High quality hybrid rice has good quality, loved by the vast number of consumers. Foliar fertilizer, as a way of applying fertilizer outside the roots of crops, has an important effect on the increase of crop yield. In order to screen out a reasonable ratio of foliar fertilizer and improve the yield of high quality rice, 'Longjing Youdizhan' (Oryza sativa) was used in this study. The effects of different foliar fertilizer ratios on the physiological indexes of high-quality hybrid rice leaves, the expression of yield-related genes enoyl-CoA hydratase (NOG1) and pale green leaf-9 (PGL-9) and yield were studied. The combination of brassinolide (BR), silicon (Si) and selenium (Se) foliar fertilizers were used to study 8 different treatments to find out the suitable foliar fertilizer combination based on the yield at harvest. The results showed that the yield of each treatment was higher than that of the control (CK), and the yield of Se treatment was the highest (8.8%), followed by BR treatment (8.3%) and Se+BR treatment (8.2%). Under Si treatment, chlorophyll content and catalase (CAT) activity were the highest at booting stage. Under the treatments of Si+BR, BR and Si, the soluble sugar content, superoxide dismutase (SOD) and peroxidase (POD) activities were the highest at heading stage. Under BR treatment, malonaldehyde (MDA) was the lowest at tillering stage. Correlation analysis showed that the expression of NOG1 and PGL-9 gene and chlorophyll content were significantly correlated with yield at heading stage. The correlation coefficients were -0.881, 0.932 and -0.940, respectively. In addition, the expression of NOG1 and PGL-9 gene were significantly correlated with chlorophyll content at heading stage, and the correlation coefficients were 0.993 and -0.971, respectively. In summary, selenium fertilizer was the best foliar fertilizer, and its application at heading stage can significantly increase the yield of high-quality rice 'Longjing Youdizhan'. This study can provide reference for high-yield cultivation techniques of high-quality rice.

VQ protein, a class of amino acid sequences containing “FxxxVQxLTG” conservative motifs, is a plant specific protein family, which participated in plant growth, development, and various stress response. In order to clarify the biological function of the VQ gene family in cabbage, 67 BoVQ genes were identified from the whole genome of cabbage, and their bioinformatics and expression patterns were analyzed. The phylogenetic analysis showed that BoVQ genes were divided into 7 subgroups. Chromosomes mapping exhibited BoVQ genes were unevenly distributed in single or cluster on 9 chromosomes of cabbage. The transcriptome data suggested that BoVQ genes expressions varied at different tissues. Eleven BoVQ genes (BoVQ10, BoVQ20, BoVQ22, BoVQ32, BoVQ36, BoVQ43, BoVQ49, BoVQ54, BoVQ59, BoVQ60 and BoVQ67) participated in the whole process of cabbage flower development. The results of real-time fluorescence quantitative PCR (qRT-PCR) showed that BoVQ2, BoVQ12, BoVQ21, BoVQ26 and BoVQ44 genes were significantly high expressed under the infection of clubroot pathogen. Prediction of cis-elements revealed that the pathogen response element W-box and the defense and stress response element TC-rich repeats were enriched in the promoter of BoVQ genes in response to Plasmodiophora brassicae. Protein interaction network prediction showed that there were complex interactions between BoVQ proteins. This study provides a reference for further exploring the function and evolution of BoVQ genes in cabbage.

Arabidopsis histidine phosphotransfer proteins (AHPs) play important roles in plant growth, development and resistance to biotic or abiotic stresses. In this study, bioinformatics, transcriptome analysis and qRT-PCR were used to explore the biological function and expression pattern of AHP in melon (Cucumis melo), including the number of AHP gene family members, chromosome location, gene structure, subcellular localization, conserved motifs, phylogenetic analysis, cis-acting elements and gene expression. Results showed that the CmAHP family had 7 members, and distributed on 7 chromosomes. The length of amino acids ranged from 109 to 412 aa, all of which were hydrophilic. All the members were localized in the nucleus or extracellular matrix. Phylogenetic analysis revealed that the melon AHP members were highly homologous to cucumber (Cucumis sativus). CmAHPs promoter mainly contained cis-acting elements such as light response and hormone response elements. Gene expression analysis showed that CmAHP5 and CmAHP7 were highly expressed in all 5 tissues, including roots, leaves, female flowers, male flowers and fruits, which might play negative roles in fruit development and ripening. Transcriptome data showed that CmAHP5 and CmAHP7 might play negative and positive roles, respectively, in defending Fusarium wilt and powdery mildew. qRT-PCR data showed that CmAHP1~CmAHP5 were involved in low temperature response, among which CmAHP4 might be a key candidate gene. The present study provides a theoretical reference for analyzing the function of AHP gene in melon.

The wall-associated kinase (WAK) is a unique class of receptor-like kinase (RLK), which plays an important role in regulating cell growth and defense against stress response. In this study, 27 CsWAK genes were identified from the tea plant (Camellia sinensis) genome using bioinformatics methods, which were mainly clustered on 2 chromosomes and 1 contig. These CsWAK were classified into 5 subgroups (Ⅰ~Ⅴ) based on the phylogenetic analysis of the protein sequences, and most members of the same subgroup had similar gene structures and conserved motifs. The transcriptome data analysis showed that most CsWAK were expressed primarily in tea leaves, followed by roots and stems. Most members of the CsWAK family had little difference in response to biotic/abiotic stresses such as hormone, low temperature, and high salt. The difference of most CsWAK gene expression were not significant, only several CsWAK gene expression had significant difference, with diversity and specificity. In addition, some differentially expressed CsWAK in response to cold treatment, PEG treatment and salt treatment were analyzed by qPCR. The results showed that under the stress of low-temperature, the selected 9 genes all showed an expression pattern that rose first and then fell, and most of the genes were significantly upregulated at low-temperature treatment for 3 h; under the stress of PEG, the expression level of the selected 9 genes was slightly different from the transcriptome data, but the expression trend was similar; and under NaCl treatment, the expression level of the selected 9 genes was significantly downregulated compared to that of untreated, among them, CsWAK14 was the most sensitive to salt stress. These results provide a reference for further analysis of the biological functions of CsWAK gene.

Phoebe bournei is a precious tree spiece unique to China. Afforestation in red loam which is very popular in south of China is prone to low phosphorus stress, affecting cultivation benefit. Exploring the gene resources in P. bournei with phosphorus tolerance is of great significance to develop P. bournei varieties with enhancing ability of low phosphorus tolerance by molecular assisted breeding. SPX gene is associated with phosphorus absorption and balance in plants. In this study, members of the SPX gene family in P. bournei were identified in the whole genome, and the expression of PbSPX gene in the roots, stems, and leaves of P. bournei were analyzed using qPCR, as well as the expression patterns in the roots and leaves of P. bournei after 10 and 60 d of phosphorus deficiency treatment. The results showed that 20 PbSPXs were distributed on chromosomes 1 to 10 of P. bournei. According to characteristics of protein and phylogenetic tree, PbSPXs were divided into 4 subfamilies, including 6 members of SPX subfamily, 7 members of SPX-EXS subfamily, 3 members of SPX-MFS subfamily and 4 members of SPX-RING subfamily. Analysis of promoter cis-acting elements showed that PbSPX genes containing elements related to phosphorus stress response, such as P1BS, W-box and MBS. Tissue specific expression analysis showed that PbSPX2, PbSPX-EXS1 and PbSPX-RING2 were mainly expressed in roots, and another 12 PbSPXs were expressed highly in leaves. In roots, when treated with phosphorus deficiency after 10 and 60 d, the expression of PbSPX-MFS1 and all PbSPX subfamily members except for PbSPX6 were induced and the expression of them were significantly up-regulated compared to the control, however, the expression of PbSPX-MFS2 was significantly down-regulated. At the same time, the transcript levels of PbSPX-RING1 and PbSPX-EXS1 were significantly increased at 10 d of phosphorus deficiency treatment, while the expressions of PbSPX-RING3 and PbSPX-EXS2 were significantly up-regulated until after 60 d of phosphorus deficiency treatment. The expression of PbSPX-EXS6 was significantly decreased when treated with phosphorus deficient for 10 d, whereas it was significantly up-regulated after 60 d of phosphorus deficiency treatment. In leaves, the expressions of PbSPX1~5, PbSPX-EXS1 and PbSPX-MFS1 were induced after 60 d of phosphorus deficiency treatments, and their expressions were significantly up-regulated compared to the control. The transcript level of PbSPX-MFS2 was significantly down-regulated after 60 d of phosphorus deficiency treatment, which was consistent with the expression trend in roots. As phosphorus transporters candidates, PbSPX-EXS1, PbSPX-EXS2 and PbSPX-EXS6 responded differently to phosphorus level signals in different tissues. The results showed that members of the SPX subfamily of P. bournei were involved in phosphorus stress response, and their expression was regulated by phosphorus levels in different tissues. This study revealed the relationship between PbSPX genes and the response of low phosphorus stress in P. bournei, providing a reference for exploiting key gene resources of low phosphorus tolerance in P. bournei and further studying function and mechanism of PbSPX.

Phytochelatin synthase (PCS) is a protease-like enzyme that catalyzes glutathione (GSH) to form phytochelatins (PCs), and plays an important role in detoxification of heavy metals in plants. The heavy metal detoxification function of plant PCS varies according to crops varieties and doses of heavy metals. In order to study the molecular mechanism of ramie (Boehmeria nivea) PCS response to heavy metal cadmium (Cd), the p426 GDP-BnPCS1 yeast (Saccharomyces cerevisiae) expression vector was converted into wild-type and cadmium-sensitive yeast respectively, and the results showed that transgenic yeast appeared stronger cadmium tolerance under 50~100 μmol/L Cd²⁺ stress. In addition, BnPCS1 was overexpressed in Arabidopsis thaliana, the root length and biomass of Arabidopsis thaliana were significantly higher than those of the wild type under 100 μmol/L Cd²⁺ stress, and the antioxidant capacity, GSH and PCs content of Arabidopsis thaliana under cadmium stress were significantly enhanced, as well as transcription levels of AtPCS1, γ-glutamate cysteine ligase 1 (AtGCL1), glutamate synthase 1 (AtGS1), heavy metal ATPase 1 (AtHMA1) and ATP-binding cassette transporter 1 (AtATM1). The above results show that ramie BnPCS1 can actively respond to cadmium stress and enhance cadmium tolerance. The study provides a reference for ramie-tolerant molecular breeding.

By connecting to its high affinity receptor tyrosine protein kinase A (TrkA), nerve growth factor (NGF) plays a crucial physiological role in the adaptive regulation of hypoxic ischemia in central nervous system tissues or cells. The purpose of this study was to investigate the expression, distribution and interaction of NGF and TrkA during adaptation to low oxygen environments in the diencephalon and brainstem of yaks (Bos grunniens). Different areas of the diencephalon and brainstem of yaks and cattles (B. taurus) were collected at different altitudes, and the expression levels and distribution features of NGF and TrkA were examined using qPCR, Western blot, and immunohistochemistry (IHC) methods. The results showed that the expression levels of NGF and TrkA genes and protein were significantly higher in the thalamus of yaks and cattles diencephalon and brainstem than in other tissues (P<0.05); Compared with cattles, NGF gene and protein expression levels in the thalamus and medulla oblongata of yaks were significantly higher than in cattles (P<0.05), but lower in the hypothalamus and pons. There was no significant difference between cattle and yaks in the hypothalamus, but there were considerably increased TrkA gene expression levels in the thalamus, pons, and medulla oblongata in yaks (P<0.05).The levels of TrkA protein in the cantamus, hypothalamus, pons, and medulla oblongata were not significantly different from those of cattle. The distribution characteristics of TrkA in the yaks diencephalon and brainstem coincided with NGF, and the intensity of the immunopositive response to TrkA protein was higher than that of NGF protein. The positive products of NGF and TrkA were primarily distributed in the cytoplasm of pleomorphic cells in the thalamus and hypothalamus, motor neurons in the pons, and neurons in the medulla oblongata. According to the aforementioned findings, the thalamus in the diencephalon and brainstem of yaks was the region of the brain that was most vulnerable to hypoxia. The above results showed that the hypothalamus and pons were more tolerant to hypoxia, while the thalamus and medulla were more sensitive, and increased tolerance to hypoxia in the yak thalamus and medulla by activating related neuroprotective mechanisms that up-regulated NGF expression, whereas TrkA did not play a significant under hypoxic conditions. NGF and TrkA were primarily found in the cytoplasm of neurons and glial cells in various regions of the diencephalon and brainstem, implying that they worked together to exert endogenous neuroprotective effects on neurons and glial cells, thereby protecting brain tissue. This study provides theoretical basis and data reference for further exploring the hypoxic adaptation mechanism of yak brain tissue

AGPAT4 (1-acylglycerol-3-phosphate O-acyltransferase 4) is a member of the 1-acylglycerol-3-phosphate O acyltransferase family of enzymes that catalyze synthesis of the triacylglycerol (TAG). In order to clarify the gene sequence and protein structure of AGPAT4 and to preliminarily analyze the effect of AGPAT4 on lipid metabolism in goat mammary epithelial cells (GMECs), In this study, the mammary epithelial cells of Xinong Saanen dairy goat (Capra hircus) were used as experimental materials, the AGPAT4 gene was cloned by reverse transcription-PCR (RT-PCR). Based on the cloned sequence, the bioinformatics analysis was conducted using online website and tools. qPCR was applied to detect the expression of GMECs lipid metabolism-related genes after AGPAT4 interference and overexpression. Then, the content of triacylglycerol in GMECs was detected after interference or overexpression of AGPAT4. The results showed that the CDS region of AGPAT4 gene was successfully cloned, with 1 137 bp (sequence ID: XM_005684959.3), encoding 378 amino acids, protein molecular weight of 43 939.35 u, theoretical pI value of 8.92, transmembrane structure and no signal peptide, which was an unstable protein with positive charge. The AGPAT4 protein interacted with AGPAT1, AGPAT2, lipin3 (LPIN3), diacylglycerol kinases (DGK), phospholipase D (PLD), cytidine diphosphate-diacylglycerol synthase (CDS) and other proteins. The downregulation of AGPAT4 via mRNA interference caused a significant reduction in acetyl coa carboxylase (ACC) gene expression (P<0.05). Overexpression of AGPAT4 significantly up-regulated the expression of stearoyl-coenzyme A desaturase (SCD1) and ACC genes (P<0.05) and significantly promoted the accumulation of intracellular TAG (P<0.05). These results indicated that AGPAT4 gene had positive regulation on lipid synthesis in GMECs. This study provides basic data for further research on the regulatory mechanism of AGPAT4 gene on milk fat metabolism of goat milk.

The number and activity of mammary epithelial cells, lactation volume and milk composition are very important indicators in the breed selection of dairy goats (Capra hircus). The growth, development and lactation of the mammary gland is an extremely complex process. circACAP2 (circular ArfGAP with coiled-coil, ankyrin repeat and PH domains 2) can promote cardiomyocyte apoptosis while being highly expressed in breast cancer tissues, accelerating the progression of breast cancer. However, the regulatory effect of circACAP2 on goats mammary epithelial cells (GMECs) is not clear. In this study, circACAP2 overexpression and interference vector were constructed and transfected into GMECs, the results of double luciferase assay showed that circACAP2 could target binding with miR-21. qRT-PCR results showed that the interference with circACAP2 could significantly increase the expression of miR-21. The Cell proliferation test kit CCK8 (Cell Counting Kit-8) and 5-ethynyl-2'-deoxyuridine (EdU) experiments showed that circACAP2 inhibited the proliferation of GMECs, and the AnnexinV-FITC/PI apoptosis assay kit and flow cytometry detection showed that circACAP2 promoted the apoptosis of GMECs. The detection of ELISA kit showed that circACAP2 reduced the secretion of β-casein and triglyceride (TG). The ERK-Bax/Bcl-2 apoptosis signaling pathway and PI3K/mTOR/S6K lactating signaling pathway were selected for Western blot detections. The results showed that circACAP2 inhibited the expression of pERK1/2 and Bcl-2 and promoted the expression of Bax. Meanwhile, the phosphorylation level of PI3K, mTOR, and S6K decreased. The above results indicate that CircACAP2 might affect the apoptosis and proliferation of GMECs through the apoptosis signaling pathway ERK-Bax/Bcl-2, and affect the secretion of β-casein and TG in GMECs through the lactating signaling pathway PI3K/mTOR/S6K. This study provides basic data for further study of the lactation mechanism of Guanzhong dairy goats.

Awang sheep (Ovis aries), as a special and precious sheep breed resource in the Tibet Plateau, has unique flavors, high-quality meat production and strong plateau adaptability. To prevent the decline of Awang sheep in stock and to protect local germplasm resources, this study analyzed the effects of different follicle-stimulating hormone (FSH) sources and its doses of FSH, superovulation times and the estrus history on the effect of superovulation in Awang sheep by synchronous estrus, superovulation and surgical embryo recovery. The results showed that domestic FSH had the best effect on superovulation in Awang sheep. The average number of corpus luteum and average effective embryos in domestic FSH group were significantly higher than those in the imported FSH group and the domestic + imported FSH group (P<0.05). There was no significant difference in the average number of available embryos and embryo recovery rate (P>0.05). The 270 and 500 IU of domestic FSH had the same effect on superovulation in Awang sheep, and had no significant effect on the superovulation effect of Awang sheep weighing(>50 kg and 40~50 kg), respectively. There was no significant effect of oestrus history on the effect of superovulation. In addition, the total number of corpus luteum and the number of recovered embryos in the first superovulation group were extremely significant higher than those in the multiple superovulation (P<0.01). In conclusion, 270 IU of domestic FSH had the best effect on superovulation in Awang sheep, which can be used as a reference for the conservation and propagation of local Awang sheep.

As an important economic fish in China, the natural germplasm resources of Coilia ectenes are declining rapidly. Exploring the correlation between genetic polymorphisms of C. ectenes fatty acid-binding proteins 7 (CeFABP7) gene with the growth traits plays an important role in the breeding and improvement of growth traits of C. ectenes. Based on the transcriptome data of C. ectenes and gene cloning technology, the cDNA and DNA sequences of CeFABP7 were obtained. The SNPs of CeFABP7 gene was screened by DNA direct sequencing method using 20 individuals that selected from the F₃ generation population of C. ectenes with extreme growth characteristics. Then 100 individuals that randomly selected from the same population of C.ectenes were genotyped by DNA direct sequencing or SnaPshot method. The relationship between these SNPs and growth traits of C. ectenes was analyzed by least squares method. The cDNA length of CeFABP7 was 1 295 bp, containing an open reading frame of 399 bp. The genomic DNA of CeFABP7 was 1 899 bp in length containing 3 introns and 4 exons. Six polymorphic sites were detected in 5′-UTR (2), intron 1 (2) and intron 3 (2). The locus g.276C>G was significantly associated with the body length, body height and body weigh traits of C. ectenes (P<0.05), the locus g.306A>C,g.747C>T and g.825C>T were significantly associated with body height and body weigh traits of C. ectenes (P<0.05), while the locus g.1447A>T and g.1532I>D were not significantly associated with 3 growth traits of C. ectenes (P>0.05). The result of population genetic analysis of cultured population of C. ectenes basing on 6 mutation locus showed that the average value of observed heterozygosity (Ho), expected heterozygosity (He) and polymorphism information content (PIC) were 0.257、0.339 and 0.275, respectively. The locus g.276C>G in the CeFABP7 gene was expected to be an important candidate molecular marker for eliminating inferior breeding parents of C. ectenes. This study has theoretical instructive significance for molecular marker assisted breeding in C. ectenes.

Pomacea canaliculata is one of the 16 most harmful invasive alien species in China. SRY-related HMG box gene 9 (SOX9) is a subgroup of SOXE genes associated with early embryonic development. In order to explore the function of the SOX9 gene of snails, the cDNA sequence of the SOX9 like gene from snail P. canaliculata (named as PcSOX9L, GenBank No. OP564918) was cloned by RT-PCR on the basis of transcriptome sequencing. Using P. canaliculata of different developmental stages (rankⅡ~ⅣP. canaliculata) as materials, the bioinformatics as well as expression characteristics of PcSOX9L were analyzed. The results showed that PcSOX9L gene contained a 1 701 bp open reading frame, encoding a hypothetical protein of 566 amino acids, and contained 2 conserved domains, namely Sox-N and HMG-box-domain; the amino acids of PcSOX9L and other animal SOX9 proteins showed that the sequence similarity ranged from 52.72% to 82.84%. qRT-PCR analysis showed that PcSOX9L gene was expressed in various tissues including gonad, liver, gill, mantle, and pleopod of rank Ⅳ P. canaliculata, as well as the albumin gland of female rank Ⅳ P. canaliculata. Among them, the expression level of PcSOX9L was the highest in the liver of male rank Ⅳ P. canaliculata, and was extremely significantly higher than the same tissues of female rank Ⅳ P. canaliculata (P<0.01). No significant difference was detected between the gonad of male and female rank Ⅳ P. canaliculata. The PcSOX9L was expressed in 3 different developmental stages, with the highest expression in rank Ⅱ P. canaliculata; at the same time, the changes of PcSOX9L expression in 3 different developmental stages toward male rank Ⅳ P. canaliculata in descending order as rank Ⅱ P. canaliculata>male rank Ⅳ P. canaliculata>>rank Ⅲ P. canaliculata; and the changes of PcSOX9L expression in 3 different developmental stages toward female rank Ⅳ P. canaliculata in descending order as rank Ⅱ P. canaliculata>rank Ⅲ P. canaliculata>female rank Ⅳ P. canaliculata, which indicated that this gene might play an important role in the early embryonic development and sex determination of P. canaliculata. The present results provide basic data for further analysis of the mechanism of SOX9 gene in the gonad development and sex determination of P. canaliculata.

Monopartite geminiviral V2 protein plays an important role in the process of virus infection, while the research on the function of bipartite geminiviral AV2 protein is relatively few. To reveal the function of the AV2 protein of bipartite geminiviral Tomato leaf curl Hsinchu virus (ToLCHsV) by using the technique of proximity-dependent biotin identification (BioID)-mass spectrometry, several host proteins proximal to AV2 were preliminarily screened in Nicotiana benthamiana. The interaction between mevalonate kinase a (NbMVKa) and ToLCHsV-AV2 was verified by yeast two hybrid (YTH) and bimolecular fluorescence complementation (BiFC). Using the Tobacco rattle virus (TRV)-induced gene silencing technique, ToLCHsV infectious clone was inoculated in N. benthamiana plants. The results showed that 10 d post inoculation (dpi), NbMVKa silenced N. benthamiana plants showed no obvious symptoms, while the non-silenced plants showed leaf curl. Meanwhile, using the Southern blot analysis, the accumulation of viral DNA in silenced plants was relatively decreased. After 20 dpi, symptoms of NbMVKa silenced N. benthamiana plants were basically the same as those of the non-silenced plants. The results revealed that NbMVKa might play a positive role in regulating the pathogenesis of the virus by interacting with ToLCHsV-AV2. These results provide theoretical basis for further elucidating the function of bipartite geminiviral AV2 protein.

In recent years, Papaya leaf curl Guangdong virus (PaLCuGDV) and Euphorbia leaf curl virus (EuLCV) were usually identified in Passiflora edulis. The full-length sequences of the 2 viruses, 2 732 and 2 745 bp respectively, were amplified from Passiflora edulis showing typical geminiviral damage symptoms in Fujian province. In order to determine the pathogenicity of PaLCuGDV and EuLCV, the infectious clones of 2 viruses were constructed by homologous recombination technology and Nicotiana benthamiana plants were inoculated with Agrobacterium-mediated method. The results showed that the symptoms of N. benthamiana inoculated with EuLCV were obviously dwarfed and the top leaves were deformed and curled, which was significantly heavier than that of PaLCuGDV, while the latter only showed slightly curly new leaves and yellowing leaves. Further more, the symptoms of mixed infection of the 2 viruses was much heavier than any of the symptoms induced by PaLCuGDV or EuLCV alone. Southern blot results showed that N. benthamiana inoculated with PaLCuGDV and EuLCV alone or in combination had high virus accumulation. In this study, the infectious clone of PaLCuGDV was constructed for the first time, and it was found that the mixed infection of EuLCV and PaLCuGDV could aggravate the induced symptoms. This study provides a reference for understanding the pathogenic mechanism of PaLCuGDV and EuLCV.

Senecavirus A (SVA), an emerging RNA virus in recent years, which seriously endangers the development of global pig industry. Its viral structural protein 2 (VP2) plays an important role in inducing the body's immune response. To prepare a soluble structural protein VP2 of SVA and systematically analyze its immunogenicity, firstly, the SVA-VP2 protein recombinant expression plasmid pET28a-SVA-VP2 was constructed based on the gene sequence of the SVA/CH-FJ-2017 strain, and then transferred the pET28a-SVA-VP2 recombinant plasmid and the molecular chaperones pTf16 plasmid into Escherichia coli BL21 (DE3) competent cells. The soluble recombinant SVA-VP2 protein was induced to express by using the characteristics of molecular chaperones trigger factor (TF) to promote the soluble expression of protein in the prokaryotic expression system. BALB/c mice (Mus musculus) was immunized by the purified target protein which was identified by SDS-PAGE and Western blot. Serum antibody levels was detected by indirect ELISA, and neutralizing antibody titers was measured by virus neutralization experiments, and the CD4⁺ and CD8⁺ T lymphocyte subpopulations in the spleen and the proliferation and cytokine production of spleen lymphocytes were detected by flow cytometry. The results showed that the soluble recombinant VP2 protein was successfully prepared by using molecular chaperones TF16. The recombinant VP2 protein could react specifically with SVA positive serum. The serum antibody titer level of immunized mice could reach 1:32 000, and the neutralizing antibody level could reach 1:128. The percentage of CD4⁺ and CD8⁺ T lymphocytes in the spleen of immunized mice was very extremely significantly higher than that of the control group (P<0.001), and the expression levels of interferon-γ (IFN-γ), interleukin-2 (IL-2), IL-4 and IL-10 were very extremely significantly higher than those in the control group (P<0.001). In conclusion, this study successfully prepared soluble recombinant VP2 protein by co-expression of molecular chaperones TF, and the recombinant VP2 protein had good immunogenicity both in vivo and in vitro, which can trigger strong humoral immune response and cellular immune response of the body. This study provided experimental materials and theoretical basis for the development of SVA subunit vaccine and diagnostic agent.

The reproductive performance and semen quality of male livestock and poultry are of great significance to animal husbandry, and are closely related to the conception rate, litter size and survival rate of female livestock and poultry. As an important reproductive organ, the epididymis is responsible for sperm concentration, maturation (including sperm vitality acquisition and fertilization ability), protection and storage. MicroRNA (miRNA) is a kind of non coding single stranded RNA molecule with a length of about 22 nt, which is closely related to a series of physiological processes in epididymis of livestock and poultry. miRNA participates in regulating different biological processes, including sperm vitality, sperm maturation, sperm cell apoptosis, ion channel activity, copper ion detoxification, etc. This paper summarized the similarities and differences in the structure and function of epididymis between mammals and poultry, reviewed the research progress of miRNA in regulating sperm motility, maturation, ion transport and other biological functions in epididymis of livestock and poultry. This review provides reference for the research of miRNA regulation mechanism in epididymis and genetic breeding of breeding male reproductive performance.

The diseases caused by phytopathogens seriously impact crop yield and quality during the interaction between plant and microbes, whereas the beneficial endopytes in plant tissues play important roles in plant growth promotion and disease resistance, among which seed endophytes are gaining more and more attentions. Previous studies have shown that through the long-term co-evolution, endophytes are capable of colonizing seeds and being vertically transmitted between generations, to form plant-endophyte holobiont thus to benefit each other, and eventually play essential roles in growth promotion and disease resistance throughout entire life of host plant. Compared to rhizosphere microorganisms, it has not been thoroughly explored overall about the mechanism underlying the impact by host genetics and environmental conditions, as well as the function and relative mechanisms of seed endophytes in plant growth promotion and disease resistance. This paper summarized the current research progress of endophytes, focusing on the diversity and transmission of seed endophytes, as well as the mechanisms of interactions between seed endophytes and phytopathogens. Meanwhile, the functions and potential applications of seed endophytes in plant growth promotion and disease management were fully discussed. Furthermore, the article discussed several issues on current research of seed endophytes, while solutions and advices on the future directions were provided. This review provides theoretical references to explore and utilize seed endophytes for crop improvement in the future.

Fusarium wilt caused by Fusarium oxysporum f. sp. melongenae seriously restricts the production of eggplant (Solanum melongena). However, its pathogenic mechanism is still unclear. In this study, a transformation system of F. oxysporum f. sp. melongenae was established and optimized, in which GFP expression vector pCAMsgfp was transformed into conidia using , ATMT (Agrobacterium tumefaciens-mediated transformation) method. The results showed that a satisfactory transformation with an efficiency of 319±10.21 transformants per 1×10⁶ spores could be achieved in a system of 1×10⁶ spores per milliliter of F. oxysporum f. sp. melongenae spore suspension which were co-cultured with Agrobacterium tumefaciens suspension at a concentration of OD₆₀₀ 0.5 under the co-culture medium containing 200 μmol/mL acetosyringone at 25~28 ℃ for 48 h. PCR identification of the GFP gene and fluorescence observation showed that GFP gene could be expressed normally. In this study, the genetic transformation system of F. oxysporum f.sp. melongenae mediated by ATMT was established and optimized, and the GFP-labeled strain was successfully obtained, which could provide powerful technic for further explore the infection process and the pathogenic mechanism of F. oxysporum f. sp. melongenae.

Fritillaria thunbergii is a genuine medicinal material which is one of the important cash crops in Jinhua city, Lishui city and other places in Zhejiang Province. In recent years, fungal diseases have occurred frequently, including root rot, which affected the yield and quality of F. thunbergii. In this study, pathogenic fungi of root rot disease of F. thunbergii were isolated and purified on potato dextrose agar (PDA) medium by diseased tissue isolation method. The pathogenic fungi were preliminarily identified by morphological observation of colony, mycelium and spore. Using the genomic DNA of fungi, the DNA fragments of internal transcribed spacer (ITS), glyceraldehyde 3-phosphate dehydrogenase (G3PDH), translation elongation factor 1 alpha (TEF-1α), RNA polymerase Ⅱ beta subunit (RPB2), heat shock protein (HSP60), and ITS-specific fragment of Fusarium solani (SOL) were amplified by PCR and sequenced, respectively. The homology of each DNA fragment was aligned in GenBank and the phylogenetic adjacent trees were constructed to identify the species of the fungi. The pathogenicity of the fungi was determined by inoculation of host plant with acupuncture and smear method. A total of 14 fungal isolates were isolated by PCR amplification and electrophoresis, and 4 fungi species causing the root rot disease of F. thunbergii were isolated and identified. By constructing a developmental tree and pathogenicity experiments, it was proved that the pathogenic bacteria were F. solani, F. oxysporum, F. incarnatum and F. equiseti, respectively. The partial sequence of the G3PDH gene of F. incarnatum (MN386064) and F. equiseti (MN386065) was obtained by PCR amplification for the first time. The results enrich the gene sequences of the 2 Fusarium species, and also provid a basis for subsequent related research.