羊抗病育种候选基因进展

Progress in research on candidate genes for disease resistance breeding in sheep

摘要

本文聚焦羊抗病育种领域的候选基因与育种技术两大方向，旨在为畜牧业抗病遗传改良提供理论依据与技术支撑。文章系统解析了天然免疫基因（TLR、MHC），宿主－病原互作基因（IFN、ILs），抗病毒基因（TMEM154、APOBEC3、Mx1）及抗菌基因（SBD）四类关键基因的分子机制；同时评估了全基因组关联分析（GWAS）、单细胞测序和CRISPR/Cas9基因编辑的育种应用价值，通过整合多组学数据与精准编辑技术，抗病育种正从传统表型选择向分子设计育种转型，为畜牧业可持续发展开辟了新路径。

Abstract

This paper reviews the progress in research on the field of sheep disease resistance breeding, focusing on the two major directions of disease resistance candidate genes and breeding technology, with an aim of providing theoretical basis and technical support for genetic improvement of disease resistance in animal husbandry. The molecular mechanisms of four key genes are systematically analyzed: innate immune genes (TLR, MHC), host pathogen interaction genes (IFN, ILS), antiviral genes (tmem154, APOBEC3, MX1) and antibacterial genes (SBD). At the same time, the application value of genome-wide association analysis (GWAS), single cell sequencing and CRISPR/cas9 gene editing in breeding is evaluated. By integrating multi-omics data and precise editing technology, disease resistance breeding is transforming from traditional phenotypic selection to molecular design breeding, opening up a new path for sustainable development of animal husbandry.

关键词

羊 / 候选基因 / 育种技术 / 抗病

Key words

sheep / candidate genes / breeding technology / disease resistance

参考文献

[1] 王引, 张秋桐, 樊平． 养羊业发展趋势及针对性防疫措施[J]． 中国动物保健, 2024, 26(4)： 87-88．

[2] BINPARK W, KIM S, SHIM S, et al. Identification of dendritic cell maturation, TLR, and TREM1 signaling pathways in the Brucella canis infected canine macrophage cells, DH82, through transcriptomic analysis[J]. Front Vet Sci, 2021, 8: 619759.

[3] ZHANG E, MA Z, LU M. Contribution of T-and B-cell intrinsic toll-like receptors to the adaptive immune response in viral infectious diseases[J]. Cell Mol Life Sci, 2022, 79(11): 547.

[4] SATO T, IKEGAMI I, YANAGI M, et al. Interleukin 9 mediates T follicular helper cell activation to promote antibody responses[J]. Front Immunol, 2024, 15: 1441407.

[5] DOMINGUEZ-FLORES A, RODRÍGUEZ LÓPEZ G M, SORIA-CASTRO R, et al. Brucella abortus induces mast cell activation through TLR-2 and TLR-4[J]. Microb Pathog, 2023, 176: 106005.

[6] LIN Y T, CHEN Y P, FANG C H, et al. Capsid proteins of foot-and-mouth disease virus interact with TLR2 and CD14 to induce cytokine production[J]. Immunol Lett, 2020, 223: 10-16.

[7] RHODES J, ZIPFEL C, JONES J D G, et al. Concerted actions of PRR-and NLR-mediated immunity[J]. Essays Biochem, 2022, 66(5): 501-511.

[8] XU X L, ZHAO Y, CHEN M M, et al. Shifts in intestinal microbiota and improvement of sheep immune response to resist Salmonella infection using Toll-like receptor 4 (TLR4) overexpression[J]. Front Microbiol, 2023, 14: 1075164.

[9] LUCAS R M, LUO L, STOW J L. ERK1/2 in immune signalling[J]. Biochem Soc Trans, 2022, 50(5): 1341-1352.

[10] BARRIONUEVO P, DEL PINO M V, VELÁSQUEZ L N, et al. Brucella abortus inhibits IFN-γ-induced FcγRI expression and FcγRI-restricted phagocytosis via toll-like receptor 2 on human monocytes/macrophages[J]. Microbes Infect, 2011, 13(3): 239-250.

[11] ZHANG Y, WANG X, LI Z, et al. Brucella melitensis outer membrane protein 25 interacts with ferritin heavy polypeptide 1 in human trophoblast cells[J]. Mol Med Rep, 2022, 26(1). DOI: 10.3892/mmr.2022.12740.

[12] GARCÍA-MARTÍNEZ M, CORTEZ L M, OTERO A, et al. Distinctive toll-like receptors gene expression and glial response in different brain regions of natural scrapie[J]. Int J Mol Sci, 2022, 23(7): 3579.

[13] HASAN M W, HASEEB M, EHSAN M, et al. The immunogenic maturation of goat monocyte-derived dendritic cells and upregulation of toll-like receptors by five antigens of Haemonchus contortus in vitro[J]. Res Vet Sci, 2021, 136: 247-258.

[14] KUMAR R, MEENA A S, BARAIYA T, et al. Expression of Toll-like receptors in Haemonchus Contortus resistant sheep: an innate immune parameter for host defense against gastrointestinal nematode infection[J]. Vet Immunol Immunopathol, 2024, 275: 110813.

[15] PÉREZ-HERNÁNDEZ T, HERNÁNDEZ J N, MACHÍN C, et al. Variability in the response against Teladorsagia circumcincta in lambs of two Canarian sheep breeds[J]. Int J Mol Sci, 2023, 24(1): 29.

[16] SALIM B, NAKAO R, CHATANGA E, et al. Exploring genetic diversity and variation of Ovar-DRB1 gene in Sudan Desert Sheep using targeted next-generation sequencing[J]. BMC Genomics, 2024, 25(1): 160.

[17] ŚWIDEREK W P, GRUSZCZYŃSKA J, WINNICKA A. Polymorphism of selected regions of ovar-MHC and the health status of the ovine mammary gland[J]. Animals, 2020, 10(12): 2325.

[18] MALIK M, KHAN S, ULLAH A, et al. Proteome-wide screening of potential vaccine targets against Brucella melitensis[J]. Vaccines, 2023, 11(2): 263.

[19] DING H, WANG G, YU Z, et al. Role of interferon-gamma (IFN-γ) and IFN-γ receptor 1/2 (IFNγR1/2) in regulation of immunity, infection, and cancer development: IFN-γ-dependent or independent pathway[J]. Biomed Pharmacother, 2022, 155: 113683.

[20] RODRÍGUEZ-MARTÍN D, LOULOUDÉS-LÁZARO A, AVIA M, et al. The interplay between bluetongue virus infections and adaptive immunity[J]. Viruses, 2021, 13(8): 1511.

[21] ROJAS J M, AVIA M, MARTÍN V, et al. Inhibition of the IFN response by bluetongue virus: the story so far[J]. Front Microbiol, 2021, 12: 692069.

[22] SARRY M, CAIGNARD G, DUPRÉ J, et al. Host-specific interplay between foot-and-mouth disease virus 3D polymerase and the type-I interferon pathway[J]. Viruses, 2023, 15(3): 666.

[23] RANJITHA H B, AMMANATHAN V, GULERIA N, et al. Foot-and-mouth disease virus induces PERK-mediated autophagy to suppress the antiviral interferon response[J]. J Cell Sci, 2020, 134(5): jcs240622.

[24] TANG J, TANG A, DU H, et al. Peste des petits ruminants virus exhibits cell-dependent interferon active response[J]. Front Cell Infect Microbiol, 2022, 12: 874936.

[25] BRAHMA A, JAS R, PATRA A K, et al. Characterization of interferon gamma gene in relation to immunological responses in Haemonchus contortus resistant and susceptible Garole sheep[J]. Vet Res Commun, 2023, 47(2): 599-614.

[26] SOLIMAN S M, ALJAHDA LI N H, ALI MAJRASHI K, et al. Immune cell reaction associated with coenurus cerebralis infection in sheep with particular reference to ELISA as a diagnostic tool[J]. Life, 2022, 12(10): 1515.

[27] YANG J, ZHAO Y, FU Y, et al. Recombinant antigen P29 of Echinococcus granulosus induces Th1, Tc1, and Th17 cell immune responses in sheep[J]. Front Immunol, 2023, 14: 1243204.

[28] WANG C, YANG S H, NIU N, et al. lncRNA028466 regulates Th1/Th2 cytokine expression and associates with Echinococcus granulosus antigen P29 immunity[J]. Parasit Vectors, 2021, 14(1): 295.

[29] RUIZ-CAMPILLO M T, BARRERO-TORRES D M, ABRIL N, et al. Fasciola hepatica primo infections and reinfections in sheep drive distinct Th1/Th2/Treg immune responses in liver and hepatic lymph node at early and late stages[J]. Vet Res, 2023, 54(1): 2.

[30] KUMAR K, KUMAR P, SINDHOORA K, et al. Detection and immune cell response of natural maedi visna virus (MVV) infection in Indian sheep and goats[J]. Microb Pathog, 2022, 165: 105467.

[31] BEASLEY A M, KAHN L P, WINDON R G. The periparturient relaxation of immunity in Merino ewes infected with Trichostrongylus colubriformis: parasitological and immunological responses[J]. Vet Parasitol, 2010, 168(1-2): 60-70.

[32] ALBUQUERQUE A C A, BASSETTO C C, ALMEIDA F A, et al. Differences in immune responses to Haemonchus contortus infection in the susceptible Ile de France and the resistant Santa Ines sheep under different anthelmintic treatments regimens[J]. Vet Res, 2019, 50(1): 104.

[33] FTHENAKIS G C, MAVROGIANNI V S, GALLIDIS E, et al. Interactions between parasitic infections and reproductive efficiency in sheep[J]. Vet Parasitol, 2015, 208(1-2): 56-66.

[34] DONSKOW-ŁYSONIEWSKA K, MARUSZEWSKA-CHERUIYOT M, KRAWCZAK-WÓJCIK K, et al. Nematode galectin binds IgE and modulates mast cell activity[J]. Vet Parasitol, 2022, 311: 109807.

[35] LALRAMLUNA M, BORDOLOI G, PANDIT S, et al. Parasitological and immunological response to Haemonchus contortus infection: comparison between resistant Garole and susceptible Sahabadi sheep[J]. Vet Parasitol Reg Stud Reports, 2020, 22: 100477.

[36] GUTIÉRREZ-AMÉZQUITA R A, MORALES-MONTOR J, MUÑOZ-GUZMÁN M A, et al. Progesterone inhibits the in vitro L3/L4 molting process in Haemonchus contortus[J]. Vet Parasitol, 2017, 248: 48-53.

[37] ALKALAL DEH M, GIBSON J, LEE S H, et al. Detection of genomic regions underlying resistance to gastrointestinal parasites in Australian sheep[J]. Genet Sel Evol, 2019, 51(1): 37.

[38] NARANJO-LUCENA A, CORREIA C N, MOLINA-HERNÁNDEZ V, et al. Transcriptomic analysis of ovine hepatic lymph node following Fasciola hepatica infection-inhibition of NK cell and IgE-mediated signaling[J]. Front Immunol, 2021, 12: 687579.

[39] DICKEY A M, SMITH T P L, CLAWSON M L, et al. Classification of small ruminant lentivirus subtype A2, subgroups 1 and 2 based on whole genome comparisons and complex recombination patterns[J]. F1000Res, 2020, 9: 1449.

[40] RAMÍREZ H, ECHEVERRÍA I, BENITO A A, et al. Accurate diagnosis of small ruminant lentivirus infection is needed for selection of resistant sheep through TMEM154 E35K genotyping[J]. Pathogens, 2021, 10(1): 83.

[41] MATERNIAK-KORNAS M, ROPKA-MOLIK K, PIÓRKOWSKA K, et al. Identification of new single nucleotide polymorphisms potentially related to small ruminant lentivirus infection susceptibility in goats based on data selected from high-throughput sequencing[J]. Pathogens, 2024, 13(10): 830.

[42] XIE S, ISAACS K, BECKER G, et al. A computational framework for improving genetic variants identification from 5,061 sheep sequencing data[J]. J Anim Sci Biotechnol, 2023, 14(1): 127.

[43] RODRIGUES C S, DE FARIA D A, LACERDA T S, et al. Lentivirus susceptibility in Brazilian and US sheep with TMEM154 mutations[J]. Genes, 2022, 14(1): 70.

[44] MORETTI R, SARTORE S, COLITTI B, et al. Susceptibility of different TMEM154 genotypes in three Italian sheep breeds infected by different SRLV genotypes[J]. Vet Res, 2022, 53(1): 60.

[45] MATERNIAK-KORNAS M, PIÓRKOWSKA K, ROPKA-MOLIK K, et al. First report of SNPs detection in TMEM154 gene in sheep from Poland and their association with SRLV infection status[J]. Pathogens, 2025, 14(1): 16.

[46] YAMAN Y, BAY V, AYMAZ R, et al. A novel 2 bp deletion variant in Ovine-DRB1 gene is associated with increased Visna/maedi susceptibility in Turkish sheep[J]. Sci Rep, 2021, 11(1): 14435.

[47] HU Y, GUDNADÓTTIR R B, KNECHT K M, et al. Structural basis for recruitment of host CyPA and E3 ubiquitin ligase by maedi-visna virus Vif[J]. Sci Adv, 2023, 9(2): eadd3422.

[48] LUPERCHIO A M, JÓNSSON S R, SALAMANGO D J. Evolutionary conservation of PP2A antagonism and G2/M cell cycle arrest in maedi-visna virus vif[J]. Viruses, 2022, 14(8): 1701.

[49] ARCANGELI C, LUCARELLI D, TORRICELLI M, et al. First survey of SNPs in TMEM154 TLR9 MYD88 and CCR5 genes in sheep reared in Italy and their association with resistance to SRLVs infection[J]. Viruses, 2021, 13(7): 1290.

[50] BENBRAIEK M, SZYM CZAK S, ANDRÉ C, et al. A single base pair duplication in the SLC33A1 gene is associated with fetal losses and neonatal lethality in Manech Tête Rousse dairy sheep[J]. Anim Genet, 2024, 55(4): 644-657.

[51] ZHANG H, WANG X, QU M, et al. Foot-and-mouth disease virus structural protein VP3 interacts with HDAC8 and promotes its autophagic degradation to facilitate viral replication[J]. Autophagy, 2023, 19(11): 2869-2883.

[52] SUN P, ZHANG S, QIN X, et al. Foot-and-mouth disease virus capsid protein VP2 activates the cellular EIF₂S1-ATF4 pathway and induces autophagy via HSPB1 [J]. Autophagy, 2018, 14(2): 336-346.

[53] ZHANG R, QIN X, YANG Y, et al. STING1 is essential for an RNA-virus triggered autophagy[J]. Autophagy, 2022, 18(4): 816-828.

[54] ZHANG L, XIAO H, HUANG J, et al. Identification and expression analysis of the β-defensin genes in the goat small intestine[J]. Gene, 2021, 801: 145846.

[55] GE L, ZOU S, YUAN Z, et al. Sheep β-defensin 2 regulates Escherichia coli F17 resistance via NF-κB and MAPK signaling pathways in ovine intestinal epithelial cells[J]. Biology, 2021, 10(12): 1356.

[56] MI X, WU L, SONG Y, et al. Evolutionary dynamics and regulatory site analysis of AMP family genes in cattle and sheep[J]. Int J Biol Macromol, 2025, 290: 138922.

[57] HAQ R I U, PARRAY O R, NAZIR Q U A, et al. Immune and oxidative response against sonicated antigen of Mycoplasma capricolum subspecies capripneumonia-a causative agent of contagious caprine pleuropneumonia[J]. Microorganisms, 2022, 10(8): 1634.

[58] MA W T, GU K, YANG R, et al. Interleukin-17 mediates lung injury by promoting neutrophil accumulation during the development of contagious caprine pleuropneumonia[J]. Vet Microbiol, 2020, 243: 108651.

[59] 马宗武． 基于全基因组关联分析筛选与湖羊性情相关的候选基因[D]． 兰州： 甘肃农业大学, 2024．

[60] ZHANG M, WANG Y, CHEN Q, et al. Genome-wide association study on body conformation traits in Xinjiang brown cattle[J]. Int J Mol Sci, 2024, 25(19): 10557.

[61] ARZIK Y, KIZILASLAN M, WHITE S N, et al. Genomic analysis of gastrointestinal parasite resistance in akkaraman sheep[J]. Genes, 2022, 13(12): 2177.

[62] THORNE J W, REDDEN R, BOWDRIDGE S A, et al. Genome-wide analysis of sheep artificially or naturally infected with gastrointestinal nematodes[J]. Genes, 2023, 14(7): 1342.

[63] MOHAMMADI H, FARAHANI A H K, MORADI M H, et al. Weighted single-step genome-wide association study uncovers known and novel candidate genomic regions for milk production traits and somatic cell score in Valle del belice dairy sheep[J]. Animals, 2022, 12(9): 1155.

[64] ARGYRIADOU A, MICHAILIDOU S, VOURAKI S, et al. A genome-wide association study reveals novel SNP markers associated with resilience traits in two Mediterranean dairy sheep breeds[J]. Front Genet, 2023, 14: 1294573.

[65] YANG H, ZHU M, WANG M, et al. Genome-wide comparative analysis reveals selection signatures for reproduction traits in prolific Suffolk sheep[J]. Front Genet, 2024, 15: 1404031.

[66] MA C, LI M, PENG H, et al. Mesomycoplasma ovi pneumoniae from goats with respiratory infection: pathogenic characteristics, population structure, and genomic features[J]. BMC Microbiol, 2023, 23(1): 220.

[67] HUI W, JIANG S, LIU X, et al. Gene expression profile in the liver of sheep infected with cystic echinococcosis[J]. PLoS One, 2016, 11(7): e0160000.

[68] NICIURA S C M, CARDOSO T F, IBELLI A M G, et al. Multi-omics data elucidate parasite-host-microbiota interactions and resistance to Haemonchus contortus in sheep[J]. Parasit Vectors, 2024, 17(1): 102.

[69] SHI X, ZHANG Y, CHEN S, et al. Differential gene expression and immune cell infiltration in maedi-visna virus-infected lung tissues[J]. BMC Genomics, 2024, 25(1): 534.

[70] ZHANG H, WANG F, CHEN H, et al. Comparative RNA sequencing analysis of three Capripoxvirus infections in an immortalized hTERT-bOEC cell model[J]. Virology, 2025, 603: 110352.

[71] MAI J, QIAN Q, GAO H, et al. scTWAS Atlas: an integrative knowledgebase of single-cell transcriptome-wide association studies[J]. Nucleic Acids Res, 2025, 53(D1): D1195-D1204.

[72] LIM J, PARK C, KIM M, et al. Advances in single-cell omics and multiomics for high-resolution molecular profiling[J]. Exp Mol Med, 2024, 56(3): 515-526.

[73] KE M, ELSHENAWY B, SHELDON H, et al. Single cell RNA-sequencing: a powerful yet still challenging technology to study cellular heterogeneity[J]. BioEssays, 2022, 44(11): 2200084.

[74] WANG Y, YANG S, HAN B, et al. Single-cell landscape revealed immune characteristics associated with disease phases in brucellosis patients[J]. iMeta, 2024, 3(4): e226.

[75] JIA G X, MA W J, WU Z B, et al. Single-cell transcriptomic characterization of sheep conceptus elongation and implantation[J]. Cell Rep, 2023, 42(8): 112860.

[76] SUN S, SHEN X, LI Y, et al. Single-cell RNA sequencing provides a high-resolution roadmap for understanding the multicellular compartmentation of specialized metabolism[J]. Nat Plants, 2023, 9(1): 179-190.

[77] DAVIES C J, FAN Z, MORGADO K P, et al. Development and characterization of type I interferon receptor knockout sheep: a model for viral immunology and reproductive signaling[J]. Front Genet, 2022, 13: 986316.

[78] YU Y, ZHANG F, DUAN X, et al. ORFV can carry TRAP gene expression via intracellular CRISPR/Cas9 gene editing technology[J]. J Virol Meth, 2023, 312: 114652.

[79] YANG H, LIU Z, LÜ W, et al. Construction and validation of sheep VASA gene knock-in vector based on CRISPR/Cas9 system[J]. Chin J Biotechnol, 2023, 39(10): 4219-4233.

[80] 马秀玲, 张欣如, 陈莹, 等． 阿勒泰羊胚胎PDGFD基因编辑研究[J]． 畜牧兽医学报, 2025, 56(4)： 1700-1711．

基金资助

国家科技重大专项(2023ZD0405104)

中央引导地方科技发展专项(ZYYD2023B14)

国家重点研发计划项目(2021YFD1600701-3)