With this approach, we aim to characterize more PRRs against different pathogens & pests in the future.
September 4, 2025 at 7:33 PM
With this approach, we aim to characterize more PRRs against different pathogens & pests in the future.
To summarize, we reported the strategies to discover and engineer plant immune receptors.
As usual, we have uploaded the data, including RCM analysis, receptor & peptide sequences, raw data…etc etc on Zenodo
(zenodo.org/records/1546... )
As usual, we have uploaded the data, including RCM analysis, receptor & peptide sequences, raw data…etc etc on Zenodo
(zenodo.org/records/1546... )
September 4, 2025 at 7:33 PM
To summarize, we reported the strategies to discover and engineer plant immune receptors.
As usual, we have uploaded the data, including RCM analysis, receptor & peptide sequences, raw data…etc etc on Zenodo
(zenodo.org/records/1546... )
As usual, we have uploaded the data, including RCM analysis, receptor & peptide sequences, raw data…etc etc on Zenodo
(zenodo.org/records/1546... )
Together with structural prediction, phylogenomics data, and lots of in-planta validations, we engineered synthetic SCORE that perceive CSPs from a range of plant pathogens and pests.
September 4, 2025 at 7:33 PM
Together with structural prediction, phylogenomics data, and lots of in-planta validations, we engineered synthetic SCORE that perceive CSPs from a range of plant pathogens and pests.
With the polymorphic CSP recognition specificity, we wonder if SCOREs can be engineered.
We predicted receptor-peptide structures to narrow down the region of interest.
We predicted receptor-peptide structures to narrow down the region of interest.
September 4, 2025 at 7:33 PM
With the polymorphic CSP recognition specificity, we wonder if SCOREs can be engineered.
We predicted receptor-peptide structures to narrow down the region of interest.
We predicted receptor-peptide structures to narrow down the region of interest.
They do!
Across the 22 SOCRE orthologs we tested, most exhibit unique recognition specificity towards different CSP peptides.
Across the 22 SOCRE orthologs we tested, most exhibit unique recognition specificity towards different CSP peptides.
September 4, 2025 at 7:33 PM
They do!
Across the 22 SOCRE orthologs we tested, most exhibit unique recognition specificity towards different CSP peptides.
Across the 22 SOCRE orthologs we tested, most exhibit unique recognition specificity towards different CSP peptides.
Bioinformatic analyses indicate that CSPs are very diverse across organisms (not just found in bacteria).
We wonder if SCORE orthologs across the flowering plant lineage can perceive different CSP peptides.
We wonder if SCORE orthologs across the flowering plant lineage can perceive different CSP peptides.
September 4, 2025 at 7:33 PM
Bioinformatic analyses indicate that CSPs are very diverse across organisms (not just found in bacteria).
We wonder if SCORE orthologs across the flowering plant lineage can perceive different CSP peptides.
We wonder if SCORE orthologs across the flowering plant lineage can perceive different CSP peptides.
Unlike the previously identified PRR CORE, "181" exhibits a more selective recognition towards csp15 variants, so we named it “Selective CORE” (SCORE).
In addition, CORE and SCORE are not closely related; these receptors convergently evolved to perceive CSPs.
In addition, CORE and SCORE are not closely related; these receptors convergently evolved to perceive CSPs.
September 4, 2025 at 7:33 PM
Unlike the previously identified PRR CORE, "181" exhibits a more selective recognition towards csp15 variants, so we named it “Selective CORE” (SCORE).
In addition, CORE and SCORE are not closely related; these receptors convergently evolved to perceive CSPs.
In addition, CORE and SCORE are not closely related; these receptors convergently evolved to perceive CSPs.
We screened for chimeric receptors activated by the plant pathogen Agrobacterium, and identified the receptor “181” from Pomelo.
Following biochemical analyses, we discovered that 181 perceives cold shock protein (CSP) peptides.
Following biochemical analyses, we discovered that 181 perceives cold shock protein (CSP) peptides.
September 4, 2025 at 7:33 PM
We screened for chimeric receptors activated by the plant pathogen Agrobacterium, and identified the receptor “181” from Pomelo.
Following biochemical analyses, we discovered that 181 perceives cold shock protein (CSP) peptides.
Following biochemical analyses, we discovered that 181 perceives cold shock protein (CSP) peptides.
To characterize immune receptors (PRRs) in plants, we cluster PRRs into subgroups based on conservation of their inner LRR residues.
We obtained >1000s subgroups with this approach, and cloned ~210s into receptor chimeras for further characterization.
We obtained >1000s subgroups with this approach, and cloned ~210s into receptor chimeras for further characterization.
September 4, 2025 at 7:33 PM
To characterize immune receptors (PRRs) in plants, we cluster PRRs into subgroups based on conservation of their inner LRR residues.
We obtained >1000s subgroups with this approach, and cloned ~210s into receptor chimeras for further characterization.
We obtained >1000s subgroups with this approach, and cloned ~210s into receptor chimeras for further characterization.
To summarize, we:
-Designed a system to characterize the pathogen recognition landscape of LRR-RLK-XIIs in plants
-Identified an ancient but lineage-specific PRR that exhibits remarkable polymorphism in MAMP recognition
-Designed PRRs to perceive specific MAMPs from multiple crop pathogens
-Designed a system to characterize the pathogen recognition landscape of LRR-RLK-XIIs in plants
-Identified an ancient but lineage-specific PRR that exhibits remarkable polymorphism in MAMP recognition
-Designed PRRs to perceive specific MAMPs from multiple crop pathogens
March 18, 2025 at 2:01 AM
To summarize, we:
-Designed a system to characterize the pathogen recognition landscape of LRR-RLK-XIIs in plants
-Identified an ancient but lineage-specific PRR that exhibits remarkable polymorphism in MAMP recognition
-Designed PRRs to perceive specific MAMPs from multiple crop pathogens
-Designed a system to characterize the pathogen recognition landscape of LRR-RLK-XIIs in plants
-Identified an ancient but lineage-specific PRR that exhibits remarkable polymorphism in MAMP recognition
-Designed PRRs to perceive specific MAMPs from multiple crop pathogens
Through this mini scan, we designed SCORE variants that robustly perceives CSPs from many devastating crop pathogens. Hopefully these would improve disease resistance against pathogens in crops
(XXI)
(XXI)
March 18, 2025 at 2:01 AM
Through this mini scan, we designed SCORE variants that robustly perceives CSPs from many devastating crop pathogens. Hopefully these would improve disease resistance against pathogens in crops
(XXI)
(XXI)
We did a mini-mutational scan on these 3 AAs, generated 37 SCORE variants and tested them against 104 csp15 peptides. To a certain extent, the surface charges in this region determine csp15 recognition specificity in SCORE
(XX)
(XX)
March 18, 2025 at 2:01 AM
We did a mini-mutational scan on these 3 AAs, generated 37 SCORE variants and tested them against 104 csp15 peptides. To a certain extent, the surface charges in this region determine csp15 recognition specificity in SCORE
(XX)
(XX)
Due to the AF prediction of ionic bonds between csp15 & 10th LRR of SCORE, we noticed that the surface charges in this region are different between SCORE orthologs. We can also drastically modify the surface charges, simply by mutating 3 AA residues in the 10th LRR
(XIX)
(XIX)
March 18, 2025 at 2:01 AM
Due to the AF prediction of ionic bonds between csp15 & 10th LRR of SCORE, we noticed that the surface charges in this region are different between SCORE orthologs. We can also drastically modify the surface charges, simply by mutating 3 AA residues in the 10th LRR
(XIX)
(XIX)
This indicates that 8th-11th LRR determines csp15 recognition specificity in SCOREs. We perform a bunch of LRR swaps between orthologs and narrow it further down to the 10th LRR. csp15 recognition specificity can, in an extend, be transferred between SCORE orthologs
(XVIII)
(XVIII)
March 18, 2025 at 2:01 AM
This indicates that 8th-11th LRR determines csp15 recognition specificity in SCOREs. We perform a bunch of LRR swaps between orthologs and narrow it further down to the 10th LRR. csp15 recognition specificity can, in an extend, be transferred between SCORE orthologs
(XVIII)
(XVIII)
These predictions became convincing when combined with phylogenomics. While the AAs that interacts with csp15 in 3rd-7th & 12th-16thLRR are relatively conserved between SCORE orthologs, the LRR 8th-11th AAs that interacts with the variable region of csp15 are also variable
(XVII)
(XVII)
March 18, 2025 at 2:01 AM
These predictions became convincing when combined with phylogenomics. While the AAs that interacts with csp15 in 3rd-7th & 12th-16thLRR are relatively conserved between SCORE orthologs, the LRR 8th-11th AAs that interacts with the variable region of csp15 are also variable
(XVII)
(XVII)
We generated AF models of SCORE-csp15. AF predictions indicate that the N-terminus of csp15 binds to the 12th-16th LRR of SCORE, and C-terminus of csp15 binds to the 3rd-7th LRR of SCORE. The variable central region of csp15 interacts with the 8th-11th LRR of SCORE
(XVI)
(XVI)
March 18, 2025 at 2:01 AM
We generated AF models of SCORE-csp15. AF predictions indicate that the N-terminus of csp15 binds to the 12th-16th LRR of SCORE, and C-terminus of csp15 binds to the 3rd-7th LRR of SCORE. The variable central region of csp15 interacts with the 8th-11th LRR of SCORE
(XVI)
(XVI)
I was very excited to discover that SCOREs exhibit highly diverse recognition specificity towards CSP variants, suggesting dynamic evolutionary arms races between this PRR & the MAMP. This led us to our next question: How do SCOREs achieve their specificity in CSP recognition?
(XV)
(XV)
March 18, 2025 at 2:01 AM
I was very excited to discover that SCOREs exhibit highly diverse recognition specificity towards CSP variants, suggesting dynamic evolutionary arms races between this PRR & the MAMP. This led us to our next question: How do SCOREs achieve their specificity in CSP recognition?
(XV)
(XV)
& SCORE evolution: We found functional SCORE orthologs in Magnoliids, Oxalidales, Myrtales & Sapindales. SCORE is thus relatively ancient (>150MYA) & subsequently lost in multiple angiosperm lineages. Here's some photos of plants (most taken by myself) with SCOREs
(🍁🍋🍊🌳)
(🍁🍋🍊🌳)
March 18, 2025 at 2:01 AM
& SCORE evolution: We found functional SCORE orthologs in Magnoliids, Oxalidales, Myrtales & Sapindales. SCORE is thus relatively ancient (>150MYA) & subsequently lost in multiple angiosperm lineages. Here's some photos of plants (most taken by myself) with SCOREs
(🍁🍋🍊🌳)
(🍁🍋🍊🌳)
Before that, here's some CSP bioinformatics: CSPs are found across multiple kingdoms. We selected 104 highly conserved csp15 peptides that represents 73% (35881) of all searched species with CSPs. Some csp15s are particularly enriched in some kingdoms
(🦠🥒🍄🌿🪱🐈)
(🦠🥒🍄🌿🪱🐈)
March 18, 2025 at 2:01 AM
Before that, here's some CSP bioinformatics: CSPs are found across multiple kingdoms. We selected 104 highly conserved csp15 peptides that represents 73% (35881) of all searched species with CSPs. Some csp15s are particularly enriched in some kingdoms
(🦠🥒🍄🌿🪱🐈)
(🦠🥒🍄🌿🪱🐈)
We were interested in the selectivity of csp22 by SCOREs. How selective are SCOREs? Does CSP selectivity by SCOREs evolve? To answer these questions, we investigate the interaction between 104 csp15 peptides and 21 SCORE orthologs (2163 peptide-receptor interactions)
(XIV)
(XIV)
March 18, 2025 at 2:01 AM
We were interested in the selectivity of csp22 by SCOREs. How selective are SCOREs? Does CSP selectivity by SCOREs evolve? To answer these questions, we investigate the interaction between 104 csp15 peptides and 21 SCORE orthologs (2163 peptide-receptor interactions)
(XIV)
(XIV)
CORE & SCORE are not closely related & have convergently evolved to perceive csp15/22. They also exhibit differential preferences for csp22 variants. csp22 perception by CORE heavily rely on conserved aromatic AAs, while SCORE rely on variable AAs in the middle of csp22
(XIII)
(XIII)
March 18, 2025 at 2:01 AM
CORE & SCORE are not closely related & have convergently evolved to perceive csp15/22. They also exhibit differential preferences for csp22 variants. csp22 perception by CORE heavily rely on conserved aromatic AAs, while SCORE rely on variable AAs in the middle of csp22
(XIII)
(XIII)
Surprisingly, 181 responses to csp22ᵐᶦᶜ but not CSPᶜᵘᵖ, while CORE responses to both. BAK1-interaction & in-vivo peptide binding assays also suggest that 181 perceives csp22ᵐᶦᶜ, but not csp22ᶜᵘᵖ. So, we named 181 ‘Specific COld-shock protein REceptor’ (SCORE)
(XII)
(XII)
March 18, 2025 at 2:01 AM
Surprisingly, 181 responses to csp22ᵐᶦᶜ but not CSPᶜᵘᵖ, while CORE responses to both. BAK1-interaction & in-vivo peptide binding assays also suggest that 181 perceives csp22ᵐᶦᶜ, but not csp22ᶜᵘᵖ. So, we named 181 ‘Specific COld-shock protein REceptor’ (SCORE)
(XII)
(XII)
High conservation & abundance of CSPs make them good candidates as MAMPs. A Solanaceae-specific PRR, "CORE", perceives a conserved peptide within CSP (csp22/15). We decided to test 181’s response against full-length CSP from Cupriavidus (cup) and csp22 from Micrococcus (mic)
(XI)
(XI)
March 18, 2025 at 2:01 AM
High conservation & abundance of CSPs make them good candidates as MAMPs. A Solanaceae-specific PRR, "CORE", perceives a conserved peptide within CSP (csp22/15). We decided to test 181’s response against full-length CSP from Cupriavidus (cup) and csp22 from Micrococcus (mic)
(XI)
(XI)
What MAMP does 181 perceive? We combined BRI1-chimera system with anion-exchange + size chromatography and identified 2 active protein fractions. 3 of the top 5 hits from MS data are cold-shock proteins (CSPs). CSPs are conserved nucleic acid-binding proteins that protects cells from❄️damage
(X)
(X)
March 18, 2025 at 2:01 AM
What MAMP does 181 perceive? We combined BRI1-chimera system with anion-exchange + size chromatography and identified 2 active protein fractions. 3 of the top 5 hits from MS data are cold-shock proteins (CSPs). CSPs are conserved nucleic acid-binding proteins that protects cells from❄️damage
(X)
(X)
We cloned 181 from the Pomelo cultivar "banpeiyu" (晩白柚; Citrus maxima). Banpeiyu produces the world’s largest citrus fruits.
Here's a photo of me holding a farm-grown banpeiyu from Kumamoto, versus the only single banpeiyu produced by my home-grown baby tree🍊
Here's a photo of me holding a farm-grown banpeiyu from Kumamoto, versus the only single banpeiyu produced by my home-grown baby tree🍊
March 18, 2025 at 2:01 AM
We cloned 181 from the Pomelo cultivar "banpeiyu" (晩白柚; Citrus maxima). Banpeiyu produces the world’s largest citrus fruits.
Here's a photo of me holding a farm-grown banpeiyu from Kumamoto, versus the only single banpeiyu produced by my home-grown baby tree🍊
Here's a photo of me holding a farm-grown banpeiyu from Kumamoto, versus the only single banpeiyu produced by my home-grown baby tree🍊