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Monthly, Started in1993
ISSN 1674-7968  CN 11-3342/S
Administrated by: Ministry of Education of People's Republic of China
Sponsored by: China Agricultural University and Chinese Society of Agricultural
                          Biotechnology
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WU Wei-Hua
Associate editor-in-chief:CHEN Hua-Lan   LI Kui   LI Yi   LIN Min   PENG Yu-Fa
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农业生物技术学报
2025 Vol. 33, No. 6  Published: 01 June 2025
 
Articles and Letters
Analysis on the Basic Characteristics of the Effector Pt31812 of Puccinia triticina
WEN Xiao-Lei, LI Jian-Yuan, LI Ming-Yuan, LI Hao, ZHANG Na, YANG Wen-Xiang
2025 Vol. 33 (6): 1181-1191   DOI:10.3969/j.issn.1674-7968.2025.06.001 |  Full text (HTML) (1 KB)  |    PDF  (10919 KB)  ( 7 )
Abstract
Effectors play an important role in the process of pathogen invasion of plants. Understanding their functions is the key to analyzing the pathogenic mechanism of pathogenicity. In order to reveal the pathogenic mechanism of Puccinia triticina (Pt), based on the analysis of the biological information of effector Pt31812, the basic characteristics of the effector Pt31812 were analyzed using the secreting system of saccharase- deficient yeast, qRT-PCR and bacterial type Ⅲ secreting system. The results showed that Pt31812 CDS was 627 bp, encoding 208 amino acid, with 7 serine, 3 threonine and 2 tyrosine potential phosphorylation sites. The N-terminus has a hydrophobic structure, 1~20 amino acids were signal peptides, and had a secretory function, had no transmembrane domain, and did not act on mitochondria. It could inhibit the programmed cell death (PCD) induced by the Pseudomonas syringae pathovar. tomato strain DC3000 in wheat leaves. The gene began to express in the early stage of leaf rust infection, and the expression level was the highest at 96 and 36 h in the compatible and incompatible interactions, respectively. It could inhibit the deposition of callose and accumulation of reactive oxygen species in wheat both 'Thatcher' ('Tc') and 'TcLr1', as well as the expression of pathogenesis-related proteins gene 1 (TaPR1) and TaPR2, interfering with the host defense response. The results provide theoretical basis for further study of the pathogenic mechanism of wheat leaf rust and have guiding significance for durable resistant breeding.
Genome-wide Identification of PR-1 Gene Family in Capsicum annuum and Analysis of Its Expression Pattern in Response to Phytophthora capsici
HE Wei, ZHANG Hui, WANG Jia-Lin, HUANG Yue, WANG Xing-Liang, SHEN Xiao-Yu
2025 Vol. 33 (6): 1192-1205   DOI:10.3969/j.issn.1674-7968.2025.06.002 |  Full text (HTML) (1 KB)  |    PDF  (9695 KB)  ( 3 )
Abstract
Pathogenesis-related protein 1 (PR-1), which are defense proteins in plant-pathogen interactions, play an important role in the resistance and defense of plants against diseases. The present study aimed at genome-wide identification and bioinformatics analyses of PR-1 genes in pepper (Capsicum annuum). The analyses resulted in the identification of 19 novel CaPR-1 genes, each of which produced a protein belonging to the CAP superfamily. Most of the CaPR-1 proteins contained an N-terminal signal peptide. Phylogenetic tree analysis of a total of 54 PR-1 protein sequences from pepper, tomato (Solanum lycopersicum) and Arabidopsis thaliana showed that the PR-1 family proteins were divided into 3 categories, and the pepper PR-1was closely related to tomato PR-1. The structural analysis revealed similar 3D structures of all CaPR-1 proteins except CaPR-1.1 and CaPR-1.12. In the promoter of CaPR-1 gene, hypothetical cis-elements related to defense stress response, plant hormones, light and transcription factor were detected. The GO and KEGG functional annotations predicted their function in the defense stress response, plant hormones and plant- pathogen interactions. Microarray data analysis showed that CaPR-1 not only responded to cold, salinity, drought and heat stresse, but also affected by exogenous hormones such as salicylic acid, methyl jasmonate, ethylene and abscisic acid. CaPR-1.4 and CaPR-1.15 genes were up-regulated under salinity or drought stress, and CaPR-1.17 gene was up-regulated inducted by 4 exogenous hormones. The expression pattern of CaPR-1 transcript was altered under infection with Phytophthora capsici, with up-regulation of alkaline CaPR-1 genes such as CaPR-1.4, CaPR-1.15 and CaPR-1.17. The drastic variation in the transcript levels of CaPR-1.17 gene was further validated through qRT-PCR and it showed a significant up-regulation in infected samples (P<0.05). The result showed that the CaPR-1 family might be important in plant stress resistance. This study provides a reference for in-depth exploration of the mechanism of pepper PR-1 family involved in P. capsic resistance.
Identification and Expression Characterization Analysis of Trihelix GeneFamily in Mung Bean (Vigna radiata)
LI Meng-Jiao, XING Zhan-Ning, YU Qing, LIU Fei, XING Bao-Long, LIU Zhi-Ping
2025 Vol. 33 (6): 1206-1219   DOI:10.3969/j.issn.1674-7968.2025.06.003 |  Full text (HTML) (1 KB)  |    PDF  (7546 KB)  ( 3 )
Abstract
Trihelix transcription factor is a plant-specific transcription factor that plays an important role in plant seed formation, flower development, light regulation and stress resistance. In order to analyze Trihelix transcription factor role in the growth and development process and stress response, 38 Trihelix family members (VrGT) were identified in the genome of Mung bean (Vigna radiata) via bioinformatic analyses, which were unevenly distributed on 1~10 chromosome and classified into 5 sub-families. Members of the same sub-family shared the similar motifs types and quantities, and contained specific conserved motifs. Most of the VrGT genes contained 1~2 introns. Fragment duplication was the main driver of the expansion of the VrGT genes. There were 12 and 30 VrGT genes that were interspecific collinearity with Arabidopsis thaliana and soybean (Glycine max) genomes, respectively. Numerous of cis-acting elements related to plant growth and development, light responsiveness, hormones responsiveness and stress responsiveness were predicted in the promoter regions (-2000 bp) of the VrGT genes. Multiple transcription factor families, such as WRKY, MYB and bHLH, were coexpressed with VrGT genes. The relative expression patterns of 38 VrGT genes were different at different growth stages, in different tissues of plants and under different abiotic stresses, among which the expression level was the highest in germinated seeds and 4 genes had high expression levels during the growth and development process of mung bean. There were 3 genes that had a high response level in the early and late stages of drought stress, and 8 genes that had a significant regulatory effect under salinity- alkalinity stress. This study provides a reference for further understanding the biological function of VrGT family in mung bean.
Genome-wide Identification of MaMKK Family in Banana (Musa spp.)and Their Expression Analysis in Response to Low Temperature Stress
LIN Wei, WEI Mi, LI Yue-Sen, WU Shui-Jin
2025 Vol. 33 (6): 1220-1233   DOI:10.3969/j.issn.1674-7968.2025.06.004 |  Full text (HTML) (1 KB)  |    PDF  (7508 KB)  ( 2 )
Abstract
The mitogen-activated protein kinase (MAPK) cascade is crucial to plant growth, development, and stress response, with MAPK kinase (MKK) serving a critical intermediary role in the cascade. Low temperature is one of the main reasons affecting banana (Musa spp.) yield. To explore the key genes for cold resistance in bananas, systematic evolutionary, gene structure assessment, chromosome localization andcollinearity analysis were conducted on the MaMKK gene family, clarifying the expression of the MaMKKgene family induced by low temperature in the 'Cavendish' (Musa AAA Cavendish) and 'Fenjiao' (MusaABB Pisang Awak). Subsequent analyses included sequence alignment and promoter element identification of the candidate genes. The result showed that A total of 13 members of the MaMKK gene family were identified and divided into 4 subclasses. Collinearity analysis showed that the members of the MaMKK gene family homologous had a strong connection, which had been purified and selected. After exposure to 7 ℃ , theexpression levels of MaMKK2a, MaMKK2b, and MaMKK5a significantly increased in both varieties, with agreater upregulation in 'Fenjiao' than in 'Cavendish'. However, the expression levels of MaMKK1, MaMKK2c, MaMKK3, and MaMKK5d showed a negative or unrelated relationship in the 2 varieties. Further analysis of these 7 candidate genes showed that MaMKK2a, MaMKK2b, and MaMKK5a responded more rapidly to low temperatures in 'Cavendish' than in 'Fenjiao'. The expression trends of MaMKK1, MaMKK2c, MaMKK3 and MaMKK5b showed significant differences within 48 h of low temperature treatment in 'Cavendish'/'Fenjiao'.The tissue expression results showed that there were significant differences in the tissue distribution of the 7 candidate genes in 'Cavendish'/'Fenjiao'. Analysis of the functional domains of 7 candidate genes revealed that they all contained phosphorylation targets for MEKK and MAPK, as well as multiple stress-related promotercis-acting elements. Among them, MaMKK2a and MaMKK5a contained low-temperature promoter cis-actingelements. The results indicated that the MaMKK gene in banana responded to low temperature stress, and the7 candidate genes screened may be related to the differences in cold resistance between 'Cavendish' and'Fenjiao'. This study provides a theoretical basis for developing cold-resistant banana varieties.
Identification and Expression Analysis of DNAJ Gene Family in Melon(Cucumis melo) Based on Transcriptome Sequencing
WANG Tao, I Ze-Lin, DU Qing-Jie, LI Juan-Qi, WANG Hu, WANG Ji-Qing, XIAO Huai-Juan, LI Meng
2025 Vol. 33 (6): 1234-1245   DOI:10.3969/j.issn.1674-7968.2025.06.005 |  Full text (HTML) (1 KB)  |    PDF  (8552 KB)  ( 11 )
Abstract
DNAJ protein usually plays an important role in plant growth and development and resistance to biological and non-biological stress as an auxiliary companion molecule of heat shock protein70 (HSP70). A total of 10 melon (Cucumis melo) DNAJ candidate members were identified in the transcriptome sequencing data of melon seedlings after low-temperature treatment by bioinformatics methods in this study.Furthermore, chromosome position, gene structure, subcellular localization, protein secondary/tertiary structure were determined. Meanwhile, the protein interaction, systematic evolution, cis-acting element and transcription level under abiotic stresses were analyzed. Results showed that 4 CmDNAJ members were distributed on 3 chromosomes, and were named CmDNAJ1~CmDNAJ4. The length of amino acids ranged from161 to 503 aa, all of which were hydrophilic, and located in the nucleus (CmDNAJ2/3), cytoplasmic (CmDNAJ12/3/4) and endoplasmic reticulum (CmDNAJ2). Systematic evolutionary analysis found that members of the melon DNAJ gene family were closely related to cucumber (C. sativus). The predictiveanalysis found that the 4 DNAJ promoters mainly contained cis-acting elements such as hormone response, light response and abiotic stress response. There were significant differences in tissue-specific expression of the 4 genes, only CmDNAJ1 and CmDNAJ2 showing highly expressed in male flowers, female flowers, roots,fruits and leaves of melons. CmDNAJ1, CmDNAJ2 and CmDNAJ3 might play a positive regulatory role in the process of fruit development and maturity, while CmDNAJ4 might play a negative regulatory role. The results of qRT-PCR confirmed that CmDNAJ1~CmDNAJ4 all responded to low temperature, high salt, abscisic acid (ABA), and drought induction. CmDNAJ1 and CmDNAJ2 were the key gene for responding to cold stress; CmDNAJ2 and CmDNAJ3 were the key genes for responding to salt stress, ABA stress and drought stress. Thisstudy provides a theoretical reference for analyzing the DNAJ gene function of melon.
Cloning and Prokaryotic Expression of the UDP-glycosyltransferaseGene PpUGT2 from Paris polyphylla var. chinensis
WANG Li-Ya, WANG Jing-Han, ZHOU Hui-Min, ZHANG Yao, CHEN Wen-Qiang, LU Zhi-Guo, ZHU Li-Na, TANG Qi
2025 Vol. 33 (6): 1246-1258   DOI:10.3969/j.issn.1674-7968.2025.06.006 |  Full text (HTML) (1 KB)  |    PDF  (13663 KB)  ( 4 )
Abstract
Saponins are important active components of Paris polyphylla chinensis, but their biosynthetic pathway has not been fully elucidated. Research suggests that UPD-glycosyltransferases (UGTs) are key enzymes in the downstream biosynthetic pathway of polyphyllin. This study based on the correlation analysisbetween the transcriptome of P. polyphylla var. chinensis and the total saponins content, a glycosyltransferasecandidate gene PpUGT2 (GenBank No. pq588702) was screened out, which might be involved in the biosynthesis of polyphyllin. The PpUGT2 gene was cloned using PCR technology, and an expression vector was constructed for prokaryotic expression. Expression was induced using IPTG, followed by purification ofthe recombinant protein. Concurrently, qPCR was employed to analyze the expression patterns of PpUGT2across different parts of P. polyphylla var. chinensis. The results indicated that PpUGT2 had a total length of1 773 bp, encoded a protein consisting of 590 amino acids with a relative molecular weight of 64.6 kD. This hydrophilic protein was unstable and exhibited good lipid solubility. SDS-PAGE analysis confirmed the expression of the PpUGT2 recombinant protein at approximately 64.6 kD. Furthermore, qPCR resultsdemonstrated expression of the PpUGT2 gene in roots, stems, leaves, and young fruits of P. polyphylla var.chinensis, with a distinct expression hierarchy of roots>stems>leaves>young fruits, suggesting a predominant role of this gene in root tissues. This study enriches the study of UGT in P. polyphylla var. chinensis and provides a theoretical basis for further analysis of the biosynthesis pathway of polyphyllin.
Identification and Expression Analysis of TCP Transcription FactorFamily Members in Dontostemon elegans
GU Yu-Feng, LIU Xin-Xin, Huerxidan•TUERXUN, GE Feng-Wei
2025 Vol. 33 (6): 1259-1269   DOI:10.3969/j.issn.1674-7968.2025.06.007 |  Full text (HTML) (1 KB)  |    PDF  (4392 KB)  ( 3 )
Abstract
Dontostemon elegans is an important species in the desert ecosystem of Dabancheng, with excellent characteristics of drought resistance and salt tolerance. TCP (teosinte branched 1/cycloidea/proliferating cell factor) as a unique type of transcription factor in plants, the family is widely involved in the regulation of plant growth, development, and stress response. In this study, bioinformatics methods were used to identify the members of the TCP cell factor and analyzed their expression patterns. Twenty-two DeTCPs were identifiedfrom the transcriptome of D. elegans, and the proteins encoded by these members all contained a TCP domain. Phylogenetic analysis showed that DeTCPs could be divided into 2 categories and 3 subfamilies. The members of the same group had similar gene structures, and the distribution of conserved domains of the encodedproteins was similar. The expression pattern analysis showed the expression of the 5 DeTCPs members was tissue-specific in the D. elegans, 5 DeTCPs were significantly up-regulated under NaCl stress(P<0.05). UnderPEG stress, 3 DeTCPs were significantly up-regulated, and the relative expression of 2 DeTCPs was significantly down-regulated (P<0.05). In summary, this study showed that the members of the DeTCPs contained the basic characteristics, had tissue expression specificity, and responded to drought and salt stress.The members of the family might be involved in the regulation of drought and salt stress resistance. This study provides a data basis and basis for further mining the function of TCPs.
Deoxynivalenol Induces Inflammation in IPEC-J2 Cells by ActivatingTXNIP/NLRP3 Inflammasome Pathway
ZHOU Chuang, XIA Ying, DING Shuo, WANG Xing-Di, WEI Xiao-Jun, SHI Lu, ZHANG Hua
2025 Vol. 33 (6): 1270-1279   DOI:10.3969/j.issn.1674-7968.2025.06.008 |  Full text (HTML) (1 KB)  |    PDF  (5082 KB)  ( 2 )
Abstract
Deoxynivalenol (DON) is one of the most serious mycotoxins which severely contaminates grains and their products. Exposure to DON causes animal poisoning, and its primary target organ is the intestine. To explore the connection and mechanisms involved in the intestinal inflammatory response triggered by DONand its interaction with the NOD-like receptor signaling pathway, in this study, porcine intestinal epithelial cells (IPEC-J2) were cultured with 1 μg/mL DON. ELISA was used to quantify the concentration of interleukin-1β (IL-1β) in the cell supernatants. The expression levels of thioredoxin interaction protein (TXNIP), cysteinyl aspartate specific proteinase-1 P20 (Caspase-1 P20) and NOD-like receptor protein 3 (NLRP3) were detected by Western blot. TXNIP siRNA transfection was used to further verify the regulation of NLRP3 by TXNIP. Antioxidant N-acetyl-cysteine (NAC) was used to detect the interaction among reactive oxygen species (ROS) and NLRP3 inflammasome activation. The results showed that compared with blank control group, the secretion of IL-1β in the cell supernatant was extremely significantly elevated in the DON-treated group. (P<0.01), and the protein expressions of TXNIP/NLRP3 inflammasome pathway-relatedproteins TXNIP, NLRP3 and Caspase-1 P20 in the cells were significantly up-regulated (P<0.05). Compared with negative transfection in DON-treated group, the secretion of IL-1β in the supernatant of TXNIP siRNA transfection in DON-treated group was extremely significantly decreased (P<0.01), and the proteinexpression of Caspase-1 P20 in the cells was significantly down-regulated (P<0.05). Compared with DON- treated group, the NAC intervention group exhibited significantly reduced cellular ROS levels (P<0.05), decreased secretion of IL-1β in the supernatant, and significantly downregulated expression of TXNIP andCaspase-1 P20 protein levels (P<0.05) in the cells. This study showed that DON induced the inflammatory response of IPEC-J2 cells through the TXNIP/NLRP3 inflammasome pathway, and the antioxidant NAC could alleviate the activation of NLRP3 induced by DON. This study identifies novel therapeutic targets and offers new insights for the effective prevention and management of DON-induced toxicity.
Expression Analysis of IGF-1, IGF-2 and TGF- β2 in the Lung of Yaks(Bos grunniens) in Different Ages
DENG Yan-Jiang, ZHONG Dong-Lan, CUI Yan, YU Si-Jiu, WEI Peng-Qiang, ZHANG Qian, NIU Yue-Yue, XIAN Kang-Le, WANG Deng-Hui, CHEN Chun-Yan
2025 Vol. 33 (6): 1280-1289   DOI:10.3969/j.issn.1674-7968.2025.06.009 |  Full text (HTML) (1 KB)  |    PDF  (13229 KB)  ( 4 )
Abstract
Insulin like growth factor (IGF) IGF-1 and IGF-2, as hypoxia target factors, can cooperate with transforming growth factor- β2 (TGF- β2) to participate in lung growth and development, immune regulation and injury repair. The purpose of this study was to investigate the expression and distribution characteristics of IGF-1, IGF-2 and TGF- β2 in the lungs of yaks (Bos grunniens) in different ages. The distribution and expression levels of IGF-1, IGF-2 and TGF- β2 in the lungs of yaks at different ages were investigated by immunohistochemical staining, qRT-PCR and Western blot. Immunohistochemical analysis showed that the expression positions of the three growth factors in the yak lungs were similar, and they were mainly distributed in all levels of bronchial epithelial cells and alveolar epithelial cells. In addition, IGF-1 and IGF-2were also expressed in pulmonary artery endothelial cells. The results of qRT-PCR showed that the expression levels of IGF-1 and TGF-β2 genes in the lungs of adult yaks were significantly higher than those of newborn, juvenile and elderly yaks (P<0.05). The expression of IGF-2 gene in the lung of newborn yak was significantly higher than that in other age groups (P<0.05). The results of Western blot showed that theexpression of IGF-1 and IGF-2 protein was the highest in the primary stage and significantly higher than that in other age groups (P<0.05). The expression of TGF- β2 protein was the highest in adulthood, and was significantly higher than that in other age groups (P<0.05). The expression levels of the 3 factors were similar with age, and the expression levels were higher in newborn and adulthood, suggesting that IGF-1, IGF-2, and TGF-β2 might be involved in the development of lung and the formation of hypoxic adaptive structure in yak at this age. This study provides the basis for exploring the specific adaptation mechanism of yak lung in hypoxic environment.
Evaluation of the Effect of Progesterone Packaging in Yak (Bos grunniens) Uterine Fluid Exosomes and Its Effect on Mouse (Mus musculus) Uterine Receptivity
HE Yong-Gang, YU Tian-Jun, HE Yu-Long, WANG Xin-Qiang, LI Zhi-Jie, WANG Jing-Lei, WANG Meng, LI Tian-Hao, PAN Yang-Yang, XU Geng-Quan
2025 Vol. 33 (6): 1290-1300   DOI:10.3969/j.issn.1674-7968.2025.06.010 |  Full text (HTML) (1 KB)  |    PDF  (13985 KB)  ( 3 )
Abstract
Progesterone (P4) is an important reproductive hormone that regulates the synchronisation of endometrial tolerance and embryonic development, and ensures successful follicle implantation; exosomes have good biocompatibility, low immunogenicity and are excellent carriers for drug transport. In order toexplore whether yak (Bos grunniens) uterine fluid-derived exosomes could be used as delivery vehicles fordrugs such as hormones, proteins or RNAs. In this study, exosomes of yak uterine fluid were obtained by ultra- high-speed centrifugation, and the morphological and marker proteins (CD81 and TSG101) were identified by transmission electron microscopy and Western blot, respectively, and the histological changes of mouse (Musmusculus) organs were analyzed by intraperitoneal injection of mice to evaluate their safety. Yak uterine fluid exosomes were combined with progesterone ultrasonication packaging, and the effect of exosome packaging on progesterone was analyzed by comparing the endometrial thickness and the expression levels of leukemia inhibitory factor (LIF) and interleukin-6 (IL-6) in mice. The results showed that the exosomes of yak uterine fluid obtained by ultracentrifugation were typical in morphology, and the labeled proteins of CD81 and TSG101 were clearly expressed. The expression levels of exosome marker proteins CD81 and TSG101 decreased by sonication, which were 0.74 and 0.83 times higher than those in the control group, respectively. The packaging rate of exosome-packed progesterone was (48.4±1.29)%. Through histological comparison, the optimal concentration of dimethyl sulfoxide (DMSO) as a progesterone co-solvent for exosome packaging was0.5%, and yak exosomes had no damage to mouse organs and tissues. Progesterone after exosome packaging significantly increased endometrial thickness and up-regulated the mRNA and protein expression levels of LIF in mice (P<0.05). The mRNA and protein expression levels of endometrial IL-6 in the single progesterone-treated group were lower than those in the control group, while the expression levels of IL-6 in the exosome- packaged progesterone-treated group increased. The results showed that yak uterine fluid exosomes could be used as effective drug delivery vehicles for progesterone, which could significantly improve progesterone bioavailability. This study provides a theoretical reference for the application of reproductive tract exosomes in the regulation of animal reproduction.
Protective Effect of Ellagic Acid Against Inflammatory Injury Induced byHigh Concentration of LPS in Ovarian Granulosa Cells in vitro
LU Qing-Mei, PAN Zhi-Ren, WEN Xiao-Yan, LIU Bin, ZHANG Xi-Lu, CHEN Yi, XIAO Yang, ZHAO Jia-Fu
2025 Vol. 33 (6): 1301-1311   DOI:10.3969/j.issn.1674-7968.2025.06.011 |  Full text (HTML) (1 KB)  |    PDF  (1920 KB)  ( 2 )
Abstract
Proliferation, apoptosis and steroid hormone synthesis of ovarian granulosa cells (OGCs) play important roles in follicle development, lipopolysaccharide (LPS) -induced inflammatory responses and oxidative stress have been recognized as important factors affecting ovarian function. As a natural polyphenolic compound, ellagic acid (EA) exhibits multiple biological activities, including antioxidant, anti- cancer, anti-inflammatory, and anti-viral properties. In this study, OGCs were isolated and cultured bycollecting ovarian tissues from healthy and sexually mature Guizhou black goats (Capra hircus). The effects of EA and LPS on the survival rate of OGCs were detected by CCK-8 assay, and the relative expression of genes related to the synthesis of inflammation factors and steroid hormones were detected by qRT-PCR. The content of estradiol (E2), progesterone (P4) and related antioxidant indexes in the cell supernatant were detected by ELISA to evaluate the functional changes of OGCs. The results showed that the concentration of EA at 1 μmol/L had the best effect on the survival rate of OGCs, after 24 h of LPS treatment, the half maximalinhibitory concentration (IC50) for OGCs was 200 μg/mL. qRT-PCR results showed that, compared with the control group, the inflammatory factors in the LPS group, interleukin-1β (IL-), IL-6 and tumor necrosis factor α (TNF- α) were extremely significantly higher in relative expression (P<0.01); cytochrome P450 family 11 subfamily A1 member (CYP11A1), CYP19A1, 3β -hydroxysteroid dehydrogenase ( -HSD) and steroidogenic acute regulatory protein (STAR), steroid hormone synthesis genes were extremely significantly reduced in expression (P<0.01). Compared with the LPS group, the relative expression of IL- and TNF- α was extremely significantly lower in the LPS+EA group (P<0.05), IL-6 had a tendency to decrease; CYP11A1 and CYP19A1 expression was extremely significantly up-regulated (P<0.01), and the results of ELISA showed that the secretion of E2 in the LPS group was significantly reduced (P<0.05), and the secretion of P4 was extremely significantly reduced (P<0.01), malondialdehyde (MDA) concentration was significantly increased (P<0.05), while the concentrations of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and reactive oxygen species (ROS)extremely significantly increased (P<0.01); while E2 and P4 were significantly up-regulated (P<0.05) and GSH-Px, SOD and ROS concentrations were extremely significantly decreased (P<0.01) in the LPS+EA group compared with the LPS group, and the concentration of MDAtended to decrease. Exogenous addition of EA could serve to alleviate LPS-induced inflammatory responses and oxidative damage in OGCs and could moderate the functional disorders of OGCs caused by LPS to some extent. This study revealed the potential mechanism of ellagic acid in protecting OGCs from inflammatory responses and oxidative damage, which provides a theoretical basis for the development of safe and effective natural anti-inflammatory antioxidants. Meanwhile, it also provides new ideas for research on improving the reproductive performance of livestock and coping with reproductive system-related diseases.
Expression Characteristics Analysis of Goat (Caprahircus) Phosphodiesterase 4B (PDE4B) and Its Effect on the Differentiation ofIntramuscular Precursor Adipocytes
LI Jin-Lan, SUN Shi-Yu, HE Xiao-Fei, ZHANG Li-Yi, LIN Ya-Qiu, LIU Wei, XING Jia-Ni
2025 Vol. 33 (6): 1312-1323   DOI:10.3969/j.issn.1674-7968.2025.06.012 |  Full text (HTML) (1 KB)  |    PDF  (10100 KB)  ( 6 )
Abstract
Intramuscular fat (IMF) in goat (Capra hircus) is an important factor in affecting meat quality traitssuch as flavor and juiciness. This study focused on the cloning and functional analysis of phosphodiesterases4B (PDE4B), to delineate its regulatory role in modulating the differentiation dynamics of caprine intramuscular preadipocytes. In this study, the intramuscular precursor adipocytes of JianZhou big-eared goats were used to amplify the target gene fragments by RT-PCR, and their sequences were analyzed by onlinebioinformatics tools. The expression level of PDE4B gene in different tissues and different differentiation stages of intramuscular precursor adipocytes in goats was detected using qPCR. The PDE4B overexpression plasmid (pEGFP-N1-PDE4B) was successfully engineered via molecular subcloning and subsequentlytransfected into caprine intramuscular preadipocytes. Subsequent Oil red O staining and qPCR assays were employed to assess its regulatory effects on intracellular lipid droplet deposition, coupled with transcriptional profiling of key adipogenic differentiation, triglyceride biosynthesis, and lipolytic metabolism markers. Theresults showed that the length of the cloned PDE4B gene was 1 815 bp, including the coding sequence (CDS)region of 1 629 bp, which encoded a total of 542 amino acids. Furthermore, the expression of PDE4B gene was significantly higher in goat lung than in other tissues examined (P<0.01), and reached the highest expression after 48 h induction of differentiation in intramuscular adipocytes (P<0.01). Oil red O staining and the quantitative results after PDE4B overexpression showed that there were significant increase in lipid droplet aggregation (P<0.05). The mRNA expression level of differentiation marker genes CCAAT/ enhancerbindingprotein α (C/EBPα), C/EBPβ and triglyceride synthesis marker genes activator protein 2 (AP2) and fatty acid synthase (FASN) were extremely significantly up-regulated after overexpression of PDE4B gene (P<0.01). Meanwhile, the expression of triglyceride catabolism marker genes adipose triglyceride lipase (ATGL), hormone-sensitive lipase (HSL) and lipoprotein lipase (LPL) were significantly down-regulated (P<0.01). In conclusion, PDE4B gene might positively regulate the process of intramuscularprecursor adipocyte differentiation in goats, and this effect might be realized by up-regulating the expression of C/EBPα, C/EBPβ, AP2, and FASN. This study provides data support for revealing the molecular mechanism of intramuscular precursor adipocyte differentiation regulated by PDE4B in goats.
Screening of Candidate Genes and Their nsSNPs for Coat Color in Arctic White Fox (Vulpes lagopus) and Arctic Blue Fox Based on Whole- genome Resequencing Technology
GUO Jian-Xu, WANG Dong-Xian, XIE Yu-Chun, QIAO Xian, PENG Yong-Dong, GONG Yuan-Fang, LIU Zheng-Zhu
2025 Vol. 33 (6): 1324-1335   DOI:10.3969/j.issn.1674-7968.2025.06.013 |  Full text (HTML) (1 KB)  |    PDF  (3033 KB)  ( 2 )
Abstract
The diversity of coat color in Arctic foxes (Vulpes lagopus) provides an important model for studying the genetic regulation of coat color, and understanding its regulatory mechanisms is crucial for the breeding of fur-bearing animals. In this study, whole-genome resequencing was performed on Arctic white foxes and Arctic blue foxes. Nonsynonymous single nucleotide polymorphism (nsSNP) sites located in exon regions were selected as candidate loci, and the corresponding candidate genes were analyzed. Candidate genes involved in coat color regulation were identified through functional enrichment analysis. The epidermalgrowth factor (EGF) and adenomatous polyposis coli (APC) genes containing homozygous nsSNP mutationswere further validated using genomic DNA extracted from skin tissues of 42 Arctic foxes (21 white and 21 blue foxes). The results revealed that 8 860 718 SNPs and 21 150 nsSNPs were identified through resequencing, with 5 787 genes annotated. The nsSNP results of candidate genes were consistent with those obtained from whole-genome resequencing. Association analysis demonstrated that the c. 3571C>A variant inthe EGF gene and the c. 6812G>A variant in the APC gene were significantly (P<0.05) and extremelysignificantly (P<0.01) associated with coat color traits in Arctic white and blue foxes. These results indicated that specific variant sites in the APC and EGF genes identified through whole-genome resequencing and functional enrichment analysis might influence the formation of coat color in Arctic foxes. This study providescritical data for understanding the genetic mechanisms underlying coat color formation in Arctic white and blue foxes. Furthermore, it provides a theoretical freference for molecular breeding and the conservation of coat color diversity in fur animals.
Identification of PPP3CA Gene SNP and Its Association Analysis withGrowth Traits in Weining Chickens (Gallus gallus)
ZHAO Zhong-Long, RAN Guang-You, ZHANG Yong, ZHANG Xi-Ben, WANG Zhi-Wei, AI Zhao-Bi, YANG Run-Qian, LI Wei-Bo, CHEN Ze-Lin, YE Hong-Ying
2025 Vol. 33 (6): 1336-1345   DOI:10.3969/j.issn.1674-7968.2025.06.014 |  Full text (HTML) (1 KB)  |    PDF  (1925 KB)  ( 3 )
Abstract
The Weining chicken (Gallus gallus) is one of the valuable local poultry genetic resources inGuizhou province. It is of great significance for the genetic improvement and molecular marker-assisted breeding of Weining chickens to study their growth traits and uncover their genetic effects using candidate genes. In this study, a total of 113 Weining chickens (58 males and 55 females) aged 300 days were selected asthe study subjects. PCR amplification and direct sequencing were used to identify the SNP locus of PPP3CAgene. Excel was used to calculate genotype frequency, allele frequency, effective allele number and polymorphism information content of each site, and to analyze whether each mutation site was in Hardy- Weinberg equilibrium state. The secondary structure analysis of mRNAs was performed using RNA fold online software. SPSS 27.0 was used to analyze the association between genotypes of different polymorphicloci and body size indexes. The results showed that 3 SNP sites were found on exon 6 of PPP3CA gene inWeining chickens, namely g. 151368C>T, g. 151389T>C and g. 151402T>G, among which missense mutation occurred at g.151368C>T site, resulting in the conversion of arginine at position 206 on mRNA to tryptophan. g. 151402T>G site missense mutation resulting in the conversion of isoleucine at position 216 to tryptophan. Linkage disequilibrium analysis showed that there was a strong linkage disequilibrium effect between g.151389T>C and g. 151368C>T, g. 151402T>G. The χ2 test showed that g. 151368C>T and g. 151389T>C matched Hardy-Weinberg equilibrium, while g. 151402T>G deviated from Hardy-Weinberg equilibrium. At the g.151402T>G site, association analysis showed that TT and GG individuals had significantly higher bodyweight, body slope length, and chest width than TG individuals (P<0.05), TT individuals had significantlyhigher chest depth than GG and TG individuals (P<0.05), GG individuals had significantly higher chest depth than TG individuals (P<0.05), and TT and GG individuals had extremely significantly longer keel and shank length than TG individuals (P<0.01). Individuals of the TT and GG types had considerably larger tibia girth than those of the TG type (P<0.05). Haplotype and diplotype analysis of 3 SNPs showed that there were 4haplotypes and 8 diplotypes. Among them, diplotype H2H3 (TCTCTT) and H1H1 (TTCCGG) could be used as molecular marker sites to improve keel length, shank length and shank girth. This study provides a scientific basis for molecular marker-assisted selection for growth traits of Weining chickens.
Effects of Different Pesticides on the Levels of Prostaglandin Synthetase, Isoprostaglandin and Reactive Oxygen Species in Adult Zebrafish (Danio rerio)
ZHOU Xin-Zong, WU Sheng-Gan, LI Shao-Bin, CHANG Ju-Hua
2025 Vol. 33 (6): 1346-1357   DOI:10.3969/j.issn.1674-7968.2025.06.015 |  Full text (HTML) (1 KB)  |    PDF  (4624 KB)  ( 2 )
Abstract
Prostaglandins (PGs) are important signaling molecules that play an important role in normal and pathophysiological processes. Many endocrine disruptors (EDCs) inhibit PG synthesis. However, there are few studies on the effect of pesticide EDCs on PG in zebrafish. The effects of 10 conventional pesticides on prostaglandin synthetase—cyclooxygenase (COX) (COX-1, COX-2) and isoprostaglandin (iso-PG) (8-iso PGF1α, 8-iso PGF2α, and iPF2α-), reactive oxygen species (ROS) and its related proteins (GST, CAT, SOD and POD) of adult zebrafish were detected by enzyme-related immunosorbent assay (ELISA). Then the correlation between different proteins and ROS or iPF2α - was analyzed. The ELISA results showed significant differences between male and female zebrafish treated with different pesticides, indicating that their effects on the levels of iPF2α- and ROS in zebrafish were sex-dependent. Correlation analysis showed that GST was significantly correlated with the levels of ROS (F=8.517; P=0.0092; R2=0.3212), iPF2α- was significantly positively correlated with ROS levels (F=75.51; P<0.0001; R2=0.8075), other proteins had no significant correlation with ROS. There was no significant correlation between each protein and iPF2α - . This study provides basic information for the screening and detection of pesticide endocrine disruptors.
Analysis of Genetic Characteristics of Cellulase in Potato Scab Pathogen in China
HUANG Yi-Fan, DING Yu, YANG Yun-Shan, ZHANG Cai-Hua, ZHAO Chuan-Xiang, CHEN Chen, HAN Zhi-Ruo, YU Xiu-Mei, ZHAO Wei-Quan
2025 Vol. 33 (6): 1358-1368   DOI:10.3969/j.issn.1674-7968.2025.06.016 |  Full text (HTML) (1 KB)  |    PDF  (6556 KB)  ( 3 )
Abstract
Potato scab is a soil-borne disease caused by several scab-related Streptomyces spp., its' pathogenic mechanism remains unclear. Cellulase is reported as a key enzyme in many pathogens to infect the plant cell wall, the present study analyzed genetic characteristics of cellulase in the pathogenic Streptomyces spp. A total of 14 cellulases were identified from a typical potato common scab strain 87.22, although they had diversefunctional domain patterns, similar three-dimensional structures were detected in most of these cellulases. 7 pairs of gene specific primer were designed based on the sequence similarity of 14 cellulases, and used to amplify genomic DNA of 18 potato common scab strains and 5 potato scab pathogenic species collected fromdifferent potato (Solanum tuberosum) growing areas of China, it was found that 4 pairs of primers obtainedtarget bands in the 2 potato common scab strains No. MZ-4 and H2, and exhibited amplification polymorphism within the potato common scab strains as well as among potato scab pathogenic species, which indicated there was significant polymorphism in cellulases from different potato scab pathogens in China. Thisstudy provides a new support for in-depth analyzing the genetic diversity of the scab-related Streptomyces spp.
Invited Reviews
Research Progress on the Regulation of Host MEK/ERK SignalingPathway by Viral Infection
ZENG Jian-Yu, GAO Peng, ZHOU Lei, GE Xin-Na, ZHOU Qiong-Qiong, HAN Jun, GUO Xin, ZHANG Yong-Ning, YANG Han-Chun
2025 Vol. 33 (6): 1369-1381   DOI:10.3969/j.issn.1674-7968.2025.06.017 |  Full text (HTML) (1 KB)  |    PDF  (5496 KB)  ( 2 )
Abstract
The mitogen-activated protein kinase kinase/extracellular regulated protein kinases (MEK/ERK) signaling pathway is a crucial intracellular signal transduction system that plays a pivotal role in regulating fundamental cellular processes including cell proliferation, differentiation, and apoptosis and so on. Recent studies have demonstrated that this signaling pathway also serves important regulatory functions in virus-host interactions. This paper comprehensively reviews the research progress on how viruses modulate biological processes such as apoptosis, autophagy, and the production and secretion of inflammatory factors through the MEK/ERK signaling pathway. Furthermore, it also provides an in-depth analysis of the current research status and potential therapeutic applications of MEK/ERK signaling pathway inhibitors as promising antiviral strategies. This review offers significant theoretical foundations and new research directions for the development of antiviral drugs.
Research Progress on the Application of Antimicrobial Peptides
GAO Nan, LI Yi-Han, SUN Jia-Qi, BI Chong-Peng, SHAN An-Shan, WANG Jia-Jun
2025 Vol. 33 (6): 1382-1397   DOI:10.3969/j.issn.1674-7968.2025.06.018 |  Full text (HTML) (1 KB)  |    PDF  (1742 KB)  ( 2 )
Abstract
The inappropriate utilization of antibiotics has precipitated a multitude of concerns, prominently featuring drug residues and the proliferation of drug-resistant bacteria. Drug residues, by way of the food chain, can inflict deleterious effects on human health and instigate ecological environmental contamination. Concurrently, drug-resistant bacteria present formidable challenges in the treatment of infectious diseases, engender a substantial escalation in medical expenditures, and, in severe cases, imperil human lives. Antimicrobial peptides (AMPs), with their multiple biological functions and unique mechanisms of action, have emerged as promising alternatives to antibiotics. Furthermore, their low toxicity and reduced likelihood of resistance development have driven research beyond theoretical studies to practical applications in production systems. This review summarizes the mechanisms of action of AMPs and their research and applications in clinical practice, the food industry, and agricultural production. The review provides a theoretical foundation for developing novel anti-infection strategies, ensuring food safety, and promoting sustainable agriculture. This work holds critical value for addressing global antimicrobial resistance challenges and achieving sustainable development goals.
Resources and Updated Technology
Visual and Highly Sensitive Serological Detection Technology for Pepper mild mottle virus Based on Monoclonal Antibody
LIU Nai-Ru, ZHANG Xi, ZHANG Cui, DONG Jie, MAO Wei-Jia, WANG Ya-Qin, ZHOU Xue-Ping, FU Shuai, WU Jian-Xiang
2025 Vol. 33 (6): 1398-1408   DOI:10.3969/j.issn.1674-7968.2025.06.019 |  Full text (HTML) (1 KB)  |    PDF  (8465 KB)  ( 3 )
Abstract
Pepper mild mottle virus (PMMoV) seriously damages the pepper (Capsicum annuum) industry worldwide. The establishment of sensitive and practical virus detection technology is the key to the prevention and control of this viral disease. In this study, using purified PMMoV virions as the immunogen, six monoclo‐ nal antibodies (i. e. 15A3, 15G7, 17D1, 11E1, 6A10 and 3B2) that specifically and sensitively recognize PM‐MoV were finally obtained through the immunization of BALB/c mice (Mus musculus), cell fusion, screeningof hybridoma cells, antibody selection, continuous cell subcloning and preparation of monoclonal antibody as‐ cites. Two serological techniques, dot enzyme-linked immunosorbent assay (Dot-ELISA) and colloidal gold immunochromatographic strip (CGICS), were established to detect PMMoV using the produced monoclonal antibodies as the detection antibodies. Both Dot-ELISA and CGICS could specifically and broadly detect dif‐ ferent isolates of PMMoV, and showed negative reactions when detecting Tobacco mosaic virus (TMV), Cucum- ber green mottle mosaic virus (CGMMV), Tomato brown rugose fruit virus (ToBRFV), Tomato mosaic virus (ToMV), Cucumber mosaic virus (CMV) and healthy pepper plants. Moreover, the sensitivities of Dot-ELISA and CGICS for detecting diseased leaf homogenates were separately up to 1∶25 600 and 1∶204 800 times dilu‐ tion (W/V, g/mL), which was 2 times and 16 times more sensitive than RT-PCR, respectively. Additionally, the monitoring results of field samples by the two serological detection techniques were 100% consistent with those from RT-PCR monitoring, indicating that the two established serological techniques can be used to accu‐ rately, broadly, sensitively and quickly detect PMMoV. This study provides technical support for the detection and diagnosis of PMMoV, and its scientific prevention and control.
Establishment of the Micro-agglutination Test for Antibody Detection ofCitrobacter freundii from Sheep (Ovis aries)
LI Jing, LIU Yi-Ting, YANG Jin-Qian, DING Lue-Xuan, XUE Yun, DING Ke, SI Li-Fang, ZHAO Zhan-Qin
2025 Vol. 33 (6): 1409-1416   DOI:10.3969/j.issn.1674-7968.2025.06.020 |  Full text (HTML) (1 KB)  |    PDF  (1007 KB)  ( 2 )
Abstract
Ovine Citrobacter freundii was an emerging infectious disease affecting sheep (Ovis aries), in order to establish a rapid detection method for the disease, this study used the inactivated whole-cell of C. freundii LK-1 strain as the antigen, establishing an antibody detection method of micro-agglutination test (MAT) byoptimizing the reaction conditions. The optimal antigen concentrations of inactivated whole-cell of C. freundiiLK-1 strain were 2.0×; 109 cfu/mL and the incubation temperature was 37 ℃ for 7~9 h. The MAT was specificity for C. freundii of sheep, demonstrating negative reactivity (titer≤; 1∶; 2) against serum samplespositive for 8 common ovine pathogens. The antibodies in sheep could be detected at 14 d post vaccinationwith inactivated C. freundii LK-1 vaccines. The values at day 21 and 35 were more than 1∶; 16 and 1∶; 64,respectively. The intra-assay coefficients of variation (CV) were consistently 0%, while the inter-assay CV ranged from 5.43% to 6.60%, indicating acceptable reproducibility. 374 serum samples, which were detected by the developed MAT and the ELISA with the same antigen. The results of tests showed that the total positive rates were 9.9% and 10.7%, respectively. The coincidence of the positive and negative results was good(Kappa value was 0.897), showing that the sheep were infected by C. freundii in a certain area of Henan Province. This study provides a basis for vaccine antibody evaluation and epidemiological investigation of ovine C. frendii.
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