In a previous study, by high-throughput sequencing, receptor interacting serine/threonine kinase 2 (RIPK2) was demonstrated to be highly up-regulated in the immune and inflammatory response of chickens (Gallus gallus). To investigate the function of RIPK2-mediated NOD/RIPK2 signaling pathway in chicken macrophage cell line (HD11) for immunity and inflammation, a chicken HD11 stably interfering with RIPK2 gene was established by using Lentivirus-mediated RNA interference approach. According to the sequence of chicken RIPK2 gene, three RNA interference target sequences and one negative control sequence were designed for RIPK2 gene, recombined with the pLVshRNA-EGFP(2A) Puro interference vector, and then transiently transfected into HD11. Interference efficiency of each target on RIPK2 was tested by qRT-PCR and the recombinant vector with high interference efficiency was packaged with Lentivirus to transfect HD11. qRT-PCR and Western blot were used to detect the expression changes of RIPK2 and the downstream key genes IKKα (component of inhibitor of nuclear factor kappa B kinase complex), IKKβ (inhibitor of nuclear factor kappa B kinase subunit beta), NFκB (nuclear factor kappa B subunit 1) and IL1β (interleukin 1 beta) of NOD/RIPK2 signaling pathways in different experimental groups. Meanwhile the RIPK2 overexpression vector (pcDNA3.1-RIPK2) was constructed for the rescue experiment of RIPK2 gene. Three recombinant plasmid of RIPK2-shRNA1 (small hairpin RNA 1), RIPK2-shRNA2 and RIPK2-shRNA3 were successfully constructed. The recombinant plasmids of RIPK2-shRNA1 and RIPK2-shRNA3 had the most obviously inhibitory effect of (77.4±0.61)% and (90.21±0.68)%, respectively on the RIPK2 gene. The RIPK2-shRNA3 recombinant plasmid was packaged into retrovirus with a virus titer of 2×10⁸ TU/mL. After the RIPK2-shRNA3 Lentivirus was transfected into HD11, the results of qRT-PCR and Western blot showed that RIPK2 mRNA and protein expression level in the interference group were significantly low compared to control group (P<0.05). The expression of downstream key genes IKKα, IKKβ, NFκB and IL1β of the NOD/RIPK2 signaling pathway were significantly down-regulated (P<0.05). The interference group was transfected with RIPK2 overexpression vector (pcDNA3.1-RIPK2), and the expression level of RIPK2 was recovered. In this study, a lentiviral vector expressing shRNA targeting chicken RIPK2 was successfully constructed, which can effectively silence the expression of RIPK2 gene in HD11. Moreover, HD11 stably expressing RIPK2-shRNA3 can interfere with NOD/RIPK2 signal transduction. The results will provide a theoretical basis for further research on the function of chicken RIPK2 gene and its mediated NOD/RIPK2 signaling pathway.

Late embriogenesis abundant protein-2 (LEA-2) gene family play important role in plant growth,development and abiotic stresses. In order to investigate the response of ZmLEA-2 family proteins to drought stress, the LEA-2 protein sequence in maize (Zea mays), Sorghum bicolor, and Arabidopsis thaliana were aligned, also analyzed the gene structure,conserved motifs and position on the chromosome. The collinearity of ZmLEA-2 gene family in maize species were analyzed. Collinearity analysis between the maize, sorghum and Arabidopsis thaliana were also performed. The expression pattern of ZmLEA-2 family genes were analyzed based on the transcriptome data, and the function of ZmLEA53 was preliminary verificated.The results showed that 81 ZmLEA-2 genes were unevenly distributed on 10 chromosomes; the phylogenetic tree analysis showed that 81 ZmLEA-2 genes were divided into 7 groups, and genes in the same group had similar gene structure and conserved motifs. 81 ZmLEA-2 genes exhibited different expression patterns. Under drought stress, the expression of ZmLEA53, ZmLEA7 and ZmLEA64 was up-regulated and decreased after rewatering. It was speculated that these genes were involved in the regulation of drought tolerance. qPCR results showed that the expression of ZmLEA53 was positively regulated by drought, salt and high temperature. Overexpression of ZmLEA53 could improve the survival rate of yeast (Saccharomyces) cells under drought (PEG), salt and high temperature stress, respectively. The results provide a basis for further functional identification of ZmLEA-2 gene family under drought stress.

Mung bean (Vigna radiata) is the main coarse grain crop in China, and the flowering time plays an important impact on the production, quality and fertile period of mung bean. FLOWERING LOCUS T (FT) genes play an important role in the flowering pathway of plant, which are also the key genes in the flowering regulatory network. In order to clarify the regulatory role of FT family homologous genes in the photoperiod-related flowering of mung bean, the cDNA sequence of VrFT2a (GenBank NO. XM_014651110) was cloned from the photoperiod-sensitive mung bean variety 'Xinzheng'. The coding region, encoded proteins characteristics, and its expression patterns of VrFT2a in different tissues and photoperiod conditions were analyzed by bioinformatics. Nine light responsive cis-acting elements were found in the promoter region of VrFT2a, the full-length of cDNA was 531 bp, which encoding 176 amino acids wherein the 27th to 162th amino acids were highly conserved PEBP (phosphatidylethanolamine-binding protein) family domain. The encoding protein of VrFT2a was a non-secretory hydrophilic protein with none signaling peptide. Subcellular localization analysis indicated that VrFT2a localized in plasma membrane and nucleus. Phylogenetic analysis showed that the amino acids of VrFT2a had a higher homology with other FTs. VrFT2a, GmFT2a, and AtFT had similar three-dimensional protein structures, and the conserved flowering-related amino acids of FT (Tyr85/Gln140). qRT-PCR analyses revealed that VrFT2a was expressed in different tissues, with its highest expression in trifoliolates of mung bean. The expression level of VrFT2a exhibited an obvious diurnal rhythm under long-day and short-day conditions, and it also was specifically induced by the short-day conditions significantly. The results implicated that VrFT2a might involve in the photoperiod regulatory pathway of mung bean. This study will provide a theoretical reference for further deconstruction of the molecular mechanisms of FT family genes in photoperiodic regulation pathways of mung bean.

Fusarium wilt seriously endangers the growth and development of sea islands cotton (Gossypium barbadense). Previous study based on the transcriptome data of sea island cotton inoculated with Fusarium oxysporum, the resistance-related gene uridine diphosphate glycosyltransferase 73C1 (GbUGT73C1) was screened and identified. In this study, the full length of the GbUGT73C1 gene CDS was found through the sea island cotton genome, and the gene was cloned from '06-146' with high resistance to Fusarium wilt. The cloned nucleic acid and amino acid sequences were performed with bioinformatics analysis, and the gene expression was analyzed after inoculation with disease resistance and susceptible material. The results showed that the full length of CDS of GbUGT73C1 was 1 486 bp, a total of 476 amino acids were encoded. The coding protein included hydrophobic, fat-soluble and unstable protein, without transmembrane structure and signal peptide. With glycocyltransferase-B (GT-B) structure, it belonged to the glycosyltransferase GT-B superfamily. GbUGT73C1 protein was predicted to be localized in the cytoplasmic. The prediction of molecular phylogenetic relationship was closest to that of G. barbadense and G. raimondii. GbUGT73C1 was expressed rapidly in disease-resistant material, and the expression level was extremely significantly higher than that in disease-susceptible material (P<0.01), indicated that resistant '06-146' resisted Fusarium wilt infection by expressing GbUGT73C1 gene more rapidly and in a higher amount .This study lays a foundation for further investigation on the mechanism of GbUGT73C1 response to sea island cotton's resistance to Fusarium wilt, and provides important gene resources for cotton disease-resistance molecular breeding.

Celery (Apium graveolens) is an important leafy vegetable with high nutritional values and numerous medicinal functions. Chlorophyll content is one of the important indexes to evaluate the quality of celery during storage after harvest. The influence on the content of chlorophyll in celery under different postharvest environmental conditions is still unclear. In this study, 'Sijixiaoxiangqin' and 'Liuhehuangxinqin' were stored at 3 different storage conditions of room temperature (25 ℃), low temperature (4 ℃) and darkness at room temperature (25 ℃). The chlorophyll contents and the expression levels of metabolism-related genes (AgMPE, AgPPH, AgPAO, AgCAO, AgNOL and AgHCAR) were detected at 0, 6, 24, 30, 48 and 54 h after treatment. The results showed that the average contents of chlorophyll a and chlorophyll b were the highest under low temperature treatment, and total accumulation of chlorophyll was the highest at low temperature storage except for 24 and 30 h, and the peak of total chlorophyll content appeared at low temperature. The total chlorophyll content under darkness at room temperature condition was higher than that under room temperature condition. The transcription level of AgCAO gene changed significantly with the increase of storage time, which was consistent with the accumulation pattern of chlorophyll b. The results indicated that low temperature condition inhibited the degradation of chlorophyll content and could better maintain the chlorophyll content of celery. Low temperature could maintain the nutritional and sensory quality of celery. This study provides a reference for exploring the regulation pattern of chlorophyll accumulation and related genes in celery after harvest.

Calmodulin-binding transcription factor (CAMTA) plays an important role in the regulation of plant growth and development and response to abiotic and biological stress. Jute (Corchorus capsularis) is an important commercial crop of bast fiber. In order to study the role of CAMTA in response to drought stress in jute, the 'Corchorus capsularis 179' was used as the material, primers were designed based on the sequence of CAMTA gene (CC.02G0003260) from the jute genome. The CAMTA gene was obtained by RT-PCR and named as CAMTA1 (OK415793). Sequence analysis showed that the CAMTA1 gene contained 13 exons and 12 introns, Its open reading frame was 3 051bp and encoded a protein containing 1 016 amino acids, which had the CG-1 domain, TIG domain, ANK repeat sequence and IQ motif of CAMTA proteins. Protein sequence alignment indicated that CAMTA1 had high homology with the protein of Malvaceae plants. qPCR analysis showed that CAMTA1 gene was expressed in root, stem, leaf and pericarp. The expression of CAMTA1 gene in leaves reached the highest at 6.0 h after drought stress induction. Fluorescence enzyme protein complementation assay suggested that CaMTA1 interacted with another stress-responsive transcription factor AP2/ERF to regulate plant growth, development and respond to both abiotic and biological stresses.These results provide a theoretical basis for further elucidating the molecular mechanism of CAMTA1 gene involved in plant drought response.

The cellulose synthase (CesA) and cellulose synthase-like (Csl) gene families belonging to the cellulose synthase gene superfamily are responsible for the biosynthesis of cellulose, hemicellulose and pectin polysaccharides of the plant cell wall, and play critical roles in plant development, growth and evolution. In this study, 36 PpCesA/Csl gene family members were identified in the Prunus persica genome, encoding 394 to 1 527 amino acids, respectively. The average molecular weight of the proteins ranged from 44.85 to 171.80 kD, and the number of introns ranged from 4 to 16, containing 20 different motifs. The subcellular localization of PpCesA/ Csl protein was diverse and can be located in chloroplasts, golgi bodies, cell membranes, cytoplasm and mitochondria. Phylogenetic analysis showed that PpCesA/Csl could be divided into 7 subfamilies (CesA, CslA, CslB, CslC, CslD, CslE and CslG). All PpCesA/Csl proteins contained 7 transmembrane domains, of which 27 members contained cellulose synthase domain PF03552 and 9 members contained glycosyltransferase family 2 domain PF00535. In addition, the other 8 PpCesA subgroup members except PpCesA2 contained zinc finger domain zf-UDP, and the other 4 CslD subgroup members except PpCslD1 contained zinc finger domain zf-RING_4. The main conserved active sites of PpCesA/Csl protein were aspartic acid D, D, D residues, followed by QxxRW motif. Most PpCesA/Csl family members contained complete amino acid sequences of this active site. The results indicated that most of the CesA/Csl genes from peach were closely related to genes in Arabidopsis thaliana, but these families had unique characteristics in terms of their gene structure, chromosomal localization, phylogeny, and deduced protein sequences, suggesting that they had evolved through different processes. The transcriptome data of 'Qian Jianbai' peach during storage were used to analyze the expression characteristics of peach CesA/Csl members in mature fruit.The results showed that most peach CesA/Csl members were not expressed or very low expression level during the storage period in 'Qian Jianbai' fruit. Finally, 1 up-regulated member (PpCslE7), 3 down-regulated members (PpCesA6, PpCesA8, PpCslG1) and 1 relatively stable member (PpCslD2) were selected from the members with high expression levels in the transcriptome for analysis of expression characteristics. The qPCR analysis of PpCesA6, PpCesA8, PpCslD2, PpCslE7 and PpCslG1 in different tissues, fruits during different development and storage periods showed that the CesA/Csl genes were constitutively expressed at different levels in different organs. Furthermore, expression analysis of CesA/Csl genes in the fruit development stage showed PpCesA6, PpCesA8 and PpCslD2 all reached the peak expression at 65 d after flowering while PpCslE7 and PpCslG1 peaked at 80 d and 35 d after flowering, respectively. During fruit storage, PpCslD2 and PpCslE7 were up-regulated in both varieties, while PpCesA8 was down-regulated. PpCslG1 was highly expressed in the early storage period of 'Qinwang' fruit, but extremely low in the storage period of 'Shahong' fruit. The family analysis and expression characteristics of CesA/Csl gene in peach provide basic data for further functional study of CesA/Csl gene in peach and other Rosaceae species.

Nuclear Factor Y (NF-Y) is a transcription factor widely found in eukaryotes and involved in plant growth and development as well as biotic and abiotic stress responses. Phoebe bournei is an important economic timber tree in South China, but the growth and accumulation of wood are seriously affected by the lack of water. Exploration of drought-resistance response factors will provide support for improving the drought resistance of P. bournei. In this study, the whole genome identification and bioinformatics analysis of the NF-Y gene family in P. bournei were conducted, and the tissue expression specificity and expression pattern of PbNF-Ys under drought stress were analyzed by qPCR. The results showed that a total of 40 PbNF-Ys were identified in the whole genome of P. bournei, which were divided into 3 subfamilies and unevenly distributed on 11 chromosomes. The molecular weights of PbNF-Ys protein ranged from 10 774.45 (PbNF-YB11) to 39 576.26 D (PbNF-YA2), and all proteins were hydrophilic. The analysis of promoter elements showed that there were hormone response elements such as auxin, gibberellin and methyl jasmonate, light response elements, growth- and development-related elements and drought stress-related elements. Tissue expression analysis of PbNF-Ys in the root, stem, and leaf was carried out by qPCR. The result showed that PbNF-Ys presents tissue-specific expression patterns. Expression analysis of drought stress treatments showed that 17 PbNF-Ys were up-regulated induced by drought stress, 16 ones were down-regulated, and 7 ones presented no significant expression change. This study provides the reference basis for further study on the biological function and evolution of NF-Y gene family in P. bournei.

Myostatin (MSTN) is a negative regulator of muscle development. Studies have shown that it can regulate cell proliferation. In order to explore the further mechanism of MSTN mutant in regulating the proliferation and differentiation of muscle-derived satellite cells (MDSCs), in this study, the MSTN mutant (MT) and wild type (WT) bovine (Bos taurus) MDSCs were used to detect the cell cycle and gene expression in the process of proliferation and differentiation. Results of EdU proliferation assay showed that the proliferation index of MSTN mutant cells (0.78±0.0559) was significantly higher than that of wild-type cells (0.57±0.0366), indicating that MSTN mutant promoted the proliferation of MDSCs. When myogenic differentiation was induced, the MSTN mutant cells began to differentiate and formed the myotubes after 1day of induction, while the wild-type cells began to differentiate after 2 days of induction. Flow cytometry was used to further analyze the effect of myogenic induction on the cell cycle. Results also showed that the MT cells began to stagnate in the G1/S phase after 1 day of induction, and the cell cycle of WT cells stagnated after 2 days of induction. These results indicated that when cells were induced to differentiate, MSTN mutant cells exited from the cell cycle and entered to a differentiated state before wild-type cells. The expression of cell cycle-related genes and proteins was detected by qPCR and Western blot. Results showed that in MSTN mutant cells, the expression of CyclinA was significantly up-regulated (Relative expression=19.5), while the expression of cyclin-dependent kinase inhibitor 1C (CDKN1C) was significantly inhibited (Relative expression=0.009). These results showed that CyclinA and CDKN1C may play important roles in the regulation of MSTN mutant to the proliferation and differentiation of MDSCs. Online prediction through the JASPAR database showed that the key transcription factors downstream of MSTN, SMAD2/ SMAD3 may bind to the promoter of CDKN1C. ChIP-qPCR results demonstrated that the SMAD2/SMAD3 transcription factor combined with the promoter of CDKN1C thus to increase the expression of CDKN1C. In conclusion, MSTN mutant may inhibit the expression of CDKN1C gene by down-regulating the combination of SMAD2/SMAD3 transcription factor and CDKN1C promoter, thereby up-regulating the expression of CyclinA-CDK2, and promote DNA synthesis and cell cycle progress.This study discovered a new mechanism for MSTN mutant to promote muscle development, and provides a certain theoretical basis for the preparation of muscular gene-edited animals.

CytochromeP450c17 (CYP17A1) and cytochromeP450 aromatase (CYP19A1) were important for follicle development, ovulation and the secretion and synthesis of major reproductive hormones, and affected the reproductive performance of animals. In this study, healthy Qianbei Ma goat (Capra hircus) were taken as the research object. The tissue expression of CYP17A1 and CYP19A1 genes was detected by qPCR, and the SNP mutation sites were screened by PCR direct sequencing. The effect of SNPs on the lambing number of different parity of Qianbei Ma goat was analyzed by establishing a general linear model. The expression of CYP17A1 and CYP19A1 genes in various tissues and the correlation between SNPs and litter size were analyzed. The results showed that the expression of CYP17A1 gene was the highest in the ovary of single and multiple lambs, and was extremely significantly higher than that of pituitary, hypothalamus, uterus and fallopian tube (P<0.01). CYP19A1 gene expression was higher in the multi-lamb ovary and uterus, and both were higher than the corresponding single-lamb tissue (P<0.01). A total of 7 SNPs were detected in the CYP17A1 gene coded region. g.59G>C, g.128G>A, g.129A>G and g.164T>C were all located in the first exon, g.2045G>A was in the second exon and g.4387G>A and g.4456C>T were in the sixth exon. Correlation analysis showed that the SNPs of CYP17A1 gene were not correlated with the litter size of Qianbei Ma goat. A mutation site g.48818C>A was detected in the 9th exon of the coding region of CYP19A1 gene and the number of lambs in the AA genotype was significantly higher than that of the CA genotype (P<0.05), but not significantly different from that of the CC genotype. The results showed that the expression of CYP17A1 and CYP19A1 genes in Qianbei Ma goat was the highest in the ovarian tissue of multiple lambs (P<0.01). There were only 4 goats in 150 with AA genotype g.48818C>A site, which was not representative and needs to be verified. This study provides basic data for further exploring the relationship between CYP17A1 and CYP19A1 gene variants and animal reproduction.

The inositol 1,4,5-trisphosphate receptor type 2 (IP3R2) is an intracellular Ca²⁺-release channel located on the endoplasmic reticulum (ER), which can regulate Ca²⁺ release and affect eggshell quality. To elucidate the regulatory effect of IP3R2 protein domain on Ca²⁺ secretion of duck (Anas platyrhyncha domestica) endometrial epithelial cells and its effect on calcium transport-related genes, in this study, the IP3R2 protein domain expression vector PCDNA3.1-IP3R2 was constructed in laying duck (Sansui duck) and transfected into duck endometrial epithelial cells. The expression of IP3R2 protein domain was detected by Flag label detection kit (ELISA); Fluo-3/AM calcium fluorescence label was used to detect intracellular Ca²⁺ content; qPCR was used to detect the overexpression levels of different domains and mRNA levels of ion transport-related genes. The results showed that overexpression of MIR, RYDR_ITPR (1), RYDR_ITPR(2), RIH_assoc and Ion_trans domains all caused the decrease of Ca²⁺ concentration in duck endometrial epithelial cells; The expression levels of RYDR_ITPR(1) and IP3R2-Ion_trans were significantly negatively correlated with intracellular Ca²⁺ concentration (P<0.05); The expression levels of RYDR_ITPR(2), RIH_assoc and 5 domains were significantly negatively correlated with intracellular Ca²⁺ concentration (P<0.01); The expression of RYDR_ITPR(2) domain was significantly positively correlated with the expression of RYR2, KCNJ15 and CA2 genes (P<0.05); The overexpression levels of MIR-domain were significantly positively correlated with the gene expression levels of IP3R1, CALB1 and CA2 (P<0.05), and the overexpression levels of the 2 RYDR_ITPR domains were significantly positively correlated with the gene expression levels of KCNJ15 and RYR2 (P<0.05); The overexpression of RIH_assoc domain was significantly positively correlated with the expression of IP3R2 and ORAI1 genes (P<0.05); and the overexpression of Ion_trans domain was significantly positively correlated with the expression of CALB1, SLC8A1 and CA2 genes (P<0.05). The co-transfection level of the five domains were significantly positively correlated with CFTR gene expression, and negatively correlated with TRPV6 gene expression (P<0.05). These results suggest that overexpression of 5 IP3R2 protein domains can cause extracellular Ca²⁺ release in duck endometrial epithelial cells, and have different regulatory effects on the expression levels of 20 ion transport genes. This study provides a reference for improving eggshell quality of ducks in terms of molecular genetic marker selection.

CWI-MAPK (cell wall integrity-mitogen-activated protein kinase) cascade pathway is mainly involved in regulating the integrity of the cell wall, cell cycle and pathogenicity in phytopathogenic fungi. Swi4 is a key transcription factor downstream of the CWI-MAPK cascade. In Saccharomyces cerevisiae, the transcription factor ScSwi4 as a regulatory expression element participates in cell wall biosynthesis and cell cycle regulation, but the function of its homologous genes in filamentous pathogenic fungi is rarely reported. The northern leaf blight of corn is caused by Setosphaeria turcica. In this study, the StSwi4 gene of Setosphaeria turcica was cloned. The analysis of the gene sequence and the characteristics of the encoded protein showed that the full length of the DNA sequence of the gene was 2 247 bp, and the full length of the cDNA sequence was 2 037 bp, containing 3 exons and 2 introns, encoding 678 amino acids; StSwi4 protein contained 1 KilA-N conserved domain and 2 ANK conserved domains. The phylogenetic relationship of the proteins revealed that the StSwi4 protein of S. turcica shared closer evolutionary relationship with Alternaria gaisen, A. alternata, and S. solani. The expression results of StSwi4-His fusion protein by IPTG showed that the fusion protein could be expressed in Escherichia coli.The purified protein was further obtained by nickel column affinity chromatography. The expression patterns of StSwi4 gene at different developmental stages were analyzed by qPCR, and the highest expression level was found at conidial stage. This study clarified the structural characteristics and expression patterns of the transcription factor StSwi4 gene, and provides basic material for further revealing its molecular mechanism in the pathogenesis of pathogens.

Overuse of chemical fertilizers leads to deterioration of the soil environment and severely affects crop production. High-carbon basal fertilizer plays an important role in improving soil ecological environment of farmland, it has a greater impact on soil chemical properties and microbial diversity. This study adopted field test methods, 5 treatments were set up, including: NF (no fertilization), GCK (conventional fertilization of 111 kg/hm² pure nitrogen), G3 (high-carbon basal fertilizer 450 kg/hm²+99.9 kg/hm² pure nitrogen)(10% reduction in nitrogen), G5 (high-carbon basal fertilizer 750 kg/hm²+88.8 kg/hm² pure nitrogen)(20% reduction in nitrogen), G7 (high-carbon basal fertilizer 1 050 kg/hm²+77.7 kg/hm² pure nitrogen)(30% reduction in nitrogen). The effects on soil chemical properties were studied, and 16S rRNA high-throughput sequencing technology was used to analyze soil bacterial diversity. These results showed that: 1) High-carbon basal fertilizer combined with nitrogen reduction could increase soil pH, content of available nitrogen (N), potassium (K) and phosphorus (P). Among them, G7 had the best effect on improving soil fertility. The soil pH, contents of available nitrogen, potassium and phosphorus were increased under treatment of G7, which were 23.49%, 8.78%, 20.28% and 27.81% higher than those of CK, respectively (at 90 d after transplantation). 2) At the phylum levels, G7 increased the relative abundance of Proteobacteria, which were 9.93% and 2.28% higher than CK at 30 and 60 d after transplantation, respectively. At 60 d after transplantation, G3 increased the relative abundance of Bacteroidetes, which were 93.42% higher than CK. High-carbon basal fertilizer combined with nitrogen reduced the relative abundance of Acidobacteria, at 30 d after transplantation, which G7 decreased the most by 35.39%. At the genus levels, at 30 and 60 d after transplantation, G7 significantly increased the relative abundance of Sphingomonas and Pseudarthrobacter. At 60 d after transplantation, G7 decreased the relative abundance of Rhodanobacter. Among them, G7 had the greatest impact on bacterial diversity. 3) According to redundancy analysis, 86.64% of the change in the bacterial phylum communities was due to physical and chemical factors of the soils. There was a positive correlation between soil available N, K, P, pH and Proteobacteria, Bacteroidetes, Gemmatimonadetes. This study showed that 1 050 kg/hm² high-carbon basal fertilizer+77.7 kg pure nitrogen/hm² (30% reduction in nitrogen) had the best effect on improvement of soil microbial diversity and soil fertility. This study elaborates the mechanism underlying impact on soil microbial diversity and soil chemical properties by used high-carbon basal fertilizers.

Myelocytomatosis (MYC) transcription factors (TFs) are the core regulators of jasmonic acid (JA) signal transduction pathway. They are involved in plant growth and development, secondary metabolite synthesis, stress response, plant hormone signal transduction and the interaction between them. In this paper, research progress of plant MYC TFs from the aspects of the characteristics of sequence, structure and function were reviewed, as well as their action mechanism in biotic and abiotic stresses. This review accumulates basic data for further exploring the functional role and regulation mode of MYC TFs in plants.

In recent years, the application of traditional Chinese medicine has become more and more popular, and the market demand is increasing, but the mixed phenomenon of counterfeits is serious. The development of identification kits based on DNA detection technology is of significant practical significance for this medicinal material. This study was based on the phylogenetic tree constructed in early stage of the experiment on A. edulis and its fakes, and specific PCR primers for A. edulis were designed on the basis of internal transcribed spacers (ITS) sequence analysis to optimize the use of specific detection reagents and reaction conditions. According to the evaluation criteria of in vitro diagnostic kits, a test kit was developed for A. edulis, and the test kit was evaluated for specificity, detection limit, repeatability, shelf life and stability. According to the test results, the kit had good specificity, the detection limit was 0.03 μg/μL, and excellent stability. The specific identification kit of authentic Chinese medicinal material A. edulis was accurate, fast and effective for the identification of A. edulis, and has broad application prospects.

Sugarcane (Saccharum officinarum) is the most important sugar crop in China. Sugarcane Pokkah boeng disease poses a serious threat to the high-quality development of the sucrose industry in China. Therefore, specific primers were designed based on the nuclear ribosomal DNA internal transcribed spacer (rDNA-ITS) sequences of 2 main pathogens of sugarcane Pokkah boeng Fusarium verticillioides and Fusarium proliferatum, respectively. The results showed that primers Fv-F3/Fv-R3 could amplify a specific amplification fragment of 400 bp from F. verticillioides and its infected sugarcane, but not other pathogens of sugarcane diseases. The detection sensitivity for genomic DNA of F. verticillioides was 30 pg/µL. Primers Fp-F4/Fp-R4 could amplify a specific amplification fragment of 362 bp from F. proliferarum and its infected sugarcane, but not other pathogens of sugarcane diseases. The detection sensitivity for genomic DNA of F. proliferarum was 30 pg/µL. A total of 117 samples with typical Pokkah boeng symptoms were collected from different main cultivated sugarcane varieties in different sugarcane areas of Yunnan Province were detected by PCR with these two pairs of specific primers. And the occurrence of the two main pathogens of sugarcane Pokkah boeng in this area was clarified. The results showed that 112 (95.7%) of the 117 samples were infected by F. verticillioides, 103 (88%) were infected by F. proliferatum. 103 (88%) were complex infected by F. verticillioides+F. proliferatum. The two pathogens were not detected in other 5 samples, which may be caused by other species. The PCR products of 23 F. verticillioides and 19 F. proliferatum from different main sugarcane varieties in different sugarcane regions were selected for sequencing and analysis. The results showed that the F. verticillioides and F. proliferatum amplification product sequences had 99.45%~100% similarity with F. verticillioides (KU508286) and 99.26%~100% similarity with F. proliferatum (MK252904), respectively. Some sequences were selected to construct phylogenetic tree, they were divided into F. verticillioides group and F. proliferatum group. The results of this study showed that the established detection system could accurately and quickly detect F. verticillioides and F. proliferatum. The detection rates of F. verticillioides and F. proliferatum were high from different main sugarcane varieties in different sugarcane areas of Yunnan Province. They were important pathogens of sugarcane Pokkah boeng in Yunnan province, and the compound infection was common. Among them, F. verticillioides was the dominant species in sugarcane areas such as Puer, Lincang, Honghe, and Yuxi sugarcane areas. However, the detection rate of F. proliferatum was also high in these sugarcane areas, and maybe there were other species of sugarcane Pokkah boeng pathogens might be present. This study provides technical support and practical basis for early diagnosis and timely prevention and control of these diseases.

Foot-and-mouth disease (FMD) is one of the most important infectious diseases causing significant economic losses to the swine (Sus scrofa domesticus) industry. Vaccine immunization is still the one of main ways to prevent and control foot-and-mouth disease. Because Foot-and-mouth disease virus is easy to mutate, the antigen mismatch between candidate vaccine strains and epidemic strains will affect the immune efficacy of vaccine and the effect of disease control. According to the molecular and functional characteristics of structural protein VP1 (viral structural protein) gene, using the inactivated Porcine foot-and-mouth disease virus type O strain (O /MYA98/BY/2010) as the template, the structural protein VP1 gene was amplified and ligated into pSFV1 vector to construct the recombinant plasmid pSFV1-VP1. To obtain mRNA, the recombinant plasmid was treated by linearization and in vitro transcription, then the mRNA was transfected into baby hamster syrian kidney-21 (BHK-21) cells and verified its expression properly. After that, it was mixed with the liposome nanoparticles to immunize guinea pigs (Cavia porcellus), and the immune response and efficacy were detected. The results showed that the mRNA was expressed in BHK-21 cells after 24 h post-transfection evaluated by immunofluorescence assay (IFA) and Western blot; In guinea pigs immunized with the mRNA vaccine, specific antibody, neutralizing antibody, interleukin 4 (IL-4) and γ interferon (IFN-γ) in serum were significantly higher than those of guinea pigs. The mRNA vaccine could provide 4/4 protection against 50 times GPID₅₀ (50% Guinea pig infectious dose) virus challenge in vaccinated guinea pigs, implying humoral and cellular immune response elicited by the developed mRNA vaccine with sound immunogenicity, which provides a new route for the development of foot-and-mouth disease emergency vaccine.

Porcine eperythrozoonosis is caused by Mycoplasma suis, M. parvum and a new species Candidatus M. haemosuis (CMh) isolated from Zhejiang province,which leads to anemia, jaundice and abortion of swine (Sus Scorfa). The prevalence and transmission of pig's hemoplasmas is not clear in China. In order to differential diagnose the main pathogens prevalence of porcine eperythrozoonosis, a triplex TaqMan probe fluorescence quantitative PCR (qPCR) was established based on glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene. According to the published GAPDH gene sequences of pig's hemotrophic mycoplasma, the primers and probes were designed by Beacon Designer 4.0 software for the 3 porcine hemoplasmas. By optimizing the concentration of Mg²⁺, hot-start rTaq, primers and probes, the triplex Taqman probe qPCR method was established, and the specificity, sensitivity, reproducibility of the method were verified. The results showed that the correlation coefficient (R²) of the established triplex Taqman probe qPCR method was between 0.999 2~0.999 6, and the amplification efficiency was between 99.9%~103.2%. No amplification curve were found from DNA that had been extracted from the pig's samples of others common pathogens. The lowest detection limit of the method for Mycoplasma suis, M. parvum and CMh was 1 copy/μL. The coefficient of intra- and inter-group variations was between 0.31%~1.75%, indicating good repeatability. 148 blood samples from Zhejiang province pig farm, were detected by triplex Taqman probe qPCR and compared with double PCR, the results showed that the positive rates of M. suis, M. parvum and CMh were 50% (74/148) , 48.6% (72/148) and 64.9% (96/148), respectively, the mixed infection rate of the three pathogens was 29.7% (44/148). The positive rates of M. suis/M. parvum and CMh detected by duplex PCR were 29.1% (43/148) and 15.5% (23/148), respectively. The positive samples detected by duplex PCR were all positive by triplex Taqman probe qPCR. The method established in this study can be used for the clinical diagnosis of M. suis, M. parvum and CMh, which is a technical support for the epidemiological investigation, prevention and control of swine Hemotrophic mycoplasma.