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高通量测序揭示杨树防御过程中转录组和小RNA的调控网络

High-Throughput Sequencing Reveals the Regulatory Networks of Transcriptome and Small RNAs During the Defense Against in Poplar.

作者信息

Liao Yangwenke, Zhang Qingyue, Cui Rongrong, Xu Xin, Zhu Fuyuan, Cheng Qiang, Li Xiaogang

机构信息

Co-Innovation Center for Sustainable Forestry in Southern China, College of Biology and the Environment, Nanjing Forestry University, Nanjing, China.

出版信息

Front Plant Sci. 2021 Sep 8;12:719549. doi: 10.3389/fpls.2021.719549. eCollection 2021.

Abstract

MicroRNAs are implicated in the adjustment of gene expression in plant response to biotic stresses. However, the regulatory networks of transcriptome and miRNAs are still poorly understood. In the present study, we ascertained the induction of genes for small RNA biosynthesis in poplar defense to a hemibiotrophic fungus and afterward investigated the molecular regulatory networks by performing comprehensive sequencing analysis of mRNAs and small RNAs in -inoculated leaves. Differentially expressed genes in -infected poplar are mainly involved in secondary metabolisms, phytohormone pathways, the recognition of pathogens, and MAPK pathway in the plant, with real-time quantitative PCR (qPCR) validating the mRNA-seq results. Furthermore, differentially expressed miRNAs, such as MIR167_1-6, MIR167_1-12, MIR171_2-3, MIR395-13, MIR396-3, MIR396-16, MIR398-8, and MIR477-6, were identified. Through psRobot and TargetFinder programs, MIR167-1-6, MIR395-13, MIR396-3, MIR396-16, and MIR398-8 were annotated to modulate the expression of genes implicated in transportation, signaling, and biological responses of phytohormones and activation of antioxidants for plant immunity. Besides, validated differentially expressed genes involved in lignin generation, which were , and , were selected as targets for the identification of novel miRNAs. Correspondingly, novel miRNAs, such as Novel MIR8567, Novel MIR3228, Novel MIR5913, and Novel MIR6493, were identified using the Mireap online program, which functions in the transcriptional regulation of lignin biosynthesis for poplar anti-fungal response. The present study underlines the roles of miRNAs in the regulation of transcriptome in the anti-fungal response of poplar and provides a new idea for molecular breeding of woody plants.

摘要

微小RNA参与植物对生物胁迫响应中基因表达的调控。然而,转录组和微小RNA的调控网络仍知之甚少。在本研究中,我们确定了杨树在抵御半活体营养型真菌时小RNA生物合成相关基因的诱导情况,随后通过对接种叶片中的mRNA和小RNA进行全面测序分析来研究分子调控网络。感染真菌的杨树中差异表达基因主要参与次生代谢、植物激素途径、病原体识别以及植物中的丝裂原活化蛋白激酶途径,实时定量PCR(qPCR)验证了mRNA测序结果。此外,还鉴定出了差异表达的微小RNA,如MIR167_1 - 6、MIR167_1 - 12、MIR171_2 - 3、MIR395 - 13、MIR396 - 3、MIR396 - 16、MIR398 - 8和MIR477 - 6。通过psRobot和TargetFinder程序,MIR167 - 1 - 6、MIR395 - 13、MIR396 - 3、MIR396 - 16和MIR398 - 8被注释为可调节与植物激素运输、信号传导和生物反应以及植物免疫抗氧化剂激活相关基因的表达。此外,选择参与木质素生成的已验证差异表达基因(分别为 、 和 )作为鉴定新型微小RNA的靶标。相应地,使用Mireap在线程序鉴定出了新型微小RNA,如新型MIR8567、新型MIR3228、新型MIR5913和新型MIR6493,它们在杨树抗真菌反应中木质素生物合成的转录调控中发挥作用。本研究强调了微小RNA在杨树抗真菌反应中转录组调控中的作用,并为木本植物分子育种提供了新思路。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/c3b7/8456019/54ae0cc7373e/fpls-12-719549-g0001.jpg

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