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Title: Directed Evolution of FLS2 towards Novel Flagellin Peptide Recognition

Abstract

Microbe-associated molecular patterns (MAMPs) are molecules, or domains within molecules, that are conserved across microbial taxa and can be recognized by a plant or animal immune system. Although MAMP receptors have evolved to recognize conserved epitopes, the MAMPs in some microbial species or strains have diverged sufficiently to render them unrecognizable by some host immune systems. In this study, we carried out in vitro evolution of the Arabidopsis thaliana flagellin receptor FLAGELLIN-SENSING 2 (FLS2) to isolate derivatives that recognize one or more flagellin peptides from bacteria for which the wildtype Arabidopsis FLS2 confers little or no response. A targeted approach generated amino acid variation at FLS2 residues in a region previously implicated in flagellin recognition. The primary screen tested for elevated response to the canonical flagellin peptide from Pseudomonas aeruginosa, flg22. From this pool, we then identified five alleles of FLS2 that confer modest (quantitatively partial) recognition of an Erwinia amylovora flagellin peptide. Use of this Erwinia-based flagellin peptide to stimulate Arabidopsis plants expressing the resulting FLS2 alleles did not lead to a detectable reduction of virulent P. syringae pv. tomato growth. However, combination of two identified mutations into a single allele further increased FLS2-mediated responses to the E. amylovoramore » flagellin peptide. Furthermore, these studies demonstrate the potential to raise the sensitivity of MAMP receptors toward particular targets.« less

Authors:
; ; ORCiD logo;
Publication Date:
Research Org.:
Univ. of Wisconsin, Madison, Madison, WI (United States)
Sponsoring Org.:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
OSTI Identifier:
1346606
Alternate Identifier(s):
OSTI ID: 1287175
Grant/Contract Number:  
FG02-02ER15342
Resource Type:
Published Article
Journal Name:
PLoS ONE
Additional Journal Information:
Journal Name: PLoS ONE Journal Volume: 11 Journal Issue: 6; Journal ID: ISSN 1932-6203
Publisher:
Public Library of Science (PLoS)
Country of Publication:
United States
Language:
English
Subject:
59 BASIC BIOLOGICAL SCIENCES; leucine-rich repeat; bacterial elicitor flagellin; disease-resistance gene; plant immune-responses; elongation-factor tu; pseudomonas-syringae; arabidopsis-thaliana; innate immunity; receptor efr; nicotiana-benthamiana

Citation Formats

Helft, Laura, Thompson, Mikayla, Bent, Andrew F., and Vinatzer, ed., Boris Alexander. Directed Evolution of FLS2 towards Novel Flagellin Peptide Recognition. United States: N. p., 2016. Web. doi:10.1371/journal.pone.0157155.
Helft, Laura, Thompson, Mikayla, Bent, Andrew F., & Vinatzer, ed., Boris Alexander. Directed Evolution of FLS2 towards Novel Flagellin Peptide Recognition. United States. doi:10.1371/journal.pone.0157155.
Helft, Laura, Thompson, Mikayla, Bent, Andrew F., and Vinatzer, ed., Boris Alexander. Mon . "Directed Evolution of FLS2 towards Novel Flagellin Peptide Recognition". United States. doi:10.1371/journal.pone.0157155.
@article{osti_1346606,
title = {Directed Evolution of FLS2 towards Novel Flagellin Peptide Recognition},
author = {Helft, Laura and Thompson, Mikayla and Bent, Andrew F. and Vinatzer, ed., Boris Alexander},
abstractNote = {Microbe-associated molecular patterns (MAMPs) are molecules, or domains within molecules, that are conserved across microbial taxa and can be recognized by a plant or animal immune system. Although MAMP receptors have evolved to recognize conserved epitopes, the MAMPs in some microbial species or strains have diverged sufficiently to render them unrecognizable by some host immune systems. In this study, we carried out in vitro evolution of the Arabidopsis thaliana flagellin receptor FLAGELLIN-SENSING 2 (FLS2) to isolate derivatives that recognize one or more flagellin peptides from bacteria for which the wildtype Arabidopsis FLS2 confers little or no response. A targeted approach generated amino acid variation at FLS2 residues in a region previously implicated in flagellin recognition. The primary screen tested for elevated response to the canonical flagellin peptide from Pseudomonas aeruginosa, flg22. From this pool, we then identified five alleles of FLS2 that confer modest (quantitatively partial) recognition of an Erwinia amylovora flagellin peptide. Use of this Erwinia-based flagellin peptide to stimulate Arabidopsis plants expressing the resulting FLS2 alleles did not lead to a detectable reduction of virulent P. syringae pv. tomato growth. However, combination of two identified mutations into a single allele further increased FLS2-mediated responses to the E. amylovora flagellin peptide. Furthermore, these studies demonstrate the potential to raise the sensitivity of MAMP receptors toward particular targets.},
doi = {10.1371/journal.pone.0157155},
journal = {PLoS ONE},
number = 6,
volume = 11,
place = {United States},
year = {2016},
month = {6}
}

Journal Article:
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DOI: 10.1371/journal.pone.0157155

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