Pseudomonas syringae activates ZAT18 to inhibit salicylic acid accumulation by repressing EDS1 transcription for bacterial infection

Abstract

Phytopathogens can manipulate plant hormone signaling to counteract immune responses; however, the underlying mechanism is mostly unclear.Here, we report that Pseudomonas syringae pv tomato (Pst) DC3000 induces expression of C2H2 zinc finger transcription factor ZAT18 in a jasmonic acid (JA)-signaling-dependent manner. Biochemical assays further confirmed that ZAT18 is a direct target of MYC2, which is a very important regulator in JA signaling. CRISPR/Cas9-generated zat18-cr mutants exhibited enhanced resistance to Pst DC3000, while overexpression of ZAT18 resulted in impaired disease resistance.Genetic characterization of ZAT18 mutants demonstrated that ZAT18 represses defense responses by inhibiting the accumulation of the key plant immune signaling molecule salicylic acid (SA), which is dependent on its EAR motif. ZAT18 exerted this inhibitory effect by directly repressing the transcription of Enhanced Disease Susceptibility 1 (EDS1), which is the key signaling component of pathogen-induced SA accumulation. Overexpression of ZAT18 resulted in decreased SA content, while loss of function of ZAT18 showed enhanced SA accumulation upon pathogen infection. Furthermore, enhanced resistance and SA content in zat18-cr mutants was abolished by the mutation in EDS1.Our data indicate that pathogens induce ZAT18 expression to repress the transcription of EDS1, further antagonising SA accumulation for bacterial infection.

Ethylene‐responsive factor ERF114 mediates fungal pathogen effector PevD1‐induced disease resistance in Arabidopsis thaliana

“…Transcriptional regulation plays a key role in plant defence against pathogens (Gao et al, 2022 ; Zheng et al, 2019 ). During this process, TFs binding to DNA regulate normal growth and the defence response to pathogens in plants.…”

Identification of stably expressed reference genes for expression studies in Arabidopsis thaliana using mass spectrometry-based label-free quantification

“…The gene names and accession numbers corresponding to the proteins are listed in Supplementary Table S1 . The abundance of the proteins corresponding to 12 reference genes commonly used in previous studies ( Czechowski et al, 2005 ; Jeong et al, 2011 ; Huang et al, 2013 ; Chen et al, 2014 ; Cheong et al, 2014 ; Cuéllar Pérez et al, 2014 ; Kim and Hwang, 2014 ; Zhang et al, 2015 ; Huot et al, 2017 ; Jia et al, 2018 ; Wu et al, 2019 ; Cui et al, 2021 ; Romero-Pérez et al, 2021 ; Gao et al, 2022 ) were also investigated using the protein dataset. The gene names and accession numbers are listed in Supplementary Table S1 .…”

“…Using RT-qPCR, we analyzed the expression levels of the 18 candidate reference genes and 12 reference genes commonly used in previous studies ( Supplementary Table S1 ; Czechowski et al, 2005 ; Jeong et al, 2011 ; Huang et al, 2013 ; Chen et al, 2014 ; Cheong et al, 2014 ; Cuéllar Pérez et al, 2014 ; Kim and Hwang, 2014 ; Zhang et al, 2015 ; Huot et al, 2017 ; Jia et al, 2018 ; Wu et al, 2019 ; Cui et al, 2021 ; Romero-Pérez et al, 2021 ; Gao et al, 2022 ) at 0 and 3 dpi with Pst DC3000 inoculation. We also evaluated the expression stability of these candidate reference genes in stress hormone treatments, including JA, SA, and ABA.…”

“…Gene names in black: candidate reference genes reported in this study; gene names in red: reference genes commonly used in previous studies. # TUB2_1 and TUB2_2 refer to two different primer pairs for TUB2 ( Wu et al, 2019 ; Gao et al, 2022 ). ^ UBC21_1 and UBC21_2 refer to two different primer pairs for UBC21 ( Cuéllar Pérez et al, 2014 ; Gao et al, 2022 ) .…”

“…The primer specificity was determined by the Primer-BLAST function on the NCBI platform against the A. thaliana genome (taxid: 3702) and validated by melting curve analysis after RT-qPCR ( Supplementary Figure S2 ). For references genes commonly used in previous studies, the primer sequences were adopted from the corresponding publications ( Czechowski et al, 2005 ; Jeong et al, 2011 ; Huang et al, 2013 ; Chen et al, 2014 ; Cheong et al, 2014 ; Cuéllar Pérez et al, 2014 ; Kim and Hwang, 2014 ; Zhang et al, 2015 ; Huot et al, 2017 ; Jia et al, 2018 ; Wu et al, 2019 ; Cui et al, 2021 ; Romero-Pérez et al, 2021 ; Gao et al, 2022 ). The primer sequences are listed in Supplementary Table S1 .…”