中国农业科学 ›› 2020, Vol. 53 ›› Issue (24): 5017-5026.doi: 10.3864/j.issn.0578-1752.2020.24.005

• 植物保护 • 上一篇    下一篇



  1. 1河北农业大学生命科学学院/河北省植物生理与分子病理学重点实验室,河北保定 071001
    2河北农业大学植物保护学院,河北保定 071001
  • 收稿日期:2020-03-28 接受日期:2020-05-10 出版日期:2020-12-16 发布日期:2020-12-28
  • 通讯作者: 郝志敏,董金皋
  • 作者简介:龙凤,E-mail:。|王擎,E-mail:
  • 基金资助:

Identification and Expression Pattern Analysis of Septin Gene Family of Setosphaeria turcica

LONG Feng1(),WANG Qing1(),ZHU Hang1,WANG JianXia1,SHEN Shen1,LIU Ning1,HAO ZhiMin1(),DONG JinGao1,2()   

  1. 1College of Life Sciences/Key Laboratory of Hebei Province for Plant Physiology and Molecular Pathology, Hebei Agricultural University, Baoding 071001, Hebei
    2College of Plant Protection, Hebei Agricultural University, Baoding 071001, Hebei
  • Received:2020-03-28 Accepted:2020-05-10 Online:2020-12-16 Published:2020-12-28
  • Contact: ZhiMin HAO,JinGao DONG


【背景】隔膜蛋白Septin广泛存在于除植物以外所有真核生物中,是高度保守的GTP结合蛋白家族,被认为是继微管、微丝和中间纤维之后的第4种细胞骨架蛋白。病原真菌的Septin蛋白参与细胞极性的确定、形态塑造及与致病相关的形态转换。【目的】鉴定玉米大斑病菌(Setosphaeria turcicaSeptin基因家族,并进一步分析其在不同发育时期的表达模式,为明确隔膜蛋白Septin与真菌侵染结构发育之间的关系打下基础。【方法】以玉米小斑病菌(Bipolaris maydis)中6个Septin蛋白的氨基酸序列为探针,在玉米大斑病菌数据库在线Blastp比对和关键词搜索,获得大斑病菌的候选Septin,对其基因结构、理化性质以及跨膜区结构等方面进行生物信息学分析。收集人造疏水介质诱导下侵染结构发育不同时期以及侵染感病寄主叶片不同时间的玉米大斑病菌材料,利用实时荧光定量PCR(real-time fluorescence quantitative PCR,RT-qPCR)技术系统分析Septin基因家族在玉米大斑病菌侵染结构形成不同阶段的转录水平。【结果】获得了玉米大斑病菌6个候选Septin,其中4个核心Septin,均含有G1、G3、G4基序,分别将其命名为StSep1StSep2StSep3StSep4。在人造疏水介质诱导下,Septin表达水平均呈上升趋势。StSep1在芽管形成后期表达最活跃,其表达量达到分生孢子时期的25.69倍(P<0.01),StSep4的表达水平在附着胞发育后期到达高峰,随后表达逐渐下调。该基因家族在病菌侵染寄主叶片过程中的转录水平变化趋势与其在人造疏水介质诱导下的表现趋于一致。StSep1在接种后6 h转录水平极显著上调(P<0.01),随着时间延长,表达水平下降,StSep4在附着胞形成阶段表达活跃,表达量在接种后18 h达到高峰值,之后表达下调,但仍高于萌发初期。StSep2StSep3在接种后18 h和24 h表达活跃,高于萌发初期。【结论】玉米大斑病菌基因组中含有4个核心Septin,StSep1StSep4分别在芽管和附着胞形成时期活跃表达,结果可为进一步明确 Septin的功能及玉米大斑病菌的侵染调控机制提供依据。

关键词: 玉米大斑病菌, Septin基因家族, 实时荧光定量PCR, 基因表达


【Background】Septin, which is widely found in all eukaryotes except plants, is a highly conserved GTP binding protein family and is considered to be the fourth cytoskeletal protein after microtubules, microfilaments and intermediate fibers. Septin proteins of pathogenic fungi are involved in cell polarity determination, morphological shaping and morphological transformation associated with pathogenicity.【Objective】The objective of this study is to identify Septin gene family of Setosphaeria turcica, analyze its expression pattern at different developmental stages, and to lay a foundation for clarifying the relationship between the membrane protein Septin and fungal infection structure development.【Method】The amino acid sequences of six Septin proteins in Bipolaris maydis were used as probe sequences, online Blastp alignment and keyword search were carried out in the S. turcica database to obtain S. turcica candidate Septins. Then bioinformatics analyses including gene structure, physical and chemical properties and transmembrane region structure of the Septins were conducted. The materials of S. turcica that were induced by artificial hydrophobic media at different stages of infective structures development and infected host leaves at different times were collected, and RT-qPCR (real-time fluorescence quantitative PCR) technology was used to analyze the transcription levels of Septins at different stages of infective structures development.【Result】Six candidate Septins were obtained, and among them, four typical Septins contained G1, G3, and G4 motifs, were named by StSep1, StSep2, StSep3 and StSep4, respectively. The expression level of Septins increased slowly under the induction on artificial hydrophobic media. StSep1 was active in the late stage of germ tube formation, and its expression level was 25.69 times of that in the conidia period (P<0.01). The expression level of StSep4 reached a peak in appressorium anaphase, and then the expression level was down-regulated. The change trend of Septin expression in the process of pathogen infection was basically consistent with that under the induction of artificial hydrophobic material. StSep1 expression was significantly up-regulated at 6 h after inoculation (P<0.01). As time went on, StSep4 actively expressed during the period of appressorial formation, and the expression reached a peak at 18 h after inoculation. After that, the expression was down-regulated, but it was still higher than that in the early stage of germination. StSep2 and StSep3 were active at 18 h and 24 h after inoculation, exceeding the initial stage of germination.【Conclusion】There are 4 core Septins in the genome of S. turcica. StSep1 and StSep4 are actively expressed during the formation of germ tube and appressoria, respectively. The results will provide a theoretical basis for further clarifying the function of Septin and the infection regulation mechanism of S. turcica.

Key words: Setosphaeria turcica, Septin gene family, RT-qRCR, gene expression