Abiotic stress seriously affects the growth and yield of wheat (Triticum aestivum). Dehydrin (DHN) is a kind of protein induced by abiotic stress, and plays an important role in protecting cells from damage caused by water deficiency and temperature changes. In order to study the role of dehydrin in plants responding to abiotic stress, this study isolated a YSK₂ type dehydrin gene WDHN2 (GenBank No. OK094572) from wheat (Triticum aestivum) in homology-based cloning technology. WDHN2 gene had an open reading frame with the length of 501 bp, and encoded a protein containing 166 amino acids, which contained 1 segment of Y, 1 segment of S, and 2 segments of K. Phylogenetic analysis revealed that WDHN2 had the closest genetic relationship with Aegilops tauschii DHN (XP_020188302.1). The bioinformatics analysis showed that WDHN2 protein was a highly-hydrophilic protein, and its higher structure was mainly composed of random coil. Subcellular localization predicted that WDHN2 protein localized in the nucleus. In addition, S segment was the main phosphorylated site. The qPCR analysis indicated that the expression of WDHN2 gene was induced by drought, salinity, low temperature and abscisic acid (ABA). The prokaryotic fusion expression vector pET28a-WDHN2 was constructed and transformed into Escherichia coli BL21 (DE3). Then isopropyl β-D-1-thiogalactopyranoside (IPTG) was used for inducible expression, the 25 kD WDHN2 fusion protein and recombinant E. coli were obtained. Resistance analysis of recombinant bacteria showed that WDHN2 protein improved the tolerance to osmotic stress, high salinity, low temperature and high temperature stresses in Escherichia coli. The above results revealed that WDHN2 might be involved in the signal transduction pathway of wheat in response to abiotic stresses. The present study could enrich the research content of the molecular mechanism of abiotic stress responses in wheat, and provide basic material for further study on the function of WDHN2 gene.

The AT-hook motif nuclear localized protein (AHL) family genes contain a unique combination of 1 or 2 AT-hook domains and regulate various biological processes in plants. To explore the biological function of cassava (Manihot esculenta) AHL family gene MeAHL19 and its role in the protein interaction network, in this study, the bait vector pGBKT7-MeAHL19 was constructed using T4 ligase and used to screen the cassava cDNA library via the yeast (Saccharomyces cerevisiae) two-hybrid (Y2H) library screening system. Real-time quantitative PCR method was used to detect gene expression patterns of MeAHL19 and the interacting proteins. The results showed that 20 positive clones were obtained through Y2H screening. Eight candidate interacting proteins were identified by PCR amplification and sequencing alignment. Y2H test confirmed the interaction between MeAHL19 and ribulose bisphosphate carboxylase small chain (RBCS) protein in vitro. BiFC results further confirmed that MeAHL19 interacted with RBCS in Nicotiana benthamiana mesophyll cells. Analysis of gene expression patterns showed that MeAHL19 was mainly expressed in roots, whereas RBCS was expressed at about the same level in roots, stems and leaves; MeAHL19 and RBCS responded to abiotic stress and exogenous hormone responses in cassava. This study provides a reference for further investigation on the biological function of MeAHL19 in stress responses of cassava.

Lycoris sprengeri is multicolor special flowera with red and blue petals in Lycoris, which flower color formation mechanism is complex, and anthocyanin synthase (ANS) is a key enzyme at the end of plant anthocyanin synthesis pathway. In this study, molecular biological methods were used to analyze the functions of LsANS cDNA and promoter. The results showed, The total length of LsANS cDNA was 1 401 bp (GenBank No. MT664240), ORF 1 068 bp, encoding 355 amino acids; The phylogenetic tree analysis showed that the LsANS had highest homology with ANS gene of L. radiate, which was 98.5%. qRT-PCR showed that the expression of LsANS was the highest in clone H4 and the lowest in clone H1; The expression was the highest at large bud stage and the lowest at full-bloom stage; The consequence of HPLC showed that in different developmental stages, the content of cyanidin-3-O-glucoside and pelargonin-3-O-glucoside in the bud stage was higher than that in other stages; in different clones, pelargonin-3-O-glucose accounted for the highest proportion of anthocyanins measured in clone H4, and the lowest proportion in clone H1; It was speculated that LsANS played an important role in the anthocyanin formation pathway. A 2 114 bp long LsANS promoter sequence was cloned, which contained MYB transcription factor binding site and cis acting elements involved in light, hormone and stress. The PBI121 LsANSpro::GUS fusion expression vector was constructed and transiently transformed into Arabidopsis thaliana, which showed that LsANSpro has the function of promoting downstream genes. By GUS staining assay, promoter activity was detected in the whole roots, leaves, and flowers . The research will provide a theoretical basis for further research the role and mechanism of ANS gene in the flower color formation process of Lycoris.

1-deoxy-d-xylulose-5-phosphate synthase (DXS) and 1-deoxy-d-xylulose-5-phosphate reductoisomerase (DXR) are the first and second rate limiting enzyme of the synthesis pathway of terpene precursor 1-deoxy-d-xylulose-5-phosphate (DXP), and play important regulatory roles in the secondary metabolism of tobacco (Nicotiana tabacum). In this study, based on the DXS and DXR in Arabidopsis thaliana,the CaMV 35S promoter and NOS terminator, which are the most commonly used plant gene expression, were selected to synthesize the whole gene sequences DXS-NOS and 35S-DXR,and the co-expression vector pSH737-DXS-DXR was constructed. The tobacco transgenic plants co-expressing DXS and DXR were successfully obtained through Agrobacterium tumefacien-mediated genetic transformation. GUS chemical activity assay, PCR and Southern blot showed that the fusion genes had been successfully introduced into the genome of regenerated tobacco plants and expressed. RT-PCR showed that the DXS gene expression in transgenic plants was significantly higher than that of the control (P<0.05). The results of GC-MS showed that the neophytodiene and glandular hair secretion of transgenic plants increased in varying degrees compared with the control (P<0.05). This study provides research materials for tobacco breeding by using genetic improvement technology, and provides a reference basis for further regulating the biosynthesis of tobacco terpenoids by means of metabolic engineering.

The growth rate and carcass quality of beef cattle (Bos taurus) have important economic value in beef cattle industry. Fat acid binding proteins 4 gene (FABP4) and fatty acid synthase gene (FASN) play important role in the process of fatty acid synthesis and fat deposition in beef cattle. Titin-cap (TCAP) gene is known to be a kind of muscle silk protein and plays important role in the assembly of myofibrils. There are a g.3691G>A SNP in FABP4 gene, a g.17924G>A SNP in FASN gene and a g.346G>A SNP in TCAP gene, which were associated with growth and carcass traits in beef cattle. The aims of this study were to investigate the genetic diversity of g.3691G>A, g.17924G>A and g.346G>A SNPs in Qinchuan cattle population, and confirm their association with growth and carcass traits in Qinchuan cattle. These 3 SNPs of 3 genes in 350 samples of Qinchuan cattle were genotyped by MassARRAY technology, and then the analyses of genetic diversity and association with growth and carcass traits in Qinchuan cattle were performed. The results showed that the genotype frequency of g.3691G>A SNP in FABP4 gene, g.17924G>A SNP in FASN gene and g.346G>A SNP in TCAP gene were consistent with Hardy-Weinberg equilibrium (P<0.05) in Qinchuan cattle population. Association analysis results showed that g.17924G>A SNP of FASN gene in Qinchuan cattle population was significantly associated with rump length and hip width (P<0.05), as well as beneficial alleles (G) frequency was significantly different from that of Korean and Angus cattle breeds (P<0.01). Association analysis results showed that g.3691G>A SNP of FABP4 gene in Qinchuan cattle population was significantly associated with hip height, hip width and chest circumference (P<0.05), as well as body weight, body length, rump length and chest depth (P<0.01), beneficial alleles (G) frequency was significantly different from that of Korean cattle (P<0.01). Association analysis results showed that g.346G>A SNP of TCAP gene in Qinchuan cattle population was significantly associated with withers height, hip height (P<0.01), as well as body weight, body length, pin bone width and intramuscular fat content (P<0.05), beneficial alleles (G) frequency was significantly different from that of Korean cattle (P<0.01). Thus, these results suggested that the g.3691G>A SNP in FABP4 gene, the g.17924G>A SNP in FASN gene and the g.346G>A SNP in TCAP gene could be an effective molecular marker for marker-assisted breeding of Qinchuan cattle, and these results could provide scientific basis for genetic improvement of Qinchuan cattle.

As RNA-binding protein, YT521-b homology domains 2 (YTHDF2) can recognize m⁶A modifications and plays a pivotal role in mammalian growth and development. In order to reveal the role of yak (Bos grunniens) YTHDF2 gene in yak fat deposition, the CDS of YTHDF2 mRNA was cloned for bioinformatics analysis in perirenal adipose tissue of yak, and the spatiotemporal expression pattern of YTHDF2 mRNA was analyzed by qPCR. The result showed that the total length of YTHDF2 mRNA CDS region was 1 743 bp, encoding 580 amino acids; YTHDF2 was most closely related to wild yak; the amino acid isoelectric point was 8.87, and the instability coefficient (Ⅱ) was 50.00, grand average of hydropathicity was -0.66, YTHDF2 showed strong hydrophilicity in general, YHDF2 had a conserved domain YTH of 133 bp, no signal peptide and transmembrane structural domain; there were 69 potential phosphorylation sites. The advanced structure of YTHDF2 protein was composed of irregularly coiled (67.59%), α-helix (15.15%), extended chain (12.24%) and β-turn (4.66%) connected with each other. YTHDF2 interacted with methylation modification-related proteins and played an important role in RNA specific binding. The results of qPCR showed that YTHDF2 gene was expressed in all 7 tissues of yak, with the highest expression in the longest dorsal muscle (P<0.05); In terms of time, there was no significant difference in the expression level of YTHDF2 at 18 and 30 months in adipose tissue; Furthermore, YTHDF2 expression showed an increasing trend during the differentiation of prerenal adipocytes, and the expression of YTHDF2 at 4, 8 and 12 d was significantly higher than that 0 d (P<0.05). The results of this study showed that YTHDF2 was an unstable alkaline water-soluble protein with a YTH conserved domain, which initially indicated that YTHDF2 gene played an important role in the process of adipose deposition in yaks. This study provides a theoretical basis for further study on the regulation and function of YTHDF2 gene on adipose deposition of yak.

Litter size is an important economic trait in sheep (Ovis aries) breeding, which determines the production efficiency of mutton sheep industry. Studies have shown that the g.36938224A>T SNP in β-1,4-N-acetyl-galactosaminyl transferase 2 gene (B4GALNT2) was a major candidate marker associated with the ovulation rate of French Lacaune sheep breed, therefore, the B4GALNT2 gene was considered as a new candidate gene for fecundity in sheep. The aims of this study were to confirm the association between the known SNPs in the B4GALNT2 and litter size of Mongolia and Ujimqin sheep breeds, and identify novel breed-specific variants of Mongolia sheep as well as perform the association study with litter size of Mongolia and Ujimqin sheep breeds. A total of 480 sheep were used in this study including 250 Mongolia sheep, 110 Ujimqin sheep, 30 Hulunbuir sheep, 30 Tan sheep, 30 Small-tailed Han sheep, and 30 Hu sheep. Then, direct sequencing was used to identify novel polymorphism and detect the g.25929884A>T SNP of B4GALNT2 gene (GenBank No. NC_040262.1), the MassARRAY technology was used to genotype the g.25921552C>T and g.25935026C>T SNPs of B4GALNT2 gene and c.*803A>G SNP of distal-less homeobox 3 (DLX3) gene in 480 samples of Mongolia sheep, Ujimqin sheep, Hulunbuir sheep, Tan sheep, Small-tailed Han sheep and Hu sheep. Therefore, association studies were performed between the novel variants and litter size in Mongolia and Ujimqin sheep breeds. The results showed that the g.25929884A>T SNP of B4GALNT2 and the c.*803A>G SNP of DLX3 were absent in any sheep breed. The g.25921552C>T and g.25935026C>T SNPs of B4GALNT2 gene were not associated with litter size of Mongolia and Ujimqin sheep. Furthermore, 8 novel SNPs were found in B4GALNT2 gene, among them, the g.25929637G>A (P<0.05), g.25929679T>C (P<0.01), g.25929819A>G (P<0.01), and g.25929965A>T (P<0.05) SNPs were associated with litter size of Mongolia sheep. Except for g.25921552C>T site in Tan sheep, the other 9 polymorphic loci were under Hardy-Weinberg equilibrium. According to PIC standard, all SNPs presented low and moderate polymorphism in 6 sheep breeds. The presnet study might facilitate effective marker-assisted selection to increase litter size in Mongolia sheep, and provide scientific basis for genetic improvement of sheep.

Histone deacetylase 6 (HDAC6) is an important epigenetic modification protein and amphiregulin (AREG) is an important epithelial growth factor-like (EGF-like). HDAC6 and AREG play important roles in regulating mammalian reproduction. This study aims to explore the expression rules of HDAC6 and AREG in ovarian follicles with different development stages of Tan sheep (Ovis aries) and analyze their correlation in follicle development, in this study, the ovaries of Tan sheep were collected for detecting the localization and expression patterns of HDAC6 and AREG in ovarian follicles with different development stages of Tan sheep by immunohistochemistry, immunofluorescence and Western blot methods, and the correlation between HDAC6 and AREG in follicular development was analyzed by Pearson method. The results showed that HDAC6 was expressed in both granulosa cells and oocytes of ovarian follicles at different developmental stages of Tan sheep, and was mainly expressed in the cytoplasm of granulosa cells and oocytes. With the development of follicles, the expression level of HDAC6 increased first and then decreased in follicles with different development stages, and the expression level of HDAC6 in follicles larger than 6 mm in diameter was significantly lower than that in follicles less than 2 mm in diameter. AREG was mainly expressed in granulosa cells of different sizes of ovarian follicles of Tan sheep, and was mainly expressed in granulosa cytoplasm. With the development of follicles, the expression level of AREG was gradually increased in follicles with different development stages, and the expression level of AREG in follicles with diameters greater than 6 mm was significantly higher than that in follicles with diameters less than 2 mm. Furthermore, the correlation analysis found that the expression level of HDAC6 was negatively correlated with that of AREG in follicles with different diameters of Tan sheep. The expression level of HDAC6 in follicles with diameters of less than 2 mm was moderately negatively correlated with that of AREG, and the expression level of HDAC6 in follicles with diameters of 2~6 mm was strongly negatively correlated. The expression level of AREG was negatively correlated with HDAC6 in follicles larger than 6 mm with diameter. The results showed that HDAC6 and AREG were expressed in ovarian follicles with different development stages of Tan sheep, and the expression patterns of HDAC6 and AREG were negatively correlated in general, suggesting that HDAC6 and AREG may be involved in ovarian follicle development of Tan sheep through negative correlation regulation. This study provides an important reference for further exploring the expression relationship of HDAC6 and AREG in granulosa cells and oocytes at different development stages of Tan sheep ovarian follicles and their effects on follicular development and oocyte maturation, and possible mechanisms.

Silent mating type information regulation 2 homolog 1 (SIRT1) is involved in lipid mobilization through deacetylation and affects lipid metabolism. To investigate the polymorphism of SIRT1 gene in Guizhou white goats (Capra hircus) and to screen the SNP loci associated with serum lipid metabolism indexes in Guizhou white goats, this study conducted bioinformatics analysis of SIRT1, and analyzed the expression of SIRT1 gene in various tissues and the association of SNPs with serum cholesterol, triglycerides and free fatty acids. The results showed that SIRT1 gene was expressed in 11 tissues, among which the expression in liver and longest dorsal muscle was extremely significant higher than other tissues (P<0.01); 11 mutation sites were detected in the SIRT1 gene of Guizhou white goat, all of which were located on exons 1 and 6; genetic analysis revealed that the PIC of the 7 SNP loci (g.21206621T>C, g.21206920T>G, g.21207071A>C, g.21215871G>A, g.21216960T>C, g.21217717T>C, g.21217777A>G) were all at moderate polymorphic levels. g.21207042C>T and g.21217404T>C mutant loci population genetic balance deviated from Hardy-Weinberg equilibrium (P<0.01). Linkage disequilibrium analysis showed that g.21206621T>C, g.21206920T>G, g.21207071A>C, g.21215871G>A, g.21216960T>C, g.21217717T>C, g.21217777A>G, g.21217777A>G sites, g.21207042C>T, g.21217404T>C sites had a strong linkage disequilibrium effect with each other; bioinformatics analysis showed that the protein had the molecular formula C₃₅₁₄H₅₅₃₀N₉₆₈O₁₁₅₃S₂₆, a non-transmembrane protein with 3 N-glycosylation sites, and the secondary and tertiary structures were mainly irregular curl. The free energy of g.21206920T>G, g.21207071A>C, g.21216060C>A and g.21216960T>C sites was unchanged after mutation, while all other sites were changed. Association analysis revealed that the cholesterol, triglyceride and free fatty acid contents at the g.21206920T>G and g.21217777A>G loci differed significantly (P<0.05) between genotypes; the cholesterol and free fatty acid contents at the g.21206621T>C, g.21216960T>C and g.21217717T>C loci differed significantly (P<0.05) between genotypes. Triglyceride and free fatty acid contents at the g.21207071A>C locus differed significantly (P<0.05) between genotypes; free fatty acid contents at the g.21215871G>A locus differed significantly (P<0.05) between genotypes. This study speculates that SIRT1 gene can be used as a candidate molecular marker for fat metabolism and deposition in white goats in Guizhou, and clarifies the loci related to fat-related metabolic indexes regulated by SIRT1 gene to provide theoretical reference for carrying out genetic markers in goats.

Small intestinal epithelial cells (IECs) are important basis for the small intestine to play an important role in digestion and absorption of nutrients and intestinal barrier. microRNA 29a (miR-29a) plays an important role in cell apoptosis, differentiation and proliferation during animal growth and development. In order to clarify the effect of miR-29a on the gene expression of small intestinal epithelial cells of piglets (Sus scrofa), miR-29a mimics group, miR-29a inhibitor group, and negative control group were constructed. The IECs were collected after transfection 48 h, extracted RNA to construct libraries and performed Illumina sequencing; The differential expressed genes (DEGs) were screened and subjected to GO functional annotation and KEGG pathway analysis; Venn analysis was used to screen target differential genes and construct target gene co-expression network, 4 genes were selected randomly to verify the accuracy of transcriptome data by qPCR. A total of 9 transcriptome libraries were obtained in this study and total clean base was 55.42 Gb, and the comparison rate with the reference genome reached more than 95%. The DESeq2 software was used to analyze the expressed genes obtained by sequencing, and the results showed that 7 102 DEGs were expressed between the miR-29a inhibitor group (VPA) and the negative control group (VPC), of which 2 558 up-regulated genes and 3 544 down-regulated genes were found in VPA; there were 2 829 DEGs between the two groups, of which 1 563 genes were up-regulated genes in miR-29a mimics group (VPT), and 1 266 genes were down-regulated in the VPT; there were 503 DEGs in the VPA and VPT. There were 197 up-regulated and 306 down-regulated genes in VPA. The GO functional annotation and KEGG pathway enrichment analysis of the differential genes showed that the IECs after miR-29a treatment were in a very active stage of cell proliferation and differentiation and the obtained DEGs were mainly enriched in DNA replication and cell cycle pathways. qPCR results were consistent with the sequencing data. The above results suggested that miR-29a might be regulated cell apoptosis by affecting genes such as CDKN2C, E2F1, MCM family and POLE in IECs, thereby affecting intestinal absorption and intestinal barrier function. This study provides basic for studying the effect of miR-29a on the intestinal of Bamei piglets.

Pasteurellosis is a zoonosis caused by Pasteurella multocida (Pm), which has a serious impact on the development of livestock economy. However, the specific pathogenic mechanism of Pm infection is still not fully understood. This study aimed to explore the immune changes induced by Pm infection in the kidney tissue and the infection mechanism of Pm. The mouse (Mus musculus) model of goat-derived Pm (HN02 strain) infection was established and its kidney samples were collected for transcriptome sequencing. Compared with the control group, there were 3 836 differentially expressed genes (DEGs) in Pm infection group, of which 1 741 DEGs were significantly up-regulated and 1 426 DEGs were significantly down-regulated; Gene Ontology (GO) function enrichment and Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation results found that DEGs were mainly significantly enriched in signal transduction, energy metabolism and immune response, which mainly included transmembrane transporter activity, mitochondrion, tumor necrosis factor (TNF) signaling pathway, Human papillomavirus (HPV) infection, and mitogen-activated protein kinase (MAPK) signaling pathway in the process of Pm infecting mouse kidney tissue. After interaction analysis of immune related signaling pathways, some DEGs were found to participate in multiple different signaling pathways commonly, such as DEGs including v-rel avian reticuloendotheliosis viral oncogene homolog A (ReLA), nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha (NFKBIA), tumor necrosis factor receptor superfamily, member 1A (TNFRSF1A), epidermal growth factor (EGF) and phosphoinositide-3-kinase, regulatory subunit 3 (PIK3R3). Then they were selected for qRT-PCR validation. The results of qRT-PCR showed that the expression trends of these genes were consistent with the sequencing results. This study preliminarily proved that kidney involved in a series of immune responses in the process of resisting Pm infection. The innate immunity of kidney tissue plays an important role in the process of resisting Pm infection, and this result provides data support for further research on the interaction mechanism between Pm and the host.

Spermine accelerates the repair of intestinal mucosal injury, improves activities of intestinal digestive enzymes, polyamine homeostasis, and intestinal function. To research the effects of spermine on key genes related to polyamine metabolism and polyamine contents in the jejunum and ileum in Sichuan white goose (Anser cygnoides), the female geese were intragastrically administered with 5 and 10 mg/kg spermine (body weight). The mRNA expression levels of key genes related to polyamine metabolism and the content of polyamines in female geese in jejunum and ileum tissues were measured by qPCR and high performance liquid chromatography. The results showed that the expression levels of ornithine decarboxylase antizyme inhibitor 2 gene (AZIN2), ornithine decarboxylase antizyme 1 (OAZ1) and OAZ2 genes, and the contents of spermidine and spermine in jejunum of female geese treated with spermine at 5 mg/kg were significantly higher than that in the control group (P<0.05), while expression levels of AZIN1 and acetylpolyamine oxidase gene (APAO) were significantly decreased (P<0.05). The expression levels of AZIN2 and spermine synthase gene (SPMS) in ileum of female geese treated with spermine at 5 mg/kg were significantly higher than that in the control group (P<0.05), while the mRNA expression level of spermidine/spermine-N'-acetyltransferase gene (SSAT) was significantly decreased (P<0.05). The expression levels of AZIN2, OAZ2, ornithine decarboxylase gene (ODC), APAO and spermine oxidase gene (SMO) and spermine content in jejunum of female geese treated with spermine at 10 mg/kg were significantly higher than that in the control group (P<0.05), while the mRNA expression level of spermidine synthase gene (SPDS) and putrescine content were significantly lower (P<0.05), respectively. The expression levels of AZIN2, ODC, SPDS and APAO in ileum of female geese treated with spermine at 10 mg/kg were significantly higher than that in the control group (P<0.05), while the content of putrescine was significantly decreased (P<0.05). These results suggest that exogenous spermine regulates polyamine homeostasis in small intestinal tissues of female geese by mediating the expression levels of polyamine metabolism genes. The effects of spermine on polyamine homeostasis in jejunum and ileum of female geese were different to some extent. This work provides some basic information to polyamine metabolism researches in goose small intestine, and lays a foundation for the further studies on the polyamine metabolism in poultry intestine.

Traditional Chinese herbs are rich in active ingredients and have the properties of improving the immunity of the body, disease resistance and residue-free, and have been widely used in aquaculture. The aim of this study was to investigate the effects of compound Chinese herbs medicine (composed of Astragalus radix, Codonopsis pilosula, Angelica sinensis, Glycyrrhiza uralensis, Ophiopogon japonicas, Poria cocos, Lonicera japonica, Isatidis radix, Isatis indigotica Fortune, Hawthorn) on the non-specific immunity parameters and immune-related gene expressions of rainbow trout (Oncorhynchus mykiss) and provide scientific basis for the healthy culture. The 120 healthy rainbow trouts with an average weight (33.0 ± 2.8) g were selected and randomly assigned into 4 groups; each group repeated the experiment 3 times and 10 rainbow trout per replicate. The fish were fed a basal diet supplemented with compound Chinese herbal medicine at concentrations of 0 (control group), 10, 20 and 30 g/kg, respectively. The feeding trial lasted 35 days. Subsequently, spleen tissue (4 per group) was collected at 7, 21 and 35 d of feeding to evaluate non-specific immune parameters. The expression of fifteen immune-related genes in spleen tissue was detected by qPCR after 35 d of feeding. The results showed that dietary compound Chinese herbal medicine significantly increased the levels of total superoxide dismutase (T-SOD), catalase (CAT), acid phosphatase (ACP) and lysozyme (LZM) in the rainbow trout spleen on the 7^th, 21^th and 35^th day (P<0.05), and the 20 g/kg supplemental groups showed the best results. Furthermore, the malondialdehyde (MDA) concentration in the spleen was significantly lower in the compound Chinese herbal medicine treatment groups than that in the control group (P<0.05), and the 30 g/kg supplemental group showed the best results. Moreover, the gene expression data indicate that dietary compound Chinese herbal medicine significantly upregulated the expression of Toll-like receptor 3 (tlr3), Toll-like receptor 7 (tlr7), medullary differentiation factor 88 (myd88), interleukin-1β (il-1β), nuclear factor kappa B inhibitor alpha (nfkbia) in the 20 g/kg supplemental group (P<0.05). The tumor necrosis factor-α (tnf-α) gene expression in the treatment group was no difference compared to the control group and Toll-like receptor 8 (tlr8) gene was significantly upregulated in the 30 g/kg supplemental group (P<0.05). Additionally, immune-related genes included melanoma differentiation-associated protein 5 (mda5), RIG-I-like receptor 2 (lgp2), interferon regulatory factor 3 (irf3), interferon regulatory factor 7 (irf7), interleukin-8 (il-8), interferon-β (ifn-β), janus kinase 1 (jak1) and signal transducer and activator of transcription 1 (stat1) in the 20 g/kg supplemental group were significantly upregulated (P<0.05). In conclusion, the current results indicated that a diet supplemented with compound Chinese herbal medicine has positive effects on non-specific immune responses of rainbow trout, it suggested that the best adding level was 20 g/kg. This study provides a scientific basis for promoting the rational application and development of compound Chinese herbal immunostimulants in aquaculture.

Screening endophytic bacteria with excellent antagonistic effect against Botrytis cinerea plays important roles in controlling of B. cinerea, increasing tomato (Solanum lycopersicum) yield and reducing postharvest tomato loss. The present study adopted LB medium and gradient dilution method to isolate endophytic bacteria from the roots of Stemona japonica, and the antagonistic strains were screened by plate confrontation method. The antagonistic strains were identified by morphological characteristics, physiological and biochemical analysis, and 16S rDNA sequence analysis. The bacteriostatic ability of the volatile compounds of antagonistic strains was measured by two-sealed-base-plates method. The control effect of antagonistic strains on B. cinerea and their colonization ability on tomato fruits were analyzed by using fruit-pricking method. Seed germination rate, root length, and bud length were measured by seed germination experiment to evaluate the growth promotion effect of antagonistic strains on tomato seedlings. Results indicated there was an excellent strain BR-1 was screened and identified as Burkholderia gladioli, which had nitrogen fixation and phosphorus solubilization effects. The results showed that BR-1 can inhibit the expansion of B.cinerea by inhibiting mycelial growth, and the inhibition rate of BR-1 could reach 73.38%, and further study showed that the volatiles of BR-1 strain exhibited significant antibacterial activity. The broad-spectrum antifungal experiments showed that BR-1 had different inhibitory effects on Fusarium moniliforme, Coccinea arachniformis, Phytophthora capsici, Fusarium oxysporum, Corynosporum cucurbitum and Alternaria brassicae, especially the inhibitory rate of F. moniliforme reached 72.97%. Seed germination experiment showed that BR-1 strain significantly improved the germination rate of tomato seeds, promoted the growth of tomato seedlings and successfully colonized tomato fruits. This study had isolated and screened a biocontrol endophytic bacterium BR-1 with significant disease resistance and growth-promoting effects, and provides important biological resources for the biocontrol of tomato gray mold.

AGL17-like clade family, which belongs to MADS-box gene family, is an important highly conserved transcription factor in plant. In rice (Oryza sativa), the AGL17-like clade comprises 5 members including OsMADS23, OsMADS25, OsMADS27, OsMADS57 and OsMADS61. Researches demonstrated that AGL17-like clade family members mainly expressed in vegetative organs, and participated in multiple biological processes such as growth and development of root, response to fluctuation of nutrient supply, tiller outgrowth, and stress response. This article expounds functions of AGL17-like clade family members in rice, and miR444 which could specificly target to AGL17-like clade family members. The review may be helpful for further researches in the fields of plant growth and development regulation and stress responses of AGL17-like clade, and provides reference for molecular breeding.

As an important food crop and trade agricultural products around the world, Banana (Musa spp.) plays a key role in economic and social development. Banana Fusarium wilt is a notorious soil-borne vascular fungal disease caused by Fusarium oxysporum f.sp. cubense (Foc), causing serious harm to the banana industry worldwide. So far, cultivation of resistant varieties is the most effective way to control banana Fusarium wilt. However, there is no significant progress in disease resistance breeding. Previous study indicated that the effector is a highly important category of pathogenic toxic factors, which plays an important role in pathogenic infection, colonization and the interaction with the host plants. This study reviewed the recent research of the effector of the banana Fusarium wilt, discussed the bioinformatic prediction, transcriptome analysis of the effector, and emphatically introduced a significant effector gene family. It will offer important principle guidelines for the study of the interaction mechanism between Fusarium oxysporum and bananas, the cultivation of resistant varieties and the disease control of banana Fusarium wilt.

Tartary buckwheat (Fagopyrum tataricum) is one of the important coarse grain crops. Different shell types lead to different hulling efficiency, which has great influence on the development of Tartary buckwheat industry. At present, there are few molecular markers study on Tartary buckwheat shell types. The development of molecular markers is helpful to judge the types of Tartary buckwheat shells, and also provides reference for the improvement of Tartary buckwheat shell characteristics and selection of Tartary buckwheat germplasm. In this study, SSR markers were developed based on the loci interval of Tartary buckwheat thin shell gene traits. The results showed that a total of 286 SSRs were detected in the 1.4 Mb mapping interval of thin-shell type. Mono- and di-nucleotide repeat types were the most numerous, and there were 92 and 103, respectively. A/T and AT/AT is the most abundant motifs in mono-nucleotid and di-nucleotide repeat types, respectively. Repeat number of SSR types concentrated between 4 and 10 times. A total of 186 pairs of SSR primers were successfully designed, and 5 pairs of SSR markers, FtgGKssr-82, FtgGKssr-131, FtgGKssr-147, FtgGKssr-149 and FtgGKssr-175, showed polymorphism between the parents of RILs population. The linkage analyses of the phenotype marker for hull type and the 5 SSR markers in RILs population indicated that marker FtgGKssr-149 and FtgGKssr-175 were most closely linked to shell type. Correlation analysis between the shell rate and the 5 SSR markers by using 32 Tartary buckwheat varieties (lines) showed that the markers FtgGKssr-147, FtgGKssr-149 and FtgGKssr-175 were closely related to the shell rate. In this study, 3 SSR markers that were closely linked to the hull type and significantly associated with the shell rate of Tartary buckwheat could be used for marker-assisted selection of Tartary buckwheat germplasms with thin hull, which is of high importance in accelerating the breeding process of thin-hull Tartary buckwheat varieties.

The detection of pregnancy-associated glycoproteins (PAGs) in cow (Bos taurus) blood based on antigen and antibody reaction is the most widely used early pregnancy detection technology in the world. boPAG6 as a marker can be applied to the diagnosis of early pregnancy of cows with higher accuracy. Therefore, the preparation of high-sensitivity boPAG6 polyclonal antibodies can provide support for the development of early pregnancy diagnostic kits for dairy cows. In this study, the boPAG6 gene (GenBank No. NM_176617.2) of Holstein cow was amplified by PCR method, and then the recombinant plasmid pET30a-boPAG6 was constructed. The yield of His-boPAG6 protein was get and optimized in Escherichia coli BL21 (DE3). The recombinant protein His-boPAG6 was used as an immunogen to immunize BALB/c mice (Mus musculus) to prepare the polyclonal antibody. The titer of serum was detected by ELISA method. The specific reaction of purified antibody with recombinant protein His-boPAG6 and cow serum was detected by Western blot method. Immunofluorescence technique was used to detect the localization of boPAG6 in primary trophoblast cells. The recombinant plasmid pET30a-boPAG6 with the size of 6 534 bp was successfully constructed. The yield of His-boPAG6 was highest when induced with 0.5 mmol/L isopropyl-beta-D-thiogalactopyranoside (IPTG) for 8 h, and its protein molecular weight was 50 kD. The titers of antiserum in ELISA immunized with purified His-boPAG6 protein was 1∶102 400. The polyclonal antibody could specifically recognize His-boPAG6 antigen and effectively distinguish pregnant dairy cows's serum from non-pregnant dairy cows's serum, and detect the presence of boPAG6 protein in primary trophoblast cells. This study provides theoretical support for the exploitation of ELISA detection system in early pregnancyof dairy cows.

Cucumber green mottle mosaic virus (CGMMV) is an important quarantine plant virus and is also transmitted by seeds which often causes seriously damages to productions of watermelon (Citrullus lanatus), melon (Cucumis melo), cucumber (Cucumis sativus), and other cucurbits. To guarantee the smooth proceeding of seed trade, it is necessary to establish more rapidly and precisely detection methods. This study developed a SYBR GreenⅠreal-time RT-PCR technique for detection of CGMMV in seeds. It is crucial to design and screen a pair of specific primers based on the conserved CGMMV gene sequences. The result showed that the development of SYBR GreenⅠreal time RT-PCR method in this study could detect CGMMV specially, and there was no cross-reactivity with other common cucurbits viruses. Sensitivity assay showed that the sensitivity of this method was 1 000 times that of RT-PCR and 100 times that of TaqMan RT-PCR. And the whole detection process could be completed within 2.5 h, indicating that the SYBR GreenⅠreal-time RT-PCR was a rapid, sensitive and simple detection method. Cucurbitaceae seeds, seedlings, and fruits imported and purchased from domestic market were employed to be detected by SYBR GreenⅠreal-time RT-PCR and TaqMan real-time RT-PCR simultaneously. The result showed that the value of kappa of the two method was above 0.82, indicating that the novel SYBR GreenⅠreal-time RT-PCR method could be effective and reliable. The present study provides new technical means for rapid clearance of seeds.