Marker-assisted selection and use of molecular markers in sunflower breeding for resistance to diseases and parasites

Keywords: DNA markers, Marker-Assisted Selection, sunflower, downy mildew, sunflower rust, broomrape


Recently, the problem of phytosanitary condition of sunflower crops has been exacerbated, which is associated with violation of crop rotations and, as a consequence, spread of common diseases. Selection for resistance to biotic factors requires comprehensive research into the crop biology and pathogens. The use of starting material, which is resistant to major pathogens and environmental stressors, in selection is a prerequisite for the breeding of highly productive hybrids. Significant progress in the breeding of heterosis sunflower hybrids has been achieved primarily due to stable inbred lines. However, their creation is time-consuming, taking 8-12 years. Selection of desirable genotypes and initial forms for crossing is complicated by the fact that it is driven by a set of polygenic traits that are prone to significant modification variability. The use of molecular genetic markers is a way to accelerate breeding. Marker-assisted selection breeding (MAS) has been theoretically justified in numerous publications and implemented in most breeding institutions around the world. However, in domestic breeding programs, MAS has not become widespread compared to traditional methods. Nevertheless, this breeding trend opens new opportunities for studying genetic diversity and determining kinship at the intraspecies and genus levels. The review provides information on the status and prospects of implementation of MAS in traditional plant breeding and highlights the achievements of modern biotechnology in sunflower breeding for resistance to biotic factors owing to molecular genetic markers. The MAS principles are outlined and the advantages of this method are described. Specific examples of application of the molecular approach during the development of starting material of sunflower for breeding for resistance to common diseases and parasites are given. The main stages and components of PCR analysis are also described. Inbred sunflower lines – carriers of the gene for resistance to the downy mildew pathogen are characterized and genetic passports using STS markers to the Pl6 locus have been formalized for 13 sunflower lines.


Download data is not yet available.

Author Biographies

Ye. Kucherenko, Plant Production Institute named after V.Ya. Yuriev of the NAAS of Ukraine

Moskovskyi Ave., 142, Kharkiv, Ukraine, 61060.

A. Zviahintseva, Plant Production Institute named after V.Ya. Yuriev of the NAAS of Ukraine

Moskovskyi Ave., 142, Kharkiv, Ukraine, 61060,

L. Kobyzeva, Plant Production Institute named after V.Ya. Yuriev of the NAAS of Ukraine

Moskovskyi Ave., 142, Kharkiv, Ukraine, 61060,

V. Kolomatska, Plant Production Institute named after V.Ya. Yuriev of the NAAS of Ukraine

Moskovskyi Ave., 142, Kharkiv, Ukraine, 61060,

K. Makliak, Plant Production Institute named after V.Ya. Yuriev of the NAAS of Ukraine

Moskovskyi Ave., 142, Kharkiv, Ukraine, 61060,

N. Vasko, Plant Production Institute named after V.Ya. Yuriev of the NAAS of Ukraine

Moskovskyi Ave., 142, Kharkiv, Ukraine, 61060,

K. Zuieva, Plant Production Institute named after V.Ya. Yuriev of the NAAS of Ukraine

Moskovskyi Ave., 142, Kharkiv, Ukraine, 61060,

T. Lutsenko, Plant Production Institute named after V.Ya. Yuriev of the NAAS of Ukraine

Moskovskyi Ave., 142, Kharkiv, Ukraine, 61060,


Akhtouch B., Muñoz-Ruz J.M., Melero-Vara J.M. et al. (2002). Inheritance of resistance to race F of broomrape (Orobanche cumana Wallr.) in sunflower lines of different origin. Plant Breeding, 121, 266–269.

Alonso J.M., Ecker J.R. (2006). Moving forward in reverse: genetic technologies to enable genomewide phenomic screens in Arabidopsis. Nature Reviews Genetics, (7), 524–536.

Alonso L.C., Fernández-Escobar J., López G. et al. (1996). New highly virulent sun-flower broomrape (Orobanche cernua Loefl.) pathotype in Spain. Advances in Parasitic Plant Research. Proc. 6th Int. Symp. Parasitic Weeds. Cordoba, Spain, 639–644.

Antonova T.S., Araslanova N.M., Ramaza`nova S.A. et al. (2011). Virulence of broomrape affecting sunflower in the Volgograd and Rostov regions. Maslichnyye Kultury. Scientific and Technical Bulletin of the All-Russian Research Institute of Oil Crops, 1(146–147), 127–130. [In Russian].

Avise J.C. (2004). Molecular markers, natural history, and evolution. Sinauer Associates, Inc. Publishers Sunderland: Massachusetts, 355 рp.

Bachlava E., Radwan O.E., Abratti G. et al. (2011). Downy mildew (Pl8 and Pl14) and rust (RAdv) resistancegenesreside in close proximity to tandemly duplicated clusters ofnon-TIR-like NBS-LRR-encoding genes on sunflowerchromosomes 1 and 13. Theoretical and Applied Genetics, 122, 1211–1221.

Barrangou R., Fremaux C., Deveau H. et al. (2007). CRISPR provides acquired resistance against viruses in prokaryotes. Science, 315(5819), 1709–1712.

Beckmann J.S., Soller M. (1983). Restriction fragment length polymorphisms in plant genetic: improvement: methodologies, mapping and costs. Theoretical and Applied Genetics, 67, 35–43.

Bernardo R., Moreau L., Charcosset A. (2006). Number and fitness of selected individuals in marker-assisted and phenotypic recurrent selection. Crop Science, 46(5), 1972–1980.

Bolle C., Schneider A., Leister D. (2011). Perspectives on systematic analyses of gene function in Arabidopsis thaliana: new tools, topics and trends. Current Genomics, 12(1), 1–14.

Boutros M., Ahringer J. (2008). The art and design of genetic screens: RNA interference. Nature Reviews Genetics, 9, 554–566.

Bouzidi M.F. Badaoui S., Cambon F. et al. (2002). Molecular analysis of a major locus for resistance to downy mildew in sunflower with specific PCR-based markers. Theoretical and Applied Genetics, 104, 529–600.

Bulos, M., Ramos, M.L., Altieri, E., Sala, C.A. (2013). Molecular mappingof a sunflower rust resistance gene from HAR6. Breeding Science, 63(1), 141–146.

Bulos M., Vergani P.N., Altieri E. (2014). Genetic mapping, marker assisted selection and allelic relationships for the Pu6 gene conferring rust resistance in sunflower. Breeding Science, 64(3), 206–212.

Collard B., Mackill D. (2008). Marker-assisted selection: an approach for precision plant breeding in the twenty first century. Philosophical Transactions of the Royal Society. Biological Sciences, 363(1491), 557–572.

Concibido V.C., Denny R.L., Lange D.A. et al. (1996). RFLP mapping and molecular marker-assisted selection of soybean cyst nematode resistance in PI 209332. Crop Science, 36(6), 1643–1650.

Duble C., Hahn V., Knapp S., Bauer E. (2004). PlArg from Helianthus argophyllus is unlinked to other known downy mildew resistance genes in sunflower. Theoretical and Applied Genetics, 109, 1083–1086.

Edwards K., Logan J., Sauders N. (2004). Real-time PCR: An assential guide. Horizon Bioscience: Norfolk, United Kingdom, 346 pp.

Eisenberg D., Marcotte E.M., Xenarios I., Yeates T.O. (2000). Protein function in the post-genomic era. Nature, 405, 823–826.

Fedorenko V.O., Ostash B.O., Honchar M.V., Rebets Yu.V. (2007). Extensive workshop on genetics, genetic engineering and analytical biotechnology of microorganisms. Publishing Center of Ivan Franko LNU: Lviv, Ukraina, 277 pp. [In Ukrainian].

Fernández-Martínez J., Perez-Vich B., Akhtouch B. et al. (2004). Registration of four sunflower germplasms resistant to race F of broomrape. Crop Science, 44(3), 1033–1034.

Fernández-Martínez J.M., Domín-guez J., Pérez-Vich B., Velasco L. (2008). Update on breeding for resistance to sunflower broomrape. Helia, 31(48), 73–84.

Fernández-Martínez J.M., Melero J.M., Vara J. et al. (2000). Selection of wild and cultivated sunflower for resistance to a new broomrape race that overcomes re-sistance to Or5 gene. Crop Science, 40(2), 550–555.

Frisch M., Melchinger A.E. (2001). Marker-assisted backcrossing for simultaneous introgression of two genes. Crop Science. 41(6), 1716–1725.

Frisch M., Melchinger A.E. (2005). Selection theory for marker-assisted backcrossing. Genetics. 170(2), 909–917.

Gimelfarb A., Lande R. (1994a). Simulation of marker-assisted selection for non-additive traits. Genetical Researh, 64(2), 127–136.

Gimelfarb A., Lande R. (1994b). Simulation of marker-assisted selection in hybrid populations. Genetical Researh, 63(1), 39–47.

Gimelfarb A., Lande R. (1995). Marker-assisted selection and marker-QTL associations in hybrid populations. Theoretical and Applied Genetics, 91, 522–528.

Glazko V.I. (2003). Introduction to genetics, bioinformatics, DNA-technology, gene therapy, DNA-ecology, proteomics, metabolism. KVITs: Kiev, Ukraina. 640 pp. [In Russian]

Gong L., Gulya T.J., Markell S.G. et al. (2013a). Genetic mapping of rust resistance genes in confectionsunflower line HA-R6 and oilseed line RHA 397. Theoretical and Applied Genetics, 126, 2039–2049.

Gong L., Hulke B.S., Gulya T.J. et al. (2013b). Molecular magging of a novel rust resistance gene R(12) insunflower (Helianthus annuus L.). Theoretical and Applied Genetics, 126(1), 93–99.

Griffiths P.E., Stotz K. (2006). Genes in the postgenomic era. Theoretical Medicine and Bioethics, 27, 499–521.

Guchetl S.Z., Chelyustnikova T.V., Arslanova N.M., Antonova T.S. (2012). Marking of the OR5 resistance gene to broomrape (Orobanche Cumana Wallr.) race E in sunflower lines bred at the All-Russian Research Institute of Oil Crops. Maslichnyye Kultury. Scientific and Technical Bulletin of the All-Russian Research Institute of Oil Crops, 2, 151–152. [In Russian].

Guimaraes E.P., Ruane J., Scherf B. et al. (2007). Marker-assisted selection: current status, and future perspectives in crops, livestock, forestry, and fish. Food and Agriculture Organization of the United Nations, Rome, Italy, 495 pp.

Gupta P., Varshney R., Sharma P., Ramesh B. (2008). Molecular markers and their application in wheat breeding. Plant Breeding, 118(5), 369–390.

Hirochika H. (2010). Insertional mutagenesis with Tos17 for functional analysis of rice genes. Breed. Science, 60(5), 486–492.

Hospital F. (2002). Marker-assisted back-cross breeding: a case study in genotype building theory. Quantitative Genetics. Genomics and Plant Breeding, 10, 135–141.

Hospital F., Charcosset A. (1997). Marker-assisted introgression of quantitative trait loci. Genetics, 147(3), 1469–1485.

Hospital F., Goldringer I., Openshaw S. (2000). Efficient marker-based recurrent selection for multiple quantitative trait loci. Genetical Researh, 75(3), 357–368.

Hsiao A., Kuo M.D. (2009). High-throughput biology in the postgenomic era. Journal of Vascular Interventional Radiology, 20 (7), 488–496.

Imerovski I., Dimitrijevic A., Miladinovic D. et al. (2016). Mapping of a new gene for resistance to broomrape races higher than F. Euphytica, 209, 281–289.

Inge-Vechtomov S.G. (2010). Genetics with breeding basics. Publishing house N-L: SPb, Russia, 30 pр. [In Russian].

Jocić S., Cvejić S., Hladni N. et al. (2010). Development of sunflower genotypes resistant to downy mildew. Helia, 33(53), 173–180.

Jocić S., Miladinović D., Imerovski I. et al. (2012). Towards sustainable downy mildew resistance in sunflower. Helia, 35(56), 61–72.

Kalachniuk M.S., Kalachniuk L.H., Melnychuk D.O. (2012). Conditions for polymerase chain reaction in the laboratory practice (methodological aspects). Biolohiia Tvaryn, 14(1), 660–667. [In Ukrainian].

Kalendar R.N., Glazko V.I. (2002). Types of molecular genetic markers and their applications. Fiziologiya i Biokhimiya Kulturnykh Rasteniy, 34(4), 279–296. [In Russian].

Korovaeva I., Popova N. (2015). Kary Mullis – bright name in the history of science. Annals of Mechnikov Institute, 3, 72–74. [In Russian].

Kozhukhova N.Ye. (2011). Molecular marker-assisted selection in plant breeding. Visnyk Kharkivskoho Natsionalnoho Ahrarnoho Universytetu. Series Biolohiia, 1(22), 35–43. [In Ukrainian].

Kutyrev V.V., Sharova I.N., Osina N.A. et al. (2010). Organization of the work in laboratories using nucleic acidamplification methods on materials containing RG1-4 microorganisms. Guidelines 1.3.2569-09. Gossanepidnadzor: Moskow, Russia, 51 pp. [In Russian].

Lande R., Thompson R. (1990). Efficiency of marker-assisted selection in the improvement of quantitative traits. Genetics, 124(3), 743–756.

Lawson W.R., Goulter K.C., Henry R.J. et al. (1998). Marker-assisted selection for two rust resistance genes in sunflower. Molecular Breeding, 4, 227–234.

Lawson W.R., Goulter K.C., Henry R.J. et al. (1996). RAPD markers for a sunflower rust resistance gene. Australian Journal of Agricultural Research, 47(3), 395–401.

Lesk A. (2009). Introduction to bioinformatics. Binom: Moscow, Russia, (2009). 320 pp. [In Russian].

Lisova H.M. (1999). Markers of wheat resistance genes to brown rust and their use in creating disease-resistant varieties. Zakhyst Roslyn, 11, 10–11. [In Ukrainian].

Liu L., Fan X.D. (2014). CRISPR-Cas system: a powerful tool for genome engineering. Plant Molecular Biology, 85, 209–218.

Lopukhov L.V., Eldeinshtein M.V. (2000). Polymerase chain reaction in microbiological diagnostics. Klinicheskaya Mikrobiologiya i Antimikrobnaya Khimioterapiya, 2(3), 96–105. [InRussian].

Lu Y.H., Melero-Vara J.M., Garcia-Tejada J.A., Blanchard P. (2000). Development of SCAR markers linked to the gene Or5 conferring resistance to broomrape (Orobanche cumana Wallr.) in sunflower. Theoretical and Applied Genetics, 100, 625–632.

Mahony J.B. (2008). Detection of respiratory viruses by molecular methods. Clinical Microbiology Revies, 21(4), 716–747.

Malyshev S.V., Kartel N.A. (1997). Molecular markers in genetic mapping of plants. Molekulyarnaya Biologiya, 31(2), 197–208. [In Russian].

Melero-Vara J., Domingues J., Fernandes-Martinez J. (1989). Evaluation of different lines in a collection of sun-flower parental lines for resistance to broomrape (Orobanche cernua) in Spain. Plant Breeding, 102(4), 322–326.

Oleshchuk O.M., Mudra A.Ye, Zozuliak N.B. (2014). PCR diagnostics: principles, achievements and prospects. Experymentalna i Klinichna Biokhimiia, 16(3), 97–103.[In Ukrainian].

Pacureanu J.M., Raranciuc S., Sava E. et al. (2009). Virulence and ag-gressiveness of sunflower broomrape (Orobanche cumana Wallr.) populations in Romania. Неlia, 32(51), 111–118.

Patrushev L.I. (2004). Artificial genetic systems. Nauka: Moskow, Russia, 526 pp. [In Russian].

Qi L.L., Hulke B.S., Vick B.A., Gulya T.J. (2011). Molecular mapping of the R4 rust resistance gene for large NBS-LRRcluster on connecting group 13 of sunlight. Theoretical and Applied Genetics, 123, 351–358.

Qi L.L., Gulya T.J., Hulke B.S., Vick B.A. (2012a). Chromosome llocation, DNA markers and rust resistance of fl owergene R5. Molecular Breeding, 30, 745–756.

Qi L.L., Seiler G.J., Hulke B.S. et al. (2012b). Genetics and mapping of R11gene conferring resistance to recently emergent rusty races closely associated with restoration of male fertility in the sun (Helianthus annuus). Theoretical and Applied Genetics, 125, 921–932.

Qi, L.L., Long, Y.M., Ma, G.J., Markell, S.G. (2015a). Map saturation and SNP marker development for the rust resistance genes (R4, R5, R13a, and R13b) in sunflower (Helianthus annuus L.). Molecular Breeding, 35, 196.

Qi L.L., Ma G.J., Long Y.M. et al. (2015b). Relocation of a rust resistance gene R2 and its marker-assisted gene pyramiding in confection sunflower (Helianthus annuus L.). Theoretical and Applied Genetics, 128, 477–488.

Radwan O., Gandhi S., Heesacker A. et al. (2008). Genetic diversity and genomic distribution of homologs encodimg NBC-LRR disease resistance proteins in sunflower. Molecular Genetics and Genomics, 280, 11–15.

Ramazanova S.A., Antonova T.S. (2018). Marking of Pl5, Pl6 и Pl8 loci controlling resistance to Plasmopara halstedii in sunflower lines developed in VNIIMK. Oil Crops, 3 (175), 19–27. [In Russian].

Resk R. (1998). Physcomitrella and Arabidopsis: the David and Goliath of reverse genetics. Trends in Plant Science, 3(6), 209–210.

Romanenko V.N., Svistunov I.V., Lavrinenko O.A. (1998). Polymerase chain reaction: principles, achievements, prospects for use in the diagnosis of infections. Laboratornaya Diagnostika, 4, 45–51. [In Russian].

Rybicki E.A. (2005). PCR. In Manual of Online Molecular Biology Techniques. University of Cape Town: South Africa, 38 pp.

Servin B., Martin O., Mezard M., Hospital F. (2004). Toward a theory of marker-assisted gene pyramiding. Genetics, 168(1), 513–523.

Shindrova P. (2006). Broomrape (Orobanche cumana Wallr.) in Bulgaria – Distribution and race composition. Helia, 29 (44), 111–120.

Sivolap Yu.M., Kozhukhova N.E., Kalendar R.N. (2011). Variability and specificity of agricultural plantgenomes. Astroprint: Odesa, Ukraina, 336 pp. [in Russian]

Small I. (2007). RNAi for revealing and engineering plant gene functions. Current Opinion in Biotechnology, 18(2), 148–153.

Solodenko A.Ye., Burlov V.V., Sivolap Yu.M. (2014). Markers of the sunflower resistance gene to downy mildew. The Bulletin of Kharkiv Agrarian University. Series Biology, 3 (33), 59–65. [In Ukrainian].

Solodenko A.Ye., Fait V.I. (2016). Identification of sources of sunflower resistance to downy mildew and broomrape using DNA markers. The Bulletin of Kharkiv Agrarian University. Series Biology, 3 (39), 57–63. [In Ukrainian].

Stehnii B.T., Herilovych A.P., Lymanska O.Yu. (2010). Polymerase chain reaction in the practice of veterinary medicine and biological research. Instructional guide. NTMT: Kharkiv, Ukrain, 227 pp. [In Ukrainian].

Struhl K. (1983). The new yeast genetics. Nature, 305, 391–397.

Sukno S., Melero-Vara J.M., Fernández-Martínez J.M. (1999). Inheritance of resistance to Orobanche cernua Loefl. in six sunflower lines. Crop Science, 39(3), 674–678.

Sulima A.S., Zhukov V.A. (2015). Tilling is a modern technology of "reverse" plant genetics. Agricultural Biology, 50(3), [In Russian].

Talukder Z.I., Hulke B.S., Qi L. et al. (2014). Candidate gene association of Sclerotinia stalk rot resistance in sunflower (Helianthus annuus L.) uncovers the importance of COI1 homologs. Theoretical and Applied Genetics, 127, 193–209.

Tang S., Heesacker A., Kishore V.K. et al. (2003). Genetic mapping of the Or 5 gene for resistance to race E in sunflower. Crop Science, 43(3), 1021–1028.

Tanksley S.D. (1983). Molecular markers in plant breeding. Plant Molecular Biology Reports, 1, 3–8.

Tanksley S.D., Rick C.M. (1980). Isozyme gene linkage map of the tomato: Applications in genetics and breeding. Theoretical and Applied Genetics, 58, 161–170.

Upadhyaya N.M., Zhu Q.H., Bhat R.S. (2011). Transposon insertional mutagenesis in rice. Plant Reverse Genetics. Methods in Molecular Biology, 678, 147–177.

Whittaker J.C., Haley C.S., Thompson R. (1997). Optimal weighting of information in marker-assisted selection. Genetical Researh, 69(2), 137–144.

Wieckhorst S., Bachlava E., Duble C. et al. (2010). Fine mapping of the sunflower resistance locus PlARG introduced from the wild species Helianthus argophyllus. Theoretical and Applied Genetics, 121(8), 1633–1644.

Xu Y., Crouch J.H. (2008). Marker-Assisted Selection in Plant Breeding: From Publications to Practice. Сrop Science, 48(2), 391–407.

Yu J.K., Tang S., Slabaugh M.B. et al. (2003). Towards saturated molecular genetic bonding for the cultured sun. Crop Science, 43, 367–387.

Zaid A., Hughes H.G., Porceddu E., Nicholas F. (2007). Glossary of Biotechnology for Food and Agriculture. FAO Research and Technology Paper 9. Food and Agriculture Organization of the United Nations: Rome, Italy, 305 pp.

Zhang M., Liu Z., Jan C.-C. (2016). Molecular mapping of a rust resistancegene R14 in cultivated sunflower line PH 3. Molecular Breeding, 36(32),

Zhang W., Smith C. (1992). Computer simulation of marker assisted selection utilizing linkage disequilibrium. Theoretical and Applied Genetics, 83, 813–820.

Zhang W., Smith C. (1993). Simulation of marker-assisted selection utilizing linkage disequilibrium: the effects of several additional factors. Theoretical and Applied Genetics, 86, 492–496.

How to Cite
Kucherenko, Y., Zviahintseva, A., Kobyzeva, L., Kolomatska, V., Makliak, K., Vasko, N., Zuieva, K., & Lutsenko, T. (2022). Marker-assisted selection and use of molecular markers in sunflower breeding for resistance to diseases and parasites. The Journal of V.N.Karazin Kharkiv National University. Series «Biology», 38, 14-29.