Poster presented at EAPR Pathology & Pests Section Meeting, Arras, France

The research leading to these results has received funding from the Norwegian Financial Mechanism 2014-2021, project DivGene: UMO2019/34/H/NZ9/0055

Brurberg, May Bente

Janiszewska, Marta

Ludwiczewska, Mirella

Lysøe, Erik

Paluchowska, Paulina

Rossmann, Simeon L.

Sobkowiak, Sylwester

Yin, Zhimin

Śliwka, Jadwiga

English

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Aggressiveness test of Phytophthora infestans isolates with different effector allelesBackground: Phytophthora infestans (Mont) de Bary, is the causal agent of late blight and is oneof the main constraints of potato and tomato production worldwide. P. infestans can infectleaves, stems, potato tubers, and tomato fruits. During infection the pathogen secretes effectorproteins that suppress the defense system of the plant. Plants have evolved to recognizeeffectors, resulting in an evolutionary cycle of defense and counter-defense in plant–microbeinteractions. The pathogen has a heterothallic mating system but reproduces primarily in aclonal manner where the clonal lineages have varying aggressiveness. The lineages are routinelydefined by analysis of simple sequence repeat (SSR) markers.Mirella Ludwiczewska1, Paulina Paluchowska1, Marta Janiszewska1, Erik Lysøe2, Simeon Lim Rossmann2, Sylwester Sobkowiak1, Zhimin Yin1, May Bente Brurberg2, 3, Jadwiga Śliwka11Plant Breeding and Acclimatization Institute – National Research Institute in Radzików, Młochów Division, Poland2Norwegian Institute of Bioeconomy Research (NIBIO), Ås, Norway3Norwegian University of Life Sciences (NMBU), Ås, Norwaym.ludwiczewska@ihar.edu.plIsolate Origin Years Genotype Effector Avr-vnt1MP1976 Przecław/Poland 2021 EU13_A2 V1/V3MP1943 Węgrzce/Poland 2016 EU13_A2 V1/V3MP1960            Sulejów/Poland 2017 EU13_A2 V1/V3MP1932    Węgrzce/Poland 2020 EU13_A2 V1/V2/V3MP1995 Węgrzce/Poland 2021 EU13_A2 V1*MP1934 Karzniczka/Poland 2020 EU41_A2 V1/V2/V3MP1931 Karzniczka/Poland 2020 EU41_A2 V1/V2/V3MP1935            Węgrzce/Poland 2020 EU41_A2 V1/V2/V3MP1936 Węgrzce/Poland 2020 EU41_A2 V1/V2/V3MP1933 Karzniczka/Poland 2020 EU41_A2 V1/V2/V3MP2019 Zybiszów/ Poland 2020 EU41_A2 No dataMP1956 Węgrzce/Poland 2020 EU41_A2 No dataMP1942 Przecław/Poland 2020 EU37_A2 V1/V2MP1940 Boguchwała/Poland 2020 EU37_A2 V1/V2MP1938 Przecław/Poland 2020 EU37_A2 V1/V2MP 940 Proszowice/Poland 2008 EU34_A1 V1*MP 849 Boguchwała/Poland 2007 EU34_A1 No dataMP 938 Czaple Małe/Poland 2008 EU34_A1 V2*MP 2076 Węgrzce/Poland 2018 EU34_A1 V1*Aim: Test aggressiveness of 19 P. infestans isolates representing four genotypes prevalent inEurope: EU13_A2, EU34_A1, EU37_A2 and EU41_A2. * data from Illumina sequencing The research leading to these results has received funding from the Norwegian Financial Mechanism 2014-2021, project DivGene: UMO- 2019/34/H/NZ9/00559Table 1. P. infestans isolates used for the tests and results of sequencing of effector gene.Conclusion and future plans:• The EU37_A2 genotype of P. infestans was the most aggressive.• Based on ongoing sequencing of multiple effector genes from a large number of P. infestansisolates from Poland and Norway, we will analyse how differences in effector repertoire affectaggressiveness.• More isolates will be tasted for aggressiveness to ensure better representation of tested P.infestans genotypes.Results:Aggressiveness (Figure 2)• Isolates of EU34_A1 produced smallest amount of spores, had the longest latent period andcaused smallest lesions.• Isolates of EU37_A2 produced biggest spores, had the shortest latent period and causedbiggest lesions.• There were no statistically significant differences in the latent period between isolates ofEU13_A2 and isolates of EU41_A2 genotypes.Results of Avr-vnt1 sequencing are shown in Table 1. Diversity of Avr-vnt1 effector was notedwithin EU13_A2 and EU34_A1 genotypes.• Isolates differed in virulence between and within SSR genotypes.Methods:Aggressiveness test• 5 leaflets × 3 cultivars × 19 isolates (Table 1) × 3 replications × 2 dates• Measurement of latent period, lesion diameter, sporulation intensityVirulence test•3 leaflets × 23 cultivars × 12 isolates (Table 2)The P. infestans isolate was identified as virulent if symptoms (lesions, sporulation) wereobservedSequencing of effector gene• 12 isolates x 2 Avr-vnt1 primer pairs - were sequenced by Sanger method• 4 isolates - data from sequencing effector by Illumina sequencingStatistics• ANOVA + Tukey’s test (Statistica 13.0 software package)Figure 1. Leaflets of potato cv. Craigs Royal inoculated with isolate of P. infestans: Six days post inoculationMaterials:• Susceptible potato cultivars: Craigs Royal, Irys, Tarpan• 19 P. infestans isolates listed in Table 1Table 2. Results of virulence test.  A-avirulent isolate, V-virulent isolate. Genotypes colour-coded as in Table 1.Isolate Black’s differential set Different cultivarsBzura(homologue R2)Sarpo Mira (Rpi-Smira1, Rpi-Smira2, R3a, R3b, R4+QTL)Biogold(Rpi-abpt)99-12/8 (Rpi-mch1)99-10/36 (Rpi-rzc1)04-IX-21 (Rpi-phu1)KellyAlouette (Rpi-vnt1.3; R3a; R3b)Gardena (Rpi-phu1)MP1976 1.2.3.4.5.6.7.8.10.11 V V V V A A V V VMP1934 1.2.3.4.5.6.7.8.11 V V V V A A V A VMP1960 1.2.3.4.5.6.7.8.9.10.11 V V V V V A V V VMP1932 1.2.3.4.5.6.7.8.10.11 V A V V A A A V VMP1995 1.4.6.7.10.11 A A A A A A A A AMP1934 1.2.3.4.5.6.7.8.11 V V V V A A V A VMP1931 1.2.3.4.5.6.7.8.10.11 V A V A A A V V AMP1935 1.2.3.4.5.6.7.8.10.11 V V V V V A V A AMP1936 1.2.3.4.5.6.7.8.10.11 V A V V A A V V AMP1933 1.2.3.4.5.6.7.8.10.11 V V V A A A V V AMP2019 1.2.3.4.5.6.7.8.10.11 V A V V A A V A VMP1956 1.2.3.4..6.7.8.10.11 V A A V A A V A VMP1942 1.2.3.4.5.6.7.8.10.11 V V V A A A V A AMP1940 1.2.3.4.5.6.7.8.10.11 V A V A A A V A AMP1938 1.2.3.4.5.6.7.10.11 V A V A A A A A AMP940 1.3.4.6.7.11 A A A V A A A A AMP849 1.2.3.4.5.6.7.10.11 V A V V A A A A AMP938 1.3.4.6.7.8.10.11 V V V V A A A A AMP2076 1.4.6.7.10.11 V A A V A A A A AFigure 2. Aggressiveness of four P. infestans genotypes (EU_13A2, EU34_A1, EU37_A2 and EU41_A2) based on lesion diameter, latent period and number of sporangia produced by isolates from each genotype. Vertical bars represent 0.95 confidence intervals. The letters indicate statistically different groups, created based on ANOVA + Tukey’s test.a) Latent period (days) (Effect: F(3.163)=172.24, p=0.000)b) Number of sporangia/µl (Effect: F (3.953)=92.663, p=0.000)c) Lesion diameter (mm)-5 day (Effect: F (3.172)=66.498, p= 0.000)

Aggressiveness test of Phytophthora infestans isolates with different effector alleles

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Aggressiveness test of Phytophthora infestans isolates with different effector alleles

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