摘要
为明确N6-甲基腺苷(N6-methyladenosine,m^(6) A)修饰在棉铃虫核型多角体病毒(Helicoverpa armigera nucleopolyhedrovirus,HearNPV)侵染棉铃虫中的作用,根据基因组和转录组数据,对棉铃虫m^(6) A结合蛋白基因YTHDF1(YTH domain-containing family protein 1)进行鉴定和分析,利用实时荧光定量PCR技术检测棉铃虫YTHDF1基因的时空表达模式、HearNPV处理后的表达变化情况以及RNA干扰效率,并调查该基因下调后对HearNPV复制及感染HearNPV棉铃虫死亡情况的影响。结果显示,棉铃虫YTHDF1基因开放阅读框全长为2019 bp,编码672个氨基酸,含有1个保守的YTH结构域,其氨基酸序列与斜纹夜蛾Spodoptera litura同源物序列一致性达87.52%。YTHDF1基因在棉铃虫不同发育阶段均有表达,其中在进入5龄期24 h幼虫中的表达量最高,在2龄取食期幼虫中的表达量最低。YTHDF1基因的空间表达谱呈现一定的组织特异性,在幼虫血细胞和成虫头部的表达量最高。HearNPV侵染棉铃虫后YTHDF1基因上调表达,经RNA干扰使该基因下调后显著抑制了HearNPV多角体蛋白基因polyhedrin的表达并延迟了感染HearNPV棉铃虫的死亡时间。表明YTHDF1基因在HearNPV侵染棉铃虫的过程中发挥着重要作用。
In order to investigate the function of N6-methyladenosine(m^(6) A)modification in cotton bollworm Helicoverpa armigera infected by H.armigera nucleopolyhedrovirus(HearNPV),YTH domaincontaining family protein 1(YTHDF1),a m^(6) A binding protein,was identified and analyzed based on genome and transcriptome data.Quantitative real-time PCR was used to detect the temporal and spatial expression patterns,the expression level changes after HearNPV treatment,and the RNA interference efficiency of YTHDF1 gene in H.armigera.Furthermore,the effects of YTHDF1 knockdown on HearNPV replication and mortality of H.armigera infected by HearNPV were investigated.The results showed that the open reading frame of YTHDF1 gene in H.armigera was 2019 bp,encoding 672 amino acids,which contained a conserved YTH domain.Homology and phylogenetic analyses showed that the amino acid sequence of YTHDF1 had the highest identity(87.52%)with that of Spodoptera litura.The results of temporal transcript profiling indicated that the YTHDF1 gene was expressed in different developmental stages of H.armigera,and the expression level was the highest at 24 h after entering the 5th instar,while the expression level was the lowest in the feeding period of the 2nd instar larvae.YTHDF1 was specifically expressed in different tissues of H.armigera,mainly in larval hemocytes and adult heads.In addition,the expression of the YTHDF1 gene in H.armigera was upregulated by HearNPV infection;the expression of polyhedrin was inhibited,and the death time of H.armigera infected by HearNPV was delayed after knockdown of the gene.These results confirmed that YTHDF1 plays an important role in the process of HearNPV infecting H.armigera.
作者
申忠健
刘彦君
祝琳
田志强
刘孝明
刘小侠
Shen Zhongjian;Liu Yanjun;Zhu Lin;Tian Zhiqiang;Liu Xiaoming;Liu Xiaoxia(Department of Entomology,College of Plant Protection,China Agricultural University,Beijing 100193,China)
出处
《植物保护学报》
CAS
CSCD
北大核心
2021年第5期1026-1033,共8页
Journal of Plant Protection
基金
国家自然科学基金(31972275)。
