title: Detection of the virulent form of AVR3a from Phytophthora infestans following artificial evolution of potato resistance gene R3a.
abstract: Engineering resistance genes to gain effector recognition is emerging as an important step in attaining broad, durable resistance. We engineered potato resistance gene R3a to gain recognition of the virulent AVR3aEM effector form of Phytophthora infestans. Random mutagenesis, gene shuffling and site-directed mutagenesis of R3a were conducted to produce R3a* variants with gain of recognition towards AVR3aEM. Programmed cell death following gain of recognition was enhanced in iterative rounds of artificial evolution and neared levels observed for recognition of AVR3aKI by R3a. We demonstrated that R3a*-mediated recognition responses, like for R3a, are dependent on SGT1 and HSP90. In addition, this gain of response is associated with re-localisation of R3a* variants from the cytoplasm to late endosomes when co-expressed with either AVR3aKI or AVR3aEM a mechanism that was previously only seen for R3a upon co-infiltration with AVR3aKI. Similarly, AVR3aEM specifically re-localised to the same vesicles upon recognition by R3a* variants, but not with R3a. R3a and R3a* provide resistance to P. infestans isolates expressing AVR3aKI but not those homozygous for AVR3aEM.
authors.affiliation: Cell and Molecular Sciences, James Hutton Institute, Invergowrie-Dundee, United Kingdom.
authors.forename: Sean
authors.initials: S
authors.lastname: Chapman
authors.affiliation: Cell and Molecular Sciences, James Hutton Institute, Invergowrie-Dundee, United Kingdom; Division of Plant Sciences, University of Dundee at James Hutton Institute, Invergowrie-Dundee, United Kingdom; Dundee Effector Consortium, Invergowrie-Dundee, United Kingdom.
authors.forename: Laura J
authors.initials: LJ
authors.lastname: Stevens
authors.affiliation: Cell and Molecular Sciences, James Hutton Institute, Invergowrie-Dundee, United Kingdom; Dundee Effector Consortium, Invergowrie-Dundee, United Kingdom.
authors.forename: Petra C
authors.initials: PC
authors.lastname: Boevink
authors.affiliation: Division of Plant Sciences, University of Dundee at James Hutton Institute, Invergowrie-Dundee, United Kingdom; Dundee Effector Consortium, Invergowrie-Dundee, United Kingdom.
authors.forename: Stefan
authors.initials: S
authors.lastname: Engelhardt
authors.affiliation: Biomathematics and Statistics Scotland, Invergowrie-Dundee, United Kingdom.
authors.forename: Colin J
authors.initials: CJ
authors.lastname: Alexander
authors.affiliation: Cell and Molecular Sciences, James Hutton Institute, Invergowrie-Dundee, United Kingdom.
authors.forename: Brian
authors.initials: B
authors.lastname: Harrower
authors.affiliation: J.R. Simplot Company, Simplot Plant Sciences, Boise, Idaho, United States of America.
authors.forename: Nicolas
authors.initials: N
authors.lastname: Champouret
authors.affiliation: Cell and Molecular Sciences, James Hutton Institute, Invergowrie-Dundee, United Kingdom.
authors.forename: Kara
authors.initials: K
authors.lastname: McGeachy
authors.affiliation: Cell and Molecular Sciences, James Hutton Institute, Invergowrie-Dundee, United Kingdom; Division of Plant Sciences, University of Dundee at James Hutton Institute, Invergowrie-Dundee, United Kingdom; Dundee Effector Consortium, Invergowrie-Dundee, United Kingdom.
authors.forename: Pauline S M
authors.initials: PS
authors.lastname: Van Weymers
authors.affiliation: Cell and Molecular Sciences, James Hutton Institute, Invergowrie-Dundee, United Kingdom; Dundee Effector Consortium, Invergowrie-Dundee, United Kingdom.
authors.forename: Xinwei
authors.initials: X
authors.lastname: Chen
authors.affiliation: Cell and Molecular Sciences, James Hutton Institute, Invergowrie-Dundee, United Kingdom; Division of Plant Sciences, University of Dundee at James Hutton Institute, Invergowrie-Dundee, United Kingdom; Dundee Effector Consortium, Invergowrie-Dundee, United Kingdom.
authors.forename: Paul R J
authors.initials: PR
authors.lastname: Birch
authors.affiliation: Cell and Molecular Sciences, James Hutton Institute, Invergowrie-Dundee, United Kingdom; Dundee Effector Consortium, Invergowrie-Dundee, United Kingdom.
authors.forename: Ingo
authors.initials: I
authors.lastname: Hein
mesh_terms: D060054:Agrobacterium / Q000502:physiology; D017209:Apoptosis; D032621:DNA Shuffling; D019020:Directed Molecular Evolution; D060467:Disease Resistance / Q000235:genetics*; D011992:Endosomes / Q000378:metabolism; D017343:Genes, Plant; D006720:Homozygote; D016297:Mutagenesis, Site-Directed; D009154:Mutation / Q000235:genetics; D055750:Phytophthora infestans / Q000302:isolation & purification / Q000378:metabolism* / Q000472:pathogenicity*; D010935:Plant Diseases / Q000235:genetics / Q000382:microbiology; D010940:Plant Proteins / Q000378:metabolism; D030821:Plants, Genetically Modified; D011401:Promoter Regions, Genetic / Q000235:genetics; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D011198:Solanum tuberosum / Q000235:genetics* / Q000382:microbiology*; D014774:Virulence; D037521:Virulence Factors
chemical_list: C562170:AVR3a protein, Phytophthora infestans; D010940:Plant Proteins; D011993:Recombinant Fusion Proteins; D037521:Virulence Factors