禽源Klebsiella variicola引起宿主血流感染的SNP筛选分析  

SNP screening and analysis of host bloodstream infection caused by avian Klebsiella variicola

作　　者：尹磊 沈学怀[1] 张丹俊[1] 赵瑞宏[1] 戴银[1] 胡晓苗[1] 周学利[1] 潘孝成[1] YIN Lei;SHEN Xuehuai;ZHANG Danjun;ZHAO Ruihong;DAI Yin;HU Xiaomiao;ZHOU Xueli;PAN Xiaocheng(Anhui Province Key Laboratory of Livestock and Poultry Product Safety Engineering,Livestock and Poultry Epidemic Diseases Research Center of Anhui Province,Institute of Animal Husbandry and Veterinary Science,Anhui Academy of Agricultural Science,Hefei 230031,China)

机构地区：[1]安徽省农业科学院畜牧兽医研究所安徽省畜禽疫病研究中心,畜禽产品安全工程安徽省重点实验室,安徽合肥230031

出　　处：《中国兽医学报》2021年第4期676-682,共7页Chinese Journal of Veterinary Science

基　　金：畜禽疫病研究中心院平台资助项目(2020YL094)。

摘　　要：旨在筛选和分析禽源Klebsiella variicola引起宿主血流感染的转录组单核苷酸多态性(SNP)位点。采用转录组学测序(RNA-Seq)技术比较了Klebsiella variicola在LB肉汤培养基和SPF鸡血清中的转录组SNP,筛选出Klebsiella variicola在宿主血清中生存和发展相关的SNP位点,对筛选出的SNP的碱基变异类型和所在位置类型进行注释分析,同时对筛选出的SNP位点对应的基因进行GO富集和KEGG通路富集分析,进一步探究其功能和涉及的生物信号通路;此外,随机选取7个位于富集通路中的基因,通过荧光定量PCR比较LB组与SPF血清组各基因mRNA转录水平。结果显示,相较于LB培养条件下,在血清中的Klebsiella variicola共获得143682个SNP位点,其中检测到单个碱基的转换(transition)有100790个,颠换(transversion)42890个,转换类型总数极显著高于颠换类型总数(P<0.01)。此外,对获得的SNP位点所在区域进行统计发现SNP主要集中在外显子区域(131093个)。筛选出的SNP位点对应的基因主要富集在ABC转运蛋白,二元调控系统,群体感应系统,脂多糖的生物合成等通路。荧光定量PCR结果表明相较于LB组,SPF血清组中的基因mRNA转录水平显著上调(P<0.01)。结果表明,本研究对禽源Klebsiella variicola在宿主血液环境中生存和发展的分子机制进行了初步分析和探索,为进一步研究Klebsiella variicola的致病机制提供理论依据。The purpose of this study was to screen and analyze transcriptome single nucleotide polymorphism(SNP)sites of host bloodstream infection caused by avian Klebsiella variicola.The transcriptome SNP of Klebsiella variicolain in LB broth medium and SPF chicken serum was compared by transcriptome sequencing(RNA-Seq).The SNP sites related to the survival and development of Klebsiella variicola in host serum were screened,and the base variation types and location types of SNP were annotated.At the same time,the genes corresponding to the screened SNP sites were analyzed by GO enrichment and KEGG pathway enrichment,their function and the biological signal pathways involved were further explored.In addition,seven genes located in the enrichment pathway were randomly selected and the mRNA transcription levels of genes in LB group and SPF serum group were compared by real-time PCR.The results showed that compared with LB culture,a total of 143682SNP sites were obtained in serumKlebsiella variicola,of which 100790were detected for single base transition(transition)and 42890for transversion(transversion).The total number of conversion types was significantly higher than that of transversion types(P<0.01).In addition,according to the statistics of the region where the SNP loci were located,it was found that SNP was mainly concentrated in the exon region(131093).The genes corresponding to the selected SNP loci were mainly enriched in ABC transporter,binary regulation system,quorum sensing system,lipopolysaccharide biosynthesis and other pathways.The results of realtime PCR showed that the level of mRNA transcription in SPF serum group were significantly upregulated compared with LB group(P<0.01).This study makes a preliminary analysis and exploration on the molecular mechanism of the survival and development of avian Klebsiellavariicola in the host blood environment,which provides a theoretical basis for further study of the pathogenic mechanism of Klebsiella variicola.

关 键 词：Klebsiella variicola 血流感染 RNA-SEQ SNP位点 

分 类 号：S852.61[农业科学—基础兽医学] R535[农业科学—兽医学]