Meng Huimin, Wang Zhangxun, Wang Yulong, Zhu Hong, Huang Bo
Anhui Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei, China.
Shandong Provincial Key Laboratory of Synthetic Biology, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, China.
Appl Environ Microbiol. 2017 Mar 17;83(7). doi: 10.1128/AEM.03230-16. Print 2017 Apr 1.
RNA interference (RNAi) is a gene-silencing mechanism that plays an important role in gene regulation in a number of eukaryotic organisms. Two core components, Dicer and Argonaute, are central in the RNAi machinery. However, the physiological roles of Dicer and Argonaute in the entomopathogenic fungus have remained unclear. Here, the roles of genes encoding Dicer ( [] and ) and Argonaute ( and ) proteins in were investigated. The results showed that the Dicer-like protein MrDCL2 and Argonaute protein MrAGO1 are the major components of the RNAi process occurring in The Dicer and Argonaute genes were not involved in the regulation of growth and diverse abiotic stress response in under the tested conditions. Moreover, our results showed that the Dicer and Argonaute gene mutants demonstrated reduced abilities to produce conidia, compared to the wild type (WT) and the gene-rescued mutant. In particular, the conidial yields in the Δ and Δ mutants were reduced by 55.8% and 59.3%, respectively, compared with those from the control strains. Subsequently, for the WT and Δ mutant strains, digital gene expression (DGE) profiling analysis of the stage of mycelium growth and conidiogenesis revealed that modest changes occur in development or metabolism processes, which may explain the reduction in conidiation in the Δ mutant. In addition, we further applied high-throughput sequencing technology to identify small RNAs (sRNAs) that are differentially expressed in the WT and the Δ mutant and found that 4 known microRNA-like small RNAs (milRNAs) and 8 novel milRNAs were dependent in The identification and characterization of components in RNAi have contributed significantly to our understanding of the mechanism and functions of RNAi in eukaryotes. Here, we found that Dicer and Argonaute genes play an important role in regulating conidiation in Our study also demonstrates that diverse small RNA pathways exist in The study provides a theoretical platform for exploration of the functions of Dicer and Argonaute genes involved in RNAi in fungi.
RNA干扰（RNAi）是一种基因沉默机制，在许多真核生物的基因调控中发挥着重要作用。Dicer和Argonaute这两个核心成分是RNAi机制的核心。然而，Dicer和Argonaute在昆虫病原真菌中的生理作用仍不清楚。在此，研究了编码Dicer（[]和）和Argonaute（和）蛋白的基因在中的作用。结果表明，类Dicer蛋白MrDCL2和Argonaute蛋白MrAGO1是中发生的RNAi过程的主要成分。在测试条件下，Dicer和Argonaute基因不参与的生长调节和多种非生物胁迫反应。此外，我们的结果表明，与野生型（WT）和基因拯救突变体相比，Dicer和Argonaute基因突变体产生分生孢子的能力降低。特别是，与对照菌株相比，Δ和Δ突变体的分生孢子产量分别降低了55.8%和59.3%。随后，对WT和Δ突变体菌株进行了菌丝体生长和分生孢子形成阶段的数字基因表达（DGE）谱分析，结果表明，发育或代谢过程中发生了适度变化，这可能解释了Δ突变体中分生孢子形成减少的原因。此外，我们进一步应用高通量测序技术鉴定了WT和Δ突变体中差异表达的小RNA（sRNA），发现4种已知的微小RNA样小RNA（milRNA）和8种新的milRNA在中依赖。RNAi中成分的鉴定和表征极大地促进了我们对真核生物中RNAi机制和功能的理解。在此，我们发现Dicer和Argonaute基因在调节中的分生孢子形成中起重要作用。我们的研究还表明中存在多种小RNA途径。该研究为探索真菌中参与RNAi的Dicer和Argonaute基因的功能提供了理论平台。
