M-protein) 2). The proteins related to adipogenesis were ADIPOQ (adiponectin),CDH13 (cadherin-13),ECH1 (enoyl coenzyme A hydratase 1),CAT (catalase) and VIM (vimentin),which were all down-regulated. Nine proteins of mitochondrial NADH dehydrogenase were also down-regulated,containing NDUFC2/B2/S6/V3/A13/B5/S3/B9/S8. To illustrate the regulation information,interaction networks of DEPs and MSTN were analyzed through the STRING database. The results revealed that the 27 DEPs related to meat quality traits all interacted with MSTN,of which MYH6,MYH1 and ADIPOQ interacted with MSTN directly. By combination of the results reported in this work and previous studies,it could be concluded that MSTN might play a critical role in muscle development and energy metabolism and affect the beef quality traits by changing the types and composition of muscle fibers,reducing intramuscular fat synthesis and decreasing oxidative characteristics. The present research would facilitate further studies on the biological response of postmortem meat and provide further insight into the molecular mechanisms of proteins in the regulation of meat quality traits.

The content and components of fatty acids in milk determine the quality of milk,while miRNAs can regulate the lipid metabolism pathway in mammary glands of mammals,thereby affect lipid content and components. The purpose of this study is to reveal the effects of miR-206 on the expression of genes related to lipid synthesis,triglyceride content and fatty acid composition in bovine (Bos taurus) mammary epithelial cells (BMECs) by miRNA transfection. In this study,60 nmol/L miR-206 mimics and inhibitor were transfected into bovine mammary epithelial cells,then qRT-PCR and biochemical experiment were used to detect the mRNA level of lipid synthesis-related genes,intracellular triglyceride content,respectively. and the changes of fatty acid composition were detected by gas chromatography. The results showed that when transfected with miR-206 mimics,the mRNA level of triglyceride synthesis-related genes including mitochondrial glycerol-3-phosphate acyltransferase (GPAM),1-acylglycerol-3-phosphate O-acyltransferase 6 (AGPAT6),diacylglycerol O-acyltransferase 1 (DGAT1) and lipin 1 (LPIN1) were significantly down-regulated (P<0.01) with significant reduction of intracellular triglyceride content (P<0.05). The expression of sterol-regulatory element binding proteins 1 (SREBP1),fatty acid synthase (FASN),acetyl-CoA carboxylase α (ACACA) and long chain fatty acid elongase 6 (ELOVL6) were down-regulated and fatty acid composition changed,in which the content of C14∶0 (P<0.01) and C22∶6n3 (P<0.05) were significantly reduced,and the content of C16∶1 (P<0.01) was significantly increased. When transfected with miR-206 inhibitor in bovine mammary epithelial cells,the expression of GPAM,AGPAT6,DGAT1 and LPIN1 were significantly up-regulated (P<0.01) and the intracellular triglyceride content increased,combined with up-regulated SREBP1,FASN,ACACA and ELOVL6 (P<0.01),but the fatty acid composition did not change. In summary,miR-206 had a negative regulatory effect on the expression of lipid synthesis-related genes in bovine mammary epithelial cells,also led to decreased intracellular triglyceride content and changed fatty acid composition,which suggested that miR-206 might play an important role in bovine fat metabolism. This study provided basic information for further study on the molecular mechanism of miR-206 in regulating lipid metabolism in bovine breast epithelial cells.

Morphometric influences sports performance,new techniques of genome-wide association study (GWAS) for morphometric trait of horse introduce new possibilities for sports performance. The aim of this study was to enrich the contents of morphometric traits and sport performance of horses (Equus caballus),it can explore the regulation genes of sport performance. In this study,GWAS methods were used to seek genetic markers and functional genes that affect the morphology traits of horses. A GWAS for morphometric traits (withers height (WH),body length (BL),cannon circumference (CC),croup height (CH),body length ratio (BLR),heart girth ratio (HGR),cannon circumference ratio (CCR) were conducted in 4 breeds of 157 individual horses (Yili horses,Thoroughbred horse,Russian speed horse and Orlov horse) which were genotyped using the Affymetrix Equine SNP 670 chip. Plink1.9 software was used to quality control for SNPs,after quality control,494 137 SNPs were participated in association analysis. Databases of Ensembl were used to carry out gene annotation and gene function analyses. Through Bonferroni correction at the genomic level,3 SNPs,15 SNPs,1 SNPs,1 SNPs and 9 SNPs were potentially significantly associated with the 5 traits of WH,CC,CH,HGR,CCR,respectively (P<9.29E-06). However,no SNPs were associated with BL and BLR. It was found that the 2 genes UBE2E3 and LUM regulated the sport performance of horses,which provided a reference for the application of marker-assisted selection in horse breeding.

Post-weaning diarrhea (PWD) causes serious economic losses to large-scale pig (Sus scrofa) farms at home and abroad,of which Escherichia coli F18 (E. coli F18) is the main pathogen causing bacterial diarrhea in piglets. E. coli F18 receptor-related molecule,α-1,3-fucosyltransferase gene (FUT3) was identified by relevant studies and sequencing. In order to investigate the basic structural characteristics and functions of FUT3 gene in pigs (Sus scrofa),the coding region of FUT3 gene was amplified and cloned by PCR. Then,this study predicted its protein structure and function regions by bioinformatics methods,detected the tissue expression profile by qRT-PCR,analyzed its localization and expression distribution by immunohistochemical methods,and detected the mRNA and protein expression levels of FUT3 in E. coli F18-stimulated intestinal porcine epithelial cell line J2 (IPEC-J2). The results showed that the coding region of FUT3 gene in Meishan pigs was 1 068 bp in length,encoding 361 amino acids,which was a lipophilic hydrophilic and unstable non-secretory protein. FUT3 protein sequence included 2 glycosylation sites,29 phosphorylation sites and 2 conserved domains (Glyco_tran_10_N and Glyco_transf_10). Tissue expression profile showed that FUT3 gene was expressed in various tissues of pigs,especially intestinal tissues (duodenum,jejunum and ileum). Immunohistochemistry analysis showed that the expression level of FUT3 in sensitive duodenum was significantly higher than that in resistant duodenum,and FUT3 was mainly distributed on the mucosal surface of small intestinal epithelial cells. After E. coli F18 bacteria stimulation in IPEC-J2 cells,the expression levels of FUT3 mRNA and protein were extremely significantly up-regulated (P<0.01). The expression levels of FUT3 gene in duodenal tissues of weaned piglets in F18-sensitive group were extremely significantly higher than those in resistant group (P<0.01). In this study,the full-length coding sequence of Meishan pig FUT3 gene was successfully cloned,and its protein structure and function were understood from the bioinformatics level. Besides,the important regulatory role of porcine FUT3 expression level on E. coli F18 resistance was preliminarily verified at the individual and cellular levels,providing a certain theoretical basis for in-depth exploration of porcine FUT3 gene function and expression regulation mechanism in the future.

Prox1 (prospero-related homeobox protein 1) is a transcription factor that plays an important role in embryonic development and carcinogenesis. This study aimed to predict and identify the nuclear localization signal (NLS) in chicken (Gallus gallus) Prox1 protein. Chicken Prox1 protein was truncated to determine the NLS region by fluorescence observation and subcellular localization analysis. The online software was then used to predict the putative NLS (pNLS) in Prox1 truncates,and the NLS was confirmed by examining the subcellular localization of the pNLS deletants. In addition,the conservation and nuclear import ability of chicken Prox1 NLS were analyzed. Moreover,the key basic amino acids determining the nuclear localization of Prox1 NLS were identified by point mutation test. The results showed that the N-terminus (1~220 aa) of chicken Prox1 protein had nuclear localization characteristics and two pNLS including pNLS1 (¹⁵KRRR¹⁸) and pNLS2 (¹⁶³RAKRAR¹⁶⁸) existed in this region by NLS software prediction. The subcellular localization analysis of pNLS deletants revealed that pNLS1 and pNLS2 codetermined the nuclear localization of chicken Prox1 protein. In addition,the NLS1 and NLS2 motifs in chicken Prox1 protein were conserved among other poultry,human (Homo sapiens) and different mammals,and had the same nuclear import ability as the NLS of SV40 large T antigen. Moreover,the results of basic amino acids mutation experiments showed that K15,R16,R18 and K165,R166,R168 were key amino acids in NLS1 and NLS2,respectively,which jointly determined nuclear localization of chicken Prox1 protein. This study confirmed two highly conserved NLSs present in chicken Prox1 protein,and both NLSs determined the nuclear localization together,which provided a reference for further studying molecular mechanism and function of the nuclear localization of chicken Prox1 protein.

Bone sialoprotein (BSP) is a major component of medullary bone in chicken (Gallus gallus) and plays an important role in bone remodeling and mineralization. To explore the regulatory mechanism of BSP gene expression and the effect of variations in the promoter region on gene expression in laying hens,Hyline Grey laying hens were used as experimental animals,and the sequence variation in BSP promoter region was analyzed with gene cloning and sequencing technology. The online software JASPAR2018 was used for binding site prediction of transcription factors. Luciferase reporter gene recombinant vectors with different length of promoter fragments were constructed and transfected into chicken fibroblast line DF-1 whose dual-luciferase activity was measured. The expression of BSP gene was determined by qRT-PCR method. The results showed that there were 6 haplotypes (Hap1~6) in BSP promoter (-1257~+142 bp),and the core promoter region was located at -202~+142 bp. The difference of transcriptional activity was also found between Hap1 and Hap5 in the promoter region. The high transcriptional activity of Hap1 in the core promoter region caused an increase in the expression level of BSP gene. In conclusion,the core region in promoter of chicken BSP gene was identified and cloned successfully,which could provide basic materials for the investigation of regulatory mechanism of BSP gene expression and genetic selection of bone quality in laying hens.

The GH-IGF signal pathway plays an important role in regulating the growth of fish. In order to search for molecular markers distribution of single nucleotide polymorphisms (SNPs) of growth hormone receptor (GHR) and insulin-like growth factor-Ⅰ (IGF-Ⅰ) genes in fast-growing traits of tilapia (Oreochromis niloticus),The SNPs locus for the above genes of Nile tilapia (O. niloticus) and genetic improvement of farmed (GIFT) tilapia (O. niloticus) were screened. In this experiment,32 SNPs were screened from the promoter region and coding region of 2 tilapia GHR1,GHR2 and IGF-Ⅰ genes by Sanger sequencing technology. After functional secondary screening,a total of 9 SNPs were retained in Jifu tilapia (JF) and distributed in the GHR1 gene (7) and GHR2 gene (2),and a total of 5 SNPs were retained in Nile tilapia egyptian strain (AJ),which were distributed in GHR1 gene (4) and GHR2 gene (1). General linear model (GLM) and least significant difference (LSD) were used to analyze the correlation between SNPs and growth traits in the offspring of 2 varieties of tilapia. the results showed that 12 genotypes of 6 SNPs loci in the GHR1 and GHR2 genes of the JF population were significantly associated with fast-growing traits,while no SNPs loci and genotypes significantly associated with the fast-growing traits were found in the AJ population. The above results can be used as candidate molecular markers in molecular assisted breeding of tilapia,and provide a theoretical basis for subsequent research on growth factors in fish.

Trialeurodes vaporariorum is a worldwide pest with the characteristics of multi feeding and wide host range. Antennae is an important organ for insect information exchange,and its olfaction-related genes are an important molecular basis for regulating insect feeding behavior. However,to date,there are few reports about gene data of T. vaporariorum antennae,and little is known about the number and type of primary olfaction-related genes. The purpose of this study is to establish a transcriptomic database of T. vaporariorum antennae,and to investigate the number of genes encoding olfaction-related proteins. The antennal transcriptomes of adult T. vaporariorum were sequenced using Illumina HiSeq 4000 platform,and bioinformatic analyses were conducted. In this study,32 890 unigenes were obtained through assembling after transcriptome sequencing,with mean length of 1 616 bp and an N50 value of 3 059 bp,and 14 206 unigenes over 1 000 bp in size．After annotating for unigenes in gene function database,17 846 unigenes obtained functional annotation information. Sequences searched into NR database (non-redundant protein sequence database) accounted for 88.77% of all annotated unigenes,most of which was optimally hit to genes of Bemisia tabaci (87.12%) species. Annotated results in GO database showed that unigenes were divided into 54 subcategories included into 3 categories,and a lot of unigenes were annotated to be associated with metabolic process and catalytic activity. In KEGG database,5 964 unigenes were annotated in 228 metabolic pathways. Among them,most unigenes (189) were involved in metabolic pathways of ribosome. Based on further annotated information of unigenes,33 genes encoding olfaction-related proteins of T. vaporariorum including 21 genes encoding odorant binding proteins (OBPs),5 genes encoding chemosensory proteins (CSPs),2 genes encoding olfactory receptor proteins (OLRs),3 genes encoding odorant receptor proteins (ORs),1 gene encoding ionotropic receptor protein (IR) and 1 gene encoding sensory neuron membrane protein (SNMP). This study obtained the antennal transcriptomic data of adult T. vaporariorum,its characteristics were preliminarily elucidated,as well as genes encoding olfaction-related proteins were identified. This result provides a reference for further investigation on behavior regulation and new control strategies of T. vaporariorum at molecular levels,and also enriches the gene database of horticultural pests.

Pluripotent stem cells have the ability of self-renewing and multiple differentiation potential. Germ cells are the carriers of genetic information. Direct differentiation of germ cells from pluripotent stem cells not only lays the foundation of studying reproductive development and organogenesis,but also provides new ideas and methods of treating human (Homo sapiens) infertility and conserving excellent animal breeds. At present,the study of induced differentiation of mouse (Mus musculus) pluripotent stem cells into germ cells is the most mature,and functional germ cells can be obtained in vitro; the system of induced differentiation of human,pig,cattle,sheep and other livestock pluripotent stem cells into germ cells is not mature,which needs further optimization. In this paper,the methods and systems of inducing differentiation of mouse,human and livestock pluripotent stem cells into germ cells are reviewed,and the problems and research strategies in inducing differentiation are prospected. This review provides a theoretical basis for the differentiation of domestic animal pluripotent stem cells into germ cells.

Cow (Bos taurus) mastitis is an inflammatory disease caused by pathogenic microorganism infection,trauma or other factors in mammary gland tissue. It has complex causes,high incidence and expensive treatment cost,which seriously restricts the development of dairy industry. MicroRNA (miRNA) can regulate the expression of messenger RNA (mRNA) at the post-transcriptional level,thus regulating biological processes including body immunity. In recent years,it has been found that miRNA played an important role in the occurrence and development of mastitis in dairy cows. In this paper,the expression pattern and molecular regulation mechanism of miRNA in mammary tissue,peripheral blood,milk of different types of mastitis cows and pathogenic bacteria-induced mammary epithelial cells were reviewed,and the problems that need to be solved were analyzed and prospected,which will provide reference for researchers to further understand the related fields.

Virus-induced gene silencing (VIGS) is an efficient and convenient reverse genetics method for rapid gene function study. The application of the VIGS is often restricted due to the incompatibility of viruses and plants. Tobacco rattle virus (TRV) -mediated VIGS technology achieves gene silencing on a variety of plant species, but its application is limited by plant damage and low silencing efficiency caused by conventional inoculation operation that applies agroinfiltration in plant leaves. In this study, soybean (Glycine max) was used as a material to validate a modified TRV-mediated VIGS protocol that used Agrobacterium tumefaciens to infect from the root after soil irrigation. The results showed that the A. tumefaciens carrying the VIGS plasmids that infected soybean seedlings implemented effectively silence in all plants tested in the same treatment batch, silencing efficiency of targeted genes were above 67%, and reached 98% of the highest. Further examination on the soybean ternately compound leaves at different positions of the treated plants revealed that the first to the third-round of ternately compound leaves had a satisfactory silencing effects, of which the first-round leaves had the highest silencing efficiency. The genes that were effectively silenced in this experiment included non-specific lipid-transfer protein (LTP), pathogenesis-related protein 1 (PR1), thaumatin-like protein (TLP), dehydrin-like protein (DHN), xyloglucan endotransglucosylase (XTH), and cysteine-rich transmembrane protein (CYSTM). The method was proved to be simple and feasible, and a large number of gene silenced plants were obtained in a short period of time. This study established an efficient system for rapid gene function verification in soybean.