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中国油料作物学报 ›› 2023, Vol. 45 ›› Issue (1): 124-130.doi: 10.19802/j.issn.1007-9084.2022012

• 种质资源·遗传育种 • 上一篇    下一篇

向日葵柄锈菌效应子P2的定位及功能研究

金佳鹏(), 连小雨, 路妍, 景岚()   

  1. 内蒙古农业大学园艺与植物保护学院,内蒙古 呼和浩特,010011
  • 收稿日期:2022-01-14 出版日期:2023-02-25 发布日期:2023-03-03
  • 通讯作者: 景岚 E-mail:2655820291@qq.com;jinglan71@126.com
  • 作者简介:金佳鹏(1996- ),男,硕士研究生,主要从事植物免疫学研究,E-mail: 2655820291@qq.com
  • 基金资助:
    国家自然科学基金(32160642);内蒙古自然科学基金(2020MS03046)

Localization and function of effector P2 of Puccinia helianthi Schw

Jia-peng JIN(), Xiao-yu LIAN, Yan LU, Lan JING()   

  1. College of Horticulture and Plant Protection, Inner Mongolia Agricultural University, Hohhot 010011, China
  • Received:2022-01-14 Online:2023-02-25 Published:2023-03-03
  • Contact: Lan JING E-mail:2655820291@qq.com;jinglan71@126.com

摘要:

由向日葵柄锈菌(Puccinia helianthi Schw.)引起的向日葵锈病严重威胁着向日葵的安全生产。效应子是一类病原菌侵染过程中分泌的蛋白,促进病原菌侵染或触发寄主免疫。为明确向日葵锈菌效应子P2在锈菌侵染过程中发挥的作用,对该基因序列进行了生物信息学分析;以向日葵锈菌夏孢子cDNA为模板,克隆了P2基因的编码序列,利用qRT-PCR技术分析了该基因在侵染过程中的表达特性;采用农杆菌瞬时表达系统在烟草上验证了P2的毒性功能,并在烟草上瞬时表达对其进行了亚细胞定位。研究结果表明,P2编码58个氨基酸,N端含24氨基酸的信号肽,无核定位信号及跨膜结构域,并在向日葵锈菌侵染早期上调表达。在本氏烟中瞬时表达P2可以抑制BAX诱导的细胞坏死,亚细胞定位结果表明P2定位在细胞膜及细胞核。

关键词: 向日葵锈病, 效应子, qRT-PCR, 细胞程序性死亡, 亚细胞定位

Abstract:

Sunflower rust, caused by Puccinia helianthi, often threatens sunflower production. Effectors are proteins secreted during pathogen infection, which promote pathogen invasion or trigger host immunity. In order to clarify the role of P2 effector in rust infection process, P2 gene was analyzed by bioinformatics. cDNA of P. helianthi urediospores was used as template to clone P2 gene. Its expression characteristics during the infection were analyzed by qRT-PCR. Then function of P2 was verified in Nicotiana benthamiana by Agrobacterium transient expression system, and subcellular localization of P2 was performed by transient expression in N.benthamiana. Results showed that P2 encoded 58 amino acids (aa) with 24 aa signal peptide at N-terminal, without nuclear localization signal and transmembrane domain. P2 expression was up-regulated during early stage of P. helianthi infection. Transient expression of P2 in N.benthamiana inhibited BAX-induced cell death. Subcellular localization showed that P2 was localized in cell membrane and nucleus.

Key words: sunflower rust, effector, qRT-PCR, programmed cell death (PCD), subcellular localization

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