In recent years, numerous genes associated with wheat (Triticum aestivum) grain size and dormancy have been identified through studies on the genetic basis controlling these traits. Based on abscisic acid (ABA) induced wheat plasma membrane 19 (AWPM19) protein family member TaPM19-B1 (TraesCS5B02G566400), this study cloned the full-length TaPM19-B1 gene (543 bp) from the cDNA of the 'Chinese Spring' wheat variety, which encoded a protein of 180 amino acids. Subcellular localization prediction of wheat protoplasts indicated that the TaPM19-B1 protein was localized to the cell membrane. Analysis of the cis-acting elements in the TaPM19-B1 promoter region revealed that the presence of multiple regulatory elements associated with seed development, stress response, and phytohormone signaling. RNA-Seq transcriptome data of wheat and qPCR validation demonstrated that TaPM19-B1 was scarcely expressed in the wheat root, stem, leaf, and spike, but was significantly expressed during the later stages of grain development, with the highest expression levels observed at 10 and 25 days after anthesis (DAA). Furthermore, the expression of TaPM19-B1 was significantly upregulated under drought, salt, heat, and ABA stress conditions. Additionally, an analysis of TaPM19-B1 gene expression at different stages of grain development in 2 large-grain varieties (1000-grain weight>50 g) and 2 small-grain varieties (1000-grain weight<21 g) revealed that TaPM19-B1 expression was significantly higher in the large-grain varieties (P<0.05). This study provides theoretical insights into the molecular regulatory mechanisms of TaPM19-B1 in wheat growth, development, and response to environmental stresses.

Stigma exsertion of millet sterile lines is a key trait affecting the efficiency of hybrid seed production of foxtail millet (Setarial italica) hybrids. The longer stigma and more obvious exsertion, which is more valuable for the seed production yield of hybrid seeds. In this study, stigma exertion length was measured among sterile lines 'Jinfen line 21A', 'Jinfen line 92A', '8116A', 'EMS-1A' and 'Fen 33A', while 'Jinfen line 92A' was tested with longest stigma exsertion length (0.66 mm), 'Jinfen line 21A' had the shortest stigma exsertion (0.06 mm). A total of 12 comparable foxtail millet stigma exsertion genes were identified using homologous alignment analysis, encoding 461~1 272 amino acid residues with molecular weight ranging from 50.173 90~138.045 10 kD, isoelectric point ranging from 5.05 to 9.13, and instability coefficient ranging from 30.80 to 50.92. Phylogenetic tree analysis demonstrated that all protein could be divided into 4 groups, and members of the groups had similar motifs and conservative domains. The Seita.2G100300 was homologous to the rice stigma exsertion gene by phylogenetic analysis. Further analysis of Seita.2G100300 at the transcriptional level showed that the expression of the gene was significantly lower than the control at heading stage and significantly increased at flowering stage (P<0.05). Correlation analysis showed a significant positive correlation between the expression level of Seita.2G100300 with the stigma exsertion length in the sterile line 'Jinfen line 92A' and '8116A' (P<0.05). This study integrated the stigma exsertion length phenotype with the expression analysis of related genes, offering novel insights into the mechanism underlying stigma exsertion in male-sterile lines of foxtail millet.

Plant dehydrins (dehydrins, DHN) are a class of specific proteins expressed when plants are subjected to adversities such as drought and salinity. To investigate the function of potato (Solanum tuberosum) StDHN-2 gene in plant drought tolerance, in this study, the CDS of StDHN-2 gene was cloned from potato cultivar 'Atlantic' plantlets, and the fusion protein expression vector was constructed by homologous recombination, the subcellular localization results of StDHN-2 protein showed that the protein might be located in the nucleus. Based on the bioinformatics analysis of StDHN-2 gene, it was found that the genome had a total length of 931 bp, including a coding region sequence of 474 bp, which encoded 157 amino acids; The molecular weight of StDHN-2 protein was 16 659.06 D, and the theoretical isoelectric point was 7.23. The analysis showed that StDHN-2 protein was a hydrophilic protein without transmembrane domain. The secondary structure of the StDHN-2 protein was mainly the random coil, and there was no β-turn structure. By constructing a phylogenetic tree, it was found that StDHN-2 protein had the closest evolutionary relationship with Solanum verrucosum; The cis-acting element analysis showed that the promoter region of StDHN-2 gene contained ABRE, DRE, P-box, LTR and other elements related to abiotic stress response; The expression patterns of StDHN-2 gene were analyzed under PEG, NaCl, and abscisic acid (ABA) stress treatments by using qRT-PCR analysis, it was found that the gene was up-regulated under all 3 kinds of stress treatments. The results of the study preliminarily indicated that the StDHN-2 gene could be a good candidate gene for drought tolerance in potato and involved in the biological process of drought tolerance in plants. This study provides basic information for analyzing the drought response mechanism of potato StDHN-2 gene.

Terpenoids are the main substances for the medicinal function of jujube (Ziziphus jujuba). 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) plays an important role in the biosynthesis of plant terpenoids. In order to further study the function of jujube HMGR genes, this study used HMM to identify the jujube HMGR gene family. Bioinformatics analysis software was used to analyze the physicochemical characteristics, gene structure, and evolutionary relationship of ZjHMGR genes. qRT-PCR was used to analyze the expression pattern of HMGR genes. The results showed that there were 21 HMGR gene members in the jujube genome, which were named ZjHMGR1~ZjHMGR21 according to their positions on the chromosome. The 21 HMGR genes were located on 10 chromosomes and 2 unplaced genomic scaffolds. The open reading frame (ORF) length of the ZjHMGR genes was 271 to 3 425 bp, encoding 88 to 1 122 amino acids, and the molecular weight ranged from 9 827.05 to 124 950.93 kD. The phylogenetic analysis of jujube and arabidopsis (Arabidopsis thaliana), apple (Malus pumila), pear (Pyrus bretschneideri), tobacco (Nicotiana tabacum) and watermelon (Citrullus lanatus) showed that the ZjHMGR1 was a separate group, while the ZjHMGR11 and ZjHMGR12, ZjHMGR5 and ZjHMGR10 were classified as separate class. And these 5 gene family members had collinear relationships with arabidopsis, apple, pear and watermelon. There were hormone-related cis-elements in the 2.0 kb promoter region of ZjHMGR gene, among which 11 candidate genes had gibberellin, methyl jasmonate and abscisic acid response cis-elements, 8 candidate genes had auxin response cis-elements, and 12 candidate genes have salicylic acid response cis-elements. Compared with young fruits, the expression level of ZjHMGR gene in mature fruits was significantly increased. The expression levels of the ZjHMGR genes in 'Linhuang No.1' and 'Muzao' fruits were significantly negatively correlated with the contents of chlorophyll a, chlorophyll b, total chlorophyll and anthocyanins, but positively correlated with the contents of carotenoids.ZjHMGR played an important role in fruit ripening. This study provides reference for genetic improvement of the synthesis of jujube terpenoids.

Aldehyde dehydrogenases (ALDHs) play an important role in aldehyde stabilization under abiotic stress in plants.The members of ALDH superfamily in tea plant (Camellia sinensis) were identified by bioinformatics methods, and the biological information such as physicochemical properties, chromosome localization, gene structure, promoter cis-acting elements and phylogenetic relationship of the encoded proteins were analyzed. The expression patterns of the CsALDH superfamily members under drought, salt, and cold stresses ware analyzed by qRT-PCR. The results showed that a total of 27 ALDH superfamily members were identified in the C. sinensis genome, and phylogenetic analyses classified them into 10 families. Family members exhibited comparable motif composition and gene structure. The co-linearity analysis indicated that CsALDHs genes underwent gene duplication events in the C. sinensis genome, which experienced purifying selective pressures.The qRT-PCR results showed that 8, 10 and 9 candidate CsALDHs genes were induced by PEG, NaCl and low temperature, respectively. In particular, CsALDH2B4 and CsALDH3H2 were significantly up-regulated. This study provides a theoretical foundation for further research on the functions of ALDH genes in tea plants.

Prunus mume, a traditional famous flower in China, holds significant ornamental value. GATA transcription factors play a crucial role in the regulation of plant growth and development as well as in the response to abiotic stress. In this study, a genome-wide identification of the GATA gene family in P. mume was conducted, and their expression patterns following cold treatment were systematically analyzed, with the functional characterization of PmGATA12 being comprehensively investigated. The results indicated that a total of 20 PmGATA genes were identified through whole-genome analysis, distribute across 7 chromosomes. Phylogenetic analysis revealed that PmGATA proteins could be divided into 3 subfamilies. Members of the same subfamily have similar conserved motifs and gene structures. Furthermore, by integrating transcriptome and qRT-PCR expression analysis, 4 differentially expressed PmGATA genes (PmGATA6, PmGATA11, PmGATA12, and PmGATA14) were identified. The genetic transformation of Arabidopsis thaliana with PmGATA12 significantly enhanced the cold tolerance of the transgenic lines. This study provides valuable gene resources for the cold-resistance breeding of P. mume and offers new insights into the molecular mechanisms underlying its cold tolerance.

Pleomorphic adenoma gene 1 (PLAG1) plays a crucial role in regulating the growth and development of livestock and poultry. Our previous study identified that PLAG1 was associated with backfat thickness (BF) in pigs by genome-wide association study (GWAS), suggesting a potential regulatory effect on adipocytes. To further investigate the relationship between PLAG1 gene and adipocytes, 3T3-L1 cells was used as a research model and PLAG1 conservation was analyzed. The effects of PLAG1 gene overexpression on cell proliferation and differentiation were examined using qRT-PCR, CCK-8 assays, flow cytometry, and Oil Red O staining. Results revealed high conservation of the PLAG1 gene, with sequence similarity among different species ranging from 80% to 92%. qRT-PCR analysis showed that, compared with the control group, the expression of proliferation-related genes in 3T3-L1 cells was extremely significantly increased following PLAG1 overexpression (P<0.01). CCK-8 assays showed that PLAG1 overexpression extremely significantly enhanced cell viability (P<0.01). Flow cytometry demonstrated a significant increase in the number of cells in the S phase following PLAG1 overexpression (P<0.05). Furthermore, Oil Red O staining indicated that the expression of adipogenic differentiation-related genes was significantly decreased after PLAG1 overexpression (P<0.05), accompanied by a extremely significant reduction in lipid droplet formation (P<0.01). In summary, PLAG1 overexpression promoted the proliferation of 3T3-L1 cells while inhibiting their differentiation, indicating that PLAG1 gene might play a regulatory role in the proliferation and differentiation of adipocytes. This study provides a theoretical foundation for further investigation of the molecular mechanisms through which the PLAG1 gene regulates fat deposition in pigs.

Nitric oxide synthase (NOS), a crucial enzyme responsible for production of endogenous nitric oxide (NO) in body, has multiple effects such as anti-inflammatory or pro-inflammatory. To investigate the expression patterns and potential functions of NOS in dairy cows (Bos taurus) with healthy or clinical mastitis (CM), the mammary gland tissues were collected from dairy cows with healthy (control, Con) and CM in this study. The distribution and expression patterns of NOS were in the mammary gland tissues of the Con and CM groups using hematoxylin-eosin (HE) staining, immunohistochemistry (IHC) and real-time fluorescence quantitative PCR (qRT-PCR). The potential role of NOS was predicted using different bioinformatics methods. HE displayed that the mammary acini were collapsed and couple with infiltration inflammatory cell in the CM group. IHC indicated that the positive expression of neuronal NOS (nNOS), inducible NOS (iNOS), and endothelial NOS (eNOS) were presented in the cytoplasm of mammary epithelial cells. Compared to the Con group, the nNOS and eNOS mRNA expression levels were extremely significantly downregulated (P＜0.01), and iNOS mRNA expression levels was extremely significantly upregulated (P＜0.01) in mammary gland tissues of the CM groups. Bioinformatics analysis revealed that NOS might participate in CM occurrence in regulation of NO and amino acid synthesis and metabolism, ion transport. In conclusion, the above results provide a reference for revealing the function and regulatory mechanism of NOS in cow CM.

Maiwa yak (Bos grunniens) is a local superior seed of Qinghai-Tibet Plateau yak. Previous studies have shown that the coat color of Maiwa yak is related to its production performance and can be used as the basis for breeding. Previous research has shown that the grey coat trait in Maiwa yak is associated with the expression level of the syntaxin-17 (STX17) gene. The STX17 protein is involved in the fusion process of lysosomes and autophagosomes, playing an important role in the synthesis, transport of melanin, and the development of melanoma. To investigate the correlation between the STX17 gene and coat color formation in Maiwa yak, 3 healthy adult male black and grey Maiwa yaks aged 3 years were selected in this study, and their heart, spleen, lung, kidney and skin tissues were selected as experimental materials. The CDS sequence of the STX17 gene were cloned by PCR, and bioinformatics analysis was carried out by using on line software, the expression of the STX17 gene was detected in different tissues and coat populations of Maiwa yak by qRT-PCR. The results showed that the CDS region of the STX17 gene in Maiwa yak was 909 bp, encodimg protein contained 302 amino acids with a theoretical value of isoelectric point (pI) of 5.82, which was an acidic protein, the STX17 protein did not have a signal peptide sequence, and there was 1 target membrane-associated soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) domain and 2 tandem transmembrane domains; Subcellular localisation predicted that the STX17 protein mainly existed in the cytoplasm. The nucleotide sequence comparison showed that the STX17 gene in Maiwa yak was most closely related to Bos taurus, followed by Bubalis bubalus. The STX17 protein was closely related to autophagy-related protein 14 (ATG14), synaptosome associated protein of 29 (SNAP29), vesicle associated membrane protein 7 (VAMP7), vesicle associated membrane protein 8 (VAMP8), synaptobrevin homolog (YKT6) and other proteins. The qRT-PCR results showed that the STX17 gene had the highest expression in skin tissues, and the expression in the skin of grey Maiwa yak was higher than black Maiwa yak, it was hypothesized that the STX17 gene was related to the formation of grey and black coat color in Maiwa yak. This study provides theoretical basis for the subsequent studies on the regulatory mechanism of coat color in Maiwa yak.

Connective tissue growth factor (CTGF), CREB regulated transcription coactivator 3 (CRTC3), and transmembrane protein 18 (TMEM18) play crucial roles in the growth and development of mammals. To identify genetic markers and gene polymorphisms associated with growth traits in the populations of Datong yak (Bos grunniens) and Ashidan yak, Sanger sequencing technology was used to detect and genotype 5 SNPs in the CTGF, CRTC3 and TMEM18 genes in Datong yak and Ashidan yak populations, and the association between the genotypes of polymorphic sites and growth traits were analyzed. The results showed that there were 2 genotypes at the G2511A loci of the CTGF gene in Datong yak and Ashidan yak populations, 3 genotypes at the C86041T, G86075A loci of the CRTC3 gene, and 3 genotypes at the C1267T, C4447T loci of the TMEM18 gene. Among them, both the C86041T loci of the CRTC3 gene and the C4447T loci of the TMEM18 gene were moderately polymorphic (0.25＜PIC＜0.50).However, the Chi-square test revealed that none of the 5 SNPs reached Hardy-Weinberg equilibrium in the 2 yak populations, suggesting that the 2 populations have undergone long-term artificial selection and breeding. The association analysis of growth traits showed that the G2511A loci of the CTGF gene was significantly correlated with body height (P＜0.05); 2 SNPs of the CRTC3 gene were significantly associated with body weight (P＜0.05), and the G86075A loci was significantly correlated with both body height and chest circumference (P＜0.05); 2 SNPs of the TMEM18 gene were significantly associated with body weight (P＜0.05). Therefore, these growth trait-related SNP loci were correlated with the growth traits of Datong yaks and Ashidan yaks, and could be used as candidate genes for molecular marker-assisted selection in future breeding improvement. This study provides potential genetic markers for yak breeding improvement.

Sperm cells are highly susceptible to damage during semen cryopreservation, in which excessive free radicals cause oxidative stress that negatively affects the cellular system by increasing the production of reactive oxygen species. Therefore, the use of antioxidants in semen cryopreservation is crucial. Rosmarinic acid (RA) is a natural polyphenolic antioxidant with strong antioxidant activity. In this study, 0, 20, 40, 80 and 120 μmol/L RA was added to the semen cryopreservation diluent of dairy goats (Capra hircus) to test the sperm quality and sperm antioxidant capacity, and sperm plasma metabolic profiles of the spermatozoa of dairy goats were analyzed by LC-MS non-targeted metabolomics after freezing. The results showed that RA improved the quality of cryopreserved dairy goat spermatozoa and sperm antioxidant capacity, and 309 differential metabolites were detected between the control group and RA-treated group. Moreover, RA regulated the energy metabolism of spermatozoa through AMP-activated protein kinase (AMPK) signaling pathway, and activated or inhibited apoptosis-related proteins, such as B-cell lymphoma-2 (Bcl-2), through PI3K-Akt signaling pathway. The present study demonstrated that the addition of 40 μmol/L RA could improve the effect of semen cryopreservation in dairy goats. This study provides a new idea and theoretical basis for the optimization of semen cryopreservation technology in dairy goats.

Natural resistance-associated macrophage protein 1 (NRAMP1) has been demonstrated to be linked with resistance to various intracellular mycobacterial diseases, including tuberculosis and brucellosis. This study aimed to develop goats (Capra hircus) with NRAMP1 gene overexpression and to conduct a comparative analysis of their blood physiology, biochemistry, and growth and development indices, which provides a research model and platform for establishing a brucellosis resistance model and for the genetic breeding of transgenic animals. Based on the lentiviral vector constructed in our laboratory for goat NRAMP1 gene, NRAMP1 gene overexpression goats were produced using the techniques of supernumerary ovulation and prokaryotic embryo microinjection, and subsequently monitored and evaluated the effects of this overexpression on physiological and biochemical changes in the blood, as well as on the growth and development of the goats. The results showed that the recovery rate of prokaryotic embryos was significantly higher than that observed at a dose of 270 IU for supernumerary ovulation when administering 210 IU follicle-stimulating hormone (FSH) in adult Shaanbei white cashmere goats (P<0.05). A total of 15 goats were obtained through the transplantation of prokaryotic embryos following microinjection, 3 of which were identified as positive gene overexpression goats via PCR, and then qRT-PCR and Western blot analyses revealed that NRAMP1 expression was significantly elevated at both the mRNA and protein levels in the testicular tissues of the NRAMP1 overexpression goats (P<0.05). The results of blood physiological and biochemical measurements indicated that there were no significant differences in blood physiological and biochemical indices between NRAMP1 overexpressing goats and wild-type goats (P>0.05). Similarly, the results of growth performance measurements revealed that the differences in growth traits, including newborn weight, body weight from 1~6 months of age, and body height and length from 3~6 months of age, between NRAMP1 gene overexpressing goats and wild-type goats were also not significant (P>0.05). In this study, goat model that overexpressing of the NRAMP1 gene was successfully developed, and notably, this overexpression did not adversely affect the health of the goats. This study provides a scientific basis for the breeding of brucellosis resistant goats and the study of their anti-resistance mechanism.

There is a significant difference in the growth of male and female giant salamanders (Andrias davidianus), and it has been confirmed that the serine/arginine-rich splicing factor 3 (srsf3) gene is involved in the selective splicing of nuclear receptor subfamily 5 group A member 2 (Nr5a2), a gene related to sex differentiation in giant salamanders. To investigate the mechanism of srsf3 gene in sex differentiation of Chinese giant salamander , qPCR technology was used to analyze the changes in expression levels in different tissues, gonads at different developmental stages, and gonads induced by sex hormones; Fluorescence in situ hybridization (FISH) was used to determine the expression and localization of the srsf3 gene in the testes and ovaries of giant salamanders. Based on the analysis of the transcriptome sequencing results of the giant salamander gonads, the full-length of srsf3 gene was 1 478 bp, with an open reading frame of 504 bp, encoding 167 amino acids. Systematic evolutionary analysis showed that the amino acid sequence of the giant salamander SRSF3 was closely related to the Xenopus tropicalis. The qPCR results showed that the expression level of srsf3 gene was highest in the ovaries of giant salamanders, followed by pituitary tissue, slightly lower in muscle and heart tissues, and low expression in liver, spleen, lung, and stomach tissues. The expression of the srsf3 gene in the gonads of giant salamanders at different stages showed that the ovaries were significantly higher than the testes at ages 2~5 (P<0.05); The expression level of srsf3 gene reached its highest in the ovaries at the age of 2, and then decreased year by year in the ovaries. The expression level tended to stabilize in the ovaries at the age of 4-6. The expression level of the srsf3 gene was highest in sex reversed males, followed by sex reversed females; The expression level of srsf3 gene in the sex reversal ovaries was significantly higher than that in the ovaries (P<0.05), and the expression level in the sex reversal testes was significantly higher than that in the testes (P<0.05). The FISH results showed that the srsf3 gene was expressed in both the ovaries and testes of the giant salamander. The above studies indicated that the srsf3 gene was involved in the gonadal development of giant salamanders and plays an important role in the process of sex reversal in giant salamanders. Functional analysis of the srsf3 gene in this study helps to deepen the understanding of the mechanism of sex differentiation in amphibians.

In recent years, due to continuous cropping, the problem of pests and diseases in Atractylodes macrocephala has been severe, resulting in a decrease in yield and a decline in quality. Among them, the A. macrocephala epidemic caused by mold has caused serious losses to A. macrocephala production. As one of the most important biocontrol fungi, Trichoderma spp. has good antagonistic effects on various plant pathogens and can produce various bioactive substances with antagonistic effects. In this study, healthy rhizosphere soil and underground rhizomes of A. macrocephala continuous cropping land were taken as materials, to screen antagonistic Trichoderma strains using the pathogen AMPH-1 of Phytophthora sansomeana as an indicator bacterium. Through the inhibitory effect and antagonistic level of antagonistic, 3 Trichoderma strains with better antibacterial effects, namely TPS-87, TPS-90, and TPS-99, were selected. The antibacterial rates against AMPH-1 were 77.19%, 80.04%, and 78.49%. With morphological and molecular identification, strain TPS-87 was determined as T. afroharzianum, while TPS-90 and TPS-99 were both T. atroviride. When applying the suspension of TPS-87, TPS-90, and TPS-99 with a concentration of 1×10⁶ cfu/mL, and with the inoculation amount of 150 mL, all 3 strains of Trichoderma had a good promoting effect on A. macrocephala seedlings, and the greenhouse control effects on A. macrocephala blight disease were 61.54%, 53.85%, and 50%, respectively. This study provides biocontrol bacteria resources for the green prevention and control measures of A. macrocephala blight disease, and provides an important basis for the further development of biocontrol agents for the prevention and control of Chinese herbal medicine diseases.

Pectate lyases (Pel) play critical roles in pectin degradation and hold significant biotechnological promise across diverse application in food and textile industries. In this study, a novel pectate lyase gene derived from Hu sheep (Ovis aries) rumen microbiota, IDSPL1-20 (GenBank No. PP975428), was cloned and heterologously expressed. The enzyme activity and biochemical characterizations of recombinant rIDSPL1-20 were determined. The results showed that the optimum temperature and pH of rIDSPL1-20 were 50 ℃ and pH 10.0, respectively. The rIDSPL1-20 exhibited poor thermal stability but excellent alkaline-tolerance. The residual activity maintained over 80% after treatment at pH 9.0~11.0 for 1 h. The rIDSPL1-20 was activated by the presence of 0.25~2 mmol/L Ca²⁺ (P<0.05). Multi-substrate specificities determination indicated that the rIDSPL1-20 showed V_max values of (316.25±34.09) and (105.55±3.72) µmol/(min·mg) against polygalacturonic acid (PGA) and 60% esterified pectin (60% DE pectin). The rIDSPL1-20 mainly released unsaturated trigalacturonate (uG3) and unsaturated digalacturonate (uG2) from PGA and 60% DE pectin as the predominant products, indicating that the enzyme probably functions as a typical endo-pectate lyase (EC 4.2.2.2). In addition, the degradation efficiency of Cellic^® CTec3 HS for peanut (Arachis hypogaea)straw was dramatically boosted by rIDSPL1-20 (P<0.05). After synergistic reaction for 2 and 4 h, the total reducing sugars yielded (0.50±0.07) and (0.57±0.14) mg/mL. This study laid the groundwork for the development of novel biomass-degrading enzymes.

Newcastle disease is an acute, febrile and highly contagious disease caused by Newcastle disease virus (NDV), in which genotype Ⅶ is widespread. Hemagglutinin-neuraminidase protein (HN) is a structural protein of viruses and an important immunogenic protein for host cell receptors to recognize and promote cell membrane fusion. Rice (Oryza sativa) cells are a promising economic and effective bioreactor platform for the production of biological drugs. The rice α-amylase 3D system contains an inducer promoter, which has strong activation activity under the condition of sugar starvation and can express foreign proteins efficiently and at a high level. In this study, NDV gene Ⅶ HN sequence was screened from NCBI, HN gene was optimized according to rice codon preference, and the recombinant plant expression vector pCAMBIA1300-HN was constructed. HN gene was transferred into rice callus by Agrobacterium-mediated method, and the positive callus was identified by hygromycin screening and PCR. HN protein expression was induced in the positive callus, and then identified by Western blot. HN protein was purified by SP cationic chromatography and Hiloard 75 pg gel chromatography, and emulsified with Al(OH)₃ and 50V adjuvant at doses of 5 μg and 10 μg, respectively. BALB/c female mice (Mus musculus) aged 6 weeks were immunized, the titer of antibody was detected by hemagglutination inhibition (HI) test and neutralizing test (NT), and the level of interleukin-2 (IL-2) after 2 weeks of immunization was detected. The results showed that the recombinant plant expression vector pCAMBIA1300-HN was successfully constructed, and 85 positive calluses were identified. HN protein was successfully expressed and purified in rice cells. After immunizing mice, the antibody levels of each group except PBS group achieved immune protection (HI≥2⁴), and neutralized antibodies were produced, which had virus neutralization effect. In this study, HN protein of Newcastle disease was expressed in rice cells, and a purification method of HN protein was established. After immunizing animals with HN protein at low dose, the immune effect was good. This study provides a reference for the preparation of Newcastle disease subunit vaccine in the future.

Strigolactones (SLs) is a newly identified class of phytohormones that strongly inhibit the formation of branches/tillers in plants, and plays an important role in the process of plant growth and development and stress resistance. With the further study of SLs synthesis and signal pathway, a series of genes have been identified that can be used to regulate plant type formation. In this review, discovery process, chemical structure, synthetic pathway, metabolic pathway, distribution and transport, signaling pathway, and its function in regulating plant growth and development of SLs were reviewed. At the same time, the development of this field was prospeced, and it was believed that the regulation of plant type, resistance and parasitism by SLs would become the focus of future research. This paper provides ideas and theoretical basis for establishing ideal plant type and breeding new varieties with high yield and stress resistance in the future.

Ganoderma lingzhi is a valuable edible and medicinal fungus, at present, there are more than 200 kinds of secondary metabolites isolated from various tissues of G. lingzhi, among which G. lingzhi triterpenoids and polysaccharides are its main active ingredients, and their pharmacological effects mainly include anti-hypertensive, anti-tumor, anti-aging, The completion of G. lingzhi genome sequencing and the establishment of genetic transformation system have gradually become a model fungus for the study of the synthesis and regulation of secondary metabolites of medicinal fungi, and its molecular biology research involves the identification of G. lingzhi germplasm resources, genetic diversity, variety breeding, functional gene cloning, etc.. In this paper, the research progress of G. lingzhi genomics, functional genes involved in the regulation of G. lingzhi growth and development, metabolite synthesis, abiotic stress and signal transduction were summarized, so as to provide a reference for the further development and application of G. lingzhi and a scientific basis for elucidating the molecular mechanism of G. lingzhi efficacy.

Livestock sex control is a reproductive technology that involves human intervention to produce offspring of the desired sex, including the separation of X and Y sperm before fertilization and the identification of embryos after fertilization. Among these methods, the separation of X and Y sperm before fertilization is the most direct and effective approach. Multiple inherent differences between X and Y sperm in livestock, and establishing simple, cost-effective, and efficient sperm sorting technologies based on these differences is of significant importance for enhancing the economic benefits of the livestock industry. This article reviewed the multidimensional biological differences between X and Y sperm in livestock, including motility, protein composition, DNA content, and membrane charge. It also discussed 3 main categories of sorting techniques: 1) methods based on the motility differences of X and Y sperm, such as the swim-up method, pH method, and Toll-like receptors 7/8 (TLR7/8) activation method; 2) methods based on differences in protein composition, such as the magnetic bead-conjugated antibody method and poly-lactic acid film-conjugated antibody method; 3) methods based on differences in DNA content and membrane charge, such as flow cytometry sorting and electrophoresis. Additionally, this article analyzed and evaluated the advantages and limitations of each method, and provides an outlook on future research directions for sperm sorting technologies in livestock, offers a theoretical basis for the development of new sex control technologies.

The utilization of the CRISPR/Cas9 system for gene editing is an effective method for investigating gene functions and pursuing genetic improvement in melons (Cucumis melo). However, there lacks a method that rapidly detects the target sites of CRISPR/Cas9 vectors prior to genetic transformation. In this study, the melon phytoene desaturase (PDS) gene CmPDS was used as target gene, two sgRNAs were designed and introduced together into the CRISPR/Cas9 vector. Agrobacterium rhizogenes K599 was used to infect cotyledon explants of the thick-skinned melon 'K7-2' and the thin-skinned melon 'LB'. The induced adventitious roots were firstly identified using Bar test strips, and then the target regions were sequenced to see if the regions were edited and clarified the types of mutation. The PCR products sequencing results showed that the 2 target sites were all edited. Further sequencing of the individual PCR products demonstrated an editing efficiency of 100% for target 1 and 76.5% for target 2, with multiple mutation types observed, including insertions and deletions of single or multiple bases, as well as large deletions between 2 target sites. This study successfully established a genetic transformation system for melons that was mediated by the A. rhizogenes, and achieved rapid detection target sites of CRISPR/Cas9 vector, and also validated the effectiveness of using the CRISPR/Cas9 system in melon gene editing. This study provides an important technical support for genetic functional studies and genetic improvements of melons.

Actin associated protein 2/3 complex subunit 3 (ARPC3) is an important component of actin skeleton tissue, which plays an important role in bacterial invasion, inflammation development and apoptosis. To investigate the role of ARPC3 protein in bovine mastitis, in this study, rabbit polyclonal antibody against bovine (Bos taurus) ARPC3 was prepared and its activity was identified. The biological characteristics of bovine ARPC3 (NM 001034271.2) were analyzed by online software. pQE-80L-ARPC3 recombinant plasmid was constructed. The prokaryotic expression system was used to express ARPC3. The concentration of isopropyl-beta-D-thiogalactopyranoside (IPTG) and induction time were optimized. The expressed protein was purified by affinity chromatography, identified by SDS-PAGE and mass spectrometry, and then immunized New Zealand white rabbits (Oryctolagus cuniculus) to prepare polyclonal antibody. The titer of polyclonal antibody was detected by ELISA, and the specificity of polyclonal antibody was identified by Western blot. The results showed that the recombinant plasmid pQE-80L-ARPC3 was successfully constructed, and the recombinant protein (about 20.5 kD) was expressed in the form of inclusion bodies after IPTG induction . The optimal expression conditions were as follows: IPTG concentration was 1 mmol/L for 8 h. The titer of ARPC3 polyclonal antibody was 1∶128 000. Western blot identification of natural protein showed that ARPC3 polyclonal antiserum could specifically recognize ARPC3 in target protein and mammary tissue. Immunohistochemical (IHC) results showed that the polyclonal antibody could be effectively applied for the detection of bovine mastitis, and the expression of ARPC3 protein was positively correlated with the expression of related apoptotic proteins. In this study, the polyclonal antibody of ARPC3 with good specificity was successfully prepared, which can be used to the establishment and application of a new technology for rapid detection of bovine mastitis.