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本研究通过转录组测序技术研究猪链球菌(Streptococcus suis)经H_2O2处理后的全基因组转录水平变化，探究氧胁迫下与猪链球菌应对氧化应激过程相关的代谢通路和基因。将9型猪链球菌DN13菌株培养至对数生长后期，用PBS和H_2O2分别处理15 min,将处理后的DN13菌株进行转录组测序(RNA-seq),对转录本中的差异表达基因进行KEGG和GO分析，并用实时定量PCR(real-time quantitative PCR, RT-qPCR)验证RNA-seq的分析结果。RNA-seq结果显示，DN13共检测到2 005个基因，与对照组相比，实验组有749个基因差异表达，占比37.4%。GO富集结果表明，差异表达基因在分子功能、生物学过程和细胞组分这3大类中均显著富集。KEGG富集结果表明，差异表达基因主要集中在核糖体、 ABC转运体、氨基糖和核苷酸糖的代谢以及半乳糖代谢等代谢途径中。RT-qPCR结果与RNA-seq数据的一致性好，证明RNA-seq分析结果具有可靠性。这说明猪链球菌应对氧化应激是一个涉及细菌多个途径和一系列基因表达变化的复杂过程。通过分析实验组和对照组之间DN13菌株的基因表达水平，发现在本研究中与抗氧化相关的差异表达基因的表达变化均与其功能一致，且均注释到催化活性、细胞结构体和结合这3个GO条目中，这说明转录组分析的数据可在整体上准确反映参与抗氧化应激基因表达水平的变化。
Abstract：In this study, we investigated the changes in genome-wide transcript levels of Streptococcus suis after H_2O2 treatment by transcriptome sequencing to explore the metabolic pathways and genes associated with the process of oxidative stress response in Streptococcus suis under oxidative stress. The strain DN13 of S. suis type 9 was cultured to the late logarithmic phase and treated with PBS and H_2O2 for 15 min. The transcriptome sequencing(RNA-seq) of the treated strain DN13 was performed. The differentially expressed genes in the transcripts were analyzed by KEGG and GO, and the results of RNA-seq were verified by RT-qPCR. The RNA-seq results showed that a total of 2 005 genes were detected in DN13, and 749 genes were differentially expressed in the experimental group compared to the control group, accounting for 37.4%. The GO enrichment results showed that differentially expressed genes were significantly enriched in three major categories: molecular function, biological process and cellular component. The KEGG enrichment results showed that the differentially expressed genes were mainly concentrated in the ribosome, ABC transporter, amino sugar and nucleotide sugar metabolism, and galactose metabolism metabolic pathways. The RT-qPCR results were consistent with the RNA-seq data, which proved that the RNA-seq analysis results were reliable. This suggests that the response of Streptococcus suis to oxidative stress is a complex process involving multiple bacterial pathways and a series of changes in gene expression. Analysis of gene expression levels of DN13 strains between the experimental group and control group revealed that the expression changes of differentially expressed genes associated with antioxidant in this study were all consistent with their functions and were all annotated to the three GO entries of cataly-tic activity, cellular anatomical entity and binding, suggesting that the data from transcriptome analysis can accurately reflect the changes in expression levels of genes involved in resistance to oxidative stress as a whole.
龚非力，尹丙娇，2014.医学免疫学.第4版.北京：科学出版社：306-307.(GONG F L,YIN B J,2014.Medical immunology.4th edition.Beijing:Science Press:306-307.)
胡玉立，2019.SrtR的鉴定及其在猪链球菌中抗氧化应激及毒力的作用研究，博士学位论文.长沙：湖南农业大学.(HU Y L,2019.Identification and characterization of the regulatory roles of SrtR in oxidative stress response and virulence in Streptococcus suis,Dissertation for Ph.D.Changsha:Hunan Agricultural University.)
谭美芳，李海琴，曾艳兵，等，2022.外源糖原调控猪链球菌基因表达的转录组分析.微生物学通报，49(5):1690-1704.[TAN M F,LI H Q,ZENG Y B,et al.,2022.Transcriptome analysis of exogenous glycogen availability regulating gene expression of Streptococcus suis.Microbiology China,49(5):1690-1704.]
王硕玥，梁子敬，陈琳，等，2020.猪链球菌抗氧化应激蛋白YED和AHH的鉴定.畜牧与兽医，52(5):81-86.[WANG S Y,LIANG Z J,CHEN L,et al.,2020.Identification of YED and AHH proteins of Streptococcus suis involved in oxidative stress response.Animal Husbandry & Veterinary Medicine,52(5):81-86.]
ARANDA J,GARRIDO M E,FITTIPALDI N,et al.,2010.The cation-uptake regulators AdcR and Fur are necessary for full virulence of Streptococcus suis.Vet.Microbiol.,144(1-2):246-249.
BREKASIS D,PAGET M S,2003.A novel sensor of NADH/NAD+ redox poise in Streptomyces coelicolor A3(2).EMBO J.,22(18):4856-4865.
CHANG P X,LI W T,SHI G L,et al.,2018.The VraSR regulatory system contributes to virulence in Streptococcus suis via resistance to innate immune defenses.Virulence,9(1):681-682.
CHASTANET A,PRUDHOMME M,CLAVERYS J P,et al.,2001.Regulation of Streptococcus pneumoniae clp genes and their role in competence development and stress survival.J.Bacteriol.,183(24):7295-7307.
CUAJUNGCO M,RAMIREZ M,TOLMASKY M,2021.Zinc:multidimensional effects on living organisms.Biomedicines,9(2):208.
EZRATY B,GENNARIS A,BARRAS F,et al.,2017.Oxidative stress,protein damage and repair in bacteria.Nat.Rev.Microbiol.,15(7):385-396.
FITTIPALDI N,SEGURA M,GRENIER D,et al.,2012.Virulence factors involved in the pathogenesis of the infection caused by the swine pathogen and zoonotic agent Streptococcus suis.Future Microbiol.,7(2):259-279.
FUJISHIMA K,KAWADA-MATSUO M,OOGAI Y,et al.,2013.dpr and sodin Streptococcus mutans are involved in coexistence with S.sanguinis,and PerR is associated with resistance to H2O2.Appl.Environ.Microbiol.,79(5):1436-1443.
GARCIA Y M,BARWINSKA-SENDRA A,TARRANT E,et al.,2017.A superoxide dismutase capable of functioning with iron or manganese promotes the resistance of Staphylococcus aureus to calprotectin and nutritional immunity.PLoS Pathog.,13(1):e1006125.
HAN H M,LIU C H,WANG Q H,et al.,2012.The two-component system Ihk/Irr contributes to the virulence of Streptococcus.Microbiology (Reading),158(Pt 7):1852-1866.
HUGHES J M,WILSON M E,WERTHEIM H F L,et al.,2009.Streptococcus suis:an emerging human pathogen.Clin.Infect.Dis.,48(5):617-625.
JIANG X W,YANG Y K,ZHOU J J,et al.,2016.Roles of the putative type Ⅳ-like secretion system key component VirD4 and PrsA in pathogenesis of Streptococcus suis type 2.Front.Cell.Infect.Microbiol.,6:172.
K?ADNA A,MARCHLEWICZ M,PIECHOWSKA T,et al.,2015.Reactivity of pyruvic acid and its derivatives towards reactive oxygen species.Luminescence,30(7):1153-1158.
LI J P,WANG Y X,DU Y B,et al.,2021.mRNA-Seq reveals the quorum sensing system luxS gene contributes to the environmental fitness of Streptococcus suis type 2.BMC Microbiology,21(1):111.
LIANG M C,WANG Z X,LI H,et al.,2018.L-Arginine induces antioxidant response to prevent oxidative stress via stimulation of glutathione synthesis and activation of Nrf2 pathway.Food Chem.Toxicol.,115:315-328.
LIANG P J,WANG M L,GOTTSCHALK M,et al.,2021.Genomic and pathogenic investigations of Streptococcus suis serotype 7 population derived from a human patient and pigs.Emerg.Microbes Infect.,10(1):1960-1974.
MéNDEZ V,RODRíGUEZ-CASTRO L,DURáN R E,et al.,2022.The OxyR and SoxR transcriptional regulators are involved in a broad oxidative stress response in Paraburkholderia xenovorans LB400.Biol.Res.,55(1):7.
PENG W,YANG X A,WANG N N,et al.,2022.PerR-regulated manganese import contributes to oxidative stress defense in Streptococcus suis.Appl.Environ.Microbiol.,88(9):e8622.
RADA B,LETO T L,2008.Oxidative innate immune defenses by Nox/Duox family NADPH oxidases.Contrib.Microbiol.,15:164-187.
ROY S,CHIKKERUR J,ROY S C,et al.,2018.Tagatose as a potential nutraceutical:production,properties,biological roles,and applications.J.Food Sci.,83(11):2699-2709.
ROY S,ZHU Y C,MA J L,et al.,2019.Role of ClpX and ClpP in Streptococcus suis serotype 2 stress tolerance and virulence.Microbiol.Res.,223-225:99-109.
SOBOTA J M,IMLAY J A,2011.Iron enzyme ribulose-5-phosphate 3-epimerase in Escherichia coli is rapidly damaged by hydrogen peroxide but can be protected by manganese.Proc.Natl.Acad.Sci.USA,108(13):5402-5407.
TAN M F,LIU W Q,ZHANG C Y,et al.,2017.The involvement of MsmK in pathogenesis of the Streptococcus suis serotype 2.MicrobiologyOpen,6(2):e00433.
TAN M F,TAN J A,ZHANG F F,et al.,2022.Exogenous glycogen utilization effects the transcriptome and pathogenicity of Streptococcus suis serotype 2.Front.Cell.Infect.Microbiol.,12:938286.
VAN DER HEIJDEN J,REYNOLDS L A,DENG W Y,et al.,2016.Salmonella rapidly regulates membrane permeability to survive oxidative stress.mBio,7(4):e01238.
WANG Y,WANG Y X,LIU B B,et al.,2019.pdh modulate virulence through reducing stress tolerance and biofilm formation of Streptococcus suis serotype 2.Virulence,10(1):588-599.
WICHGERS SCHREUR P J,REBEL J M,SMITS M A,et al.,2011.TroA of Streptococcus suis is required for manganese acquisition and full virulence.J.Bacteriol.,193(19):5073-5080.
YU H,JING H,CHEN Z,et al.,2006.Human Streptococcus suis outbreak,Sichuan,China.Emerg.Infect.Dis.,12(6):914-920.
ZHENG C K,JIA M D,GAO M M,et al.,2019.PmtA functions as a ferrous iron and cobalt efflux pump in Streptococcus suis.Emerg.Microbes Infect.,8(1):1254-1264.
ZHENG C K,XU J L,LI J Q,et al.,2014.Two spx regulators modulate stress tolerance and virulence in Streptococcus suis serotype 2.PLoS ONE,9(9):e108197.
基本信息:
DOI：10.13417/j.gab.042.000518
中图分类号:S858.28
引用信息:
[1]曹锟,杨磊,杨鹏江,等.基于转录组分析猪链球菌抗氧化应激相关基因表达的变化[J].基因组学与应用生物学,2023,42(05):518-527.DOI:10.13417/j.gab.042.000518.
基金信息:
国家自然科学基金面上项目(31972713)资助
2023-02-21
2023-02-21
2023-02-21