Aldo keto reductases (AKRs) are widely distributed in plants and animals and play an important role in the metabolism of steroids, sugars, and other carbonyls. It is associated with the metabolism of exogenous and endogenous toxins, including toxic substances produced by stimulation. At present, some AKR genes involved in stress had been reported in plants such as Arabidopsis thaliana and rice (Oryza sativa), but the research of AKRs in soybean (Glycine max) are still scarce. In this study, bioinformatics methods and qPCR were used to identify the AKR family members in soybean on whole genome and analyze its expression characteristics. The results showed that 44 GmAKR genes in soybean genome, and the proteins encoded by GmAKRs all had the Aldo-ket-red domain. Phylogenetic analysis showed that GmAKRs could be clustered into 5 families, which distributed unevenly on 16 chromosomes. The promoter regions of GmAKRs had different numbers of hormone or stress-responsive cis-acting elements, suggested that GmAKRs may be involved in various growth and development regulation processes of plants. Expression pattern analysis showed that GmAKRs had different tissue expression characteristics, and was basically consistent with the Phytozome database. These results provide a reference for further revealing the biological functions of AKR family in soybean.

Chinese cabbage (Brassica rapa ssp. pekinensis) is native to China and belongs to the crucifer family Brassica genus biennial plant. It is a vegetable crop widely cultivated in our country. In this study, a genetically stable yellow cotyledon pure line (19YC-2) and normal green cotyledon near isogenic line (19GC-2) and the F₁, F₂ populations by hybridizing with 19YC-2 and 19GC-2 were used as materials, The genetic analysis and gene mapping of yellow cotyledon mutant in Chinese cabbage were carried out. Phenotypic observation showed that the cotyledon phenotype of F₁ plants was normal green as 19GC-2, and the number of green cotyledon plants to yellow cotyledon plants in F₂ population was conformed to 3∶1 proportion, which indicated that yellow cotyledon mutation was a quality trait controlled by one single recessive nuclear gene. The parental polymorphic SSR primers were further screened by bulked segregant analysis (BSA) method and the gene mapping of yellow cotyledon was made. The yellow cotyledon gene was located on the A06 linkage group of Chinese cabbage with two nearest flanking markers SSR280-23 and SSR280-42, their genetic distances are 0.04 cM and 0.96 cM, respectively, and their interval physical distance was 83.9 kb. Through genetic analysis and gene mapping of the yellow cotyledon mutants of Chinese cabbage, this study lays a foundation for the cloning and molecular mechanism of Brassica rapa yellow cotyledon (Bryc).

The Pin-formed acicular protein (PIN), one of the plant auxin transporters, affects the quality of cotton (Gossypium spp.) fibers. Understanding the expression characteristics of GbPIN1a (GenBank No. KAB2059844.1) in sea-island cotton (G. barbadense) fiber can provide a reference for exploring the mechanism of fiber quality development. In this study, the gene was cloned by PCR technology, and the bioinformatics analysis was carried out; A transient expression vector containing GFP was constructed and cell localization analysis was carried out. The results showed that the length of the coding region of GbPIN1a was 1 758 bp, and 585 amino acids were encoded; two conserved domains including Mem trans were contained in amino acid sequence; GbPIN1a gene showed a high expression trend at 15 and 20 DPA (days post anthesis) of fiber development and was significantly negatively correlated with auxin content at the same time period (P<0.05); The expression level in roots and stems was significantly higher than that in leaves (P<0.05), with tissue specificity; The expression level fluctuated when stimulated by exogenous auxin; The expressed product was localized on the cell membrane. The results of this study preliminarily indicate that the GbPIN1a gene may be involved in the regulation of cotton fiber development, provide basic data for future research on PIN1a genes.

Natural colored cottons, including brown fiber cotton and green fiber cotton which can be spun into garments directly without chemical printing and dyeing, are special upland cottons (Gossypium hirsutum) with green environmental protection and high ecological benefits. Due to the accumulation of different anthocyanins, procyanidins and their derivatives in the fibers, colored cottons show different colors. It have been confirmed that proanthocyanidins and their derivatives are thought to be responsible for the color formation of brown fibers. Based on previous transcriptome data, this study cloned a R2R3-MYB (v-myb avian myeloblastosis viral oncogene homolog, MYB) transcription factor gene GhTT2 (transparent testa 2), which was predominantly expressed during the development stage of brown cotton fibers. The structure, expression pattern and subcellular localization of GhTT2 gene and coding protein were predicted and analyzed by phylogenetic tree construction, amino acid sequence multiple alignment, prokaryotic expression, qRT-PCR, tobacco (Nicotiana benthamiana) transient transformation, and yeast (Saccharomyces cerevisiae) hybridization assay. The results showed that the open reading frame of GhTT2 gene was 765 bp, encoding 254 amino acids, with predicted relative molecular mass 28.863 kD, isoelectric point 7.423, and a typical R2R3-MYB domain. The prokaryotic expression analysis showed that expression of GhTT2 could be efficiently induced under the condition of 37 ℃ and 1 mmol/L isopropyl β-D-thiogalactoside (IPTG). The results of qRT-PCR showed that, compared with white cotton, GhTT2 was predominantly expressed during the development stage of brown cotton fibers, and its expression in fibers was significantly higher than that in other tissues. The recombinant protein GhTT2-GFP was located in the nucleus, which conformed to the characteristics of typical transcription factors. Yeast transformation test showed that GhTT2 protein had obvious transcriptional activation activity. The study provides basic material for further verifying the biological function of GhTT2 in the color formation of brown fibers and the improvement of the color diversity of colored cotton fibers.

Passion fruit (Passiflora) is an important fruit tree in southern China, but the evolutionary relationship among Passiflora species is still unclear, and there is a lack of universal molecular markers to distinguish between different species. In this study, the phylogeny, SSR loci and marker versatility of 10 germplasms from 6 passion fruit species were investigated based on restriction site-associated DNA sequencing (RAD-seq). The results showed that EcoRⅠ was a suitable enzyme for simplified sequencing of passion fruit genome. According to the number of remaining reads/read lengths, the phylogenetic tree was constructed based on the 46 451 high-quality SNPs detected in RAD-seq, the partially assembled sequence of 'Purple fruit No. 7' (P. edulis)(P4) was used as the reference genome. The results showed that the blue-crowned passion fruit (P. caerulea)(P6), safflower passion fruit (P. coccinea)(P7), Banna passion fruit (P. xishuangbannaensis)(P8), and quadrangularis passion fruit (P. quadragularis)(P9) were on the same phylogenetic branch, green-skin passion fruit (P. edulis)(P5) and Colombian passion fruit (P. ligularis)(P10) were on the same branch, and Jinling purple fruit (P. edulis)(P1), guava golden fruit (P. edulis var. flavicarpa)(P2), and Yunnan yellow fruit native species (P. edulis var. flavicarpa)(P3) were closer to 'Purple fruit No. 7' in genetic relationship. When using 12 452 high-quality SNPs to construct the phylogenetic tree and using Banna passion fruit as the reference genome, the Yunnan yellow fruit native species, 'Purple fruit No. 7', safflower passion fruit, Banna passion fruit, quadrangularis passion fruit, and Colombian passion fruit were all on separate phylogenetic branches, Jinling purple fruit and guava golden fruit were on the same branch; and green-skin passion fruit and blue-crowned passion fruit were on the same branch. A total of 2 614 SSRs were identified from the 10 passion fruit germplasms, and their core motifs were AT, GA and AAG, etc., with 2~6 bases in length, and the repeat number was 4~16 times; 2 515 pairs of SSR primers were successfully developed accordingly. 50 pairs of SSR markers were used to evaluate the rate of shared markers among different passion fruit species, and the rate was 54.22%; notably, the shared rate in cultivar passion fruit (P1~P5) was significantly higher than the other 5 species (P6~P10), while the shared rate of wild Banna passion fruit in Yunnan, China was 0. This might be due to the distant genetic relationship between Banna passion fruit and other germplasms, and there were large differences in genome sequences. This study establishes the genetic relationships among different Passiflora species, provides a theoretical basis for the genetic improvement of passion fruit, and developes a useful panel of SSR markers for molecular marker-assisted selection of passion fruit.

The expression of purified and active target protein is prerequisite for the screening of protein-specific inhibitors. In this study, the prokaryotic expression, purification and activity analysis of tryptophan aminotransferase of Arabidopsis 1 (TAA1) were carried out. First, the TAA1 coding sequence was retrieved from GenBank. After codon optimization according to the preferred codon usage in Escherichia coli the target sequence was synthesized and inserted into the prokaryotic expression vector pET24 with a His tag to construct the recombinant expression vector pET24-GST-EK-His-TAA1. The vector was then transformed into the E. coli BL21(DE3) cell line followed by induction and cultivation with different methods (IPTG induction/TB medium, IPTG induction/LB medium, and auto-induction). The products were analyzed with sodium lauryl sulfate polyacrylamide gel electrophoresis to identify the size, amount and existing form of the expressed protein. It was found that the auto-inducible expression system combined with incubation at 37 ℃for 16 h yielded the greatest amount of protein product in the soluble fraction. This optimized expression method was then used for large-scale expression and purification of the proteins. The collected fusion protein was further cleaved by the enterokinase (EK) protease and purified with a nickel ion affinity column. Finally, purified recombined TAA1 protein at a purity of 78.7% and an amount of 0.5 mg was obtained. In vitro enzymatic activity assay proved that this protein had a transaminase activity. In conclusion, a high-efficient prokaryotic expression system for the Arabidopsis TAA1 protein was established successfully, from which the obtained purified and bioactive His-TAA1 protein are very useful for the screening of TAA1-specific inhibitors based on molecular interaction.

Erianthus fulvus, a relative wild species of sugarcane (Saccharum spp.), plays an important role in the study of sugarcane stress resistance. PHD-finger (plant homeodomain finger) protein is a transcription regulator that can respond to biological and abiotic stresses.In this study, based on the transcriptome identification of the EfPHD-finger family genes in Erianthus fulvus, bioinformatics and cold stress expression patterns were analyzed. The results showed that a total of 38 EfPHD-finger genes were identified. The number of amino acids encoded by EfPHD-finger genes ranged from 145 to 2 268, the molecular weight of EfPHD-finger proteins ranged from 16.161 0 to 251.333 7 kD, and the isoelectric point ranged from 4.35 to 9.66. The average hydrophilic coefficient ranges from -1.094 to 0.035. Subcellular localization indicated that all 37 EfPHD-finger proteins were in the nucleus, and only EfPHD24 was in the nucleus and chloroplast. Phylogenetic tree analysis demonstrated that 38 EfPHD-finger proteins were divided into 7 groups. Analysis of cold stress expression patterns based on transcriptome data revealed that 38 EfPHD-finger genes were differentially expressed under different degrees of cold stress. Among them, EfPHD13, EfPHD16, EfPHD21, EfPHD25, EfPHD28, EfPHD32, EfPHD34, and EfPHD38 were up-regulated, and EfPHD13 was sustained up-regulated, with the largest up-regulated multiple. The 8 up-regulated genes were verified by qPCR, and the correlation between the verification results and transcriptome sequencing results was analyzed. The results showed that the correlation of EfPHD25 and EfPHD28 genes was weak, and the correlation of other genes was strong (R≥0.90). Furthermore, EfPHD13 gene (GenBank No. OK356615) was cloned. Its CDS length was 1 863 bp, encoding 620 amino acids, containing PHD_MMD1_like domain, multiple phosphorylation sites, no signal peptide and transmembrane structure. This study provides reference for further research on the cold stress response function of PHD-finger family genes in Erianthus fulvus.

Exercise metabolomics is a systems biology study that evaluates the overall metabolic response of the body to exercise training stimulation. Recent years, mainly researches have focused on the sports science in human (Homo sapiens), and there are few reports on the study of Chinese horse (Equus caballus) sports metabolomics. In this study, ¹H nuclear magnetic resonance (¹H-NMR) was used to analyze the characteristics of plasma metabolomics of 6 Mongolian horses and 6 hybrid horses before and after 30 km endurance exercise, and metabolic patterns and differential metabolic markers between groups were identified by partial least square discriminant analysis (PLS-DA) package of R language and one-dimensional ANOVA test of SPSS 20.0 software. There were distinct metabolic models between 2 horse groups before and after exercise. The results showed that there were significant differences in metabolic patterns between Mongolian horses and hybrid horses before and after exercise. Ten differential metabolites were identified before and after Mongolian horse exercise in whcih the concentration of glucose, ethanol, glycine, glutamine, lysine, proline, valine, and τ-methylhistidine decreased and the concentration of methanol and creatine increased. At the same time, 15 differential metabolites were identified before and after hybrid horse exercise in which the concentration of glucose, hippurate, acetate, glutamine, valine, methanol, betaine, τ-methylhistidine decreased and the concentration of lactate, glycerol, acetone, 3-hydroxybutyrate, 3-Hydroxyisobutyrate, acetoacetate and creatinine increased. The comprehensive analysis of metabolite changes showed that the energy supply mode of Mongolian horses was mainly aerobic metabolism, while the energy supply mode of hybrid horses was mainly anaerobic metabolism of phosphate source and glycolysis, as well as fatty acid ketone metabolism. Therefore, Mongolian horses had better aerobic endurance quality and explosive potential than hybrid horse, so it is feasible to breed endurance racehorses based on Mongolian horse; and glutamine and τ-methylhistidine could be developed and applied as a monitoring indicators for horse endurance training. The results of this study provides basic data for endurance training and breeding of Mongolian horses in China.

Collagen typeⅠα1 chain (COL1A1) gene as an important component of typeⅠcollagen, plays a role in regulating follicular development and ovarian cell proliferation, and is closely related to animal reproductive traits. In order to explore the relationship between COL1A1 gene and fertility of Guizhou black goats (Capra hircus) and the effects of different expression levels on related genes in lambing, in this study, gonad axis tissues (uterus, ovary, pituitary, hypothalamus, fallopian tube) of monotocous and polytocous Guizhou black goats were collected. The expression of COL1A1 gene and its protein in the gonadal axis of monotocous and polytocous Guizhou black goats were detected by qPCR, and then ovarian granulosa cells were isolated and cultured. The interference vector of the target gene was constructed by RNA interference technology and transfected into ovarian granulosa cells. The expression level of COL1A1 gene and its protein was detected by qPCR and Western blot to screen the best interference group. qPCR was used to detect the effects of COL1A1 gene interference on the expression of lambing related genes bone morphogenetic protein 15 (BMP15), bone morphogenetic protein 1B (BMPR-1B), growth differentiation factor 9 (GDF9) and follicle stimulating hormone β-subunit (FSHβ). The results showed that COL1A1 gene was expressed in various tissues of Guizhou black goats. The expression level in ovary and hypothalamus of polytocous group was the highest, and was extremely significantly higher than that of monotocous Guizhou black goats group (P<0.01), significantly higher in pituitary gland and oviduct than in monotocous group (P<0.05), there was no significant difference in utero; Immunofluorescence results showed that the cells isolated and cultured in the experiment were ovarian granulosa cells. qPCR and Western blot results showed that shRNA-COL1A1-2 was the best interference group. After inhibiting the expression of COL1A1 gene, qPCR results showed that the expressions of lambing related genes BMP15, BMPR-1B, GDF9 and FSHβ were significantly lower than those of shRNA-NC group (P<0.01). This study revealed the difference of COL1A1 gene expression in gonad axis of monotocous and polytocous Guizhou black goat. The high expression of COL1A1 gene in the ovarian tissue of polytocous lambs group and the suppression of lambing related genes by silencing COL1A1 gene indicated that COL1A1 gene was closely related to ovarian development and follicle growth. It can be used as a candidate gene for the breeding of Guizhou black goat.

Vitamin E plays a very important role in improving the reproductive function of livestock and poultry, improving immunity and protecting the liver, and its storage and metabolism mainly occurs in the liver. α-tocopherol transfer protein (α-TTP) is mainly expressed in the liver, which can specifically bind α-tocopherol and promote its transport from the liver to peripheral tissues for absorption and utilization. However, some studies have shown that α-TTP may also require other interacting factors in the process of transporting α-tocopherol. In this study, using co-immunoprecipitation (Co-IP) and Pull-down screening, the α-TTP-interacting protein-myosin 1d (MYO1D) was obtained. Immunofluorescence observation showed that MYO1D and α-TTP co-localized significantly. The Myo1d-shRNA lentivirus plasmid was used to construct the interference vector of the Myo1d gene, and the hepatocytes line stably infected with the lentiviral interference vector was obtained by screening. The results of qRT-PCR showed that the Myo1d gene expression in the interference group decreased significantly (P<0.01), and the interference efficiency was 46.83%, while the expression of α-ttp gene in the interference group had no significant change. The results of the kit test showed that the content of α-tocopherol in the Myo1d interference group increased significantly (P<0.05). The above results suggest that MYO1D could interact with α-TTP to mediate α-tocopherol transport in hepatocytes, and might provide power in the process of transporting α-tocopherol-TTP complexes. These findings provide experimental bases for screening the interaction factors in the process of α-TTP transporting α-tocopherol, and provide a reference basis for further research on the mechanism of α-TTP transporting α-tocopherol in the liver.

TRAF family member-associated NF-κB activator (TANK) is involved in retinoic acid-inducible gene I (RIG-I)-like receptor mediated signaling pathway and plays an important role in the innate immunity of vertebrate. To explore the biological properties of TANK gene and its regulatory role in the antiviral immune response of rainbow trout (Oncorhynchus mykiss), the full length cDNA sequence of TANK gene (GenBank No. OK605587) was obtained by rapid amplification of cDNA ends (RACE) technique, and bioinformatic analysis was also performed. qPCR was used to detect the gene expression pattern in tissues of healthy fish and the expression changes in 6 important immune tissues (spleen, liver, head kidney, skin, gill and intestine) at different time points (0, 6, 12, 24, 48, 72, 96, 120 and 144 h) in rainbow trout after infection with Infectious hematopoietic necrosis virus (IHNV). The results showed that the full length cDNA of TANK gene was 1 618 bp, containing 194 bp 5' untranslated region (UTR), 317 bp 3'UTR and 1 107 bp ORF. The ORF of TANK encoded 368 amino acid residues. Bioinformatics analysis showed that molecular weight of TANK protein was 41.41 kD, theoretical isoelectric point was 6.03, instability coefficient was 70.60, average hydrophily was -0.748, and fat coefficient was 65.54, which indicated that TANK protein was an unstable hydrophilic protein. Structural analysis of the TANK protein revealed that it contained a conserved TBK1/IKKi-binding domain (TBD) and a coiled-coil region without transmembrane domain. Homology matching and phylogenetic analysis showed that rainbow trout and salmon (Oncorhynchus keta) had the highest homology (95.92%) and the closest evolutionary relationship. In addition, rainbow trout and salmon had the highest amino acid sequence similarity in the TBD structural domain of the TANK protein, showing that the TBD domain had similar functions in salmon trout species. Tissue expression analysis revealed that TANK gene was expressed at different levels in all tissues of healthy fish (head kidney, skin, eye, gill, brain, intestine, liver, spleen, heart and muscle), with the highest expression level in liver, followed by spleen and heart, and the lowest expression level in skin, which implied that TANK gene may play a major role in the innate immune response of rainbow trout. After IHNV infection, the expression level of TANK gene in important immune tissues (spleen, liver, head kidney, skin, intestine and gill) was extremely significantly up-regulated (P<0.01). The significant expression changes were appeared in skin and intestine of mucosal tissues, peaked at 48 and 72 h, which were 3.14 and 3.16 fold of control group, and the expression in liver, head kidney, spleen and gill peaked at 48, 48, 72 and 96 h, respectively. In the spleen tissue, the expression level increased firstly and then decreased, while the expression level of the liver, head kidney, skin, intestine and gill all decreased firstly, then rose and fell again. Meanwhile, the liver, head kidney, skin and intestine showed different degrees of rebound at 96 and 120 h. These results indicated that TANK gene may be involved in the innate immune response and played an important role in rainbow trout against IHNV infection. The results of this study provide basic data for further investigating the antiviral immune regulatory mechanism of TANK gene in teleost fish.

Salinity is an important environmental factor affecting the growth and survival of crustaceans. The red claw crayfish (Cherax quadricarinatus) is a freshwater species. To increase the waters available for cultivating this species, saline waters and brackish waters may be potential waters for cultivating this species. Thus, understanding the osmotic regulation mechanism of the red claw crayfish is needed to provide a theoretical basis for farming feasibility in brackish water and improve the flavor by changing the growth environment (temporary culture in brackish water). In this study, 3 genes related to osmolality, Na⁺/K⁺-ATPase (NKA), Na⁺-K⁺-2Cl^--Cotransporter (NKCC) and 14-3-3, were screened by the NCBI published transcriptome database of C. quadricarinatus, and their ORF regions were confirmed by RT-PCR. The protein structure was predicted using an online bioinformatics website, and the homologous protein sequences of different species were subjected to multiple sequence alignment and evolutionary tree construction. The expression of NKA, NKCC and 14-3-3 in different tissues and the changes of expression in gill and hepatopancreas after 24, 48 and 96 h under different salinity in the red claw crayfish were analyzed by qPCR. The results showed that NKA, NKCC and 14-3-3 proteins were highly conserved among different species. NKA, NKCC and 14-3-3 genes were expressed in the gill, muscle, heart, hepatopancreas and eyestalk tissues of red claw crayfish. Under different salinity stresses, the expression of the NKA gene in gill tissues did not change significantly at 5‰ and 10‰ salinity but increased significantly in 15‰ and 20‰ salinity groups at 48 h (P<0.05). In the hepatopancreas, the expression of NKA gene was only significantly increased in the 20‰ salinity group at 48 h (P<0.05). The expression of NKCC gene in gill tissue was significantly up regulated in 10‰, 15‰ and 20‰ salinity groups at 24, 48 and 96 h, and only significantly increased in 15‰ and 20‰ salinity groups at 48 h in hepatopancreas. The expression of the 14-3-3 gene was significantly up regulated in gill tissue in 10‰, 15‰ and 20‰ salinity groups at 48 h (P<0.05), and significantly up regulated in hepatopancreas in 5‰, 10‰ and 15‰ salinity groups at 48 h (P<0.05) and the expression significantly up regulated at 24 and 48 h in 20‰ salinity group (P<0.05). The results showed that the expression of NKA, NKCC and 14-3-3 genes was related to the change of salinity. 20‰ salinity stress after 48 h may be the key point of osmotic regulation of gill and hepatopancreas under high salinity. This study provides a reference for further exploring the role of NKA, NKCC and 14-3-3 genes in the osmoregulatory mechanism of the red claw crayfish.

In recent years, solid-state ¹³C nuclear magnetic resonance (NMR) technology has become an application hotspot in the research field of soil organic carbon molecular structure. Compared with loam and clay, sandy soil was more sensitive to the feedback mechanism of soil carbon fractions changes. To reveal the effects of soil organic carbon fractions and stability under different external organic carbon input, this study, with straw (ST) returning as the main control and chemical fertilizer (CF) treatment as the secondary control, the effects of the amendment of 3 organic materials for 8 consecutive years on the molecular structure of organic carbon in sandy soil were determined by ¹³C-NMR technology. Results showed that: 1) Soil organic carbon fractions were predominant as O-alkyl C (45~110 ppm) which was dominated by carbohydrate C (60~90 ppm) in pig manure (PM), biogas residue (BR) and biochar (BC) treatments, similar to that in the 2 control groups. 2) Compared with the ST treatment (main control), the recalcitrant organic carbon (alkyl C and aromatic C) decreased by 2.96% and 3.77% in BR and PM treatments respectively, but increased by 8.12% in BC treatment. Compared with the ST treatment, the Hydrophobicity index (HI) decreased by 4.93% and 6.25% in BR and PM treatments respectively, but increased by 14.65% in BC treatment. The aromaticity of BC treatment was about 1.7 to 2.0 times that of BR and PM treatments. 3) Redundancy analysis showed that the relative proportion of aromatic C significantly impacted the stability indexes and the content of soil organic carbon (P<0.01). Due to the differences in soil organic carbon fractions under the amendment of different organic materials, BR and PM treatments increased the relative proportion of labile organic carbon (carbohydrate C) and carboxylic C, which meant that the stability of soil organic carbon was weakened. BC treatment increased the relative proportion of recalcitrant organic carbon (aromatic C), the effect of improving the stability of soil organic carbon was more significant. The present study could provide insight for further improving the stability of farmland soil organic carbon pool.

Adenylyl cyclase (AC), the key enzyme for the synthesis of 3',5'-cyclic adenosine monophosphate (cAMP), is of vital role in the cyclic adenosine monophosphate messenger system. AC has been systematically studied in animals and microorganisms. While the research of AC in plants started late and proceeded not as fast as that in animals. Nevertheless, with the continuous development and progress of science and technology, visible research progresses have been made in the field of AC enzyme activity verification, AC gene mining, structural characterization and biological function analysis of AC protein in plants, but there is no relevant systematic review report. Therefore, this study systematically summarized the progresses on plant ACs to provide references for the further study of AC metabolism in plants.

The liver, a vital functional organ of the human body, can metabolize various substances and transport them to other parts of the body for utilization and excretion. It also has a plenty of physiological functions such as detoxification and the synthesis, metabolism, digestion and storage of various substances. Among them, fat and energy metabolism are particularly important. If this process becomes abnormal, including excessive fat synthesis, reduced metabolism, and decreased transport capacity, will cause the risk of a variety of diseases such as non-alcoholic fatty liver disease, liver fibrosis and cirrhosis, cholestasis, and even liver cell carcinoma rising rapidly. Liver is also closely related to the induction of other metabolic diseases. Recent studies have declared that glycine-N-methyltransferase (GNMT) in the liver acts as a key enzyme that regulates the conversion of S-adenosylmethionine (SAM) to S-adenosyl homocysteine (SAH), which can change the in-body level of SAM and then regulates the body's methylation status. GNMT can also alter the expression of fat metabolism and transport related genes, as well as the activity of oxidative respiratory complexⅡ to regulate fat content and reduce the occurrence and further development of diseases. This review describes the regulatory effects of GNMT on lipid metabolism, glucose metabolism, and oxidative stress in the liver. The therapeutic effects of GNMT in a variety of liver diseases are also discussed. This article can lay a solid theoretical foundation for future treatment methods and targets aim to GNMT.

Vanderbylia robiniophila is an important medicinal macrofungi. However, due to the lack of wild resources, the long period and the high cost of artificial cultivation, the fruit body of V. robiniophila is limited, so the extraction of polysaccharides from fermentation broth is an important way. In order to enrich the strain resources of V. robiniophila and improve the yield of polysaccharide in fermentation broth, a pure culture of V. robiniophila WH5 was obtained through the tissue isolation, it was identified by morphology and internal transcribed spacer (ITS) sequence analysis. The liquid fermentation technology of WH5 was optimized by single factor experiment and orthogonal experiment, and the anti-tumor activities of the exopolysaccharide in the fermentation broth were analyzed by cell experiment in vitro. The results showed that the length of ITS sequence from WH5 strain was 637 bp, and it was 99% similar to V. robiniophila (GenBank No. KX081122.1), clustered on one branch in the phylogenetic tree. Combining fruit body and mycelium morphology, strain WH5 was identified as V. robiniophila. The optimum liquid fermentation conditions of strain WH5 were as follows: Soluble starch 30 g/L, peptone 15 g/L, MgSO₄•7H₂O 1.0 g/L, KH₂PO₄ 2.0 g/L, pH7.0, rotating speed 160 r/min, fermentation temperature 30 ℃, fermentation time 9 d, the yields of mycelium and exopolysaccharide were 27.52 and 1.91 g/L, respectively. It was found that exopolysaccharides from the fermentation broth could inhibit B16 and Hep-3B tumor cells with the half-inhibitory concentration (IC₅₀) 24.12 and 13.25 mg/mL, respectively. This study is of great significance to enrich the fungus resources of V. robiniophila and promote the utilization of polysaccharides in the fermentation broth of V. robiniophila.

Fusarium pseudograminearum megabirnavirus 1 (FpgMBV1) is a mycovirus found in the hypovirulence Fusarium pseudograminearum strain FC136-2A. Four proteins were encoded in FpgMBV1. Among them, FpgMBV1-P4 composed of 268 amino acids with unknown function. Specific primers were designed and used in reverse transcription PCR (RT-PCR) to amplify the FpgMBV1-P4 gene from the hyphae of the strain FC136-2A in this study. Then a prokaryotic expression vector pGEX4T-FpgMBV1-P4 was constructed and transformed into Escherichia coli BL21(DE3) strain, and the recombinant protein was induced at 25 ℃ with 0.5 mmol/L isopropyl β-D-thiogalactoside (IPTG). Results showed that FpgMBV1-P4 in length of 807 bp was successfully amplified and the fusion protein with a molecular weight of about 60 kD was highly expressed . The purified fusion protein was used to immunize Zealand white rabbit (Lepus sinensis) to obtain antiserum. Indirect ELISA results showed the titer of this antiserum reached 1∶160 000. And the antiserum was used to specifically detect the FpgMBV1-P4 protein at about 36 kD from samples of the hyphae of FC136-2A. These results laid a foundation for the structural and functional analysis of FpgMBV1-P4, which would be meaningful in deciphering the hypovirulent mechanism of FpgMBV1.

Senecavirus A (SVA), an emerging pathogen in recent years, is posing a severe danger to the healthy development of pig (Sus domesticus) industry in China. Viral structural protein 1 (VP1), the main structural protein of SVA, can induce the body to produce specific antibodies. In order to prepare monoclonal antibodies (McAbs) against SVA VP1 and establish a rapid, efficient and simple method for the detection of SVA, the SVA VP1 gene was amplified by RT-PCR and cloned into the expression vector pET-28a (+). After induction, expression and purification, the recombinant SVA VP1 protein was obtained. BALB/c mice (Mus musculus) were immunized with recombinant SVA VP1 protein, 2 hybridoma cell lines (2E10 and 3E4) stably secreting monoclonal antibodies against SVA VP1 protein were obtained. Ascites type McAbs were obtained and purified by immunizing mice with cell lines 2E10 and 3E4. McAbs were identified by indirect immunofluorescence assay (IFA) and Western blot. The prepared 2E10 McAbs were used as capture antibody, Goat anti-mouse IgG and 3E4 McAbs were used as quality control band (C band) and detection band (T band) to produce SVA colloidal gold test strip. The results showed that the recombinant SVA VP1 protein was mainly expressed in the form of inclusion bodies, with a molecular weight of about 43 kD. The results of antibody subtype identification showed that the heavy chain subtypes of 2E10 and 3E4 were IgG2b and IgG2a, respectively, and the light chains were both κ chain. The results of IFA and Western blot showed that the obtained McAbs were specific bound to SVA. The test of strip showed that colloidal gold test strips could specifically detect SVA, and had no cross reaction with Foot and mouth disease virus (FMDV), Porcine reproductive and respiratory syndrome virus (PRRSV), Classical swine fever virus (CSFV) and Porcine circovirus type 2 (PCV2); The minimum detected viral content was 5×10^1.13 TCID₅₀/mL;The positive coincidence rate between this method and qPCR was 96.55%. The above results indicate that the prepared colloidal gold test paper based on VP1 monoclonal antibody in this study can provide a new method for the point-of-care testing of SVA.