尖孢镰孢菌与番茄互作早期milRNA调控功能研究  

Study on the regulatory function of milRNA at early stage of the interaction between Fusarium oxysporum and tomato

作　　者：畅引东 贾仡伟 何瑞 张萌[1,2] 孟广军 李新凤 张作刚[1] 王建明[1] Chang Yindong;Jia Yiwei;He Rui;Zhang Meng;Meng Guangjun;Li Xinfeng;Zhang Zuo-gang;Wang Jianming(College of Plant Protection,Shanxi Agricultural University,Shanxi Key Laboratory of Integrated Pest Management in Ag-riculture,Taiyuan 030801,China;Shanxi Plant Protection and Quarantine Centre,Taiyuan 030001,China)

机构地区：[1]山西农业大学植物保护学院农业有害生物综合治理山西省重点实验室,山西太原030031 [2]山西省植物保护植物检疫中心,山西太原030001

出　　处：《山西农业大学学报（自然科学版）》2022年第6期102-112,共11页Journal of Shanxi Agricultural University(Natural Science Edition)

基　　金：山西省优秀博士来晋工作奖励资金科研项目(SXYBKY2018045);山西省重点研发计划项目(201903D211001-1)。

摘　　要：[目的]MicroRNA通过调控其靶基因在生物活动中起到重要的作用,本研究目的是为了明确尖孢镰孢菌与番茄互作早期相关milRNAs的分子功能及调控机制。[方法]采用实时荧光定量PCR法对尖孢镰孢菌番茄专化型菌株侵染寄主早期pri-milRNAs和milRNAs靶基因进行基因表达分析,利用STRING蛋白互作网络数据库和Cytoscape软件预测该菌株milRNAs与靶基因间的蛋白互作网络,并结合DAVID在线分析工具对相关基因进行生物学功能分析。[结果]实时荧光定量PCR结果表明,供试的pri-milRNA_2a和pri-milRNA_2b与FOXG_03470、pri-milRNA_3b与FOXG_04910、pri-milRNA_4与FOXG_08812、pri-milRNA_6与FOXG_00067表达趋势基本相反,4个miLRNA与相应基因可能存在靶标关系;蛋白互作网络预测结果可知,尖孢镰孢菌fox_milRNA_1、fox_milRNA_2、fox_milRNA_3和fox_milRNA_8的靶基因间互作较为紧密。其中,fox_milRNA_1的靶基因FOXG_04240与fox_milRNA_2的靶基因FOXG_03470能够同该蛋白互作网络中诸多的基因进行互作,推测为该蛋白互作网络中的枢纽基因;该蛋白互作网络主要分子功能为氧化还原酶活性,主要参与的生物学过程为小GTP酶介导的信号转导。fox_milRNA_3的靶基因FOXG_04910可通过调节Rho基因影响菌株的生长,推断其和菌株与寄主的信号识别和菌株在寄主中定植有关。[结论]本研究通过试验验证和生物信息学预测发现,尖孢镰孢菌侵染番茄初期,存在milRNAs及其下游靶基因的调控网络。上述结果为进一步深入研究尖孢镰孢菌milRNA的功能提供一个新思路。[Objective] MicroRNA plays an important role in the biology progress by regulating its target gene. The objective of this study was to clarify the molecular function and regulatory mechanisms of milRNAs during the early stage of interaction between Fusarium oxysporum f. sp. lycopersici and tomato. [Methods] In this study, quantitative real-time PCR was applied to study the gene expression of primary milRNAs and their target genes during the early stage of infection of F. oxysporum f. sp.lycopersici on tomato. STRING protein interactive network database and Cytoscape software were used to construct the protein interactie network of milRNAs and the target genes. Furthermore, DAVID online analysis tool was used to analysis the relative genes to clarify the molecular functions and regulative mechanism of milRNAs during the infection of F. oxysporum f. sp.lycopercisi to tomato. [Results] The qRT-PCR results showed the opposite relative expression trends of the tested primilRNA_2a, 2b and FOXG_03470, pri-milRNA_3b and FOXG_04910, pri-milRNA_4 and FOXG_08812, and pri-milRNA_6 and FOXG_00067, which indicated the possible target relationship between milRNAs and related genes. The constructed protein interaction network showed close interactions within the target genes of fox_milRNA_1, fox_milRNA_2, fox_milRNA_3, and fox_milRNA_8 in Fusarium. Among which, FOXG_04240, the target gene of fox_milRNA_1,and FOXG_03470, the target gene of fox_milRNA_2, could interact with many genes of the protein interaction network.Therefore, these two target genes were predicted as the hub gene;The main molecular function and biological progress of this protein interaction network were oxoreductase activity and small GTP enzyme-mediated signal transduction, respectively.FOXG_04910, the target gene of fox_milRNA_3, influenced the growth of pathogen through regulating Rho gene, which was predicted to involved in recognition of pathogen and host, and colonization of pathogen in the host. [Conclusion] In this study, the regulatory network of milR

关 键 词：microRNA-like RNA 初级milRNA 蛋白互作网络 尖孢镰孢菌 靶基因 

分 类 号：S432[农业科学—植物病理学] S641.2[农业科学—农业昆虫与害虫防治]