【摘要】 Argonaute蛋白(AGO)介导的沉默复合体在RNA干扰(RNAi)中起着至关重要的作用。为了探究AGO1在尖孢镰刀菌RNAi中的作用机制,本文以粘团专化型尖孢镰刀菌生理1号小种FOX-A8野生型和其AGO1缺失突变体(FOX-A8-△Ago1)的菌丝和孢子为材料,分别进行了RNA提取、Illumina Hi Seq 2000高通量转录组测序、差异表达基因(DEGs)的显著富集分析;选择菌丝和孢子中的DEGs进行实时荧光定量PCR(Quantitative real-time PCR,qRT-PCR)验证。GO通路注释结果显示,相对于野生型菌株,AGO1缺失突变体菌丝中的醇脱氢酶(NADP~+)、孢子中的CAMK/CAMKL/CHK 1蛋白激酶均显著上调; KEGG通路注释结果显示,相对于野生型菌株,AGO1缺失突变体菌丝中与MAPK信号通路相关的基因、孢子中与PLD信号通路相关的基因均显著下调;另外,相对于野生型菌株,编码AGO2的基因下调,但是下调不显著。qRT-PCR检测DEGs的表达模式与RNA-Seq分析结果一致,证实了RNA-Seq结果的可靠性。更多还原

【Abstract】 The Argonaute protein(AGO) mediated silencing complex plays a crucial role in RNA interference(RNAi). In order to explore the function of AGO1 in Fusarium oxysporum,F. oxysporum f. sp. Conglutinas race 1 FOX-A8 strain and the Ago1 deletion mutant(FOX-A8-△Ago1) were used as materials for RNA extraction from spores and mycelia. Transcriptome sequencing was conducted via Illumina Hi Seq 2000 platform and significant enrichment analysis of differentially expressed genes(DEGs) was performed by the RNA-seq analysis pipeline; The selected DEGs in hyphae and spores were validated by quantitative real-time PCR(qRT-PCR) approach. The results of GO annotation showed that the alcohol dehydrogenase(NADP~+) in mycelia of the AGO1 deletion mutant and the CAMK/CAMKL/CHK 1 protein kinase in the spore of the AGO1 deletion mutant were significantly up-regulated; The results of the KEGG pathway annotation showed that the genes associated with the MAPK in mycelia of the AGO1 mutant and the PLD signaling pathway in the spore of the AGO1 deletion mutant were significantly down-regulated compared to the wild-type strain; In addition,the gene encoding AGO2 was relatively down-regulated in compare with the wild-type strain,but not significant. The expression pattern of DEGs detected by qRT-PCR was consistent with the results of RNA-Seq analysis,confirming the reliability of the transcriptome data.更多还原