Validation of microsatellite markers to identify Pl6, Pl8 and Plarg genes that control resistance to Plasmopara halstedii in sunflower

Document Type : Research Paper


V.S. Pustovoit All-Russian Research Institute of Oil Crops, 17 Filatova str., Krasnodar, 350038, Russia


Downy mildew caused by the oomycete Plasmopara halstedii (Farl.) Berl. et de Toni is one of the most harmful sunflower diseases. Among the various measures to control it, the most economical is the development of resistant genotypes. At present, Pl6, Pl8, and Plarg loci are promising for use in breeding, providing resistance to all known P. halstedii races. Microsatellite markers (SSR) help to control the transfer of genes that control resistance in breeding material. However, validation of the marker is needed to prove its reliability in gene detection. There was studied the polymorphism of 9 microsatellite loci in 196 sunflower lines with different resistance to downy mildew. The ORS328 microsatellite locus was chosen as a marker of the Pl6 gene. Amplified fragment with 271 bp allows identifying genotypes resistant to the race 330. The lines that are the sources of the Pl8 gene did not differ from the others in the allelic composition of the ORS781 locus. Among the analyzed breeding samples, no polymorphism was revealed at this locus. To identify the Plarg gene, SSR markers ORS662 and ORS509 were selected. The analysis of 12 samples of the F2 generation from RHA-419 × I3BC2 (VK585 × VK195) crossing at these loci showed that both markers are inherited codominantly. The studied DNA markers can be used in marker-assisted selection (MAS) of sunflower for resistance to downy mildew pathogen.


Agriculture in Russia 2019: Statistical collection / Rosstat - p. 29 Moscow,  91 p.
Albourie, JM & Tourvieille, J 1998, Resistance to metalaxyl in isolates of the sunflower pathogen Plasmopara halstedii. European Journal of Plant Pathology, 104: 2335-2342, DOI: 10.1023/A:1008691123239.
Gharibkhani, M, Pourkazemi, M, Rezvani Gilkolai, S, Tatina, M and Azizzadeh, L 2014, Genetic analysis of pike-perch, Sander lucioperca L., populations revealed by microsatellite DNA markers in Iran. Caspian Journal of Environmental Sciences, 12: 99-108.
Gulya, TJ, Draper, M, Harbour, J, Holen, C, Knodel, J, Lamey, A & Mason, P 1999, Metalaxyl resistance in sunflower downy mildew in North America. In: Proceedings of 21st Sunflower Research Workshop. Fargo, ND. January 14-15, pp. 118-123.
Ime rovski, I, Dimitrije vic, D, Mila dinovic, A , Jocic, S, De dic, B et al. 2014, Ide ntifica tion a nd va lida tion of bre e de r-frie ndly DNA ma rke rs for Pla rg ge ne in sunflowe r. Mole cula r Bre e ding, 34: 779-788. DOI: 10.1007/s11032-014-0074-7.
Iwebor, M, Antonova, T & Saukova, S 2018, Occurrence and distribution of races 713, 733 and 734 of sunflower downy mildew pathogen in the Russian Federation. Helia, 41: 141-151. DOI: 10.1515/helia-2018-0015.
Markell, SG, Humann, RH, Gilley, M, Gulya, TJ et al. 2016, Downy mildew pathogen. In: Harveson RM, Markell SG, Block CC, Gulya TJ (eds). Compendium of sunfower diseases and pests. American Phytopathology Press, St. Paul, pp. 15-117. ISBN: 978-0-89054-509-6.
Molinero-Ruiz, ML, Melero-Vara, JM & Dominguez, J 2003, Inheritance of resistance to two races of sunflower downy mildew (Plasmopara halstedii) in two Helianthus annuus L. lines. Euphytica, 131: 47-51. DOI:  10.1023/A:1023063726185.
Norouzi, M & Samiei, MH 2015, Cross-species amplification of Clupeidae microsatellite DNA markers in common kilka, Clupeonella cultriventris from the Caspian Sea.  Caspian Journal of Environmental Sciences, 13: 167-171.
Qi, LL, Ma, G, Talukder, ZI, Seiler, GJ, Hulke, BS, Jan, CC et al. 2016b, Molecular mapping of the disease resistance gene and its impact on sunflower breeding. In: Proceedings of the 19th International Sunflower Conference, Edirne, pp. 20-30.
Qi, LL, Talukder, ZI, Hulke, BS & Foley, ME 2017, Development and dissection of diagnostic SNP markers for the downy mildew resistance genes PlArg and Pl8 and maker-assisted gene pyramiding in sunflower (Helianthus annuus L.). Molecular Genetics and Genomics, 292: 551-563. DOI: 10.1007/s00438-017-1290-8.
Ramazanova, SA, Bad'yanov, EV & Guchetl, SZ 2020. Molecular markers of the Pl6, Pl13 and Plarg genes for resistance to downy mildew in sunflower breeding. Oil crops, 3: 20-26.
Saghai-Maroof, MA, Soliman, KM, Jorgensen, RA & Allard, RW 1984, Ribosomal DNA spacer-length polymorphisms in barley: Mendelian inheritance, chromosomal location, and population dynamics. Proceedings of the National Academy of Sciences of the United States of America, 81: 8014-8018. DOI: 10.1073/pnas.81.24.8014.
Şahin, EÇ, Kalenderoğlu, A, Aydın, Y, Evci, G & Uncuoğlu, AA 2018, SSR markers suitable for marker assisted selection in sunflower for downy mildew resistance. Open Life Sciences, 13: 319-326. DOI:10.1515/biol-2018-0039.
Shirangi, SA, M. R. Kalbassi, MR & Dorafshan, S, 2011, Microsatellite Polymorphism Reveals Low Genetic Differentiation between Fall and Spring Migratory Forms of Endangered Caspian Trout, Salmo trutta caspius (Kessler, 1870). Caspian Journal of Environmental Sciences,  9: 9-16
Solodenko, A 2018, Validation of microsatellite markers of Pl resistance genes to downy mildew of sunflower. Helia, 41: 73-82. DOI:  10.1515/helia-2017-0026.
Vear, F, Serieys, H, Petit, A, Serre, F, Boudon, JP, Roche, S et al. 2008, “Origins of major genes for downy mildew resistance in sunflower”. Proceedings of the 17th International Sunflower Conference, Cordoba.
Viranyi, F, Gulya, TJ 2015, Recent changes in the pathogenic variability of Plasmopara halstedii (sunflower downy mildew) populations from different continents. Helia, 38: 149-162. DOI: 10.1515/helia-2015-0009