ATR2Cala2 from Arabidopsis-infecting downy mildew requires 4 TIR-NLR immune receptors for full recognition

Dae Sung Kim; Yufei Li; Hee Kyung Ahn; Alison Woods-Tör; Volkan Cevik; Oliver J. Furzer; Wenbo Ma; Mahmut Tör; Jonathan D.G. Jones

doi:10.1111/nph.19790

ATR2^C^ala2 from Arabidopsis-infecting downy mildew requires 4 TIR-NLR immune receptors for full recognition

Dae Sung Kim, Yufei Li, Hee Kyung Ahn, Alison Woods-Tör, Volkan Cevik, Oliver J. Furzer, Wenbo Ma, Mahmut Tör, Jonathan D.G. Jones

Research output: Contribution to journal › Article › peer-review

Abstract

Arabidopsis Col-0 RPP2A and RPP2B confer recognition of Arabidopsis downy mildew (Hyaloperonospora arabidopsidis [Hpa]) isolate Cala2, but the identity of the recognized ATR2^Cala2 effector was unknown. To reveal ATR2^Cala2, an F₂ population was generated from a cross between Hpa-Cala2 and Hpa-Noks1. We identified ATR2^Cala2 as a non-canonical RxLR-type effector that carries a signal peptide, a dEER motif, and WY domains but no RxLR motif. Recognition of ATR2^Cala2 and its effector function were verified by biolistic bombardment, ectopic expression and Hpa infection. ATR2^Cala2 is recognized in accession Col-0 but not in Ler-0 in which RPP2A and RPP2B are absent. In ATR2^Emoy2 and ATR2^Noks1 alleles, a frameshift results in an early stop codon. RPP2A and RPP2B are essential for the recognition of ATR2^Cala2. Stable and transient expression of ATR2^Cala2 under 35S promoter in Arabidopsis and Nicotiana benthamiana enhances disease susceptibility. Two additional Col-0 TIR-NLR (TNL) genes (RPP2C and RPP2D) adjacent to RPP2A and RPP2B are quantitatively required for full resistance to Hpa-Cala2. We compared RPP2 haplotypes in multiple Arabidopsis accessions and showed that all four genes are present in all ATR2^Cala2-recognizing accessions.

Original language	English
Pages (from-to)	330-344
Number of pages	15
Journal	New Phytologist
Volume	243
Issue number	1
Early online date	14 May 2024
DOIs	https://doi.org/10.1111/nph.19790
Publication status	Published - 31 Jul 2024

Data Availability Statement

All the sequence data used in this study can be found in NCBI (see the Materials and Methods section; https://www.ncbi.nlm.nih.gov/sra/SRX13788375; https://www.ncbi.nlm.nih.gov/sra/SRX13788374; https://www.ncbi.nlm.nih.gov/nuccore/ON994189.1/; https://www.ncbi.nlm.nih.gov/nuccore/ON994190.1/; https://www.ncbi.nlm.nih.gov/bioproject/PRJNA955397/).

Funding

Financial support from the Gatsby Charitable Foundation ( http://www.gatsby.org.uk/ ), and from BBSRC grants BB/K009176/1 and BB/M003809/1 to JDGJ, is gratefully acknowledged. This work is also supported in part by the grant 09 963/A from the Leverhulme Trust to MT. We thank Matthew Smoker and Jodie Taylor for their help with transformation. The authors would like to thank Dr Kenichi Tsuda for providing luciferase assay kit for DSK. Arabidopsis

Funders	Funder number
Gatsby Charitable Foundation
Leverhulme Trust
Biotechnology and Biological Sciences Research Council	09 963/A, BB/M003809/1, BB/K009176/1
Biotechnology and Biological Sciences Research Council

Keywords

Arabidopsis
ATR2
Hyaloperonospora arabidopsidis
plant immunity
RPP2
RxLR effector
TNL

ASJC Scopus subject areas

Physiology
Plant Science

Access to Document

10.1111/nph.19790Licence: CC BY

Cite this

@article{714d897742d34955af1c3f3a8d895e24,

title = "ATR2Cala2 from Arabidopsis-infecting downy mildew requires 4 TIR-NLR immune receptors for full recognition",

abstract = "Arabidopsis Col-0 RPP2A and RPP2B confer recognition of Arabidopsis downy mildew (Hyaloperonospora arabidopsidis [Hpa]) isolate Cala2, but the identity of the recognized ATR2Cala2 effector was unknown. To reveal ATR2Cala2, an F2 population was generated from a cross between Hpa-Cala2 and Hpa-Noks1. We identified ATR2Cala2 as a non-canonical RxLR-type effector that carries a signal peptide, a dEER motif, and WY domains but no RxLR motif. Recognition of ATR2Cala2 and its effector function were verified by biolistic bombardment, ectopic expression and Hpa infection. ATR2Cala2 is recognized in accession Col-0 but not in Ler-0 in which RPP2A and RPP2B are absent. In ATR2Emoy2 and ATR2Noks1 alleles, a frameshift results in an early stop codon. RPP2A and RPP2B are essential for the recognition of ATR2Cala2. Stable and transient expression of ATR2Cala2 under 35S promoter in Arabidopsis and Nicotiana benthamiana enhances disease susceptibility. Two additional Col-0 TIR-NLR (TNL) genes (RPP2C and RPP2D) adjacent to RPP2A and RPP2B are quantitatively required for full resistance to Hpa-Cala2. We compared RPP2 haplotypes in multiple Arabidopsis accessions and showed that all four genes are present in all ATR2Cala2-recognizing accessions.",

keywords = "Arabidopsis, ATR2, Hyaloperonospora arabidopsidis, plant immunity, RPP2, RxLR effector, TNL",

author = "Kim, {Dae Sung} and Yufei Li and Ahn, {Hee Kyung} and Alison Woods-T{\"o}r and Volkan Cevik and Furzer, {Oliver J.} and Wenbo Ma and Mahmut T{\"o}r and Jones, {Jonathan D.G.}",

year = "2024",

month = jul,

day = "31",

doi = "10.1111/nph.19790",

language = "English",

volume = "243",

pages = "330--344",

journal = "New Phytologist",

issn = "0028-646X",

publisher = "Wiley-Blackwell Publishing Ltd",

number = "1",

}

TY - JOUR

T1 - ATR2Cala2 from Arabidopsis-infecting downy mildew requires 4 TIR-NLR immune receptors for full recognition

AU - Kim, Dae Sung

AU - Li, Yufei

AU - Ahn, Hee Kyung

AU - Woods-Tör, Alison

AU - Cevik, Volkan

AU - Furzer, Oliver J.

AU - Ma, Wenbo

AU - Tör, Mahmut

AU - Jones, Jonathan D.G.

PY - 2024/7/31

Y1 - 2024/7/31

N2 - Arabidopsis Col-0 RPP2A and RPP2B confer recognition of Arabidopsis downy mildew (Hyaloperonospora arabidopsidis [Hpa]) isolate Cala2, but the identity of the recognized ATR2Cala2 effector was unknown. To reveal ATR2Cala2, an F2 population was generated from a cross between Hpa-Cala2 and Hpa-Noks1. We identified ATR2Cala2 as a non-canonical RxLR-type effector that carries a signal peptide, a dEER motif, and WY domains but no RxLR motif. Recognition of ATR2Cala2 and its effector function were verified by biolistic bombardment, ectopic expression and Hpa infection. ATR2Cala2 is recognized in accession Col-0 but not in Ler-0 in which RPP2A and RPP2B are absent. In ATR2Emoy2 and ATR2Noks1 alleles, a frameshift results in an early stop codon. RPP2A and RPP2B are essential for the recognition of ATR2Cala2. Stable and transient expression of ATR2Cala2 under 35S promoter in Arabidopsis and Nicotiana benthamiana enhances disease susceptibility. Two additional Col-0 TIR-NLR (TNL) genes (RPP2C and RPP2D) adjacent to RPP2A and RPP2B are quantitatively required for full resistance to Hpa-Cala2. We compared RPP2 haplotypes in multiple Arabidopsis accessions and showed that all four genes are present in all ATR2Cala2-recognizing accessions.

AB - Arabidopsis Col-0 RPP2A and RPP2B confer recognition of Arabidopsis downy mildew (Hyaloperonospora arabidopsidis [Hpa]) isolate Cala2, but the identity of the recognized ATR2Cala2 effector was unknown. To reveal ATR2Cala2, an F2 population was generated from a cross between Hpa-Cala2 and Hpa-Noks1. We identified ATR2Cala2 as a non-canonical RxLR-type effector that carries a signal peptide, a dEER motif, and WY domains but no RxLR motif. Recognition of ATR2Cala2 and its effector function were verified by biolistic bombardment, ectopic expression and Hpa infection. ATR2Cala2 is recognized in accession Col-0 but not in Ler-0 in which RPP2A and RPP2B are absent. In ATR2Emoy2 and ATR2Noks1 alleles, a frameshift results in an early stop codon. RPP2A and RPP2B are essential for the recognition of ATR2Cala2. Stable and transient expression of ATR2Cala2 under 35S promoter in Arabidopsis and Nicotiana benthamiana enhances disease susceptibility. Two additional Col-0 TIR-NLR (TNL) genes (RPP2C and RPP2D) adjacent to RPP2A and RPP2B are quantitatively required for full resistance to Hpa-Cala2. We compared RPP2 haplotypes in multiple Arabidopsis accessions and showed that all four genes are present in all ATR2Cala2-recognizing accessions.

KW - Arabidopsis

KW - ATR2

KW - Hyaloperonospora arabidopsidis

KW - plant immunity

KW - RPP2

KW - RxLR effector

KW - TNL

UR - http://www.scopus.com/inward/record.url?scp=85192901124&partnerID=8YFLogxK

U2 - 10.1111/nph.19790

DO - 10.1111/nph.19790

M3 - Article

AN - SCOPUS:85192901124

SN - 0028-646X

VL - 243

SP - 330

EP - 344

JO - New Phytologist

JF - New Phytologist

IS - 1

ER -