In wheat (Triticum aestivum), grain weight selecting is effective for improvement of grain yield as its high heredity and stable phenotype. In previous study, a stable QTL for grain weight, qTkw-2D, was identified and this QTL has been verified in several populations. In this study, the linkage marker Xcfd233 and Wheat660K SNP array were used to scan 3 000 high generation of breeding population. Finally, near isogenic lines (NILs) of qTkw-2D were achieved in the F₆ generation of 'Kenong 2011'. By comparison the pericarp cells size of the grain slices from two NILs, it was confirmed that qTkw-2D mainly affected grain width by affecting the size of inner pericarp cells. The analysis results of filling rate showed that qTkw-2D mainly affected grain weight by affecting the grain filling rate during middle and late filling stage. The grain weight in different environments at individual and population level of NILs indicated that qTkw-2D influenced grain yield in both levels and haplotype A was of great importance to maintain grain yield during poor harvest environments. This study provides reference for grain weight improvement and function gene cloning.

Wheat (Triticum aestivum) is one of the most important food crops in China. High temperature and other abiotic stresses seriously affect the yield and quality of wheat. The discovery and identification of resistance genes can provide candidate genes and theoretical foundation for wheat stress resistance breeding. Heat shock proteins (Hsps) are crucial molecular chaperones that preserving cellular function under stress conditions and regarded as significant candidate genes for heat resistance. In this study, the TaHsp17.9 gene (GenBank No. MN684335) of wheat was amplified by PCR using the cDNA of the heat tolerance wheat variety 'Pingyao Xiaobaimai' as a template. The total CDS length of TaHsp17.9 is 474 bp, encoding 157 amino acids. The protein sequence of TaHsp17.9 contains the specific α-crystallin domain (ACD) of plant small heat shock protein (sHsp). Phylogenetic tree analysis indicated that Hsp17.9 had the closest genetic distance to Triticum dicoccoides. qPCR results showed that the expression levels of TaHsp17.9 were highest in grain and lowest in root. The TaHsp17.9 was induced to express by high temperature and drought stress, and its expression level reached the peak at 1 and 2 h respectively. The expression level of TaHsp17.9 in heat tolerance varieties was significantly higher than that in heat sensitive varieties under the heat stress. The results of subcellular localization showed that TaHsp17.9 protein was localized in the cytoplasm. Identification of heat tolerance in the TaHsp17.9 overexpression strain of Arabidopsis thaliana, the result showed that, overexpression of TaHsp17.9 could significantly improve the heat tolerance of transgenic Arabidopsis thaliana. In addition, 10 candidate interacting proteins that might interact with TaHsp17.9 were obtained based on STRING database. This study provides a reference for further studying the function of TaHsp17.9 gene in wheat.

The purpose of this study was to investigate the effects of nitrogen combined application on soil-rice (Oryza sativa) microbial system under straw returning condition, and to balance the effects of straw decomposition and nitrogen competition in rice growth at the early stage of nitrogen application, this study was based on plot experiments, with no fertilization treatment as the control (CK), the base fertilizer was set as the ratio treatment test of 5 different proportions of ammonium bicarbonate+compound fertilizer,including 100% compound fertilizer (S0), 20% ammonium bicarbonate+80% compound fertilizer (S1), 40% ammonium bicarbonate+60% compound fertilizer (S2), 60% ammonium bicarbonate+40% compound fertilizer (S3) and 80% ammonium bicarbonate+20% compound fertilizer (S4) to study the effect of nitrogen fertilizer on rice growth, soil properties and microbial diversity after 6 d of nitrogen fertilizer application impact. The results showed that in the early stage of nitrogen fertilizer application, soil organic matter, pH value, available phosphorus and available potassium decreased with the increase of ammonium bicarbonate, while the contents of ammonium nitrogen and nitrate nitrogen increased; Fresh weight, root surface area, root volume and plant carbon and nitrogen accumulation increased first and then decreased. Except fresh weight under ground, treatment S2 was the highest, and was significantly different from other treatments (P<0.05). The Chao1 index and Shannon index of soil bacterial and fungal communities first decreased and then increased. The treatment S2 was the lowest, and the ammonia-oxidizing archaea (AOA) and ammonia-oxidizing bacteria (AOB) communities related to ammonia nitrogen had no significant difference among the treatments; the β diversity index, the number of soil bacteria exclusive operational taxonomic unit (OTU) was higher in treatments S2 and S4, while the number of fungi exclusive OTU was the lowest in treatment S2. The dominant soil bacterial phyla in the soil were Chloroflexi, Actinobacteriota, Proteobacteria and Acidobacteriota, among which treatment S2 improved the relative phyla of Actinobacteriota. The abundance of Chloroflexi, Proteobacteria and Acidobacteriota decreased to a significant level. The dominant fungal phyla in soil were Ascomycota and Basidiomycota. According to redundancy analysis (RDA) analysis, Actinobacteriota and Basidiomycota were significantly positively correlated with soil available nitrogen content and rice growth indexs, and Proteobacteria and Ascomycota were significantly correlated with soil available K, soil organic matter (SOM), and Olsen P, while significantly negatively correlated with rice growth indexes. Chloroflexi was significantly positively correlated with soil pH, and the treatment of S2 was the most obvious with Actinobacteriota. Thaumarchaeota was significantly positively correlated with rice growth indexs, NH₄⁺-N, soil easily oxidizes organic carbon (EOC) and soil dissolved organic carbon (DOC), and significantly negatively correlated with pH, NO₃^--N, olsen P and available K. The reasonable combination of nitrogen fertilizer method S2 under straw returning could balance the nutrient requirements of soil microorganisms and rice. This study is of great significance for soil fertility and crop growth to apply nitrogen fertilizer reasonably under straw returning.

Natural resistance associated macrophage proteins (NRAMPs) play an important role in metal transport in the transmembrane. To investigate the role of tobacco (Nicotiana tobacum) NRAMP genes in heavy metal Cd uptake and transport, a NRAMP subfamily gene, named NtNRAMP2 (GenBank No. OP972862), was cloned from cultivated tobacco 'K326' in this study. Bioinformatics analysis and subcellular localization were carried out to clarify the biological function of the NtNRAMP2, and its expression level in tissue specificity and under exogenous Cd, ABA and ethylene treatment were analyzed by qPCR. Sequence analysis showed that the full-length CDS of tobacco NtNRAMP2 gene was 1 629 bp, encoding 542 amino acids, containing 4 exons and 10 transmembrane structural domains. Evolutionary analyses revealed that NtNRAMP2 was most closely related to tomato (Lycopersicon esculentum) as well as potato (Solanum tuberosum), subcellularly localised in the endoplasmic reticulum and vesicles. qPCR results showed that NtNRAMP2 gene was expressed in different tissues of tobacco, but the expression was highest in the roots. NtNRAMP2 expression was significantly increased under different concentrations and times of Cd stress, and NtNRAMP2 gene expression was also triggered in tobacco roots by both exogenous ABA and ethylene. Meanwhile, NtNRAMP2 overexpression vector was constructed and 18 NtNRAMP2 overexpressing transgenic tobacco plants were obtained by the Agrobacterium tumefaciens mediated transformation method. This study provides a reference for further exploration to study the function of NtNRAMP2.

Kiwifruit (Actinidia chinensis) is one of the pillar industries for farmers in mountainous areas to fight poverty. Gray mold caused by Botrytis cinerea is easy to cause postharvest corruption of kiwifruit, which seriously restricts the development of kiwifruit industry. In our research group previous study, kiwifruit samples with AcTPR2 gene down-regulated were obtained by virus induced gene silencing (VIGS) method. On this basis, kiwifruit treated with sterile water (WT), transformed with empty vector Tobacco rattle virus (TRV) without AcTPR2 gene, and transgenic kiwifruit (AcTPR2-TRV) that successfully achieved the down-regulation of AcTPR2 gene expression were used as materials, By detecting the lignin content of kiwifruit infected by B. cinerea and the expression of key enzyme genes in lignin biosynthesis pathway, the relationship between AcTPR2 induced resistance of kiwifruit to B. cinerea and the lignin content was explored. The results showed that compared with the control group, a significant increase in lignin content was detected 24~48 h after B. cinerea infection, indicating that the lignin accumulation was involved in the early stage of kiwifruit resistance to B. cinerea infection, and the expression of key enzyme genes in lignin synthesis pathway like phenylalanine ammonia lyase (PAL), cinnamate 4-hydroxylase (C4H), coumaric acid 3-hydroxylase (C3H) and cinnamoyl-CoA reductase (CCR) was consistent with the increasing trend of lignin content. The lignin content and the expression of related genes in AcTPR2-TRV fruits increased significantly compared with the control group at 24 h after successful transformation, indicating that kiwifruit with AcTPR2 down-regulated activated the lignin synthesis signal pathway to accelerate lignin biosynthesis and participate in the resistance response of kiwifruit to gray mold (P<0.05). Therefore, the lignification of kiwifruit cell wall regulated by AcTPR2 is related to the resistance response of kiwifruit to B. cinerea, which is one of the mechanisms of disease resistance response in kiwifruit. The results of the present study not only enriched the disease resistance regulation mechanism of kiwifruit, but also provide important gene resource information for disease resistance molecular breeding of kiwifruit in practice.

Heat shock protein 70 (HSP70) is a kind of protein molecular chaperone widely existing in organisms, which plays an important role in growth and development. However, the study of HSP70 in Phyllostachys edulis has not been reported. In this study, HSP70 genes were identified from P. edulis by bioinformatics methods, the protein physical and chemical properties, conservative motifs and gene structure, the chromosome location and promoter cis-regulatory element were analyzed, and a phylogenetic tree was built; Based on transcriptome data and qRT-PCR, the expression level of HSP70 gene in P. edulis of different growth heights was analyzed, and the subcellular localization of key HSP70 was verified. The results showed that a total of 40 HSP70 (PeHSP70-01~40) genes were identified from P. edulis, and 39 PeHSP70 genes were hydrophilic proteins. Gene structure analysis demonstrated that 21 PeHSP70 genes contained 1 intron. Abscisic acid-related response elements were most found in the upstream 2 000 bp of the PeHSP70 genes. The phylogenetic analysis suggested that all PeHSP70 proteins were clustered into 4 subfamilies and closely related to Oryza sativa. Transcriptome data showed that the expression of 11 PeHSP70 genes were up-regulated during the rapid growth of P. edulis. The results of qRT-PCR showed that 4 PeHSP70 genes PeHSP70-23~26 as mitochondrial localization were up-regulated in the elongated shoots. Furthermore, subcellular localization results showed that the fluorescence of GFP was colocalized with mitochondrial fluorescence dye, and these 4 PeHSP70s were located to mitochondria. Together, this study provides a basis for exploring the mechanism of HSP70 genes in regulating rapid growth of P. edulis.

Nitrogen is one of the essential macronutrient for plants, thus nitrogen content in soil seriously affects plant growth and development. Nitrate as the most important source of nitrogen absorption for P. bournei and many other land plants, and the nitrate transporter/peptide transporter family (NRT/NPF) is involved in the nitrate uptake, transport and distribution. Therefore, the genome-wide identification and expression analysis of the NRT/NPF gene family are important for the studying of nitrogen uptake and utilization in P. bournei. In the present study, the genome-wide NRT/NPF genes of P. bournei were identified using NRT protein sequences of Arabidopsis thaliana and Hidden Markov Model (HMM) (PF00854, PF07690, PF16974), and 63 PbNRT/NPF members were identified, which were unevenly distributed on 11 chromosomes. The results showed that the molecular weights of PbNRT/NPF were 22.8~135.6 kD, isoelectric point were 5.44~9.41, encoding 205 to 1 235 aa. Most of the encoded proteins were basic proteins with 2~23 transmembrane domains, phylogenetic analysis revealed that 63 PbNRT/NPF genes could be divided into 3 subfamilies, of which 60 belonged to the NRT1/NPF subfamily, 2 to the NRT2 subfamily subfamily and 1 to the NRT3 subfamily. Collinearity analysis revealed that there were 9 tandem duplication events and 10 segmental duplication events in the PbNRT/NPF family. The transcriptome data and qRT-PCR analysis showed that PbNRT/NPF members were differentially expressed among 5 tissues of P. bournei, for example, PbNPF6.3 was significantly higher expressed in the root bark than that of other tissues. Furthermore, PbNRT/NPF members showed differential expression patterns under different low nitrogen treatments, e.g. the expression of PbNRT2.4 and PbNRT2.5 significantly increased under low nitrogen treatment. Overall, this study provides a basis for studying the functions of the PbNRT/NPF genes in nitrogen absorption, transport and utilization in P. bournei, and provides a theoretical basis for breeding and selection of low-nitrogen tolerant varieties in P. bournei.

Begonia semperflorens are considered as an important garden ornamental plant, with its flowers open in clusters and gorgeous colors. In order to study the mechanism of the flower color formation of B. semperflorens, in this study, the anthocyanin synthesis pathway key enzyme flavonoid glycosyltransferase gene (UDP-glycose flavonoid glycosyltransferase, UFGT), were cloned and analyzed using B. semperflorens as materials, according to the previous stage of B. semperflorens transcriptome data. Differences in the expression of BsUFGT gene in different tissues of B. semperflorens were detected by fluorescence quantitative PCR and transformed into Nicotiana tabacum 'NC89' for functional verification. The results showed that a cDNA of 1 536 bp and 511 amino acids gene sequence was successfully cloned and named BsUFGT (GenBank No. OL329828). The molecular weight was 56 385.34 D, the theoretical isoelectric point (pI) was 5.25. Domain analysis showed that the UFGT protein encoded by BsUFGT gene had a UDPGT domain. Subcellular localization results showed that BsUFGT was mainly localized in cell membrane and nucleus.The qRT-PCR analysis showed that BsUFGT expression in B. semperflorens flowers was significantly higher than that in leaves and stems (P<0.05). BsUFGT of B. semperflorens were transformed into N. tabacum 'NC89' for functional verification. The anthocyanin content of anthocyanins in transgenic tobacco corolla was higher than that in wild-type tobacco corolla. This study provides a theoretical basis for the molecular regulation mechanism of anthocyanin synthesis in B. semperflorens.

Mastitis is the most important disease affecting dairy cows (Bos taurus), but there is no efficient method for its prevention and control, and exosomes in cow milk may have an important role in the development of mastitis. To investigate how exosomes in milk of cows change in different functional states, such as healthy, subclinical mastitis and clinical mastitis, in this study, exosomes were isolated from the milk of cows by ultracentrifugation, identified by nanoparticle tracking, transmission electron microscopy and Western blot, and compared the differences in particle diameter, number, protein concentration and surface marker molecules between the groups. The results showed that the particle size of exosomes in each group was close, the morphology was consistent with the characteristics of exosomes, and there was no significant difference between the groups; Western blot results showed that the exosome molecular marker proteins cluster of differentiation 9 (CD9), CD81 and tumor susceptibility gene 101 (TSG101) were positive in each group. The most exosomal particles were found in the subclinical mastitis group, followed by the clinical mastitis group, both of which were significantly higher than the healthy group (P<0.05); the highest exosomal protein content was found in the subclinical mastitis group, followed by the clinical mastitis group, both of which were significantly higher than the healthy group (P<0.05). In this study, exosomes were isolated from cow milk, and their number, protein content and surface molecular markers changed significantly in different functional states of the mammary gland, suggesting that exosomes may have a regulatory role in the immune process against infection in the mammary gland of cows. Exosomes may play a role in the regulation of mammary gland immunity against infection in dairy cows.

Hippo signaling pathway is a key pathway regulating the growth and development of animal tissues and organs, and plays an important role in mammalian ovarian development. To investigate the Hippo signaling pathway related genes mammalian ste20-like protein kinase 1 (MST1), large tumor suppressor 1 (LATS1) and Yes-associated protein 1 (YAP1) in yak (Bos grunniens) ovaries during different reproductive cycles, in this study, the ovarian tissues of healthy female yaks (Bos grunniens) from different reproductive cycles (follicular, luteal, gestation) were selected as the test samples. qRT-PCR and Western blot were used to detect the expression differences of the MST1, LATS1 and YAP1 and their protein in yak ovaries of different reproductive cycles. Immunohistochemistry (IHC) was used to detect the distribution of genes related to Hippo signaling pathway in yak ovaries at 3 cycles. The results of qRT-PCR showed that there were significant differences in the expression of MST1 gene in ovarian tissues at different stages (P<0.05) with the highest expression in follicular stage, the second in gestation stage and the lowest in luteal stage. The expression levels of LATS1 and YAP1 genes in pregnancy were significantly higher than those in follicular and luteal phases (P<0.05). Western blot results showed that the protein expressions of MST1, LATS1 and YAP1 increased gradually, and their expression reached the peak in pregnancy phase, then decreased and maintained at a low level, which was significantly different from other stages of pregnancy phase (P<0.05). IHC results showed that MST1 was mainly distributed in granulosa cells, membrane cells, granular luteal cells, membranous luteal cells and ovarian germ epithelium of ovarian follicles. The distribution positions of LATS1 and YAP1 in different ovarian cycles were consistent with those of MST1. MST1, LATS1 and YAP1 were expressed in yak ovaries in different reproductive cycles, and the expression levels were significantly different, suggesting that MST1, LATS1 and YAP1 may be involved in the regulation of reproductive physiology related processes in yak. This study provides insights into the relationship between the expression of MST1, LATS1 and YAP1 in follicular granulosa cells of yaks at different developmental stages, which will provide a theoretical basis for further research on the reproductive performance of yaks by exploring Hippo signaling pathway.

Cryopreservation of semen is one of the effective means to effectively improve the efficiency of artificial insemination in Ovis aries. Adding antioxidants to semen preservation diluent is the key to semen preservation process. Leonurine alkaloid as an antioxidant, has rarely been reported in semen preservation studies. In this study, 20, 40, 60, 80 and 100 μmol/mL leonurine alkaloid were added to the cryopreserved diluent of Duolang sheep semen, respectively. The effect of leonurine alkaloid on the cryopreservation of sheep semen was comprehensively evaluated by detecting semen quality, antioxidant capacity, relative expression of apoptotic related gene mRNA, and conception rate. The results showed that compared with the control group (0 μmol/mL), the supplementation of leonurine alkaloid in the cryoppreservation diluent could significantly increase the viability rate, acrosome integrity rate and plasma membrane integrity rate of the cryoppreservation semen of Duolang sheep (P<0.05); It significantly increased the activities of superoxide dismutase (SOD), glutathione peroxidase (GSH-PX) and catalase (CAT), increased the level of total antioxidant eapacity (T-AOC) and decreased the content of malondialdehyde (MDA)(P<0.05); The best preservation effect of leonurine alkaloid in cryopreservation was 60 μmol/mL (P<0.05); In addition, compared with the 0 μmol/mL group, the relative mRNA expressions of spermatozoa-related apoptotic genes TNF receptor superfamily, member 6 (Fas), caspase family omega-3 (Caspase-3) and BCL2-associated X (Bax) were significantly decreased (P<0.05), and the relative expression of anti-apoptotic gene B-cell lymphoma-2 (Bcl-2) was significantly increased in the optimal concentration of leonurine alkaloid group during cryopreserved (P<0.05); At the same time, the conception rate of 60 μmol/mL group was higher than that of 0 μmol/mL group, which reached the lowest level of fresh spermatozoa vaginal instillation. This study showed that adding 60 μmol /mL leonurine alkaloid to semen diluent could significantly improve the quality of frozen preserved semen of Duolang sheep, enhance its antioxidant capacity, and play an anti-apoptotic role, so that the conception rate could reach a good level. This study provides a useful reference for exploring the effect of leonurine alkaloid on sheep semen preservation and improving the effect of sheep semen preservation.

Toll-like receptors 10 (TLR10) plays an important role in the immune system by rapidly binding to ligands after pathogenic microorganisms enter the body and promoting the activation of various immune cells. However, the expression characteristics of TLR10 in different tissues and organs of the body need to be further explored. In this study, the TLR10 gene of Camelus bactrianus (GenBank No. XM_010962682.2) in NCBI was used to construct the prokaryotic expression plasmid pET-28a-TLR10. TLR10 recombinant protein was expressed in Escherichia coli, and purified protein was used to prepare TLR10 polyclonal antibody. ELISA and Western blot were used to detect the titer and specificity of the antibody. The results showed that the recombinant protein of bactrian camel was 50 kD in size and mainly expressed in the form of inclusion bodies. The antibody titer obtained after immunization with Oryctolagus cuniculus was 1∶32 000, and the expressed TLR10 protein could specifically bind to the antibody. TLR10 was mainly expressed in cardiomyocytes, hepatocytes and Kupffer cells, monocytes/macrophages in spleen, bronchial epithelial cells, typeⅡ alveolar cells and dust cells in lung and renal tubular epithelial cells in kidney. This study provides materials for further revealing the function of TLR10 protein and its biological role in the innate and adaptive immunity of bactrian camels.

The Jiangxi province has numerous native domestic chicken breeds. This research aimed to characterize the genetic diversity and phylogeny of Jiangxi chicken (Gallus gallus domesticus) breeds. A total of 198 blood samples were collected from 6 breeds (Anyi Gray, Baier Yellow, Chongren Partridge, Silkie, Dongxiang Blue-eggshell, and Yugan Black-bone), reared in different areas of Jiangxi province. In this study, the PCR amplification method combined with nucleic acid sequencing technology was employed to detect the complete mitochondrial DNA (mtDNA) nicotinamide adenine dinucleotide (NADH) dehydrogenase subunit 6 (ND6) sequences. The results demonstrated that the full length of ND6 gene of 6 Jiangxi chicken breeds was 522 bp. A total of 7 variable sites were identified, which defined 7 haplotypes, Hap1 was the mainstream haplotype. The differences in nucleotide diversity, haplotype diversity, and the average number of nucleotide differences were 0.002 75 ± 0.000 86, 0.694 ± 0.024, and 1.437, respectively. The median network diagram exhibited that the haplotype of 6 Jiangxi chicken breeds presented a star-shaped divergent distribution. Phylogenetic analysis showed that Silkie, Dongxiang Blue-eggshell and Yugan Black-bone clustered into one branch, and Anyi Gray, Baier Yellow, and Chongren Partridge clustered into another branch. The mitochondrial ND6 gene polymorphism information of Jiangxi chicken breeds was abundant, black bone chickens may have a different maternal origin. The results of this study will provide a reference for the preservation, improvement, and identification of Jiangxi chicken breeds.

Circular RNAs (circRNAs), a novel endogenous non-coding RNA expressed in various tissues. In this paper, circ_MBNL1 encoded by MBNL1 (musblind-like 1) gene was selected for identification and functional analyses based on the results of circRNA sequencing during the differentiation of duck (Anas platyrhynchos domesticus) embryo primary cells. The conjugated sequence was obtained by PCR amplification and pyrosequencing. And the nucleoplasmic separation assay revealed that duck circ_MBNL1 was mainly localized in the nucleus. Tissue expression profiles suggested that circ_MBNL1 was mainly expressed in duck spleen, adipose and lung tissues. The bioinformatics analysis results indicated that circ_MBNL1 and the predicted target genes were mainly enriched in adipocytokine signaling pathway, FoxO signaling and other pathways, which was hypothesized that circ_MBNL1 might be involved in regulating the differentiation process of duck adipocytes. This study provides a theoretical basis for an in-depth investigation of the regulatory role of circ_MBNL1 in the differentiation process of duck adipocytes.

Cell-cell and cell-extracellular matrix adhesion depends on cell-specific adhesion proteins and their associated regulatory signals. African green monkey kidney epithelial cells (Vero) are excellent cell lines for the proliferation, purification and vaccine production of multiple important viruses such as Influenza virus, Novel coronavirus. During in vitro culture, Vero cells usually grow in the medium containing serum with two-dimensional single-cell layer attached to the wall of petri dishes. Biomedical Research Center of Northwest Minzu University obtained Vero cell lines that could be suspended for culture by using culture method of gradually lowering serumin the medium, but the cell density was low. It is difficult to achieve large-scale high density suspension culture. Gene modification techniques and serum-free culture-medium cell screening techniques have been successfully applied to the construction of suspension cell lines. To construct suspension Vero cells using genetic engineering methods, it is necessary to screen proteins related to their adhesion function and understand how these proteins affect cell adhesion. In this study, tandem mass tag (TMT) quantitative proteomics was used to analyze the types and amounts of differential expression proteins associated with cell adhesion Vero cells (Adh_Vero) and suspension Vero cells (Sus_Vero). Compared with Adh_Vero group, the results showed that there were 190 differential expression proteins in Sus_Vero cells, including 116 up-regulated proteins and 74 down-regulated proteins. Six differentially expressed proteins related to cell adhesion were obtained by GO functional annotation and KEGG pathway enrichment screening. The up-regulated 5 proteins were syndecan 4 (SDC4), enabled homolog (ENAH), myosin light chain 12β (MYL12β), myosin light chain kinas (MYLK), occludin (OCLN), and 1 protein of integrin β 2 (ITGβ2) was down-regulated. Real-time quantitative PCR (qPCR) showed that the mRNA levels of SDC4, OCLN, MYL12β, MYLK and ENAH were most significantly down-regulated (P<0.01), while the mRNA levels of ITGβ2 was most significantly up-regulated (P<0.01). Western blot analysis showed that the protein expression of SDC4 in Sus_Vero was significantly decreased compared with that in Adh_Vero. The trend of protein expression was consistent with quantitative proteomics. The findings preliminarily revealed the adhesion mechanism of Vero cells and provided an effective target proteins for the further construction of Sus_Vero culture using genetic engineering technology.

Pepper anthracnose is one of the 3 major diseases in pepper (Capsicum annuum) production, which can cause rotting fruit and seriously affect the yield and quality of pepper. At present, the use of biological control of pepper anthracnose has green environmental protection and does not produce drug resistance, which is the inevitable trend of agricultural development. In this study, a biocontrol strain ZF438 (CGMCC No. 22291) was isolated from the pepper rhizosphere soil samples. The strain was identified as Bacillus velezensis based on biological characteristics, physiological and biochemical characteristics, and multilocus sequence analysis. At the same time, the inhibitory effect of the strain on 8 pathogenic fungi and 9 pathogenic bacteria were determined by dual culture method, and the results showed that ZF438 had broad-spectrum inhibitory activity. The stability of the antibacterial active substance of strain ZF438 in different temperatures, pH, UV irradiation, ultrasonic treatment and proteases was determined by fermentation supernatant test. The fermentation supernatant showed high stability at 4~40 ℃, pH 2.0~10.0, UV treatment and ultrasonic treatment. The control efficacy of pepper fruits in vitro and potted plants in greenhouse was 68.26% and 47.93%, respectively. The incidence rate and disease index of pepper inoculated with ZF438 strain were significantly reduced. This study verifies the biological characteristics of Bacillus velezensis ZF438, and proves that the strain ZF438 has good biocontrol potential and application prospects, and is of new resources for the biological control of pepper anthracnose.

Most of mangroves grow in intertidal zones with special and changeable environments, which are conducive for the growth of rare Actinomyces and production of active compounds. In order to explore the diversity, novelty and medicinal potential of Actinomyces resources from coastal mangrove in Leizhou, Zhanjiang, China. Pour plate method and 10 different media were used in this study to isolate Actinomyces from mangrove rhizosphere soil samples. After purification and culture, PCR amplification and sequencing and phylogenetic analysis of 16S rRNA of Actinomyces strains were performed. At the same time, according to the comparison results of 16S rRNA and the colony morphological characteristics of strains, portion of Actinomyces strains were selected for small-scale fermentation, all broths without mycelia were extracted by ethyl acetate and dried by rotary evaporator before the preparation of fermentation crude extract methanol solution. Anti-tumor activity of fermentation crude extracts against human lung cancer cell line A549 and human liver cancer cell line HepG2 were analyzed by Cell Counting Kit-8 (CCK-8). According to the result, a total of 154 Actinomycetes were isolated from 8 rhizosphere soil samples collected from 3 different sampling sites in coastal areas of Zhanjiang Mangrove National Nature Reserve, Guangdong Province, distributing in 11 orders, 15 families and 19 genera, with the dominant genera were Micromonspora (22.7%, 35 strains), followed by Microbacterium (20.8%, 32 strains) and Streptomyces (10.4%, 16 strains). And there were 7 potential new species distributing in 4 genera with 16S rRNA sequence similarity less than 98.65%. In the anti-tumor activity screening of fermentation crude extracts, 53 strains of Actinomyces showed anti-tumor activity against at least one tumor cell line, accounting for 79.1% of 67 strains. The above results suggested that, the diversity of Actinomyces and potential medicinal resources were rich in coastal mangrove areas of Leizhou. This study bases a sufficient strain foundation for the further improvement of the cognition of Actinomyces distribution in Leizhou coastal areas as well as the subsequent research on other biological activities such as antibacterial activities and related mechanisms.

DNA methylation is the process of the covalent binding of a methyl group the fifth carbon position of the cytosine of CpG dinucleotide of the genome under the action of DNA methyltransferase. As one of the important epigenetic modification mechanisms, it is a hot topic in the field of life science. With the rapid development of sequencing technology, DNA methylation detection technology has also made significant progress. In terms of animal reproduction, changes in the degree of DNA methylation may affect the expression of genes related to reproductive traits, thus changing the reproductive performance of animals. In this paper, a brief review is made on the overview of DNA methylation, the detection methods of DNA methylation and the research on animal reproduction, and the apparent genetic mechanism of DNA methylation is explored. This review provides a theoretical basis for molecular breeding to give full play to the fertility of domestic animals and select high-quality new varieties or strains.

Long non-coding RNA (lncRNA) is an important epigenetic regulator affecting gene expression and cell function, and its expression pattern is tissue-specific. The biological roles of lncRNA have been widely verified in many diseases and biological phenotypes. Feed intake is one of the main factors affecting the economic benefits of livestock. A large number of studies have focused on the effects of nutrition and immune factors on feed intake of livestock, but its genetic analysis and functional research are still slow. In order to explore the biological role of lncRNA in the regulation of feeding behavior and residual feed intake (RFI) in cattle (Bos taurus), the molecular characteristics and main regulatory functions of lncRNA were summarized in this paper. In addition, the biological roles of lncRNA in regulating RFI variation were summarized from important tissues (hypothalamic-pituitary-adrenal cortex axis, liver, skeletal muscle, and brain-gut axis) that affected the phenotype changes of feed intake in cattle. This review provides a basis for subsequent studies to reveal the phenotypic regulation of RFI by lncRNA from a cellular function perspective, and provides reference for efficient production and breed improvement of cattle.

There are abundant germplasm resources of Chinese bayberry (Morella rubra) in China, and most of them are local accessions. Although a large number of SSR molecular markers have been developed based on the whole genome, a unified set of molecular markers for the identification of bayberry germplasm resources has not been established, which hampers the proper management of genetic identities of the germplasm. In this study, based on the developed SSR molecular markers and linkage map location of Chinese bayberry, 2 SSR markers with stable amplicon length and good reproducibility were selected from each linkage group. The fluorescence-labeled primers were used and the amplified fragments were analyzed by DNA sequenator. Then the data was used to construct a molecular marker database and establish unified standard for the identification of Chinese bayberry resources. Afterwards, the genetic distances of every 2 accessions were calculated, and the neighbor-joining (NJ) clustering tree was drawn. The results showed that the amplified fragments of 16 SSR molecular markers were different and could distinguish 109 variety accessions. The geographical sources of the accessions were correlated with the genetic distance to a certain extent. A molecular database containing 109 accessions was established based on 16 SSR markers with 'Biqi', 'Dongkui', 'Dingao' and 'Anhaipianzaosheng' as reference marker of amplicon length. Further selection of 5 SSR molecular markers with high polymorphism information content (PIC) could successfully distinguish 85% (94/109) of Chinese bayberry materials, indicating high identification efficiency. This study provides a more efficient and reliable method and standard for the management of bayberry germplasm resources.