The domain of unknown function 1771 (DUF1771) gene family in plants has not been described previously. This research identified 5 members of DUF1771 gene family in Oryza sativa through bioinformatics analysis, then the phylogenetic tree analysis was conducted with these 5 members and 4 members of DUF1771 gene family identified in Arabidopsis thaliana. All 9 members were divided into 2 subfamilies. Conserved motif analysis revealed that motif 1 and motif 2 were present in all DUF1771 members. The promoters of all 5 OsDUF1771 gene family members had relatively few cis-acting elements related to various stress responses and hormone responses, and OsDUF1771-1 had the least. Real-time fluorescence quantitative PCR analysis displayed that the expression levels of OsDUF1771 family members in rice before and after various plant hormone treatments showed little variation overall. When rice was treated with low temperature, drought, and Magnaporthe oryzae, the expression levels of OsDUF1771-1, 2, 3, 5 showed some changes, but the changing ranges were small. OsDUF1771-4 had the largest expression change to low temperature treatment, and the expression level was 4.54 times that of the untreated sample at 12 h of treatment. The O2-site element related to seed storage protein metabolism regulation was found in the promoters of 4 OsDUF1771 family members, but not in the promoters of OsDUF1771-1, which was consistent with the tissue expression profile detection results. OsDUF1771-2, 3, 4, 5 all had the highest expression levels in the panicles, while OsDUF1771-1 had the highest expression levels in the leave, and the encoded protein of OsDUF1771-1 was predicted to be located in the chloroplast. In addition, the promoter of OsDUF1771-3 contained the endosperm expression element GCN4-motif, further indicated that OsDUF1771-3 played a role in the metabolism of endosperm storage proteins in rice seeds. This study provides important clues for in-depth analysis of the biological functions of OsDUF1771 family members.

Heat frequently affects yield and quality in different growth stages of maize (Zea mays). Heat shock transcription factor (Hsf) plays important roles in improving plant heat tolerance through regulating the expression of thermotolerance-related and activating multiple downstream pathways. Based on previous studies on characteristics and thermotolerance function of maize A1 subclass ZmHsf06, this study investigated its function and molecular mechanism in regulating thermotolerance through transformation in maize and chromatin immunoprecipitation-seq (ChIP-seq), and verified the protein interaction and transcriptional activation activity further. The results showed that ZmHsf06 had transcriptional activation activity and could interact with itself. Compared with the wild type, the thermotolerance of maize lines overexpressing ZmHsf06 were significantly improved. Analyzing the downstream target genes and high enrichment pathways of ZmHsf06 by ChIP-seq, a total of 2 218 differentially enriched genes were found under heat stress, including 1 169 genes up-regulated by ZmHsf06 specially, which were mainly enriched in electron carrier activity, photosynthesis, endoplasmic reticulum protein processing and heat shock response. The ZmHsf06 binding motif had heat shock elements (HSE), the binding elements of other transcription factors such as C2C2, TEAD and MADS and some unknown elements. Electrophoretic mobility shift assay (EMSA) showed that ZmHsf06 could bind to the heat shock elements located in the promoters of 3 target genes (Zm00001d000260, Zm00001d000272 and Zm00001d033987). These results will provide evidences for furthering the study of thermotolerance function and regulation mechanism of maize Hsf family.

Jasmonic acid (JA) is an important plant endogenous hormone, which plays an important role in plant growth, development and defense response. NINJA (novel interactor of JAZ) protein is an inhibitor of the JA signal transduction pathway, involved in the regulation of plant growth, development, and stress response. In this study, 2 different NINJAs were identified from Oriental lily 'Sorbonne' (Lilium hybrid division Ⅶ cv. 'Sorbonne') based on transcriptome data, and named LoSorNINJA1 (GenBank No. OQ715340) and LoSorNINJA2 (GenBank No. OQ715341), respectively. Bioinformatics analysis showed that both LoSorNINJA1 and LoSorNINJA2 contained conserved EAR motif and Jas domain, which conformed to the characteristics of the NINJA gene family. Phylogenetic analysis indicated that LoSorNINJAs had the closest genetic relationship with the NINJA of Apostasia shenzhenica, Phalaenopsis equestris and Dendrobium catenatum. The results of subcellular localization in tobacco (Nicotiana tabacum) showed that both LoSorNINJA1 and LoSorNINJA2 were located in the nucleus. Expression analysis in different tissues of 'Sorbonne' showed that LoSorNINJA2 was highly expressed in the outer petals and anthers, and LoSorNINJA1 was highly expressed in the upper leaves. The shoot apical of 'Sorbonne' lily bulbs after low-temperature storage and greenhouse transplant were collected and analyzed by qRT-PCR. LoSorNINJA1 had 2 peaks of expression at 2 and 6 w cold storage, while LoSorNINJA2 had only 1 peak of expression at 6 w cold storage. Using GENIE3 to construct a potential gene regulatory network indicated that LoSorNINJA1 might be involved in tissue development and post-embryonic development, and LoSorNINJA2 might be involved in the response to the external environment. The above results suggest that LoSorNINJA1 and LoSorNINJA2 might have undergone functional differentiation, and their functional sites and developmental stages might also be different. The present study provides basic data for in-depth analysis of the function of NINJAs in lily dormancy release.

Gelsemium elegans is a valuable traditional Chinese herb in China. The whole plant is highly toxic, but has anti-tumour and analgesic effects, as well as fattening effects on pigs and other livestock, and its main active constituents are terpenoid indole alkaloids (TIAs). Among them, the most representative ingredient is gelsenicine, which has the strongest toxicity and highest activity. In this study, 209 genes related to cytochrome P450 (CYP450) were screened based on the genomic and transcriptomic data of G.elegans, combined with metabolomic data, an important candidate gene GeCYP85A1 (GenBank No. OQ653843) related to the biosynthesis pathway of Oryza japonica was screened by "genome+expression profile+metabolome". The full-length cloning of this gene was performed by PCR, and its bioinformatics analysis was carried out. The expression pattern of the GeCYP85A1 gene in different tissues of G. elegans was analysed by qPCR. The results showed that the GeCYP85A1 gene was 1 392 bp long, encoding 463 amino acids, with a theoretical relative molecular mass of 53.390 06 kD and a theoretical isoelectric point of 9.07. It was a non-secretory protein with a subcellular localization in chloroplasts, a transmembrane region and no signal peptide. It was similar to Coffea arabica, Coffea eugenioides, Erythranthe guttata, Nicotiana tabacum, Olea europaea subsp. europaea, Phtheirospermum juttata, Salvia miltiorrhiza, Sesamum indicum, and Vitis riparia, and the amino acid sequences of the related CYP85A1 of the above 9 species were highly similar and belonged to a subfamily of the CYP450 family. By qPCR, the expression pattern of GeCYP85A1 in different parts of the plant was found to be consistent with the trend of the relative content of gelsenicine in different parts of the plant, which were root>flower>stem>leaf. This study provides a reference for further elucidation of the biosynthetic pathway for the TIAs-like alkaloids in G. elegans.

Sucrose synthase family is a crucial enzyme that regulates sucrose metabolism. It catalyzes the conversion of sucrose into various metabolic pathways and plays a significant role in plant growth and development, signal transduction, stress resistance and so on. In order to explore the regulatory mechanism of sucrose synthase gene 2 from Dendrobium officinale (DoSUS2), in polysaccharide anabolism and response to abiotic stress, the DoSUS2 gene (GenBank No. OQ657188) was cloned from leaves of D. officinale using homologous sequence method, the bioinformatics and expression characteristics were analyzed. The results showed that the ORF of DoSUS2 was 2 424 bp and encoded 807 amino acids, which had 36 bases differences compared with the reference sequence (Gene ID: LOC110110417), but all of them were synonymous substitutions. The theoretical molecular weight and isoelectric point of DoSUS2 protein were 92.05 kD and 5.97, respectively. It was a hydrophobic stable protein with only one signal peptide and 69 phosphorylation sites but no transmembrane domains, which contained conserved domain with SUS protein, such as sucrose synthase and glycosyl transferase domains. Tertiary structure modeling of DoSUS2 was highly homologous with AtSUS1 of Arabidopsis thaliana (73.82%), and belongs to the SUSⅠ subgroup of monocotyledonous. The expression of DoSUS2 had obvious differences among different tissues, which were relatively high in roots and floral organs while lower in leaves and stems. The promoter element analysis revealed that DoSUS2 contained several cis-acting elements in response to stress, and it was significantly induced by cold, drought stress and abscisic acid (ABA) treatment. The expression of DoSUS2 was up-regulated significantly (P<0.01) and peaked at 6 h after cold and ABA treatment, which reached 267 and 233 times compared with untreated. Similarly, the expression was also up-regulated accompanying with drought treatment time increased, and reached its peak at 9 h, its expression reached a extremely significant difference level significantly (P<0.01) which was 18.6 times then untreated group. The results showed that the DoSUS2 gene might participate in the response process to low temperature and drought stress through the ABA signaling pathway. At last, an DoSUS2 protein interaction network was generated with several enzymes closely related to glucose metabolism. These findings above suggested that DoSUS2 might respond to abiotic stress by regulating the expression of related enzyme genes and glucose metabolism pathways, thereby improving the stress resistance of D. officinale. The results would provide valuable theoretical basis for the further investigations of DoSUS2 function and its molecular mechanism in polysaccharide biosynthesis and abiotic stress response.

The degree of proliferation and differentiation of adipocytes deposited in muscle fibers or between muscle bundles determines the level of beef marbling index directly, which affects the quality level of beef. This study was to assess the effects of conjugated linoleic acid (CLA) on the proliferation and differentiation of intramuscular preadipocytes in Luxi cattle (Bos taurus). In vitro cell model was developed for proliferation of bovine intramuscular preadipocytes which were cultured in growth medium supplemented with CLA (50, 100 and 150 μmol/L) for 4 d. 100 μmol/L CLA significantly increased the proportion of S phase cells (P<0.05, increased by 1.63 times), decreased the proportion of G0/G1 phase cells (P<0.05, decreased by 9.2%), and the other treatment groups had no significant difference (P>0.05). The results showed that 100 μmol/L CLA significantly increased the proliferation rate of bovine intramuscular preadipocyte by flow cytometry (P<0.05). Bovine intramuscular preadipocytes were cultured in differentiation media with different doses of CLA, the induction of adipogenesis were indicated by measuring the lipid droplets. Six days later, lots of lipid droplets were observed in differentiated preadipocytes, in which cytosolic lipid content remarkably increased at 100 and 150 µmol/L CLA, respectively (P<0.05). Four combination of CLA (100 µmol/L), insulin (10 μg/mL) and dexamethasone (0.25 μg/mL) was designed and cultured cells were induced for 6 d with CLA, insulin and dexamethasone. When dexamethasone, insulin or CLA were removed, fat content in the cytoplasm decreased by 74%, 68% and 31%, respectively. The combination of CLA, insulin, and dexamethasone also resulted in the greatest cytosolic lipid content. 100 μmol/L CLA can increase the expression of peroxisome proliferator-activated receptor-γ (PPARγ) gene by about 3 times, and the expression level of CCAAT/enhancer-binding protein-α (C/EBPα) gene by about 2 times (P<0.05). Taken together, the present findings indicated that CLA may promote bovine intramuscular preadipocyte proliferation and differentiation, which highlighted the potential use of CLA on bovine intramuscular preadipocyte proliferation and differentiation to improve beef quality.

High concentrate diet fattening can increase the daily weight gain of mutton sheep (Ovis aries), but also cause the rumen acid production and fermentation too fast, and the body metabolism imbalance. Complex probiotics can maintain the balance of gastrointestinal flora and promote the growth and development of ruminants through the synergistic action of various probiotics. To investigate the effects of different types and proportions of complex probiotics on rumen microbial community of Hu sheep, 60 healthy male Hu sheep lambs with similar body weight and age were randomly divided into 4 treatment groups. The control group was fed basal diet, and experimental group1, 2 and 3 were supplemented with 2% and 4% probiotic preparation FA1 and 2% probiotic preparation FA2 in the basal diet, respectively. After 60 d of feeding, the sheep were weighed, rumen fluid was collected to measure pH and detect rumen microbial composition through 16S rRNA sequencing of bacteria. The results showed as follows, compared with the control group, the average daily gain of experimental groups supplemented with probiotics showed an increasing trend, but no significant difference, and the average ratio of feed to gain was significantly decreased (P<0.05). There were no significant differences in rumen microbial Alpha diversity and Beta diversity index among groups. The bacterial abundance of Clostridia UCG-014 significantly increased in experimental group 1. The bacterial abundance of Solobacterium significantly increased in experimental group 2. The bacterial abundance of Lachnospiraceae-NK3A20-group and Acidaminococcus significantly increased in experimental group 3. The functional pathway of cellulolysis in test group 2 was significantly reduced, and the content of crude fiber in feces was significantly increased (P<0.05). The results showed that the addition of different types and proportions of complex probiotics in the diet could increase pH of rumen fluid and the abundance of anaerobic probiotics and improve the growth performance of Hu sheep. However, 4% complex probiotics FA1 had a negative effect on the apparent digestibility of crude fiber. This study provides a reference for the theoretical research and production application of complex probiotics.

Forkhead box protein O1 (FoxO1) is a key transcription factor regulating fatty acid metabolism. Although FoxO1 promoter has been shown to be regulated by upstream transcription factors in non-ruminants, the mechanism of FoxO1 regulation during lactation in dairy goats (Capra hircus) remains unclear. This study designed primers based on the genome sequence of Yunnan black goat and cloned the 5' flanking sequence of FoxO1 gene. Bioinformatic analysis of FoxO1 promoter sequence was performed using online software to predict the binding sites of transcription factors. Double luciferase reporter vectors containing different lengths of FoxO1 promoter fragments were constructed by deletion mutation method, and double luciferase reporter vectors containing mutations of cis-acting elements were constructed by overlapping extension PCR method. The recombinant constructs and reference vectors were co-transfected into goat mammary epithelial cells (GMEC), and the promoter activity was detected by dual luciferase system. The results showed that 1 500 bp FoxO1 promoter sequence was cloned from the blood genome of dairy goats. Bioinformatics analysis revealed 2 binding sites of FoxO1 on the FoxO1 promoter. Deletion mutation results showed that region from -95 to -24 bp was necessary for basal transcription activity of FoxO1 promoter. Overexpression of FoxO1 significantly increased the activity of FoxO1 promoter. Mutation of the forkhead transcription factor 1 (FKH1) (-359 bp) site showed a significant decrease in FoxO1 promoter activity compared to wild type, but unchanged after mutation of FKH1 and FKH2 sites. Above results suggest that FoxO1 might promote FoxO1 transcription by binding to the FKH elements located at the promoter. This study provides theoretical reference for the functional study of the FoxO1 gene promoter and basic data for constructing the fatty acid metabolism network in mammary gland of dairy goats.

Cell-death-inducing DNA fragmentation factor-α-like effector a (CIDEa) is a lipid droplet associated protein, which is involved in regulating lipid metabolism and maintaining body energy balance. In order to further explore the function of CIDEa in lipid metabolism in mammary gland of dairy goats (Capra hircus), in this study, dairy goat mammary epithelial cells were used as a model to study the effect of CIDEa gene on lipid synthesis by constructing pcDNA3.1-CIDEa overexpression vector. The results showed that the mRNA expression of CIDEa gene was increased nearly 10-fold after 48 h of transfection with pcDNA3.1-CIDEa in dairy goat mammary epithelial cells (P<0.01). The results of triacylglycerol (TAG) contents assay and Oil Red O staining showed that the accumulation of lipid droplets and the content of triglyceride (P<0.05) in cells were increased after CIDEa overexpression. The results of real-time quantitative PCR showed that CIDEa overexpression significantly increased the expression of acetyl-CoA carboxylase alpha (ACACA)(P<0.05), cluster of differentiation 36 (CD36)(P<0.05), sterol regulatory element binding protein 1 (SREBP1)(P<0.05) and diacylglycerol acyltransferase 2 (DGAT2)(P<0.01).These results indicated that CIDEa gene could promote lipid synthesis in mammary epithelial cells. This study provides a theoretical basis for further studying the role of CIDEa gene in the regulation of milk fat metabolism in dairy goats.

Porcine (Sus scrofa) interferon α/interleukin-2 (PoIFN-α-linker-PoIL-2) is mostly expressed as inclusion bodies in Escherichia coli. To obtain soluble expression of active recombinant (r) PoIFN-α-linker-PoIL-2 in the E. coli system, the chimeric gene PoIFN-α-linker-PoIL-2 was synthesised by soluble modification based on the codon preference of E. coli. The modified PoIFN-α-linker-PoIL-2 chimeric gene was cloned into the expression vector pET-32a(+) for prokaryotic expression, and the expressed soluble recombinant fusion protein (rPoIFN-α-linker-PoIL-2) was purified using a nickel-chromium affinity chromatography column. The proliferative activity of rPoIFN-α-linker-PoIL-2 protein on peripheral blood T lymphocytes in vitro was detected by lymphocyte proliferation assay, could also be detected by ELISA assay using anti-PoIL-2 monoclonal antibody or anti-PoIFN-α monoclonal antibody. The antiviral bioactivity of rPoIFN-α-linker-PoIL-2 protein was tested by inhibiting the 50% appearance of cytopathic effect (CPE) of different viruses on different cell lines. The results showed that the chimeric gene PoIFN-α-linker-PoIL-2 could be efficiently expressed in E. coli. The expressed rPoIFN-α-linker-PoIL-2 protein had a molecular weight of about 55 kD. The purity of rPoIFN-α-linker-PoIL-2 protein was over 90% after purification by Ni-Cr affinity chromatography, which had significant proliferative activity on peripheral blood T lymphocytes (PBLC). Specific immune response can be detected by anti-PoIL-2 and anti-PoIFN-α monoclonal antibodies. The rPoIFN-α-linker-PoIL-2 protein had different inhibitory activities on the proliferation of different viruses in different cells. The activity units of inhibiting the proliferation of Vesicular stomatitis virus (VSV) in porcine kidney cells (PK-15) and human amniotic cells WISH were were 1.8×10⁶ and 2.5×10⁶ IU/mg. The active units of inhibiting the proliferation of Pseudorabies virus (PRV) and Seneca virus (SVA) on PK-15 cells were 2.2×10⁴ and 1.3×10⁵ IU/mg, respectively. The activity unit for inhibiting the proliferation of Porcine epidemic diarrhea virus (PEDV) in Verda Reno (Vero) was 3.2×10⁴ IU/mg. The soluble rPoIFN-α-linker-PoIL-2 protein was obtained in this study, which could inhibit the proliferation of VSV, PRV, SVA and PEDV in different cells. This study provides basic data for further study on the activity of rPoIFN-α-linker-PoIL-2 protein in pigs.

During the process of growth and reproduction, poultry is susceptible to various stress factors, the most prominent of which is oxidative stress. To investigate the effects of oxidative stress on the testicular structure and expression of genes related to testosterone synthesis in chickens (Gallus gallus), 60 healthy Kangle yellow roosters aged 180 d were randomly divided into 4 groups: Control group, diquat 5 mg/kg group, diquat 15 mg/kg group and diquat 20 mg/kg group. Testicular tissue samples were collected after 21 d of treatment. qRT-PCR was used to identify the presence of oxidative stress by measuring the expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), hemeoxygenase-1 (HO-1) and glutathione peroxidase 1 (Gpx1), and Antioxidant Function Assay Kit was used to detect the enzyme activities of catalase (CAT), total superoxide dismutase (T-SOD) and the concentration of malondialdehyde (MDA). Hematoxylin-eosin staining (HE staining) was used to analyze the effect of oxidative stress on testicular tissue structure. The relative protein expressions of apoptosis-related B-cell lymphoma-2 associated X protein (Bax), B-cell lymphoma-2 (Bcl-2) and Caspase3 were detected by using Western blot. The relative mRNA and protein expressions of steroidogenic acute regulatory protein (StAR), cytochrome P450 family 11 subfamily A member 1 (Cyp11a1) and 3β-hydroxysteroid dehydrogenase (Hsd3b1), which were involved in testosterone biosynthesis, were detected by using qRT-PCR and Western blot, respectively. The results showed that the mRNA level of oxidative stress-related genes HO-1, Nrf2 and Gpx1 were significantly down-regulated in the testes of the different doses diquat groups when compared to the control group (P<0.05). CAT and T-SOD activity were significantly decreased (P<0.05), while MDA concentration was significantly increased (P<0.05). After diquat induction, the structure of chicken testes was damaged, and spermatogenesis in seminiferous tubules was significantly inhibited with severe cell apoptosis, according to HE staining. The protein expressions of Bax and Caspase3 were significantly up-regulated (P<0.05), while the expression of Bcl-2 was significantly down-regulated (P<0.05) in diquat-treated testes compared to control group. The mRNA and protein expressions of testosterone synthesis-related genes StAR, Cyp11a1 and Hsd3b1 were significantly down-regulated in diquat-treated testes compared to control group (P<0.05). These results indicated that oxidative stress could damage the tissue structure of chicken testes, promote the apoptosis of testicular cells, and significantly inhibit the expression of genes involved in testosterone synthesis. This study can provide basic data for exploring the mechanism of oxidative stress damaging reproductive performance of chickens.

The cultivation of XY type pseudo female fish is a necessary step in obtaining faster growing all male strains of channel catfish (Ictalurus punctatus). In order to investigate the ovarian development at the period of adult and transcriptomic expression during sex differentiation after XY female channel catfish was induced by 17β-estradiol. The ovarian development and oocyte development of channel catfish were compared and analyzed by histological and anatomical methods with positive genetic sex identification, and the gonad development of XY pseudo-female channel catfish at the 3-year-old (500~2000 g) was evaluated by statistical gonad index. Transcriptomic sequencing was performed on XY pseudo-female channel catfish and normal female channel catfish at the age of 60 d after hatching. The gonad index of the 3-year-old XY female channel catfish was significantly different, the ovary of the large size XY females and the normal females was larger, with long sacs, densely filled with blood vessels and full of yolk. Oocytes in vitellogenic stage were mainly accompanied by cortical follicular stage and a little number of primary growth oocytes. Transcriptome sequencing revealed 11 288 differentially expressed genes between the ovaries of 60 d after hatching XY type pseudo female fish and normal female fish, enriched in multiple immune related biological processes and signaling pathways. According to GO enrichment and KEGG signaling pathway, among the top 30 GO enriched pathways, the female gamete generation pathway was screened, in which 32 genes showed differential expression, with down regulation of progesterone receptor (pgr) gene and androgen receptor (ar) gene expression. The research provides reproductive biology techniques and theoretical basis for sex control breeding of channel catfish.

The solute carrier 6 (SLC6) gene family, known as neurotransmitter transporters, plays a crucial role in stress regulation and osmotic regulation of aquatic organisms. In the present study, the members of the SLC6 gene family of Sinonovacula rivularis (SrSLC6) were identified and analyzed by using bioinformatics methods, and expression patterns under salinity stress were investigated. The results showed that a total of 29 SrSLC6 genes were identified, which belonged to 4 subfamilies and contained 12 conserved motifs and 2 conserved domains SLC6sbd-TauT-like and SLC5-6-like_sbd. Under high and low salinity stress, 16 SrSLC6 gene family members showed different expression responses and their expression levels were up-regulated or down-regulated. Amino acid transporter 4 protein of S. rivularis (SrAAT4) related to salinity adaptation was mainly located in lateral ciliated columnar cells and flat cells on the side of gill filaments, and the expression patterns showed the same trend as transcriptome expression profile, which was significantly decreased at different time points under low salt stress (P<0.05), and significantly increased at different time points under high salt stress (P<0.05). These results indicated that the amino acid transporters (AATs) and neurotransmitter transporters (NTTs) members of SrSLC6 gene family might played important roles in osmotic regulation of S. rivularis, which provided theoretical basis for further research on the osmotic regulation and salinity adaptation of SLC6 gene family in mollusks.

Organic cultivation can improve soil fertility and biodiversity by protecting soil organic matter. Greenhouse cultivation can artificially control the growing environment of plants and protect crops from outdoor disturbances such as pests and diseases, extreme weather conditions and pesticide drift. In this study, two different land use methods (solar greenhouse and open field vegetable cultivation)(0~15) cm top soil and (15~30) cm deep soil were used as research objects, and Illumina sequencing was utilized to analyze the variations in soil microbial community structure. Key soil environmental factors affecting fungal community structure were identified through dbRDA analysis. Meantime, FUNGuild was used to analyze the response of soil fungal function to solar greenhouse cultivation.The results showed that organic cultivation in greenhouse significantly reduced (P<0.05) the richness and diversity of topsoil fungal communities. The dominant fungi of different land-use mode were Ascomycota, Glomeromycota and Basidiomycota. Although there was no change in the species of dominant fungi in the soil due to different land-use modes, their relative abundance was significantly influenced. Solar greenhouse cultivation increased the number and abundance of beneficial fungi such as Chaetomium, Mortierella, Orbilia, Acaurospora, and inhibited the growth of plant pathogenic fungus Fusarium in the surface soil. LEfSe analysis showed that 22 and 21 different species were detected in greenhouse and open soil, respectively. The Mortierellomycota, Rozellomycota, and Chaetomium, Mortierella, and Humicola fungal genera in greenhouse soil showed significant differences in microbial communities. Fusarium, Stachybotry, and Cephaliophora played a dominant role in the open field cultivation. dbRDA analysis showed that total potassium (TK), soil organic matter (SOM), pH, total nitrogen (TN) and total phosphorus (TP) were the environmental factors that significantly affected the soil fungal community in solar greenhouse. FunGuild function analysis showed that the most advantageous nutrient type in greenhouse soil was Saprotroph, while in open feiled was Pathotroph-Saprotroph.Thus, greenhouse organic vegetable cultivation could significantly affect the fungal community structure, improve the abundance of beneficial fungi in soil and reduce the content of pathogenic fungi, and adaptation of changes in fungal ecological functions to environmental influences caused by different land-use patterns. This study revealed the effect of vegetable cultivation in solar greenhouse on the structure and function of soil fungal community under long-term organic cultivation mode, and will provide theoretical basis for the composition of soil fungal community and long-term organic cultivation under greenhouse conditions.

Enzymatic deglycosylation is an important means to study the structure and function of glycoproteins. Traditional enzymatic deglycosylation mostly uses free enzymes, so that the residual deglycosylase becomes a potential source of contamination signals for subsequent glycoprotein analysis (such as mass spectrometry analysis). To explore new utilization methods of deglycosylation is of great value for the analysis of glycoprotein glycan chain structure. In this study, the recombinant endo-β-N-acetylglucosaminidase (EndoEf) of Enterococcus faecaliswas expressed and purified, using sepharose CL 6B activated by epichlorohydrin as the carrier to immobilize, and the catalytic characteristics of immobilized EndoEf were analyzed. The results showed that EndoEf could be efficiently expressed in Escherichia coli BL21 Star (DE3), and 186.5 mg of enzyme protein could be purified per liter of bacterial solution. The optimal conditions for immobilization of EndoEf were 4.0 mg enzyme per gram of carrier in 50 mmol/L phosphate buffer at pH 8.0, 12 h conjugated at 4 ℃. Immobilized enzymes could remove N-glycan chains of ribonuclease B (RNase B) and ovalbumin (Ova), remaining active when stored at 4 ℃ for 60 d. Immobilized EndoEf had higher activity in the range of 30~40 ℃ and pH 6.0~7.0. The tolerance to DL-dithiothreitol (DTT) and sodium dodecyl sulfate (SDS) was significantly improved than that of the free enzyme, which could tolerate denaturation conditions of 12.0 mmol/L DTT and 2.0% SDS, and tolerate 1.0 mol/L NaCl. Matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS) analysis showed that centrifugation completely removed immobilized EndoEf from the reaction system. The establishment of immobilization method and the analysis of catalytic characteristics could provide a basis for the application of immobilized EndoEf in glycoprotein research.

Madin Darby Canine Kidney (MDCK) cell line is one of the most commonly used cell lines to produce Influenza virus vaccine, and miR-2779-x, as a non-coding RNA, can inhibit spontaneous tumor transformation in MDCK cells. In this study, stable cell lines with overexpression and knockdown of miR-2779-x were constructed based on the lentivirus system, and the transfected cells were detected by cell proliferation detection kit CCK-8 (Cell Counting Kit-8), cell apoptosis and cell cycle detection kit, as well as cell scratch test and soft agar cloning test. The results showed that overexpression of miR-2779-x significantly decreased cell proliferation and relative mobility (P<0.01), and increased cell invasion (P<0.01), while knockdown cell lines showed opposite results. The early apoptosis rate of overexpressed cell lines and the late apoptosis rate of knockdown cell lines were significantly increased (P<0.01). miR-2779-x overexpression caused cell block in S phase, and knockdown cell lines were blocked in G0/G1 phase. Measured at 50% tissue culture infective dose (TCID₅₀), the sensitivity of MDCK cells with miR-2779-x lentivirus overexpression and knockdown did not change significantly to Influenza virus A H1N1. The expression changes of miR-2779-x target genes were verified through qRT-PCR and Western blot, and multiple genes related to transformation, tumorigenesis, and growth were found to be negatively regulated by miR-2779-x. It is speculated that the target gene CASP9 (caspase 9) might act as an apoptotic pathway factor and co-regulate MDCK cell proliferation with PI3K/AKT signaling pathway. This study provides a new approach for establishing a novel MDCK cell line, which can provide a better cell matrix for influenza vaccine production.

Pseudoenzymes are proteins that have sequence homology with enzyme families but are proven or predicted to lack enzyme activity due to mutations in otherwise conserved catalytic amino acids. However, pseudoenzymes may be involved in promoting or inhibiting the activity of enzymes, competing with active enzymes for substrates, regulating signal pathways, or serving as protein scaffolds to form protein complex, etc. Although pseudoenzymes have been extensively studied in the medical field, research on the role of pseudoenzymes in plants is still scarce. Recent studies have shown that pseudoenzymes play important roles in stress response, growth, and development in plant. In this paper, multidimensional introduction to pseudoenzymes was provided, including their definition, identification methods, evolutionary processes, and biological roles in plants. Furthermore, the potential roles of pseudoenzymes in plant development regulation were also discussed. This review provides new insights for plant developmental regulation by using pesudoenzymes.

To understand the global patent landscape and industrial development trends of agricultural animal gene editing technology, in present study, patent bibliometrics were used to analyze patent data in the gene editing technology field from 2015 to 2022. A systematic analysis was conducted, including major research institutions, research teams, research frontiers, and the overview of gene editing technology for different agricultural animal species. The results showed that gene editing technology patents had been on the rise globally over the past 8 years. China ranked first in the world in terms of the number of gene editing technology patents applied and published, but overall, the United States was leading the world in gene editing technology. Particularly in terms of core technology innovation, the quality of papers and patents, top research institutions and teams, transformation of knowledge achievements, and industrialization mechanisms, China still needs to improve further. In China, the number of gene editing technology patents related to agricultural animal breeding was relatively small. However, some related companies had already completed first-round financing at the level of 10 millions. This study provides a systematic analysis of the global industrialization trends of agricultural animal gene editing technology, and provides a reference for China to use gene editing technology to address major problems in agricultural animal breeding.

The cytoplasmic male sterile line (CMS) is one of the important prerequisites for breeding superior hybrid rice (Oryza sativa) varieties, cultivating excellent CMS is of great significance for hybrid rice breeding. In this study, the high-resistant and high-quality rice variety 'Huahang Simiao' was used as the donor parent, and it was crossed with the Indica three-line maintainer line 'Fudao B', which had good comprehensive agronomic traits but poor grain quality and rice blast resistance. In the F₃ generation, superior plants were selected as male parents and crossed with 'Gufeng A' as the female parent. The progeny from the crosses were continuously subjected to shuttle breeding and selection in Sanya and Shaxian bases. After multiple generations of elimination and selection, a stable CMS line named 'Funong A' was finally bred, and its corresponding maintainer line was named 'Funong B'. Comprehensive analysis of agronomic traits revealed that 'Funong A' had good leaf morphology, sturdy stems, and erect flag leaves, with a morning flowering ratio of 85%. The stem diameter and thickness of 'Funong A' were significantly larger than those of the parent lines. Field tests and microscopic observations showed that the sterility rate of 'Funong A' was 100%, pollen sterility ratio was 99.99%, and stigma exsertion rate was 57.12%. 'Funong A' exhibited a significantly larger basal internode diameter and thicker stem wall, as well as higher content of hemicellulose and silicon in stems. High-density chip analysis revealed the presence of semi-dwarf gene Sd-1, spikelet development-related gene qNGR9, and grain weight gene OsSPL16 in 'Funong A'. Additionally, Funong A successfully combined 3 rice blast resistance genes, Pi5, Pid2, and Pid3, with an indoor identification rate of resistant strains reaching 81.82%. Moreover, the 7 rice varieties developed using 'Funong A' exhibited high yield, good grain quality, and intermediate resistance to rice blast disease. It is evident that 'Funong A' possesses strong development potential and can provide support for the combination and breeding of hybrid rice with good resistance, high yield, and superior rice quality.

Morel (Morchella spp.) is a world-famous rare edible and medicinal mushroom. In the artificial cultivation process morels must be covered with soil to produce fruit. At present, the mechanism of soil microorganisms affecting the growth and development of morel is still unknown. 132 strains of bacteria were isolated and purified from morel soil. Through the plate confrontation culture test, it was found that 15 strains had an antagonistic effect on the hyphae of Morchella importuna and 4 strains had the effect of promoting growth. Among them, when strain X14 was co-cultured with morel on plate for 4 and 8 d, the growth rate of morel hyphae was significantly increased by 7.6% and 11.6% compared with the control (P<0.05). Morel and strain X14 were inoculated into wheat medium in test tubes and co-cultured. The growth rate of morel hyphae was measured at different culture time. The results showed that the growth rate of morel hyphae treated by strain X14 was very extremely significantly increased by 28.3%, 24.8% and 24.0% compared with the control at 4, 6 and 7 d, respectively (P<0.001). Morel and strain X14 were inoculated into stock flasks and co-cultured. The results showed that the growth rate of morel hyphae treated by strain X14 was not significantly different from the control at 5 d of co-culture (P>0.05). After 10 d of co-cultivation, the growth rate of morel hyphae treated with strain X14 was extremely significantly increased by 14.1% compared with the control (P<0.01). The culture period of the stock seed was shortened by 4~6 d compared with the control. Based on the comprehensive analysis of colony morphology, physiological and biochemical characteristics and 16S rDNA sequence, the strain X14 was identified as Pseudomonas putida. In this study, a strain of Pseudomonas putida X14 was screened from the soil of morel, which could promote the hyphae growth of Morchella importuna and provide technical support for the development of microbial agents that promote the growth of morel.

Fusarium oxysporum is one of the dominant pathogens causing Astragalus membranaceus var. mongholicus (AMM) root rot. In order to rapidly and accurately detect and quantify F. oxysporum in AMM plants and soil, primers were designed based on the elongation factor-1α (EF-1α) sequence and verified for their specificity. The real-time quantitative PCR (qRT-PCR) detection system was established for the pathogen. The constructed qRT-PCR system was used to detect F. oxysporum in AMM plants and soil from AMM planting sites. The results showed that primer pair FaeF2/FaeR2 was highly specific, and the sensitivities of qRT-PCR detection were 0.896 pg/μL for F. oxysporum genomic DNA in AMM and 10² conidia/g for F. oxysporum conidia in soil. The correlation coefficients of the constructed standard curves for F. oxysporum detection in AMM and soil were 0.998 and 0.973, respectively. Repeatability evaluation showed that the system was stable and reliable. The AMM plants and soil samples were detected by the qRT-PCR detection system. The detection rate of the pathogenic F. oxysporum was 100% in F. oxysporum-inoculated samples, 66.7% in suspected diseased samples and 43.75% in soil samples; the pathogen colonization in AMM was positively related with the disease index and the number of conidia in diseased soil was significantly greater than that in healthy soil at the same AMM cultivation years. The qRT-PCR system established in this study was suitable for the detection of the pathogenic F. oxysporum causing AMM root rot, and will provide technical support for the rapid diagnosis, accurate evaluation and soil warning of AMM root rot.

Goose circovirus (GoCV) is a potential immunosuppressive pathogen that mainly infects host lymphocytes, leading to a decline in immune function, and easily causes secondary infection by other pathogens, which greatly harms the goose (Anser cygnoides orientalis) breeding industry. To establish an indirect ELISA method for the detection of GoCV antibodies, the ORF3 gene from GoCV was synthesized and cloned into the pET32a(+) vector. The recombinant plasmid pET-32a-ORF3 was constructed and used to express ORF3 protein in a prokaryotic expression system. After conditional optimization, the recombinant ORF3 protein was purified by NI-NTA affinity chromatography and showed good biological activity after SDS-PAGE and Western blot analysis. An indirect ELISA to detect GoCV antibodies was established using the obtained ORF3 protein. The optimized reaction conditions were as follows: the best coating antigen (the obtained ORF3 protein) concentration was 2.00 μg/mL, the dilution of serum was 1∶80, and goat anti-duck IgG-HRP (or goat anti-chicken IgG-HRP) was 1∶4 000 (or 1∶8000). Specificity results showed that only GoCV-positive sera were positive, with other goose origin pathogen-positive sera (such as GPV, ATmV, AIV, GRV, GoAstV) being negative, indicating good specificity. The results of repeatability tests within and between batches showed that the coefficients of variation were both less than 5%, indicating good repeatability. The optimized indirect ELISA conditions were used to carry out epidemiological investigation, and the data showed that GoCV sera positivity was found in ducks (Anatinae) collected in Fujian and Guangdong, indicating GoCV cross-species infection from geese to ducks. In conclusion, the established indirect ELISA method for the detection of GoCV antibodies in this study has good reproducibility and specificity and provides a practical method for serological investigation and surveillance of Goose circovirus infection.