J Integr Plant Biol. ›› 2020, Vol. 62 ›› Issue (6): 761-776.DOI: 10.1111/jipb.12859

Special Issue: Protein kinases

• New Technology • Previous Articles     Next Articles

Engineering plants to secrete affinity-tagged pathogen elicitors for deciphering immune receptor complex or inducing enhanced immunity

Shuang Miao, Jiuer Liu, Jianhang Guo and Jian‐Feng Li*   

  1. Guangdong Provincial Key Laboratory of Plant Resources, State Key Laboratory of Biocontrol, MOE Key Laboratory of Gene Function and Regulation, School of Life Sciences, Sun Yat‐sen University, Guangzhou 510275, China

    Email: Jian-Feng Li(lijfeng3@mail.sysu.edu.cn)

  • Received:2019-06-17 Accepted:2019-07-26 Online:2019-07-30 Published:2020-06-01


Plant cells mount plenty of pattern‐recognition receptors (PRRs) to detect the microbe‐associated molecular patterns (MAMPs) from potential microbial pathogens. MAMPs are overrepresented by proteinaneous patterns, such as the flg22 peptide from bacterial flagellin. Identification of PRR receptor complex components by forward or reverse genetics can be time/labor‐consuming, and be confounded by functional redundancies. Here, we present a strategy for identifying PRR complex components by engineering plants to inducibly secrete affinity‐tagged proteinaneous MAMPs to the apoplast. The PRR protein complexes bound to self‐secreted MAMPs are enriched through affinity purification and dissected by mass spectrometry. As a proof of principle, we could capture the flg22 receptor FLS2 and co‐receptor BAK1 using Arabidopsis plants secreting FLAG‐tagged flg22 under estradiol induction. Moreover, we identified receptor‐like kinases LIK1 and PEPR1/PEPR2 as potential components in the FLS2 receptor complex, which were further validated by protein–protein interaction assays and the reverse genetics approach. Our study showcases a simple way to biochemically identify endogenous PRR complex components without overexpressing the PRR or using chemical cross‐linkers, and suggests a possible crosstalk between different immune receptors in plants. A modest dose of estradiol can also be applied to inducing enhanced immunity in engineered plants to both bacterial and fungal pathogens.

Editorial Office, Journal of Integrative Plant Biology, Institute of Botany, CAS
No. 20 Nanxincun, Xiangshan, Beijing 100093, China
Tel: +86 10 6283 6133 Fax: +86 10 8259 2636 E-mail: jipb@ibcas.ac.cn
Copyright © 2022 by the Institute of Botany, the Chinese Academy of Sciences
Online ISSN: 1744-7909 Print ISSN: 1672-9072 CN: 11-5067/Q