Subtype-selective agonists of plant hormone co-receptor COI1-JAZs identified from the stereoisomers of coronatine

Hayashi, Kengo; Kato, Nobuki; Bashir, Khurram; Nomoto, Haruna; Nakayama, Misuzu; Chini, Andrea; Takahashi, Satoshi; Saito, Hiroaki; Watanabe, Raku; Takaoka, Yousuke; Tanaka, Maho; Nagano, Atsushi J.; Seki, Motoaki; Solano, Roberto; Ueda, Minoru

doi:10.1038/s42003-023-04709-1

Download PDF

Article
Open access
Published: 25 March 2023

Subtype-selective agonists of plant hormone co-receptor COI1-JAZs identified from the stereoisomers of coronatine

Communications Biology volume 6, Article number: 320 (2023) Cite this article

3035 Accesses
1 Citations
9 Altmetric
Metrics details

Subjects

Abstract

Severe genetic redundancy is particularly clear in gene families encoding plant hormone receptors, each subtype sharing redundant and specific functions. Genetic redundancy of receptor family members represents a major challenge for the functional dissection of each receptor subtype. A paradigmatic example is the perception of the hormone (+)-7-iso-jasmonoyl-L-isoleucine, perceived by several COI1-JAZ complexes; the specific role of each receptor subtype still remains elusive. Subtype-selective agonists of the receptor are valuable tools for analyzing the responses regulated by individual receptor subtypes. We constructed a stereoisomer library consisting of all stereochemical isomers of coronatine (COR), a mimic of the plant hormone (+)-7-iso-jasmonoyl-L-isoleucine, to identify subtype-selective agonists for COI1-JAZ co-receptors in Arabidopsis thaliana and Solanum lycopersicum. An agonist selective for the Arabidopsis COI1-JAZ9 co-receptor efficiently revealed that JAZ9 is not involved in most of the gene downregulation caused by COR, and the degradation of JAZ9-induced defense without inhibiting growth.

A plant NLR receptor employs ABA central regulator PP2C-SnRK2 to activate antiviral immunity

Article Open access 13 April 2024

Spatial co-transcriptomics reveals discrete stages of the arbuscular mycorrhizal symbiosis

Article Open access 08 April 2024

Combinatorial biosynthesis for the engineering of novel fungal natural products

Article Open access 18 April 2024

Introduction

Sub-functionalization or neo-functionalization of genes occurs during evolution through duplication events in the genome. Gene duplications leading to genetic redundancy can allow the acquisition of new functions, interactions, and expression patterns to benefit plant adaptive responses to the changing environmental stimuli^1,2,3. Examples of severe genetic redundancy can be observed in plant hormone signaling modules^1,2. Genetic redundancy in plant hormone receptors makes it difficult to study and characterize their complex signaling networks³. A paradigmatic example can be found in the case of a lipid-derived plant hormone, (+)-7-iso-jasmonoyl-L-isoleucine (JA-Ile, 1, Fig. 1a), which is perceived by COI1-JAZ co-receptors consisted of F-box protein CORONATINE INSENSITIVE 1 (COI1) and JASMONATE-JIM DOMAIN (JAZ) repressor proteins. Subsequent degradation of JAZ through the 26S-proteasome induces the expression of jasmonate (JA)-responsive genes, including defenses against necrotrophic pathogens and herbivorous insects in Arabidopsis thaliana and several crop plants^4,5,6. Concurrently, the COI1-JAZ-mediated JA signaling regulates growth inhibition, fertility, and secondary metabolite production. The molecular basis of the multiple effects of JA-Ile partially lies in the severe genetic redundancy of the 13 JAZ gene family members encoded in the Arabidopsis genome^3,7,8.

**Fig. 1: Chemical library of coronatine stereoisomers.**

Differences in spatiotemporal expression of JAZ genes during growth and development lead to redundant and complementary JA signaling in Arabidopsis. Genetic redundancy seriously challenges canonical genetic approaches to functionally dissect JAZ gene function as the effect of a single JAZ knock-out in Arabidopsis can be easily complemented by other members of the JAZ gene family³. Tremendous efforts have been made to dissect the redundant function of JAZ genes in jasmonate signaling. Genetic studies using Arabidopsis mutant lines lacking multiple JAZ genes (jaz1/3/4/9/10 quintuple mutant as jazQ, jaz1/2/3/4/5/6/7/9/10/13 decuple mutant as jazD, jaz1/2/3/4/5/6/7/8/9/10/13 as jazU, and recently reported jaz1/2/3/4/5/6/7/9/10/11/12 undecuple mutant) are pioneering works to understand the function of JAZ family genes^9,10,11,12. Recently, genetic studies using the model liverwort Marchantia polymorpha L have attracted attention as an alternative approach to analyze the jasmonate signaling because no genetic redundancy is observed in the key JA signaling pathway components, COI1, JAZ, and MYC^{13,14,15,16,17}. In addition, chemical genetics is a powerful and promising alternative. Chemical genetics approaches have been successful in analyzing genetically redundant signaling in other hormonal pathways, such as those of abscisic acid and strigolactones, in which specific hormonal agonists activated distinctive receptor subtypes triggering specific responses^{18,19,20,21,22,23,24}. Transient inhibition/degradation of a single JAZ protein in a fully developed plant by using chemicals may therefore disclose the specific function of a particular JAZ gene. Thus, the chemical genetic approach can complement the genetic approach^25,26.

Here we report that the analysis of the chemical library (Fig. 1b) harboring all the stable stereochemical isomers of JA-Ile mimic, coronatine (COR, 2, Fig. 1a), provided the agonists of selective affinity among the COI1-JAZ co-receptor pairs in two plant species, A. thaliana and Solanum lycopersicum. Significantly, the stereoisomer 2e (Fig. 1b) caused protein degradation of the JAZ9 repressor to activate JAZ9-dependent jasmonate signaling in A. thaliana. The use of 2e enabled the analysis of a unique gene expression pattern governed by a single JAZ repressor, JAZ9, among the genetically redundant JAZ family genes.

Results

Chemical synthesis of all possible stereochemical isomers of coronatine

The use of a focused chemical library of 2-stereoisomers (Fig. 1b) can be a promising approach to look for novel, specific ligands of COI1-JAZ co-receptor pairs. Stereochemical isomers of 2 have the same size exclusion volume as naturally occurring 2 and slightly different 3D orientation of functional groups in a molecule, thus they can be accommodated in the ligand-binding pocket of COI1 with modulation of interaction network to each co-receptor pair leading to selectivity among JAZs (Fig. 1c).

The chemical library of all the stable 2-stereoisomers was constructed by coupling stereoisomers of coronafacic acid (CFA; 3) and coronamic acid (CMA; 4) (Fig. 1a). Enough amount of the stable stereoisomers of 3 was achieved by gram-scale racemic CFA synthesis and subsequent optical resolution, epimerization, and separation of diastereomers (Supplementary Fig. 1a)^27,28,29. The optical purities of four CFAs (3-3c) were determined by chiral HPLC analyses³⁰. Four CFAs (3-3c) were coupled with four CMAs (4-4c) which were synthesized by the reported method³¹, and subsequent deprotection provided all the stable 16 stereochemical isomers of coronatine (2 and 2a-2o in Fig. 1a, b, and Supplementary Figs. 1b and 2).

Stereochemical isomers demonstrated unique selectivity for COI1-JAZ co-receptor pairs of two plant species, Arabidopsis thaliana and Solanum lycopersicum

We examined the affinities of 16 stereochemical isomers of 2 with COI1-JAZ co-receptors of two plant species, Arabidopsis thaliana (encoding 13 JAZ proteins), as a representative model dicot, and Solanum lycopersicum (encoding 13 JAZ proteins), as a model plant belonging to the Solanaceae family (Supplementary Fig. 3). Chemical screening using in vitro fluorescence anisotropy (FA) assay was performed for all of the functional combinations of COI1-JAZ co-receptor pairs^8,32.

For Arabidopsis COI1-JAZ pairs, in vitro FA assay was carried out for all of the functional combinations of AtCOI1-AtJAZ1/2/3/4/5/6/9/10/11/12 (Supplementary Fig. 3)³³. AtCOI1-AtJAZ7/8/13 pairs were excluded in all of the following assays because AtJAZ7/8/13 lack the short conserved canonical degron sequence and thus cannot form the AtCOI1-1/2-AtJAZ complex^34,35. Instead of full-length AtJAZ proteins, we used the fluorescein-labeled AtJAZ degron short peptides (Fl-AtJAZPs), including the ligand-interacting domain of full-length AtJAZ proteins^33,36. FA assay provided the binding affinity (K_d) of each stereochemical isomer of 2 against AtCOI1-AtJAZ pairs (Fig. 2a, and Supplementary Figs. 4 and 5). The results of the FA assay were summarized as an affinity’heatmap’ profile of 16 stereochemical isomers (Fig. 2a). Based on their affinity, stereochemical isomers were categorized into four groups; no affinity (2a, 2b, 2g, 2j, 2k, 2m, 2n, and 2o), non-selective affinity (2, 2d, 2h, 2i, 2l), moderately selective affinity (2c), and highly selective affinity (2e, 2f) to some of the AtCOI1-AtJAZ combinations (Fig. 2a and Supplementary Fig. 5). In vitro pull-down assays using Fl-AtJAZP and GST-AtCOI1 protein confirmed the highly selective affinity of 2e and 2f (Supplementary Fig. 6). Surprisingly, 2e has a selective affinity to the AtCOI1-AtJAZ9 pair in FA and pull-down assays. Docking and subsequent in silico molecular dynamics (MD) simulations demonstrated that 2e, as well as 2, can be accommodated in the binding pocket of COI1-InsP₈-JAZ9 complex (Supplementary Fig. 7). In this MD simulation, we used inositol octakisphosphate (InsP₈) instead of inositol pentakisphosphate (InsP₅) because previous reports showed that InsP₈ also formed stable complex with COI1-2-JAZ instead of phosphates (PO₄)^37,38. In planta AtJAZ9-selectivity of 2e was tested by using a GUS reporter assay using A. thaliana reporter lines, 35S: JAZ1-GUS, 35S: JAZ9-GUS, 35S: JAZ10-GUS, and 35S: JAZ12-GUS (Fig. 2b and Supplementary Fig. 8). All JAZ-GUS fusion proteins were degraded in 2-treated seedlings. In contrast, 2e caused protein degradation in the AtJAZ9-GUS reporter line, whereas no degradation was observed in the AtJAZ1-GUS and AtJAZ10-GUS reporter lines. In addition, 2e-treatment caused degradation of JAZ9-GFP protein in 35S: JAZ9-GFP, but not of JAZ1-GFP protein in 35S: JAZ1-GFP (Supplementary Fig. 9). These results confirmed that 2e-treatment caused the selective ‘protein knock-out (degradation)’ of JAZ9 protein in A. thaliana.

**Fig. 2: Affinity heatmaps of coronatine stereoisomers on COI1-JAZ co-receptors of *A. thaliana* and *S. lycopersicum*.**

For SlCOI1-SlJAZ pairs of S. lycopersicum, in vitro FA assay was also carried out for SlCOI1-SlJAZ1-11/13 (Fig. 2c). Phylogenetic characterization and functional annotation of SlJAZs were reported previously (Supplementary Fig. 3)³². Based on this result, we reported the affinity between 1/2 and SlCOI1-SlJAZ pairs using GST-SlCOI1 and fluorescein-labeled SlJAZ degron short peptides (Fl-SlJAZPs)³⁹. According to this previous report, SlJAZ12 was excluded in the FA assay because it does not have a conserved JAZ degron sequence³². The stereochemical library showed distinct selectivity against SlCOI1-SlJAZ pairs (Fig. 2c and Supplementary Figs. 10 and 11), as with the case of AtCOI1-AtJAZ pairs. Among them, remarkable selectivity was found for the stereochemical isomers 2e and 2f. 2e has selectivity against SlCOI1-SlJAZ5/6/8, and 2f has selectivity against SlCOI1-SlJAZ5/6/8/13. Unfortunately, we cannot examine the affinity of stereoisomers with the SlCOI1-SlJAZ7P pair in the FA assay because the severe non-specific binding was observed in the absence of ligand. Interestingly, isomers 2e and 2f also showed selectivity against AtCOI1-AtJAZ pairs (Fig. 2a).

Through these examinations, we found two stereochemical isomers, 2e and 2f, showing distinct selectivity for COI1-JAZ co-receptor pairs of two plant species. These results demonstrated that the stereochemical library of 2 is a promising chemical tool for discovering ligands with selective affinity for some subtypes of the genetically redundant COI-JAZ co-receptor family in higher plants.

A stereochemical isomer induced defenses without growth inhibition through selective activation of jasmonate signaling in A. thaliana

To examine the power of stereochemical isomers to dissect the jasmonate signaling pathway, we focused on 2e, with selective affinity to the AtCOI1-AtJAZ9 pair (Fig. 2a). We explored the biological effect of 2e and its mode of action on A. thaliana.

2e showed no growth inhibition on 4-week-old A. thaliana, whereas 2 caused severe growth inhibition (Fig. 3a, b, and Supplementary Fig. 12). In addition, no growth inhibition was observed on root by the addition of 2e (Supplementary Fig. 13). However, 2e, as well as 2, moderately activated resistance response against the infection of fungal pathogen Alternaria brassicicola compared to the mock-treatment (Fig. 3c, d, and Supplementary Fig. 14)³⁶. This 2e-mediated defense trait was impaired in jaz9-1, in which other members of the JAZ family protein likely complemented the original function of JAZ9 protein in presence of non-selective ligand, and 2e cannot cause their degradation (Fig. 3e and Supplementary Fig. 15) This result was in clear contrast to 2, which caused a close defense trait in both Col-0 and jaz9-1 (Fig. 3c–e and Supplementary Fig. 15). These effects of 2e on A. thaliana strongly suggested that 2e can induce a resistance response by activating the AtJAZ9-mediated partial jasmonate signaling. Reverse transcription-qPCR (RT-qPCR) analyses demonstrated that 2e-treatment in 4-week-old A. thaliana (without pathogen infection) caused the upregulation of defense-related genes, such as ORA59, ERF1, PDF1.2, and HEL, without affecting the expression of JA marker genes, such as MYC2 and VSP1/2 (Fig. 3f). In the downstream of JA signaling, ORA59, ERF1, PDF1.2, and HEL belong to ERF/ORA branch, and MYC2 and VSP1/2 belong to the MYC2-branch (Supplementary Fig. 16). Thus, 2e selectively upregulated the ERF/ORA branch of JA-signaling without affecting MYC-branch. In addition, 2e-mediated expression of ORA59 and PDF1.2 was impaired in coi1-1, which confirmed the COI1-dependent mode-of-action of 2e (Supplementary Fig. 17).

**Fig. 3: In planta analyses of stereoisomers in *Arabidopsis thaliana*.**

Chemical and genetic AtJAZ9 knock-out showed distinct gene expression profiles

Previous reverse genetic studies demonstrated that jaz9-1 has no significant phenotype regarding root-growth inhibition and defense response^10,11. In contrast, our 2e-induced protein knock-out of JAZ9 caused activation of the defense trait. To examine the distinct difference between chemically induced protein degradation or loss-of-function of JAZ9 by 2e and genetic knock-out in jaz9-1, RNA sequencing (RNAseq) analyses were carried out for a comprehensive comparison of gene expression profiles between 6-day-old seedlings of WT (Col-0) treated by 2/2e and jaz9 mutant (jaz9-1) treated by mock (Supplementary Data 1).

To analyze the entire gene expression profiles, we performed principal component analysis, Venn diagram analyses, and heatmap analyses of upregulated genes with >2.5-fold changes (Fig. 4a–c). In principal component analysis, no significant difference was observed between gene expression profiles of the mock-treated jaz9-1 and the mock-treated Col-0 (Fig. 4a), suggesting that in the jaz9-1 mutant, most functions of JAZ9 as a transcriptional repressor would be complemented by other partially redundant members of the JAZ family. In contrast, a clear difference was found between the gene expression profiles of 2e-treated Col-0 and mock-treated jaz9-1 (Fig. 4a). This result showed a distinct difference in gene expression profiles between genetic knock-out and chemically induced protein degradation (or loss-of-function).

**Fig. 4: Gene expression analyses of 2- and 2e-treated Col-0 and jaz9-1 seedlings.**

We further examined the unique gene expression caused by 2e. Detailed analyses of the gene expression profiles shown in the Venn diagram and heatmap also showed that 2e caused a part of 2-mediated gene expression: 2 upregulated expressions of 950 genes, of which 2e upregulated 103 genes (Fig. 4b). A significant difference was observed between the results of 2e-induced chemical degradation (or knock-out) and genetic knock-out of JAZ9: 91 of 103 gene upregulations caused by 2e were not observed in jaz9-1 (Fig. 4b and Supplementary Fig. 18 as a list of 91 genes). Heatmap analysis (Fig. 4c) showed that 2 upregulated major JA signaling-related genes, such as JAZ, JA-responsive genes, defense-related genes, and metabolism-related genes, whereas 2e primarily upregulated expressions of ethylene-mediated defense-related genes (see also Supplementary Figs. 19 and 20).

To our surprise, a remarkable difference in expression profiles was observed in downregulated genes: 2e caused downregulation of gene expression only for eight genes, whereas 2 caused downregulation for 614 genes (Fig. 4d and Supplementary Fig. 19). GO analysis of 2-repressed genes showed that GO terms related to cell wall organization/biogenesis were significantly repressed (Fig. 4d). RT-qPCR analyses showed that 2 repressed the gene expression of cell wall organization/biogenesis-related genes such as EXLA2, XTH19, and XTH23, which were previously reported as essential genes for plant root growth^40,41, in a COI1-dependent manner, but 2e did not affect their expression (Fig. 4e). The reason why 2e did not cause growth inhibition could be explained by the lack of gene downregulation of these cell-wall-related genes.

A stereoisomer caused AtJAZ9-dependent gene expressions

The fact that mode of action of 2e is dependent mainly on JAZ9 was confirmed by a comparison of RNAseq data on 2e-treated Col-0 and 2e-treated jaz9-1 (Fig. 4f). In jaz9-1, 2e activated the expression of 100 genes, and 52 of 100 genes were upregulated in 2e-treated Col-0, of which three were upregulated in mock-treated jaz9-1 (Fig. 4f). Thus, expressions of 49 genes (Supplementary Fig. 20a as a list of 49 genes) appear to be induced independently of JAZ9 in the 2e-treated jaz9-1. The 49 genes include JA-marker genes such as JAZ1, MYC2, and AOS, and defense-related genes belonging to the ERF/ORA branch, such as ORA59, ERF1, PDF1.3, and PDF1.2 (Supplementary Fig. 21a). However, the RT-qPCR analysis showed that the expression levels of most of the 49 genes found in 2e-treated jaz9-1 were not statistically significant compared to those in 2e-treated Col-0, while the expression levels of ERF/ORA-genes which are upregulated by 2e in Col-0 were significantly suppressed in 2e-treated jaz9-1 (Fig. 4g and Supplementary Fig. 21b). These results showed that 2e upregulated the expression of defense-related genes in a mostly JAZ9-dependent manner. This result agrees with the result in Fig. 3e, in which 2e caused no remarkable resistance response against the pathogen in the jaz9-1 mutant (Fig. 3e and Supplementary Fig. 15). To test if 2e activity may also require JAZ10, as previously described for the JA-Ile selective agonist N-coronafacyl ent-coronamic acid O-phenyloxime (NOPh)³⁶, the requirement for JAZ10 in 2e-induced responses was analyzed. 2e did not affect the growth and upregulated defense-related genes belonging to the ERF/ORA branch in the jaz10-1 mutant (Supplementary Fig. 22). Moreover, no growth inhibition and expressions of marker genes belonging to ERF/ORA branch were observed in 2e-treated jaz12-1 (Supplementary Fig. 23). These results rule out the role of JAZ10 in 2e-mediated regulation and they suggested that 2e upregulated the gene expressions mostly in a JAZ9-dependent manner.

We successfully developed the chemical tool 2e which mediates JAZ9-selective degradation in planta. This 2e-mediated activation leads to the expression of genes normally repressed by JAZ9 in A. thaliana. Examination of differential gene expression induced by JAZ9 protein degradation (or loss-of-function) revealed the unique function of JAZ9, which did not affect the downregulation of genes, including cell-wall-related ones which play important roles in the plant root growth, and caused upregulation of defense-related genes through the EIN3/EIL1-ERF/ORA59 pathway^40,41,42,43. The results highlight the power of a stereochemical library of 2 to screen the JAZ-selective activator of jasmonate signaling in other plant species.

Discussion

The chemical genetic approach is a promising way to dissect the plant hormone signaling of high genetic redundancy^25,26. Previous studies have reported success in abscisic acid and strigolactone using small molecules screened from chemical libraries to elucidate the function of receptor subtypes belonging to genetically redundant receptor families^18,23,24. Chemical libraries composed of planar molecules have been successfully used to screen hit compounds in drug discovery and plant chemical biology^26,44,45. Chini et al. recently discovered a general antagonist of AtCOI1-AtJAZ using a chemical library⁴⁶. To the best of our knowledge, this was the only successful example of the chemical library screening of COI1-JAZ co-receptor agonist/antagonist. By contrast, we have screened a targeted library, based on stereoisomers of 2, for highly subtype-selective ligands of 10 AtCOI1-AtJAZ and 11 SlCOI1-SlJAZ co-receptor pairs. 2e/2f, in addition to high selectivity for AtJAZ9, also showed selective affinity with SlCOI1-SlJAZ5/6/8, which are the closest tomato homologs of AtJAZ9 belonging to Group V (Supplementary Fig. 3). The chemical biology community focused on the conventional chemical libraries being composed of planar molecules, whereas we envisioned a chemical library composed of the stereoisomers of three-dimensional structures. These two approaches are complementary, and we demonstrated that stereochemical libraries could provide hit compounds that cannot be obtained from a chemical library of planar molecules.

In the present study, we have successfully identified AtCOI1-AtJAZ9-selective agonist 2e from the stereochemical library of 2. Taking advantage of the JAZ9-selective mode-of-action of 2e, we successfully find that JAZ9 plays a unique function in jasmonate signaling: JAZ9 does not participate in the gene downregulation caused by 2. Among the genes specifically repressed by 2, but not by 2e, we focused on genes related to cell wall formation and biosynthesis, such as EXLA2, XTH19, and XTH23 (Fig. 4e). Previous reports suggested that these genes play important roles in root growth: overexpression lines of EXLA2 and double knock-out mutant of XTH19 and XTH23, which affect the root growth and lateral root formation^40,41,47. Thus, the current results answered the question, why 2e did not affect the growth inhibition (Fig. 3a, b). In addition, using 2e, we demonstrated that pathogen-resistance through ERF/ORA-branch JA-signaling is controlled by JAZ9-meditated jasmonate signaling. These findings agree with our previous results, in which NOPh has a selective affinity with AtCOI1-AtJAZ9/10 pairs to enhance resistance response against pathogen A. brassicicola without causing growth inhibition³⁶. NOPh selectively activates ERF/ORA-branch to unpair growth and defense through COI1/JAZ9-EIN3/EIL1-ERF/ORA59 signaling pathway⁴². In contrast to NOPh, 2e mostly caused the selective degradation of single JAZ repressor, thus enabling analysis of gene expression profiles regulated by a single JAZ. 2e is specific mainly for JAZ9, although RNAseq suggest that it might target other JAZ at least partially and weakly. However, since the 2e-mediated expression level of JAZ9-independent JA-responsive genes is low (Fig. 4g and Supplementary Fig. 21b), we concluded that 2e induces gene expression mainly in a JAZ9-selective manner. 2e effectively reduces the redundancy of the COI1-JAZ co-receptors and it led to the identification of specific genes most likely regulated by the JAZ9 repressor (Fig. 4b and Supplementary Fig. 18 as a list of genes).

Previous achievements demonstrated that the limitations of the genetic approach, such as redundancy or lethality of essential genes, could be overcome by chemical genetics^48,49. We hypothesized that transient inhibition/degradation of a single JAZ protein in fully developed plants may disclose the function of a single JAZ gene within the redundant JAZ gene family. Our results demonstrated the uniqueness of “chemical mutants” in which gene redundancy does not seem to compensate the transiently degraded protein. We showed that this chemical genetic approach is complementary to genetic approaches^25,26. The current result paves the way for the rational design of JAZ-subtype selective agonists in crops and edible plants, which will elucidate the role of each JAZ gene in the highly redundant JAZ family of these plants.

Methods

Chemical syntheses

Details of chemical syntheses are described in Supporting Information.

Fluorescence anisotropy measurement

Fluorescence anisotropy titration experiments were performed at 25 °C in 50 µL of Tris buffer (50 mM Tris-HCl, pH 7.8, 100 mM NaCl, 10% Glycerol, 0.1% Tween20, 20 mM 2-mercaptoethanol (2-ME) and 1 µM inositol-1,2,4,5,6-pentakisphosphate, IP₅) containing 100 nM COI1 and 100 nM Fl-JAZPs using a 96-well clear-bottom plate unless otherwise noted. Each ligand was added to the solution and incubated at 25 °C until the r-value was not-fluctuated, and anisotropy intensities were measured (λ_ex/λ_em = 485 nm/516 nm). r values were calculated using the following equation: r = (I_VV – G × I_VH)/(I_VV + 2G × I_VH), I_VV and I_VH are the fluorescence intensities observed through polarizers parallel and perpendicular to the polarization of the exciting light, respectively, and G is a correction factor to account for instrumental differences in detecting emitted compounds. Fluorescence anisotropy was recorded on an EnVision 2105 (PerkinElmer, USA).

Pull-down assay of AtCOI1-AtJAZ peptide

Purified GST-COI1 (5 nM) with ASK1 and each compound (5 µM) are dissolved in 350 µL of Tris buffer and added Fl-JAZPs (10 nM). After incubation on ice for 1 h, the mixture was combined with an anti-Fluorescein antibody (GENETEX, GTX26644, 0.2 µL) and incubated at 4 °C for 5 h with rotation. After incubation, the mixture was combined with Protein G beads (Bio-Rad, 1614023, 10 µL in 50% Tris buffer slurry) and incubated at 4 °C for 1 h with rotation. After incubation, the sample was washed five times with PBS-T. The washed beads were resuspended in 35 μL of SDS-PAGE loading buffer containing dithiothreitol (DTT, 100 mM). After heating for 10 min at 60 °C, the samples were subjected to SDS-PAGE and analyzed by western blotting. SDS-PAGE and western blotting were carried out using a Mini-Protean III electrophoresis apparatus (Bio-Rad, Hercules, CA). Chemiluminescence was observed on Amersham Imager 680 (Cytiva, USA).

Plant materials and growth conditions

Arabidopsis thaliana Col-0 is the genetic background of wild-type and mutant lines used in this study. For biological assay using seedlings, surface-sterilized seeds were grown under a 16-h-light (75 μmol m⁻¹ s⁻¹; cool-white fluorescent light)/8-h-dark cycle at 22 °C on a half-strength Murashige-Skoog (MS) plate (2% sucrose, 0.4% gellan gum) or liquid medium (0.5% sucrose) in a Biotron LPH-240SP growth chamber (Nippon Medical & Chemical Instruments Co., Ltd., Osaka, Japan) after vernalization in the dark at 4 °C for 2 days. For biological assay using adult plants, seeds were grown directly in soil or 1:1 soil/vermiculite under a 12-h-light (75 µmol m⁻¹ s⁻¹, cool-white fluorescent light)/12-h-dark cycle at 22 °C.

For the 35S-JAZ1-GFP line, Amplified 35S-JAZ3-GFP from pGWB5-JAZ2^5,50 was inserted into the SalI-XhoI site of pET-32a (Invitrogen). pET-32a-35S-JAZ1-GFP was created by an in-fusion reaction between amplified ORF of JAZ1 and pET-32a-35S-GFP amplified by inverse PCR of pET-32a-35S-JAZ3-GFP. The adapter (EcoRI-SalI-XhoI) was inserted into the HindIII-SacI site of pGWB5-JAZ2. And then, 35S-JAZ1-GFP in prepared pET-32a-35S-JAZ1-GFP was introduced into the SalI-XhoI site of the adapted pGWB5. For the 35S-JAZ9-GFP line, amplified JAZ9 was introduced into pET-32a-35S-GFP by the in-fusion reaction. Amplified 35S-JAZ9-GFP was introduced into the SalI-XhoI site of the adapted pGWB5.

Agrobacterium GV3101, containing these constructs, was used to transform Col-0 plants by floral dipping (Clough & Bent, 1998, conducted by INPLANTA INNOVATIONS Inc., Japan). Transgenic lines were checked for Hygromycin resistance, and lines carrying one insertion were driven to homozygous and used for further experiments.

The mutant lines have been previously described: coi1-1, jaz9-1 (SALK_004872), and jaz10-1 (SAIL_92_D08)^51,52,53,54. The marker lines 35S: JAZ1-GUS, 35S: JAZ9-GUS, and 35S: JAZ10-GUS were previously described^4,55.

GUS-reporter assay

Four-day-old seedlings on a half-strength MS plate were transferred to a half-strength MS liquid medium in the presence or absence of each compound. After treatment for 2 h, each seedling was fixed with 90% aqueous acetone for 20 min on ice. Each seedling immersed in GUS staining buffer (50 mM phosphate buffer; pH 7.0; 1 mM K₄Fe(CN)₆; 1 mM K₃Fe(CN)₆; 0.1% Triton X-100; 2 mM 5-bromo-4-chloro-3-indolyl β-D-glucuronic acid (X-Gluc, Biomedical Science, Japan)). After staining at 37 °C for two days, each seedling was pictured with an E-520 digital camera (Olympus Corp., Japan).

To quantify the GUS activity, 9–11 roots of ligands-treated seedlings were flash-frozen, and homogenized with extraction buffer (50 µL; 50 mM phosphate buffer, pH 7, 10 mM EDTA, 10 mM 2-mercaptoethanol, 0.1% sarcosyl (N-lauroylsarcosine sodium salt; >94%, Sigma-Aldrich) and 0.1% Triton X-100). After centrifugation (15,000 × g, 4 °C, 10 min), each supernatant was collected. 10 µL extracts were incubated with a 40 µL extraction buffer containing 1 mM 4-MUG (methylumbelliferyl-β-D-glucuronide hydrate; ≥98%, Sigma-Aldrich). 10 µL samples were collected at t = 0 and t = 1 h. Then the reaction was quenched with 90 µL of 0.2 M Na₂CO₃. Fluorescence (Ex./Em. = 365/460 nm) was detected using the spectrophotometer Infinite M200Pro (TECAN, Switzerland). Each fluorescence intensity was normalized by the amounts of proteins determined by the Bradford assay (595 nm).

Chemical treatment for biological assay using adult plants

For growth inhibition assay, 2 µL of an aqueous solution containing each compound were dropped on the rosette center of a leaf of 2-week-old plants. After incubation for 6 days, each compound was applied again similarly. Pictures of plants were taken after 6 days of definitive treatment. The fresh weight of the aerial part was measured, and then metabolite analyses were performed in the same way as seedlings.

For gene expression analysis, leaves of 3-week-old plants were treated with each compound. After incubation for 6 days, each compound was applied again similarly. After treatment for 8 h, each leaf was flash-frozen, and then total RNA extraction and RT-qPCR were performed in the same way as seedlings.

Root growth inhibition analysis using seedlings

Two-day-old seedlings on half-strength MS plate were transferred on half-strength MS plate in the presence or absence of each compound. After treatment for 4 days, each seedling was pictured with an E-520 digital camera (Olympus Corp., Japan), and the root length of these seedlings was measured using Image J 1.45S software (http://imagej.net/Welcome).

Fungal infection analyses

Seeds were grown directly in the soil. As previously described, three leaves (leaf numbers 4, 5, and 6) of 3-week-old plants were inoculated with A. brassicicola suspension of 5 × 10⁶ spores/ml PDB (Difco)⁵⁶. Compound was applied twice at 5 ho before pathogen inoculation and at the same time of pathogen inoculation. Pictures of infected plants were taken 5 days after inoculation. Quantification of spores was carried out in a hemocytometer under a light microscope (Leica). Five to eight inoculated leaves of at least six different plants were pooled for each biological sample, and four to six independent biological replicates were measured for each treatment. Results were analyzed by one-way ANOVA/Tukey HSD post hoc test (p < 0.05). The infection assay was repeated three times with similar results. Data are shown as mean ± SEM.

RNAseq analysis

Six-day-old seedlings (5–8 seedlings/sample) grown on half-strength MS liquid medium were treated with half-strength MS liquid medium in the presence or absence of each compound. After treatment for 8 h, each sample was flash-frozen, and then total RNA was extracted using RNeasy Mini Kit (Qiagen Co. Ltd., Germany). RNAseq libraries were prepared using the low-cost and easy RNA-SEQ (Lasy-Seq) method described previously⁵⁷. Paired-end sequencing was performed using HiSeq X Ten (Illumina, San Diego, CA). RNA-seq analyses were performed with three biological replications, and the data was analyzed using DESeq2.

Low-quality reads and adapter were trimmed using trimmomatic version 0.39 (http://www.usadellab.org/cms/?page=trimmomatic) with ‘ILLUMINACLIP:TruSeq3-SE.fa:2:30:10 LEADING:3 TRAILING:3 SLIDINGWINDOW:4:15 MINLEN:36’ options. HISAT2 (http://daehwankimlab.github.io/hisat2/) version 2.2.1 was used to align the reads for reference genome (https://phytozome.jgi.doe.gov/pz/portal.html#!info?alias=Org_Athaliana) with ‘–max-intronlen 5000’ option. Aligned reads within gene models were counted using featureCounts version 2.0.1 (http://subread.sourceforge.net/) with ‘--fracOverlap 0.5 -O -t gene -g ID -s 1 --primary’ options. Differentially expressed genes were identified using R version 4.0.4 (https://www.r-project.org/) and DESeq2 version 1.30.1 (https://bioconductor.org/packages/release/bioc/html/DESeq2.html) package.

Heat map analyses were performed with an online tool heatmapper⁵⁸. The normalized log₂ values were used to compare the transcriptomic changes using MapMan 3.6.0RC1 (Supplementary Fig. 19)⁵⁹. Gene ontology (GO) enrichment analyses were carried out using the TAIR website (https://www.arabidopsis.org/tools/go_term_enrichment.jsp).

RT-qPCR of Arabidopsis thaliana seedlings

six-day-old seedlings (5–8 seedlings/sample) grown on half-strength MS liquid medium were treated with half-strength MS liquid medium in the presence or absence of each compound. After treatment for 2 or 8 h, each sample was flash-frozen, and then total RNA was extracted using ISOGEN (NIPPON GENE, Japan). Then first-strand cDNA was gained with ReverTra Ace reverse transcriptase (Toyobo, Japan) with oligo-dT primers. A StepOnePlus Real-Time PCR System (Life Technologies, USA) was used for quantitative PCR (all primers sequences for RT-qPCR in Supplementary Data 2). Polyubiquitin 10 was used as a reference gene. To select homozygous coi1-1, grown 3-day-old heterozygous coi1-1 on a half-strength MS plate containing 10 µM JA was transferred to a half-strength MS liquid medium. After 3 days of culturing, each seedling was treated with each compound.

Statistics and reproducibility

All the experiments were performed three or more times with independent samples and reproduced with similar results. Quantitative values are expressed as the mean values ± S.D. or S.E. Student’s t-test was performed between two groups using Microsoft Office Excel. Analysis of variance (ANOVA) was performed among more than two groups using the CoStat (version 6.400) software.

Reporting summary

Further information on research design is available in the Nature Portfolio Reporting Summary linked to this article.

Data availability

The data supporting the findings of this study are available in the paper and its Supplementary Information. All data generated or analyzed are available from the corresponding authors on reasonable request. The raw RNAseq data can be downloaded from the Gene Expression Omnibus repository (GSE184730). All source data to create all bar and box plots in the main text are described in Supplementary Data 3.

References

Hanada, K., Zou, C., Lehti-Shiu, M. D., Shinozaki, K. & Shiu, S. H. Importance of lineage-specific expansion of plant tandem duplicates in the adaptive response to environmental stimuli. Plant Physiol. 148, 993–1003 (2008).
Article CAS PubMed PubMed Central Google Scholar
Panchy, N., Lehti-Shiu, M. & Shiu, S. H. Evolution of gene duplication in plants. Plant Physiol. 171, 2294–2316 (2016).
Article CAS PubMed PubMed Central Google Scholar
Chini, A., Gimenez-Ibanez, S., Goossens, A. & Solano, R. Redundancy and specificity in jasmonate signalling. Curr. Opin. Plant Biol. 33, 147–156 (2016).
Article CAS PubMed Google Scholar
Thines, B. et al. JAZ repressor proteins are targets of the SCF(COI1) complex during jasmonate signalling. Nature 448, 661–665 (2007).
Article CAS PubMed Google Scholar
Chini, A. et al. The JAZ family of repressors is the missing link in jasmonate signalling. Nature 448, 666–671 (2007).
Article CAS PubMed Google Scholar
Wasternack, C. & Feussner, I. The oxylipin pathways: biochemistry and function. Annu. Rev. Plant Biol. 69, 363–386 (2018).
Article CAS PubMed Google Scholar
Pauwels, L. & Goossens, A. The JAZ proteins: a crucial interface in the jasmonate signaling cascade. Plant Cell 23, 3089–3100 (2011).
Article CAS PubMed PubMed Central Google Scholar
Howe, G. A., Major, I. T. & Koo, A. J. Modularity in jasmonate signaling for multistress resilience. Annu. Rev. Plant Biol. 69, 387–415 (2018).
Article CAS PubMed Google Scholar
Campos, M. L. et al. Rewiring of jasmonate and phytochrome B signalling uncouples plant growth-defense tradeoffs. Nat. Commun. 7, 12570 (2016).
Article CAS PubMed PubMed Central Google Scholar
Major, I. T. et al. Regulation of growth-defense balance by the JASMONATE ZIM-DOMAIN (JAZ)-MYC transcriptional module. N. Phytol. 215, 1533–1547 (2017).
Article CAS Google Scholar
Liu, B. et al. Functional specificity, diversity and redundancy of Arabidopsis JAZ family repressors in jasmonate and COI1-regulated growth, development and defense. N. Phytol. 231, 1525–1545 (2021).
Article CAS Google Scholar
Guo, Q. et al. JAZ repressors of metabolic defense promote growth and reproductive fitness in Arabidopsis. Proc. Natl Acad. Sci. USA 115, E10768–E10777 (2018).
Article CAS PubMed PubMed Central Google Scholar
Monte, I. et al. Ligand-receptor co-evolution shaped the jasmonate pathway in land plants. Nat. Chem. Biol. 14, 480–488 (2018).
Article CAS PubMed Google Scholar
Monte, I. et al. A single JAZ repressor controls the jasmonate pathway in Marchantia polymorpha. Mol. Plant 12, 185–198 (2019).
Article CAS PubMed Google Scholar
Gimenez-Ibanez, S., Zamarreno, A. M., Garcia-Mina, J. M. & Solano, R. An evolutionarily ancient immune system governs the interactions between Pseudomonas syringae and an early-diverging land plant lineage. Curr. Biol. 29, 2270–2281.e2274 (2019).
Article CAS PubMed Google Scholar
Penuelas, M. et al. Jasmonate-related MYC transcription factors are functionally conserved in Marchantia polymorpha. Plant Cell 31, 2491–2509 (2019).
Article CAS PubMed PubMed Central Google Scholar
Monte, I. et al. An ancient COI1-independent function for reactive electrophilic oxylipins in thermotolerance. Curr. Biol. 30, 962–971.e963 (2020).
Article CAS PubMed Google Scholar
Park, S.-Y. et al. Abscisic acid inhibits type 2C protein phosphatases via the PYR/PYL family of START proteins. Science 324, 1068–1071 (2009).
Article CAS PubMed PubMed Central Google Scholar
Mosquna, A. et al. Potent and selective activation of abscisic acid receptors in vivo by mutational stabilization of their agonist-bound conformation. Proc. Natl Acad. Sci. USA 108, 20838–20843 (2011).
Article CAS PubMed PubMed Central Google Scholar
Vaidya, A. S. et al. A rationally designed agonist defines subfamily IIIA abscisic acid receptors as critical targets for manipulating transpiration. ACS Chem. Biol. 12, 2842–2848 (2017).
Article CAS PubMed Google Scholar
Vaidya, A. S. et al. Dynamic control of plant water use using designed ABA receptor agonists. Science 366, eaaw8848 (2019).
Article CAS PubMed Google Scholar
Mikame, Y. et al. Synthesis of all stereoisomers of RK460 and evaluation of their activity and selectivity as abscisic acid receptor antagonists. Chemistry 25, 3496–3500 (2019).
Article CAS PubMed Google Scholar
Uraguchi, D. et al. A femtomolar-range suicide germination stimulant for the parasitic plant Striga hermonthica. Science 362, 1301–1305 (2018).
Article CAS PubMed Google Scholar
Okamoto, M. et al. Activation of dimeric ABA receptors elicits guard cell closure, ABA-regulated gene expression, and drought tolerance. Proc. Natl Acad. Sci. USA 110, 12132–12137 (2013).
Article CAS PubMed PubMed Central Google Scholar
Halder, V. & Russinova, E. Understanding the language of drugged plants. Nat. Chem. Biol. 15, 1025–1028 (2019).
Article CAS PubMed Google Scholar
Dejonghe, W. & Russinova, E. Plant chemical genetics: from phenotype-based screens to synthetic biology. Plant Physiol. 174, 5–20 (2017).
Article CAS PubMed PubMed Central Google Scholar
Littleson, M. M. et al. Scalable total synthesis and comprehensive structure-activity relationship studies of the phytotoxin coronatine. Nat. Commun. 9, 1105 (2018).
Article PubMed PubMed Central Google Scholar
Kato, N. et al. A scalable synthesis of (+)-coronafacic acid. Chirality 32, 423–430 (2020).
Article CAS PubMed Google Scholar
Miyamoto, K. et al. Facile preparation of optically active jasmonates and their biological activities in rice. Biosci. Biotechnol. Biochem. 83, 876–881 (2019).
Article CAS PubMed Google Scholar
Watanabe, R. et al. Stereoselective syntheses of all the possible stereoisomers of coronafacic acid. ChemistryOpen 9, 1008–1017 (2020).
Article CAS PubMed PubMed Central Google Scholar
Toshima, H. & Ichihara, A. Practical stereoselective syntheses of all four stereoisomers of coronamic acid (2-Ethyl-1-aminocyclopropane-1-carboxylic acid). Biosci. Biotechnol. Biochem. 59, 497–500 (1995).
Article CAS Google Scholar
Chini, A., Ben-Romdhane, W., Hassairi, A. & Aboul-Soud, M. A. M. Identification of TIFY/JAZ family genes in Solanum lycopersicum and their regulation in response to abiotic stresses. PLoS ONE 12, e0177381 (2017).
Article PubMed PubMed Central Google Scholar
Takaoka, Y. et al. A comprehensive in vitro fluorescence anisotropy assay system for screening ligands of the jasmonate COI1-JAZ co-receptor in plants. J. Biol. Chem. 294, 5074–5081 (2019).
Article CAS PubMed PubMed Central Google Scholar
Shyu, C. et al. JAZ8 lacks a canonical degron and has an EAR motif that mediates transcriptional repression of jasmonate responses in Arabidopsis. Plant Cell 24, 536–550 (2012).
Article CAS PubMed PubMed Central Google Scholar
Thireault, C. et al. Repression of jasmonate signaling by a non-TIFY JAZ protein in Arabidopsis. Plant J. 82, 669–679 (2015).
Article CAS PubMed Google Scholar
Takaoka, Y. et al. A rationally designed JAZ subtype-selective agonist of jasmonate perception. Nat. Commun. 9, 3654 (2018).
Article PubMed PubMed Central Google Scholar
Cui, M., Du, J. & Yao, X. The binding mechanism between inositol phosphate (InsP) and the jasmonate receptor complex: a computational study. Front. Plant Sci. 9, 963 (2018).
Article PubMed PubMed Central Google Scholar
Cui, M., Zhang, K., Wu, R. & Du, J. Exploring the interaction mechanism between antagonist and the jasmonate receptor complex by molecular dynamics simulation. J. Comput. Aided Mol. Des. 36, 141–155 (2022).
Article CAS PubMed Google Scholar
Saito, R. et al. Extended JAZ degron sequence for plant hormone binding in jasmonate co-receptor of tomato SlCOI1-SlJAZ. Sci. Rep. 11, 13612 (2021).
Article CAS PubMed PubMed Central Google Scholar
Xu, P., Fang, S., Chen, H. & Cai, W. The brassinosteroid-responsive xyloglucan endotransglucosylase/hydrolase 19 (XTH19) and XTH23 genes are involved in lateral root development under salt stress in Arabidopsis. Plant J. 104, 59–75 (2020).
Article CAS PubMed Google Scholar
Boron, A. K. et al. Over-expression of AtEXLA2 alters etiolated arabidopsis hypocotyl growth. Ann. Bot. 115, 67–80 (2014).
Article PubMed PubMed Central Google Scholar
Zhu, Z. et al. Derepression of ethylene-stabilized transcription factors (EIN3/EIL1) mediates jasmonate and ethylene signaling synergy in Arabidopsis. Proc. Natl Acad. Sci. USA 108, 12539–12544 (2011).
Article CAS PubMed PubMed Central Google Scholar
Mielke, S. & Gasperini, D. Interplay between plant cell walls and Jasmonate production. Plant Cell Physiol. 60, 2629–2637 (2019).
Article CAS PubMed Google Scholar
Laraia, L., Robke, L. & Waldmann, H. Bioactive compound collections: from design to target identification. Chem 4, 705–730 (2018).
Blackwell, H. E. & Zhao, Y. Chemical genetic approaches to plant biology. Plant Physiol. 133, 448–455 (2003).
Article CAS PubMed PubMed Central Google Scholar
Chini, A. et al. A small molecule antagonizes jasmonic acid perception and auxin responses in vascular and non-vascular plants. Plant Physiol. 187, 1399–1413 (2021).
Article CAS PubMed PubMed Central Google Scholar
Yokoyama, R. & Nishitani, K. A comprehensive expression analysis of all members of a gene family encoding cell-wall enzymes allowed us to predict cis-regulatory regions involved in cell-wall construction in specific organs of Arabidopsis. Plant Cell Physiol. 42, 1025–1033 (2001).
Article CAS PubMed Google Scholar
Toth, R. & van der Hoorn, R. A. L. Emerging principles in plant chemical genetics. Trends Plant Sci. 15, 81–88 (2010).
Article CAS PubMed Google Scholar
Hicks, G. R. & Raikhel, N. V. Small molecules present large opportunities in plant biology. Annu. Rev. Plant Biol. 63, 261–282 (2012).
Article CAS PubMed Google Scholar
Mita, S., Suzuki-Fujii, K. & Nakamura, K. Sugar-inducible expression of a gene for beta-amylase in Arabidopsis thaliana. Plant Physiol. 107, 895–904 (1995).
Article CAS PubMed PubMed Central Google Scholar
Xie, D. X., Feys, B. F., James, S., Nieto-Rostro, M. & Turner, J. G. COI1: an Arabidopsis gene required for jasmonate-regulated defense and fertility. Science 280, 1091–1094 (1998).
Article CAS PubMed Google Scholar
Alonso, J. M., Hirayama, T., Roman, G., Nourizadeh, S. & Ecker, J. R. EIN2, a bifunctional transducer of ethylene and stress responses in Arabidopsis. Science 284, 2148–2152 (1999).
Article CAS PubMed Google Scholar
Yang, D. L. et al. Plant hormone jasmonate prioritizes defense over growth by interfering with gibberellin signaling cascade. Proc. Natl Acad. Sci. USA 109, E1192–E1200 (2012).
Article CAS PubMed PubMed Central Google Scholar
Sehr, E. M. et al. Analysis of secondary growth in the Arabidopsis shoot reveals a positive role of jasmonate signalling in cambium formation. Plant J. 63, 811–822 (2010).
Article CAS PubMed PubMed Central Google Scholar
Monte, I. et al. Rational design of a ligand-based antagonist of jasmonate perception. Nat. Chem. Biol. 10, 671–676 (2014).
Article CAS PubMed Google Scholar
Chini, A. et al. An OPR3-independent pathway uses 4,5-didehydrojasmonate for jasmonate synthesis. Nat. Chem. Biol. 14, 171–178 (2018).
Article CAS PubMed Google Scholar
Kamitani, M., Kashima, M., Tezuka, A. & Nagano, A. J. Lasy-Seq: a high-throughput library preparation method for RNA-Seq and its application in the analysis of plant responses to fluctuating temperatures. Sci. Rep. 9, 7091 (2019).
Article PubMed PubMed Central Google Scholar
Babicki, S. et al. Heatmapper: web-enabled heat mapping for all. Nucleic Acids Res. 44, W147–W153 (2016).
Article CAS PubMed PubMed Central Google Scholar
Thimm, O. et al. MAPMAN: a user-driven tool to display genomics data sets onto diagrams of metabolic pathways and other biological processes. Plant J. 37, 914–939 (2004).
Article CAS PubMed Google Scholar

Download references

Acknowledgements

We thank Dr. Takuya Kaji (Tohoku University) for his support in preparing Figures. We also thank Dr. Yasuhiro Ishimaru, Mr. Wataru Kozaki, and Ms. Hikaru Hoshikawa (Tohoku University) for their technical assistance. The computations in this study were performed using the TSUBAME3.0 supercomputer at Tokyo Institute of Technology, Research Center for Computational Science of Institute for Molecular Science (IMS), and Research Center for Advanced Computing Infrastructure of Japan Advanced Institute of Science and Technology (JAIST). This work was financially supported by a Grant-in-Aid for Scientific Research from JSPS, Japan (nos. 22KK0076, 21K19037, 20H00402, JPJSBP120229905, and JPJSBP120239903 for M.U., nos. 18H02101, 19H05283, and 21H00270 for Y.T., no. 19K06968 for N.K., 19K05378 and 20H04791 for H.S.), SUNBOR GRANT (N.K.), JSPS Core-to-Core Program Asian Chemical Biology Initiative (M.U.), Nagase Science and Technology Foundation (M.U.), and the Spanish Ministry of Science and Innovation AEI/FEDER (no. PID2019-107012RB-I00 for R.S. and A.C.).

Author information

Authors and Affiliations

Department of Chemistry, Graduate School of Science, Tohoku University, Sendai, 980-8578, Japan
Kengo Hayashi, Nobuki Kato, Haruna Nomoto, Misuzu Nakayama, Yousuke Takaoka & Minoru Ueda
Plant Genomic Network Research Team, RIKEN Center for Sustainable Resource Science, Yokohama, 230-0045, Japan
Khurram Bashir, Satoshi Takahashi, Maho Tanaka & Motoaki Seki
Department of Life Sciences, SBA School of Science and Engineering, Lahore University of Management Sciences, 54792, Lahore, Pakistan
Khurram Bashir
Plant Molecular Genetics Department, National Centre for Biotechnology (CNB), Consejo Superior de Investigaciones Cientificas (CSIC), Campus University Autonoma, 28049, Madrid, Spain
Andrea Chini & Roberto Solano
Faculty of Pharmaceutical Sciences, Hokuriku University, Kanazawa, 920-1181, Japan
Hiroaki Saito
Department of Molecular and Chemical Life Sciences, Graduate School of Life Sciences, Tohoku University, Sendai, 980-8578, Japan
Raku Watanabe & Minoru Ueda
Faculty of Agriculture, Ryukoku University, Shiga, 520-2194, Japan
Atsushi J. Nagano
Institute for Advanced Biosciences, Keio University, Yamagata, 997-0017, Japan
Atsushi J. Nagano

Authors

Kengo Hayashi
View author publications
You can also search for this author in PubMed Google Scholar
Nobuki Kato
View author publications
You can also search for this author in PubMed Google Scholar
Khurram Bashir
View author publications
You can also search for this author in PubMed Google Scholar
Haruna Nomoto
View author publications
You can also search for this author in PubMed Google Scholar
Misuzu Nakayama
View author publications
You can also search for this author in PubMed Google Scholar
Andrea Chini
View author publications
You can also search for this author in PubMed Google Scholar
Satoshi Takahashi
View author publications
You can also search for this author in PubMed Google Scholar
Hiroaki Saito
View author publications
You can also search for this author in PubMed Google Scholar
Raku Watanabe
View author publications
You can also search for this author in PubMed Google Scholar
Yousuke Takaoka
View author publications
You can also search for this author in PubMed Google Scholar
Maho Tanaka
View author publications
You can also search for this author in PubMed Google Scholar
Atsushi J. Nagano
View author publications
You can also search for this author in PubMed Google Scholar
Motoaki Seki
View author publications
You can also search for this author in PubMed Google Scholar
Roberto Solano
View author publications
You can also search for this author in PubMed Google Scholar
Minoru Ueda
View author publications
You can also search for this author in PubMed Google Scholar

Contributions

Conceptualization: M.U.; methodology: M.U., K.H., N.K., H.S., K.B., and Y.T.; validation: K.H., Y.T., R.S., M.S., R.S., and M.U.; formal analysis: K.H., H.N., M.N., A.C., H.S., Y.T., K.B., S.T., M.T., and A.N.; investigation: K.H., H.N., M.N., A.C., H.S., Y.T., K.B., S.T., M.T., A.N., R.S., M.S., and M.U.; resources: N.K., K.M., and K.O.; data curation: A.C., Y.T., K.B., A.N., M.S., R.S., and M.U.; writing—original draft preparation: M.U.; writing—review and editing: K.H., N.K., A.C., K.B., M.S., R.S., and M.U.; visualization: K.H., H.N., M.N., A.C., Y.T., and K.B.; supervision: M.U.; project administration: M.U.; funding acquisition: M.U., Y.T., and N.K. All authors have read and agreed to the published version of the manuscript.

Corresponding author

Correspondence to Minoru Ueda.

Ethics declarations

Competing interests

The authors declare no competing interests.

Peer review

Peer review information

Communications Biology thanks Carlos Figueroa and the other, anonymous, reviewer(s) for their contribution to the peer review of this work. Primary Handling Editors: José Estevez and Gene Chong. Peer reviewer reports are available.

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Peer Review File

Supplementary Information

Description of Additional Supplementary Files

Supplementary Data 1

Supplementary Data 2

Supplementary Data 3

Reporting Summary

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

Reprints and permissions

About this article

Cite this article

Hayashi, K., Kato, N., Bashir, K. et al. Subtype-selective agonists of plant hormone co-receptor COI1-JAZs identified from the stereoisomers of coronatine. Commun Biol 6, 320 (2023). https://doi.org/10.1038/s42003-023-04709-1

Download citation

Received: 19 October 2022
Accepted: 14 March 2023
Published: 25 March 2023
DOI: https://doi.org/10.1038/s42003-023-04709-1

Comments

By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.