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Title: Directed evolution of FLS2 towards novel flagellin peptide recognition

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 wild type 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.more » amylovora flagellin peptide. As a result, these studies demonstrate the potential to raise the sensitivity of MAMP receptors toward particular targets.« less
 [1] ;  [2] ;  [3]
  1. Univ. of Wisconsin, Madison, Madison, WI (United States). Dept. of Plant Pathology, Cellular and Molecular Biology Program; Howard Hughes Medical Institute, Chevy Chase, MD (United States)
  2. Univ. of Wisconsin, Madison, Madison, WI (United States). Dept. of Plant Pathology; Yale Univ., New Haven, CT (United States)
  3. Univ. of Wisconsin, Madison, Madison, WI (United States). Dept. of Plant Pathology
Publication Date:
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Accepted Manuscript
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Additional Journal Information:
Journal Volume: 11; Journal Issue: 6; Journal ID: ISSN 1932-6203
Public Library of Science
Research Org:
Univ. of Wisconsin, Madison, Madison, WI (United States)
Sponsoring Org:
USDOE Office of Science (SC), Basic Energy Sciences (BES) (SC-22)
Country of Publication:
United States
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; flagellin; seedlings; MAMP-triggered immunity; genetically modified plants; leaves; plant pathogens; sensory receptors