Pyramiding potato resistance genes against late blight Jadwiga Śliwka Instytut Hodowli i Aklimatyzacji Roślin – Państwowy Institut Badawczy Plant Breeding and Acclimatization Institute – National Research Institute The G2P-SOL project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 677379
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Pyramiding potato resistance genes against late blight · Pyramiding potato resistance genes against late blight Jadwiga Śliwka. Instytut Hodowli i Aklimatyzacji Roślin – Państwowy
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Pyramiding potato resistance genes against late blight
Jadwiga Śliwka
Instytut Hodowli i Aklimatyzacji Roślin – Państwowy Institut Badawczy
Plant Breeding and Acclimatization Institute – National Research Institute
The G2P-SOL project has received funding from the European Union’s Horizon 2020 research and innovation programme under grant agreement No 677379
Potato Pathogens Laboratory
Phytophthora infestans Solanum tuberosum
Resistance gene mapping
Wild Solanum sp. as resistance sources
Resistance QTL analysis
Resistance gene expression study
Marker-assisted selection
Testing breeding lines
Collection of isolates
Population study
Mating type, metalaxyl resistance, mitochondrial haplotype, virulence, SSR markers
Effector diversity and expression
Resistance gene cloning
Poster 1,1 Marta Brylińska
Presentation: Emil Stefańczyk
Hot spots of resistance to P. infestans on potato genetic map
• 1970 S. stenotomum x S. phureja comes from CIP, Lima, Peru Crosses with S. tuberosum 2x • 1992 first resistant tetraploid (unreduced gametes) • 2006 mapping the gene (Śliwka et al. 2006, TAG) • 2009 sequencing, virulent isolate P. infestans EC1; MP1162? Syn. Rpi-vnt1.1 (Foster et al. 2009, MPMI) • 2010 MAS (Śliwka et al. 2010, JAG) • 2012 expression of the Rpi-phu1? • 2012 pyramiding with Sárpo Mira • 2014 GMO field tests in The Sainsbury Laboratory [Jones et al.] • ???? Cultivar???
Rpi-phu1
11.05.2016
• Originates from: Bolivia, Peru
habitat: inter-Andean valleys, barren, eroded or cultivated soil
2400-3000 m above sea level
• Syn. S. sparsipilum, re-identified as • S. brevicaule
• Selected clones:
- resistant to P. infestans – low tendency to enzymatic blackening
Rpi-phu1: 12 isolates virulent (1%) Rpi-rzc1: 8 isolates virulent (0.7%) Virulent on both plants: none
The goal: Rpi-phu1 + Rpi-rzc1
• Virulence is easily lost in storage of the isolate • Virulence test: laborious and variation in results • Marker-Assisted Selection • Tetraploid breeding lines
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Resistance assessment
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leaflet tests performed on individuals from the G2P populations • P. infestans isolates
• MP324 on G2P-1, G2P-2 and G2P-3 • MP324x (virulent on plants with Rpi-phu1) on G2P-2
susceptible-to-resistant ratio consistent with the expected ratio • 1:1 (G2P-1 and G2P-3; gene Rpi-rzc1)
• 3:1 (G2P-2; Rpi-rzc1 and Rpi-phu1 genes) or 1:1
We cannot distinguish plants with Rpi-phu1 + Rpi-rzc1 from plants with Rpi-rzc1
Crosses
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resistance plants
population origin gene 1 gene 2 grown secured
G2P-1 DG 06-508 x DG 08-238 Rpi-rzc1 239 124
G2P-2 DG 97-943 x DG 99-10/36 Rpi-phu1 Rpi-rzc1 290 194
• Sequencing of the homologs • Development of more specific markers for Rpi-phu1 • Validation on diploid and tetraploid level (five F1, 500-750 plants)
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Rpi-phu1 homologs in the Rpi-rzc1 plants
primers based on the Rpi-phu1 sequence (Rpi-vnt1.1; FJ423044.1) a) covering the whole open reading frame b) spanning shorter fragments of the gene
Rpi-phu1 2676 bp
PCR
reamplification
matrix: DNA
template: purified PCR product
Sequencing, sequence comparison and design of better marker in progress
• Introgression of Rpi-rzc1 to new breeding lines and populations
• Pyramiding Rpi-rzc1+Rpi-phu1 on diploid level • MAS is not trivial, marker optimization • Transfer of Rpi-rzc1 to tetraploid level • Pyramiding Rpi-rzc1+Rpi-phu1 on tetraploid level
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Potato Pathogens Laboratory
Jadwiga Śliwka Marta Brylińska Małgorzata Frączak Anna Jarzyńska Sylwester Sobkowiak Emil Stefańczyk Paulina Smyda-Dajmund Jarosław Plich