The OsGASR6 (GA-stimulated transcript-related 6) gene of rice (Oryza sativa) pertains to the plant gibberellic acid-stimulated transcript (GAST) gene superfamily. Existing research has demonstrated that this gene family regulates the growth of plant cells and influences the morphology and size of plant organs. In this study, bioinformatics prediction and analysis of the sequence of the OsGASR6 gene disclosed that the OsGASR6 gene possessed typical sequence characteristics of GAST-like genes. The promoter region contained numerous cis-acting elements responsive to hormones, stress, light, and tissue-specific expression. The subcellular localization prediction and localization results of fusion expression vector showed that OsGASR6 was located in the extracellular space. The promoter activity of OsGASR6 gene was analyzed using the β-glucuronidase (GUS) reporter gene, and it was discovered that the OsGASR6 promoter was higher in rice endosperm, and it was also active in roots and stems. Phenotypic identification and analysis of rice lines with overexpression and RNAi interference of OsGASR6 gene expression were carried out. It was found that overexpression of the OsGASR6 gene extremely significantly reduced the height of rice stems (P<0.01), while the seed thickness of RNAi interference lines extremely significantly increased (P<0.01). This study provides a critical target for molecular breeding aimed at improving rice plant architecture and grain quality.

The diversity of color in mung bean (Vigna radiate) seed coat is one of the important phenotypic characteristics. Flavonoids are the key substances affecting the color of mung bean seed coat, and they have important medicinal value. There are great differences in nutritional and economic value for mung beans with different seed coat color. 'Zhangjiakou yellow mung bean' (ZH), 'Jin mung bean 8' (JL8) and 'Ji black green 0506' (JH) were used as experimental materials. The phenotypic parameters of mung bean were determined by non-contact spectrophotometer. The metabolites of mung bean seeds were detected and analyzed by widely targeted metabolomics technology. The key differential metabolites were screened by K-means clustering analysis and the correlation analysis was done with phenotypic parameters. The molecular mechanism of the nutritional value and colour formation were explored for the mung beans with different seed coat color. The 537 metabolites, 218 differential metabolites and 15 common differential metabolites were detected in the 3 mung beans. ZH shows significant differences in the distribution and accumulation of secondary metabolites, especially flavonoids, compared to JL8 and JH. Metabolome data analysis showed that selphinidin 3-glucoside and quercetin 3-O-β-D-glucosyl-(1→2)-rhamnoside might be the substances that cause the black color of the seed coat of JH. The luteolin 7-rutinoside, vitexin 2''-glucoside, nudifloramide and i-inositol were the causes of green accumulation in mung bean seed coat by the analysis of K-means clustering and correlation analysis. This study provides essential data for further revealing the pigments and the difference of nutrients of mung beans with different seed coat colors at the molecular level.

Heat shock transcription factor (HSF) plays an important role in regulating heat response and tolerance in plants. In order to further investigate the function of HSF genes in flowering Chinese cabbage (Brassica rapa var. parachinensis), this study used bioinformatics methods to systematically identify and analyze the HSF gene family and its molecular characteristics, and analyzed the gene expression patterns under high temperature stress. The results showed that a total of 39 HSF family genes were identified at the whole genome level of flowering Chinese cabbage, with protein sequence lengths ranging from 140 to 487 aa, molecular weights ranging from 16.10~53.82 kD, and theoretical isoelectric points ranging from 4.63 to 10.08. Protein subcellular localization prediction showed that 38 HSF proteins in flowering Chinese cabbage were located in the nucleus. The gene structure of most members of the HSF gene family in flowering Chinese cabbage was relatively conserved, with most genes contained 2 exons and 1 intron, as well as highly conserved DBD domain, conserved motif, and HSF domain. The 39 HSF genes in flowering Chinese cabbage were unevenly distributed on 10 chromosomes, with chromosome 3 had the most members. In synteny analysis, there were 32 genes in flowering Chinese cabbage that had a synteny relationship with Arabidopsis thaliana. The comparative analysis of the phylogenetic relationship between the HSF genes in flowering Chinese cabbage and A. thaliana indicated that these members of the HSF gene family could be divided into 3 categories: A, B, and C, with varying numbers of groups, and there were 9 pairs of orthologous genes and 2 pairs of paralogous genes. In addition, a large number of cis-acting elements related to plant hormones and stress response were found to be widely distributed in the promoter regions of HSF genes in flowering Chinese cabbage. qRT-PCR analysis showed that high temperature stress significantly induced the specific expression of most HSF genes in flowering Chinese cabbage (P<0.05), with 37 HSF genes showed up-regulation and 2 HSF genes showed down-regulation. Ten HSF genes highly expressed in the leaves or roots of flowering Chinese cabbage seedlings were screened, suggested that these genes play an important role in regulating the response of flowering Chinese cabbage to heat shock. This study provides a theoretical basis for understanding the characteristics of the HSF gene family in flowering Chinese cabbage and analyzing the functions of HSF family members in response to high temperature stress.

The swollen stem was induced by the infection of Ustilago esculenta in Zizania latifolia, following with the inhibition of polar growth of plants and the down-regulation of expression of auxin (IAA) polar transport related genes. In this research, the calcium-dependent protein kinase (CDPK) related kinase 5 (CRK5) gene of Z. latifolia was cloned with the full-length gDNA sequence of 4 224 bp, encoding 453 amino acids and containing the typical STKc_CAMK conformal domain of CRK kinase family. It was closely related to Oryza brachyantha. qPCR analysis showed that Zl-CRK5 was highly expressed in stem and leaf sheath of Z. latifolia (P<0.05), but lower in root and leaf (P<0.05). The expression of Zl-CRK5 in stems was significantly down-regulated (P<0.05) after 7 d infection of U. esculenta in plantlets of Z. latifolia. It was significantly down-regulated during the initial swolling stem of normal Z. latifolia (P<0.05), and contained lower expression level following the subsequent swollen and development (P<0.05), which was similar to the reported gene of Zl-PIN1b in Z. latifolia. Further analysis showed that the Zl-CRK5 gene and IAA polar transport-related genes PIN (PIN-formed) of Zl-PIN1b and Zl-PIN1c showed a coordinated expression relationship in the stems of Z. latifolia after treatment with IAA polar transport-inhibitor of N-1-Naphthylphthalamic (NPA). It was speculated that Zl-CRK5 might be involved in the regulation of polar growth of stems by activating the expression of IAA polar transport-related genes Zl-PIN1b and Zl-PIN1c. Further research would help to reveal the regulatory mechanism of stem expansion induced by infection of U. esculenta.

SVP (short vegetative phase) is one of the important genes in the regulation of plant flowering. In this study, the AfSVP gene (GenBank No. AfisC4G02946) of green onion (Allium fistulosum) was cloned. Its CDS was 744 bp and encoded for 247 amino acids. Bioinformatics analysis revealed that the AfSVP protein had a molecular weight of 60.35 kD and was a hydrophobic protein. Sequence alignment across multiple species demonstrated it was high degree of conservation; Following the introduction of the AfSVP gene into Nicotiana benthamiana protoplasts, laser-confocal microscopy was employed to visualize the protein's subcellular localization. The results showed that AfSVP was present in both the cytoplasm and the nucleus. Tissue-specific expression analysis further indicated that the expression level of AfSVP was significantly higher in flowers compared to other tissues (P<0.05). The analysis of high, low temperature and drought stress response showed that the expression of AfSVP reached the highest at 12 h under high temperature stress, at 72 h under low temperature stress and at 12 h under drought stress (P<0.05). This study offers a reference for further research on how plants adapt to environmental changes.

Anthocyanin is one of the key substances determining the color of lily (Lilium brownii var. viridulum) flowers, and its biosynthesis is susceptible to various environmental factors. SPA1 (suppressor of phyA-105) transcription factor plays a crucial role in regulating plant light signal transduction. To further understand the role of LhSPA1 in the regulation of anthocyanin induction in oriental hybrid lily (Lilium oriental hybrid), this study utilized 'Sunny Martinique' cultivar as material. The differences of anthocyanin content in lily buds treated with different light treatments were compared, and ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) was used to detect the perianth slices of flower buds in the complete color development period. Combined with the previous transcriptome data, LhSPA1 was cloned and then subjected to bioinformatics analysis; and the expression levels of genes related to anthocyanin biosynthesis under different light conditions were analyzed using qRT-PCR. The results showed that the main color components in the tepals of 'Sunny Martinique' were cyanidin and pelargonidin. Shading treatment reduced the accumulation of anthocyanin in lily buds. The results of gene expression analysis showed that the expression of anthocyanin biosynthesis-related genes was down-regulated under shading treatment, but the expression of LhSPA1 was significantly higher than that under normal light conditions. The coding sequence length of LhSPA1 (GenBank No. PQ499086) was 2 931 bp, encoding 976 amino acids. Phylogenetic tree analysis showed that LhSPA1 protein was closely related to ZmSPA1 protein in monocotyledonous plant. Sequence analysis revealed that LhSPA1 protein shared conserved domains with SPA1 protein from other species. Subcellular localization results revealed that the fluorescence signal of LhSPA1 protein was located in the nucleus of Nicotiana tabacum. Transient overexpression of LhSPA1 and constitutive photomorphogenic 1 (LhCOP1) in lily petals resulted in light lily flower color. In conclusion, it was speculated that LhSPA1 played an important role in light-induced anthocyanin biosynthesis in lily. This study lays a foundation for analyzing the molecular mechanism of light signal regulating lily anthocyanin biosynthesis pathway.

SQUAMOSA promoter binding protein-like (SPL) is a transcription factor that plays a crucial role in plant growth and development, as well as stress response. In this study, the DoSPL gene family was identified based on the whole genome of Dendrobium officinale through the bioinformatics approach. Their chromosome position, composition of promoter elements, gene structure, phylogeny, and subcellular localization were analyzed. Meanwhile, their expression patterns in different tissues and under stresses were detected using real-time quantitative reverse transcription PCR (qRT-PCR). Results showed that D. officinale harbored a total of 16 SPL family members, all contained SBP zinc finger structure, and divided into 7 subfamilies. Majority of them were hydrophilic proteins, located in the nucleus, anchored on 10 chromosomes, and possessed 3 tandem repeat gene pairs. Many regulatory elements associated with growth and development, and stress response were presented in the upstream promoter regions of DoSPLs. Multi-species phylogenetic analysis revealed that D. officinale was phylogenetically close to rice (Oryza sativa) SPL proteins. Gene expression analysis showed that DoSPLs were significantly and differentially expressed in different tissues of D. officinale, and exhibited high expression in flowers. Most DoSPLs genes were evidently upregulated or downregulated under the induction of low temperature, methyl jasmonate, or salt stress. Specially, DoSPL12 was significantly upregulated under the above stresses, and might play a crucial role in stress responses. This study provides fundamental data for further exploration the function of the DoSPL gene family.

The root of Sophora flavescens is rich in flavonoid secondary metabolites. Chalcone synthase (CHS) is the first rate-limiting enzyme in the flavonoid biosynthesis pathway, and its characteristics and activity directly affect the biosynthesis and accumulation of secondary metabolites. To analyze the phylogenetic relationship of CHS genes in S. flavescens and their expression profiles, and explore their roles in flavonoid biosynthesis, this study identified 21 CHS family members in S. flavescens using bioinformatics methods. Gene duplication contributed greatly to this family expansion, and the duplicate SfCHS genes underwent strong purifying selection. Then, these CHS members were subdivided into 3 groups (GroupⅠ,ⅡandⅢ) based on phylogenetic analysis. CHS family were not conserved during the evolution, but these members belonging to the same group were conserved in gene structure and motif distribution. The SfCHS04 promoter region contained the highest number of cis-elements for stress resistance. Moreover, based on the expression level of SfCHS genes among different tissues (root, stem, leaf, and flower), majority genes were specifically expressed in flowers. Based on the co-expression network analysis for transcriptome and metabolome data of the pod development, SfCHS04 and SfCHS20 were located in the center of the network, and the main regulatory transcription factors included members of ERF, MYB, WRKY, and NAC families, which played important roles in the development of S. flavescens pods and the accumulation of flavonoids. Finally, the quantitative of SfCHS genes using qPCR technology, which revealed that distinct SfCHS genes showed significant variations in expression levels during root development and among multiple tissues at different growth stages. In summary, the results not only clarify the phylogeny and expression profile of SfCHS genes, but also provide reference value for understanding the molecular mechanism of flavonoid biosynthesis and improving the medicinal value of S. flavescens through molecular breeding methods.

Nuclear receptor subfamily 4 group A member 1 (NR4A1) is widely expressed in tissues of sheep (Ovis aries) and plays multiple biological functions, participating in the regulation of muscle and bone development. This study aimed to investigate the sequence characteristics and polymorphisms in the 3' untranslated regions (3'UTR) of NR4A1 and its correlation with early growth traits in Hu sheep. The 3'UTR sequence of NR4A1 was obtained by PCR, rapid amplification of cDNA ends (RACE) and Sanger sequencing, and analyzed by bioinformatics method. The mutation sites in NR4A1 3'UTR were screened by Pooled-DNA sequencing, and genotyped by DNA sequencing. Then, the polymorphism of the sequence and its correlation with early growth traits was analyzed. Results showed that NR4A1 3'UTR sequence of Hu sheep with a length of 556 bp was obtained. Two mutations, g.26473 C>T and g.26656 G>A, were found in the 3'UTR of NR4A1 of Hu sheep. The 2 loci were completely linked with 3 genotypes, named CCGG, CTAG and TTAA, and CCGG was the dominant genotype (88.4%). Association analysis showed that there was no significant difference in birth weight among the 3 genotypes (P>0.05), while weaning weight and average daily gain from birth to weaning with CTAG genotype were significantly higher than those with CCGG genotype in Hu sheep (P<0.05). In conclusion, the 3'UTR of NR4A1 of Hu sheep has 2 completely linked mutation sites with 3 genotypes CCGG, CTAG and TTAA, and Hu sheep with CTAG genotype in the 3'UTR of NR4A1 has advantages in early growth, which is expected to become a potential marker for the molecular breeding of sheep.

Growth factor receptor-bound protein 2 (GRB2)-associated binding protein 1 (GAB1) is an adaptor protein that plays a role in the intracellular signaling cascade triggered by activated receptor type kinases. It plays an important role in maintaining the functional integrity of osteoblasts. The aim of this study was to analyze the association between single nucleotide polymorphisms (SNP) of GAB1 gene and body size traits in Hu sheep (Ovis aries). A total of 506 healthy Hu sheep with clear pedigree, accurate experimental data records and physical fitness were selected as the experimental subjects. Three genotypes (CC, TC and TT) of g.14514496 T>C polymorphism in exon 5 of GAB1 gene were detected by PCR amplification and AQP^TM genotyping. The general linear model was used to analyze the association between body size traits and genotypes of sheep GAB1 gene. qPCR was used to detect the expression level of GAB1 gene in different tissues of Hu sheep. The result showed that sheep GAB1 gene was widely expressed in various tissues of Hu sheep, and its expression level was the highest in the rumen. The results of association analysis showed that this polymorphic locus was significantly associated with body height, body length and chest circumference of Hu sheep (P<0.05). For individuals carrying CC genotype, the body height of 140, 160 and 180 d of age, and the body length 160 and 180 d of age, and the chest circumference at 120 and 160 d of age were significantly higher than those of individuals carrying TT genotype (P<0.05), which suggested that GAB1 g.14514496 T>C might affect the body size traits of Hu sheep. Therefore, this locus can be used as a potential molecular marker. This study provides theoretical support for breeding studies of Hu sheep.

Phytosterol esters (PE) are active substances formed by esterification of phytosterols with fatty acids. In order to investigate the effects of PE on skeletal muscle development in mice (Mus musculus), 3-week-old mice were randomly divided into control and experimental groups (i.e., the 0.3% PE group), and the effects of PE on muscle development were detected by feeding test, body composition, grip strength and weight lifting test, changes in myofibre area and type as well as myogenic factors were detected by immunofluorescence and qPCR, and finally, the interleukin content and its receptor downstream signaling pathway were analysed by ELISA and Western blot. The results showed that, 1) Dietary supplementation with 0.3% PE significantly increased the average daily intake of mice (P<0.05), and the gastrocnemius index was significantly up-regulated (P<0.05), while the subcutaneous fat index showed a down-regulation trend. 2) Dietary supplementation with 0.3% PE significantly improved the grip strength and weight lifting scores of mice (P<0.05), while there was no significant change in the running time and the distance of the run. 3) Dietary supplementation with 0.3% PE significantly increased the mean muscle fibre cross-sectional area of gastrocnemius and soleus muscles (P<0.05), significantly up-regulated myogenic factor 5 (Myf5) mRNA levels (P<0.05), and there was a tendency for up-regulation in the levels of myogenic differentiation (MyoD) and myogenin proteins as well. (4) Dietary supplementation of 0.3% PE significantly increased serum levels of interleukin-11 (IL-11)(P<0.05), Janus kinase 2 (JAK2) phosphorylation level had a tendency to increase, and signal transducers and activators of transcription 3 (STAT3) phosphorylation level was significantly increased (P<0.05). In conclusion, dietary addition of phytosterol esters could promote skeletal muscle development, increase muscle fibre diameter and enhance muscle function, which might be related to IL-11 and its downstream JAK2/STAT3 signaling pathway. This study provides a theoretical basis for phytosterol esters to promote muscle development in production.

In fish, muscle weight is a key trait reflecting growth performance. The myogenic regulatory factors (MRFs) family of genes plays an important role in muscle generation and development. This study aims to investigate the correlation between the polymorphism of MRFs family genes and the growth traits of adult Nile tilapia (Oreochromis niloticus). PCR amplification and multiple sequence alignment were performed on the coding regions of myogenic factor 5 (myf5), myf6, myogenic determination gene 1 (myod1), myod2, and myogenin (myog). A total of 26 SNPs were screened from the 5 MRFs family genes, including 7 SNPs in myf5, 8 SNPs in myf6, 6 SNPs in myod1, 2 SNPs in myod2, and 3 SNPs in myog. The correlation analysis between the SNPs and growth traits showed that the N7 (C2543T) site of myf5 was significantly related to body weight (P<0.05), and the N9 (A11440G) site of myf6 was significantly related to body weight, total length, body length, and head length (P<0.05). No other SNP significantly associated with growth traits were found in other gene. Additionally, haplotype analysis identified 3 haplotypes in myf5 significantly associated with body thickness (P<0.05) and 4 haplotypes in myf6 significantly associated with growth traits (P<0.05), while no haplotypes significantly associated with growth traits were found in the other genes. These results expand the evaluation of molecular marker applicability and identify 2 candidate markers for molecular-assisted breeding in adult Nile tilapia, providing a theoretical basis for Nile tilapia breeding and quality improvement.

Liver-expressed antimicrobial peptide 2 (LEAP-2) is a cationic antimicrobial peptide that plays a crucial role in the innate immunity of animals. The cDNA sequences of TmLEAP-2A and TmLEAP-2C were identified through transcriptome sequencing in black scraper (Thamnaconus modestus) in this study. Sequence analysis indicated that TmLEAP-2A and TmLEAP-2C encoded 103 and 79 amino acids, respectively, with molecular weights of 11.18 and 9.14 kD. TmLEAP-2A and TmLEAP-2C exhibited the highest amino acid homology with LEAP-2A and LEAP-2C from the large yellow croaker (Larimichthys crocea), at 72.8% and 62.5% respectively. Phylogenetic tree analysis showed that TmLEAP-2A and TmLEAP-2C had the closest evolutionary correlation with LEAP-2A and LEAP-2C in large yellow croaker respectively. qPCR analysis showed that both TmLEAP-2A and TmLEAP-2C were predominately expressed in the liver. Further, their expression levels were significantly induced by infection with Vibrio alginolyticus in liver of black scraper. TmLEAP-2A mature peptide (TmLEAP-2AP46) only exhibited inhibitory activity against V. vulnificus, while TmLEAP-2C mature peptide (TmLEAP-2CP39) exhibited inhibitory activity against both V. alginolyticus and V. vulnificus. Further research demonstrated that TmLEAP-2CP39 exhibited a hydrolytic effect on the genome of V. alginolyticus. Following incubation with TmLEAP-2CP39, V. alginolyticus showed increased uptake of propidium iodide, indicating a significant reduction in the proportion of viable cells. Scanning electron microscopy (SEM) observations revealed that treatment with TmLEAP-2CP39 resulted in altered morphology of V. alginolyticus, characterized by surface wrinkles and the presence of depressions or holes on most bacterial cells. Additionally, transmission electron microscopy (TEM) analysis revealed that, after treatment with TmLEAP-2CP39, the cell membrane of V. alginolyticus exhibited pronounced wrinkling, significant separation from the cytoplasm, rupturing of the membrane, and notable leakage of cellular contents. Overall, TmLEAP-2A and TmLEAP-2C were found to be closely associated with V. alginolyticus infection; however, only TmLEAP-2C demonstrated antibacterial activity against V. alginolyticus in vitro. TmLEAP-2C exerted direct antibacterial effects by disrupting the integrity of the bacterial cell membrane and hydrolyzing genomic DNA (gDNA) within V. alginolyticus cells. These findings not only enhance the understanding of fish antimicrobial peptides but also provide a theoretical foundation for the research and development of fish-derived antimicrobial peptide drugs.

The frequent occurrence of summer heatwaves and visceral white-nodules disease caused by Pseudomonas plecoglossicida has emerged as critical constraints on the sustainable development of small yellow croaker (Larimichthys polyactis) aquaculture. Notably, trypsin, a key enzyme in amino acid metabolism, may play a pivotal role in mediating the physiological responses of small yellow croaker to both thermal stress and bacterial infection. To investigate the response characteristics of the trypsin gene in L. polyactis under high temperature stress and P. plecoglossicida infection, the CDS sequence of the trypsin gene (Lp_try) in L. polyactis was cloned in this study. The sequence of Lp_try spaned 744 bp, encoding 247 amino acids and containing the conserved Tryp_SPc domain. Expression analysis revealed that the Lp_try gene showed pan-tissue expression characteristics, but it showed a specific high expression pattern in the digestive system (intestine, liver) and circulatory system (heart). The expression characteristics of Lp_try gene in the liver of L. polyactis subjected to 32 ℃ high temperature stress and P. plecoglossicida infection were analyzed. The results showed that high temperature stress initially induced a significant increase of Lp_try expression (P<0.05), followed by a swift decreased with prolonged exposure. Additionally, during P. plecoglossicida infection, Lp_try expression exhibited a dynamic trend of decrease-increase-decrease over time, suggesting its regulatory role in the processes of L. polyactis in response to both high temperature stress and bacterial infection. This study provides an important foundation for further understanding of the physiological regulatory mechanisms in fish in response to high temperature stress and pathogen infection.

Ustilago esculenta exclusively infected Zizania latifolia and could enlarge the host stems to form edible Jiaobai. Its pathogenicity was closely related to the formation and phenotypic differentiation of Jiaobai. It was found that synthetic high osmolarity- sensitive protein (Sho1), as an environmental signal receptor upstream of the high osmolarity glycerol (HOG) pathway, plays an important role in the regulation of fungal pathogenicity. In order to study the role of Sho1 in the regulation of the pathogenicity of U. esculenta, the UeSho1 gene (GenBank No. MW768948.1) was cloned and identified. Its open reading frame was 1 137 bp in length with one 126 bp intron, encoding 336 amino acids including a SH3 conserved domain. It was obviously induced expressed during in vitro mycelial growth and infection process of U. esculenta. Further in vitro growth and infection experiments were carried out after knocking out UeSho1 in U. esculenta, and the results showed that the deletion of UeSho1 did not affect the haploid morphology, growth rate, or mating process of U. esculenta in vitro, but it inhibited the filamentous growth of mycelia, slowed down the infection process, and suppressed the expression of b signaling pathway genes related to pathogenicity. Additionally, the absence of UeSho1 did not affect the growth state of U. esculenta in vitro under osmotic, temperature or oxidative stress. This study reveals the critical role of UeSho1 in the regulation of U. esculenta pathogenicity and provides a theoretical basis for further exploration of its role in the interaction between U. esculenta and Zizania latifolia, particularly in inducing host stem swelling.

The ATP-binding cassette transporter protein (ABCT) in cells plays a crucial role in the tolerance of toxic substances, which participates in bacterial growth and the elimination of toxic compounds. Bacillus safensis ST7 has strong manganese oxidation ability, and the ABCT (Gene ID: 46572106) was significantly upregulated under manganese stress, which may be related to the manganese tolerance of Bacillus safensis. In order to analyze the role of ABCT in manganese stress in B. safensis, PCR, qPCR, and SDS-PAGE methods were used to clone the ABCT gene in B. safensis ST7. Overexpression vectors were constructed and transformed into Escherichia coli BL21(DE3). The growth curve, biofilm formation ability, manganese tolerance, and manganese oxidation activity of the recombinant strain were measured to investigate the effect of heterologous expression of the ABCT gene on the manganese tolerance and manganese oxidation activity of E. coli. The results showed that the recombinant strain carried the complete ABCT gene, and the expression level of the recombinant plasmid pET28a-ABCT was significantly upregulated in BL21(DE3) cells (P<0.05). Its growth was not affected, and its biofilm formation ability was enhanced. Under manganese stress, the expression of ABCT gene increased by 1.41 times, with time and concentration dependence. Compared with the control strain, the recombinant strain showed an increase of over 1.5 times in manganese tolerance concentration and manganese oxidation activity. The results suggested that the ABCT gene of B. safensis ST7 was involved in the manganese transport and biofilm formation process in E. coli. Overexpression of this gene could enhance the oxidative activity and tolerance of E. coli cells to manganese (Ⅱ). This study is important for the development and application of manganese-oxidizing bacteria.

Moschus chrysogaster viral hemorrhagic disease (McVHD) is an acute and highly lethal infectious disease caused by Moschus chrysogaster hemorrhagic disease virus (McHDV) in alpine musk deer. Currently, there are no preventive or therapeutic measures available for this disease. In order to develop a safe and effective vaccine against McVHD, in this study, the full-length gene of McHDV VP60 was amplified by PCR. The VP60 gene was then inserted into the vector pFastBac1, followed by transformation into DH10Bac competent cells containing the Bacmid plasmid. A recombinant bacmid, rBacmid-McHDV-VP60, carrying the VP60 gene, was constructed. Subsequently, Sf9 insect cells were transfected with the recombinant bacmid to obtain the recombinant baculovirus rBac-McHDV-VP60. The expression of the target protein in the recombinant baculovirus rBac-McHDV-VP60 was identified by Western blot and SDS-PAGE analyses. The VP60 protein was identified by transmission electron microscopy and erythrocyte agglutination assay. The results indicated that the bacmid rBacmid-McHDV-VP60 was successfully constructed and the recombinant baculovirus rBac-McHDV-VP60 was obtained. SDS-PAGE and Western blot confirmed that Sf9 cells infected with the recombinant baculovirus produced a soluble McHDV-VP60 protein, approximately 63 kD in size. Transmission electron microscopy showed that the McHDV-VP60 protein self-assembled into virus-like particles (VLPs), which were approximately 35 nm in size, spherical in shape, and smooth on the surface. Erythrocyte agglutination assay showed that the recombinant McHDV-VP60 protein solution had a hemagglutination titer of 1∶2¹⁵ against 1% human (Homo?sapiens) O erythrocytes. In this study, a recombinant baculovirus expressing McHDV-VP60 protein was successfully constructed using an insect cell-baculovirus expression system, and the McHDV-VP60 protein could self-assemble into VLPs. This study provides technical support for the development of a vaccine for McVHD virus-like particles.

The creation and selection of superior new germplasm resistant to herbicides is a highly cost-effective and environmentally friendly method of weed control. CRISPR/Cas gene editing technology, characterized by simplicity, efficiency, and precision, holds tremendous potential for application in crop breeding. This study summarizes the research progress of CRISPR/Cas gene editing technology in herbicide-resistant crop breeding, with a focus on the types of crop herbicides, the mutant sites of resistance gene targets, and the development of CRISPR/Cas-mediated precision editing technology. It also provides a detailed introduction to the application of gene editing technology in the creation of superior new herbicide-resistant crop germplasm. Finally, by elaborating on the existing challenges and future development directions of CRISPR/Cas gene editing technology, this study provides a theoretical basis and technical support for the creation of new herbicide-resistant crop germplasm.

RNA interference (RNAi) is a conserved mechanism that regulates gene expression in eukaryotes. Within the domains of plants and microbes, this mechanism not only governs intracellular gene expression but also transcends species boundaries, mediating gene silencing across distinct biological kingdoms. This phenomenon, termed Cross-kingdom RNAi (ckRNAi), enables RNAi signals to operate bidirectionally between organisms from different taxonomic groups. Notably, ckRNAi is widely observed in plant-fungal systems, where it reciprocally regulates growth and behaviors of both interacting partners. This review summarizes recent advances in ckRNAi research within plant-microbe interactions and explores its application scenarios, aiming to provide reference for developing ckRNAi-based technologies or products to combat plant diseases.

Muscovy duck-origin goose parvovirus (MDGPV) is a novel waterfowl parvovirus generated by genetic recombination of Classical Goose parvovirus (cGPV) and Muscovy Duck parvovirus (MDPV). It is one of the main pathogens causing severe diarrhea and exudative enteritis in young Muscovy ducks (Cairina moschata), leading to a decline in their immunity and an increase in morbidity and mortality. The study aimed to establish a three-primer PCR method for simultaneous detection and differentiation of MDGPV and MDPV and monitor the current epidemic situation of the 2 viruses. Based on the recombination characteristics of the viral capsid protein (VP1) gene of MDGPV and MDPV published in GenBank, 3 differential primers were designed and synthesized using the Oligo 7.0 software. Two primers were combined to form 2 sets of PCR reaction systems for amplification, with the target fragment sizes of 493 bp (MDGPV) and 827 bp (MDPV). Standard positive recombinant plasmids of MDGPV and MDPV were constructed as reference materials for the amplification test. The results showed that this study successfully established a three-primer duplex PCR discrimination method capable of simultaneously detecting MDGPV and MDPV. The three-primer duplex PCR method had good repeatability, high specificity and sensitivity, with the minimum detection limits for MDGPV and MDPV being 99.6 and 96.1 copies/μL of total DNA, respectively. The detection results for 102 clinical samples suspected of being infected with waterfowl Parvovirues were as follows: the positive detection rate of MDGPV was 31.37% (32/102), the positive detection rate of MDPV was 7.84% (8/102), the positive detection rate of mixed infection was 1.96% (2/102), which were consistent with the results of SYBR Green Ⅰ-based PCR detection and virus isolation for diagnosing waterfowl Parvovirus infections. In summary, the three-primer PCR diagnostic method established in this study has high specificity, good sensitivity, and is a valuable addition to the detection methods for avian Parvovirus diseases. It is helpful for the clinical diagnosis and epidemiological surveillance of MDGPV and MDPV.