title: Multiple candidate effectors from the oomycete pathogen Hyaloperonospora arabidopsidis suppress host plant immunity.
abstract: Oomycete pathogens cause diverse plant diseases. To successfully colonize their hosts, they deliver a suite of effector proteins that can attenuate plant defenses. In the oomycete downy mildews, effectors carry a signal peptide and an RxLR motif. Hyaloperonospora arabidopsidis (Hpa) causes downy mildew on the model plant Arabidopsis thaliana (Arabidopsis). We investigated if candidate effectors predicted in the genome sequence of Hpa isolate Emoy2 (HaRxLs) were able to manipulate host defenses in different Arabidopsis accessions. We developed a rapid and sensitive screening method to test HaRxLs by delivering them via the bacterial type-three secretion system (TTSS) of Pseudomonas syringae pv tomato DC3000-LUX (Pst-LUX) and assessing changes in Pst-LUX growth in planta on 12 Arabidopsis accessions. The majority (~70%) of the 64 candidates tested positively contributed to Pst-LUX growth on more than one accession indicating that Hpa virulence likely involves multiple effectors with weak accession-specific effects. Further screening with a Pst mutant (ΔCEL) showed that HaRxLs that allow enhanced Pst-LUX growth usually suppress callose deposition, a hallmark of pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI). We found that HaRxLs are rarely strong avirulence determinants. Although some decreased Pst-LUX growth in particular accessions, none activated macroscopic cell death. Fewer HaRxLs conferred enhanced Pst growth on turnip, a non-host for Hpa, while several reduced it, consistent with the idea that turnip's non-host resistance against Hpa could involve a combination of recognized HaRxLs and ineffective HaRxLs. We verified our results by constitutively expressing in Arabidopsis a sub-set of HaRxLs. Several transgenic lines showed increased susceptibility to Hpa and attenuation of Arabidopsis PTI responses, confirming the HaRxLs' role in Hpa virulence. This study shows TTSS screening system provides a useful tool to test whether candidate effectors from eukaryotic pathogens can suppress/trigger plant defense mechanisms and to rank their effectiveness prior to subsequent mechanistic investigation.
authors.affiliation: The Sainsbury Laboratory, John Innes Centre, Norwich, United Kingdom.
authors.forename: Georgina
authors.initials: G
authors.lastname: Fabro
authors.forename: Jens
authors.initials: J
authors.lastname: Steinbrenner
authors.forename: Mary
authors.initials: M
authors.lastname: Coates
authors.forename: Naveed
authors.initials: N
authors.lastname: Ishaque
authors.forename: Laura
authors.initials: L
authors.lastname: Baxter
authors.forename: David J
authors.initials: DJ
authors.lastname: Studholme
authors.forename: Evelyn
authors.initials: E
authors.lastname: Körner
authors.forename: Rebecca L
authors.initials: RL
authors.lastname: Allen
authors.forename: Sophie J M
authors.initials: SJ
authors.lastname: Piquerez
authors.forename: Alejandra
authors.initials: A
authors.lastname: Rougon-Cardoso
authors.forename: David
authors.initials: D
authors.lastname: Greenshields
authors.forename: Rita
authors.initials: R
authors.lastname: Lei
authors.forename: Jorge L
authors.initials: JL
authors.lastname: Badel
authors.forename: Marie-Cecile
authors.initials: MC
authors.lastname: Caillaud
authors.forename: Kee-Hoon
authors.initials: KH
authors.lastname: Sohn
authors.forename: Guido
authors.initials: G
authors.lastname: Van den Ackerveken
authors.forename: Jane E
authors.initials: JE
authors.lastname: Parker
authors.forename: Jim
authors.initials: J
authors.lastname: Beynon
authors.forename: Jonathan D G
authors.initials: JD
authors.lastname: Jones
mesh_terms: D017360:Arabidopsis / Q000235:genetics / Q000276:immunology* / Q000382:microbiology; D058947:Bacterial Secretion Systems; D029688:Brassica napus / Q000276:immunology / Q000382:microbiology; D002478:Cells, Cultured; D018506:Gene Expression Regulation, Plant; D005936:Glucans / Q000096:biosynthesis / Q000378:metabolism; D054884:Host-Pathogen Interactions; D009868:Oomycetes / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D010935:Plant Diseases / Q000235:genetics / Q000276:immunology* / Q000382:microbiology; D030821:Plants, Genetically Modified; D011506:Proteins / Q000378:metabolism*; D044224:Pseudomonas syringae / Q000201:enzymology / Q000254:growth & development* / Q000276:immunology; D011993:Recombinant Fusion Proteins / Q000378:metabolism
chemical_list: D058947:Bacterial Secretion Systems; D005936:Glucans; D011506:Proteins; D011993:Recombinant Fusion Proteins; C048306:callose
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