MOLECULAR METHODS OF RESEARCH AND MONITORING OF LATE BLIGHT PATHOGEN PHYTOPHTHORA INFESTANS

The oomycete Phytophthora infestans (Mont.) de Bary, the casual agent of the late blight, still remains the main cause resulting in economic problems for potato growers. Molecular screening of (a)virulence genes of the pathogen (Avr genes) and plant genes for resistance to P. infestans (Rpi genes) would help monitor the changes in the pathogen populations and predict the most vulnerable potato varieties. To assess variation in isolates due to pathogen migration, we compared P. infestans genotypes in the isolates sampled in 2013-2015 in the Pushkin collection of interspecific hybrids (St. Petersburg) and in the commercial potato stands in several regions of Russia. The presence of two mating types, A1 and A2, also promotes pathogen variation due to sexual recombination. Molecular methods were employed for genotyping pathogen samples and assessing their mating type and Avr gene profiles. These molecular indices were related to such phenotypic characteristics as mating type, metalaxyl resistance, virulence factors tested with the differential plants and aggressiveness in the potato tuber test.

Phytophthora infestans, фитофтороз картофеля, SSR генотипирование, A1/A2 тип спаривания, факторы вирулентности, Avr гены вирулентности P. infestans, Rpi гены устойчивости к P. infestans

1. Кузнецова М. А., Козловский Б. Е., Бекетова М. П. и др. Фитопатологическая и молекулярная характеристика изолятов Phytophthora infestans, собранных с устойчивых и восприимчивых генотипов картофеля // Микология и фитопатология. 2016. Т. 50. С. 175-184.

2. Фадина О. А., Бекетова М. П., Соколова Е. А. и др. Упреждающая селекция: использование молекулярных маркеров при создании доноров устойчивости картофеля к фитофторозу на основе сложных межвидовых гибридов // Сельскохозяйственная биология. 2017. Т. 52. № 1. С. 84-94.

3. Чижик В.К., Соколова Е.А., Мартынов В.В. Применение метода SSCP для анализа генетического полиморфизма Phytophthora infestans // Эпидемии болезней растений: мониторинг, прогноз, контроль / под ред. С.С. Санина. Б. Вяземы: ВНИИФ. 2017. С. 280-287.

4. Armstrong M. R., Whisson S. C., Pritchard L. etc. An ancestral oomycete locus contains late blight avirulence gene Avr3a, encoding a protein that is recognized in the host cytoplasm // Proceedings of National Academy of Sciences of USA. 2005. Vol. 102. Pp. 7766-7771.

5. Bradshaw J.E. Potato breeding at the Scottish Plant Breeding Station and the Scottish Crop Research Institute 1920-2008. // Potato Research. 2009. Vol. 52. Pp. 141-172. [Электронный ресурс]. URL: http://dx.doi.org/10.1007/s11540-009-9126-5 (дата обращения: 27.04.2018).

6. Brylińska M., Sobkowiak S., Stefańczyk E. etc. Evaluation of PCR markers for Phytophthora infestans mating type determination // European Journal of Plant Pathology. 2018. [Электронный ресурс]. URL: https://doi.org/10.1007/s10658-018-1445-4 (дата обращения: 27.04.2018).

7. Champouret N., Bouwmeester K., Rietman H. etc. Phytophthora infestans isolates lacking class I ipiO variants are virulent on Rpi-blb1 potato // Molecular Plant-Microbe. Interactions. 2009. Vol. 22. Pp. 1535-1545. doi: 10.1094/MPMI-22-12-1535.

8. Chmielarz M., Sobkowiak S., Dezbski K. etc. Diversity of Phytophthora infestans from Poland // Plant Pathol. 2014. Vol. 63. Pp. 203-211. DOI: 10.1111/ppa.12076

9. Cooke D.E.L., Cano L.M., Raffaele S. etc. Genome analyses of an aggressive and invasive lineage of the Irish potato famine pathogen // PloS Pathog. 2012. 8(10): e1002940. doi:10.1371/journal.ppat.1002940

10. Du, J., Vleeshouwers, V. G. New strategies towards durable late blight resistance in potato // The Potato Genome. Springer, Cham. 2017. Pp. 161-169. doi.org/10.1007/978-3-319-66135-3_10

11. Fry W. E. Phytophthora infestans: New Tools (and Old Ones) Lead to New Understanding and Precision Management. Annu. Rev. Phytopathol. 2016. 54:22.1-22.19. doi:10.1146/annurev-phyto-080615-095951

12. Gilroy E. M., Breen S., Whisson S. C. etc. Presence⁄absence, differential expression and sequence polymorphisms between PiAVR2 and PiAVR2-like in Phytophthora infestans determine virulence on R2 plants // New Phytologist. 2011. Vol. 191. Pp. 763-776. doi: 10.1111/j.1469-8137.2011.03736.x.

13. Judelson H. S., Spielman L. J., Shattock R. C. Genetic mapping and non-Mendelian segregation of mating type loci in the oomycete, Phytophthora infestans // Genetics. 1995. Vol. 141. Pp. 503-512.

14. Kim H.-J., Lee H.-R., Jo K.-R., etc. Broad spectrum late blight resistance in potato differential set plants MaR8 and MaR9 is conferred by multiple stacked R genes // Theoretical and Applied Genetics. 2012. Vol. 124. Pp. 923-935. doi: 10.1007/s00122-011-1757-7.

15. Li Y., Cooke D. E. L., Jacobsen E. etc. Efficient multiplex simple sequence repeat genotyping of the oomycete plant pathogen Phytophthora infestans // Journal of Microbiological Methods. 2013. Vol. 92. Pp. 316-322. doi: 10.1016/j.mimet.2012.11.021.

16. Malcolmson J.F., Black W. New R genes in Solanum demissum Lindl. and their complementary races of Phytophthora infestans (Mont.) de Bary // Euphytica. 1966. Vol. 15. Pp. 199-203.

17. Sokolova E. A., Kuznetsova M. A., Ulanova T.I. etc. Pathogenecity of East European strains of Phytophthora infestans vs. resistance of colonized potato plants: the profiles of AVR genes vs. R gene pyramids // Schepers H.T.A.M. (ed.). PAGV-Special Report. Wageningen, DLO Foundation. 2017. No. 18. Pp. 259-267. [Электронный ресурс]. URL: www.wur.eu/AAVR (дата обращения: 27.04.2018).

18. Statsyuk N. V., Kuznetsova M. A., Kozlovskaya I. N. etc. Characteristics of the Phytophthora infestans population in Russia // PPO-Special Report, Wageningen, 2010. No 14. Pp. 247-254. [Электронный ресурс]. URL: www.wageningen.nl.UR/ppo (дата обращения: 27.04.2018).

19. van Poppel P. M. J. A., Guo J., van de Vondervoort P. J. I. etc. The Phytophthora infestans avirulence gene Avr4 encodes an RXLR-dEER effector // Molecular Plant-Microbe. Interactions. 2008. Vol. 21. Pp. 1460-1470. doi: 10.1094/MPMI-21-11-1460.

20. Vleeshouwers V. G. A. A., Raffaele S., Vossen J. H. etc. Understanding and exploiting late blight resistance in the age of effectors // Annual Review of Phytopathology. 2011. Vol. 49. Pp. 507-531. doi: 10.1146/annurev-phyto-072910-095326.

21. Vossen J. H., Jo K. R., Vosman B. Mining the genus Solanum for increasing disease resistance // R. Tuberosa, ed., Genomics of Plant Genetic Resources. Springer Netherlands. 2014. Pp. 27-46. doi: 10.1007/978-94-007-7575-6.

22. Young, G. K., Cooke, L. R., Watson, S., Kirk, W. W., Perez, F. M., & Deahl, K. L. The role of aggressiveness and competition in the selection of Phytophthora infestans populations. Plant Pathology. 2018 () doi.org/10.1111/ppa.12856.