Journal of Integrative Agriculture ›› 2023, Vol. 22 ›› Issue (3): 799-811.DOI: 10.1016/j.jia.2022.08.117

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JIA-2021-0026 南方根结线虫效应子 Minc03329 抑制植物免疫反应并促进寄生

  

  • 收稿日期:2022-01-10 接受日期:2022-03-21 出版日期:2023-03-20 发布日期:2022-03-21

A Meloidogyne incognita effector Minc03329 suppresses plant immunity and promotes parasitism

ZHOU Jing-jing, ZHANG Xiao-ping, LIU Rui, LING Jian, LI Yan, YANG Yu-hong, XIE Bing-yan, ZHAO Jian-long#, MAO Zhen-chuan#    

  1. Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing 100081, P.R.China

  • Received:2022-01-10 Accepted:2022-03-21 Online:2023-03-20 Published:2022-03-21
  • About author:#Correspondence MAO Zhen-chuan, Tel: +86-10-82109545, E-mail: maozhenchuan@caas.cn; ZHAO Jian-long, E-mail: zhaojianlong@caas.cn
  • Supported by:

    This work was funded by the National Natural Science Foundation of China (31672010 and 31871942), the Beijing Natural Science Foundation, China (6222054), the China Agriculture Research System (CARS-23), and the Natural Science Foundation of Inner Mongolia Autonomous Region of China (2018MS03083).

摘要:

根结线虫病是农业生产上的毁灭性病害,每年造成巨大的经济损失。南方根结线虫是一种寄生范围广、危害严重、防治困难的根结线虫,其防治主要依赖化学农药,不仅污染环境、危害人类健康,而且大大增加线虫的抗药性。解析根结线虫的致害分子机制,对于制定环保、经济、高效的防治策略具有重要研究价值。研究发现,根结线虫在寄生过程由食道腺表达、通过口针分泌出许多效应子,在线虫侵染和寄生阶段发挥重要作用。不同种类效应子与寄主植物之间产生错综复杂的相互作用,功能机制有待深入解析。在本研究中,我们鉴定了一种新的南方根结线虫效应子Minc03329,对氨基酸序列分析发现,其包含用于分泌的信号肽序列和一个C型凝集素结构域。酵母信号序列捕获实验表明Minc03329的信号肽是有功能的,具有分泌功能;原位杂交实验结果表明Minc03329在南方根结线虫亚腹食道腺中特异表达;实时荧光定量PCR结果证实 Minc03329 在线虫寄生初期表达量显著升高;病毒介导的基因沉默干扰线虫 Minc03329 表达,显著降低了南方根结线虫的致病性;相反,Minc03329转基因拟南芥接种南方根结线虫后根结数和卵块数显著增加,表明效应子Minc03329在植物细胞中表达,可以显著增加植物对南方根结线虫的敏感性;Minc03329 在本氏烟草叶片细胞中瞬时表达能抑制由小鼠促凋亡蛋白BAX引发的细胞程序性死亡;通过对Minc03329转基因拟南芥和野生型拟南芥进行转录组数据比较分析,发现Minc03329转基因拟南芥中许多防御相关基因表达显着下调;此外,一些差异表达基因可能参与了南方根结线虫摄食位点的形成,但是其分子机制有待深入解析。本研究是在揭示凝集素效应子MiCTL1功能机制后,解析的第二个南方根结线虫凝集素类效应子的功能。验证了凝集素类效应子在线虫与植物互作过程通过抑制植物免疫反应,帮助线虫寄生的分子机制。研究结果为揭示根结线虫致病分子机理,以及根结线虫防治分子靶标利用提供了重要理论依据。

Abstract:

Meloidogyne incognita is a devastating plant-parasitic nematode.  Effectors play important roles during the stages of nematodes infection and parasitism, but their molecular functions remain largely unknown.  In this study, we characterized a new effector, Minc03329, which contains signal peptide for secretion and a C-type lectin domain.  The yeast signal sequence trap experiments indicated that the signal peptide of Minc03329 is functional.  In situ hybridization showed that Minc03329 was specifically expressed in the subventral esophageal gland.  Real-time qPCR confirmed that the expression level of Minc03329 transcript was significantly increased in pre-parasitic and parasitic second-stage juveniles (pre-J2s and par-J2s).  Tobacco rattle virus (TRV)-mediated gene silencing of Minc03329 in host plants largely reduced the pathogenicity of nematodes.  On the contrary, ectopic expression of Minc03329 in Arabidopsis thaliana significantly increased plant susceptibility to nematodes.  Transient expression of Minc03329 in Nicotiana benthamiana leaves suppressed the programmed cell death triggered by the pro-apoptotic protein BAX.  Moreover, the transcriptome analysis of Minc03329-transgenic Arabidopsis and wild type revealed that many defense-related genes were significantly down-regulated.  Interestingly, some different expressed genes were involved in the formation of nematode feeding sites.  These results revealed that Minc03329 is an important effector for Mincognita, suppressing host defense response and promoting pathogenicity.

Key words: Meloidogyne incognita , effector ,  C-type lectin ,  pathogenicity ,  plant immunity