bioselБиотехнология и селекция растенийPlant Biotechnology and Breeding2658-62662658-6258VIR10.30901/2658-6266-2024-4-o8biosel-259Research ArticleИЗУЧЕНИЕ ГЕНЕТИЧЕСКИХ РЕСУРСОВ РАСТЕНИЙ С ИСПОЛЬЗОВАНИЕМ МЕТОДОВ МОЛЕКУЛЯРНОЙ ГЕНЕТИКИSTUDY OF PLANT GENETIC RESOURCES USING MOLECULAR GENETICS METHODSГенотипирование линий генетической коллекции подсолнечника ВИР с использованием аллель-специфичных маркеров локуса Rf1Genotyping of lines from the VIR sunflower genetic collection using allele-specific markers of the Rf1 locushttps://orcid.org/0000-0003-0474-8860АнисимоваИ. Н.AnisimovaI. N.

Ирина Николаевна Анисимова, доктор биологических наук, ведущий научный сотрудник, ВИР

190000 Россия, Санкт-Петербург, ул. Б. Морская, 42, 44

Irina N. Anisimova, Dr. Sci. (Biology), Leading Researcher, VIR

42, 44, Bolshaya Morskaya Street, St. Petersburg, 190000 Russia

irina_anisimova@inbox.ruhttps://orcid.org/0000-0002-5531-2728АлпатьеваН. В.AlpatievaN. V.

Наталья Владимировна Алпатьева, кандидат биологических наук, старший научный сотрудник, ВИР

190000 Россия, Санкт-Петербург, ул. Б. Морская, 42, 44

Natalia V. Alpatieva, Cand. Sci. (Biology), Senior Researcher, VIR

42, 44, Bolshaya Morskaya Street, St. Petersburg, 190000 Russia

alpatievanatalia@mail.ruhttps://orcid.org/0000-0003-4444-3658РязановаМ. К.RyazanovaM. K.

Мария Константиновна Рязанова, аспирант, ВИР

190000 Россия, Санкт-Петербург, ул. Б. Морская, 42,44

Maria K. Ryazanova, PhD Student, VIR

42, 44, Bolshaya Morskaya Street, St. Petersburg, 190000 Russia

m.ryazanova@vir.nw.ruhttps://orcid.org/0009-0008-1706-0022БердиганР. Д.BerdiganR. D.

Роман Дмитриевич Бердиган, студент, СПбГУ

199034 Россия, Санкт-Петербург, Университетская набережная, 7–9

Roman D. Berdigan, Student, SPbSU

7-9, Universitetskaya Embankment, St. Petersburg, 199034 Russia

rberdigan@mail.ruhttps://orcid.org/0000-0002-3019-0306РадченкоЕ. Е.RadchenkoE. E.

Евгений Евгеньевич Радченко, доктор биологических наук, главный научный сотрудник, заведующий отделом, ВИР

190000 Россия, Санкт-Петербург, ул. Б. Морская, 42, 44

Evgeny E. Radchenko, Dr. Sci. (Biology), Chief Researcher, Head of Department, VIR

42, 44, Bolshaya Morskaya Street, St. Petersburg, 190000 Russia

eugene_radchenko@rambler.ruhttps://orcid.org/0000-0002-8110-9168ГавриловаВ. А.GavrilovaV. A.

Вера Алексеевна Гаврилова, доктор биологических наук, главный научный сотрудник, ВИР

190000 Россия, Санкт-Петербург, ул. Б. Морская, 42, 44

Vera A. Gavrilova, Dr. Sci. (Biology), Chief Researcher, VIR

42, 44, Bolshaya Morskaya Street, St. Petersburg, 190000 Russia

v.gavrilova@vir.nw.ruФедеральный исследовательский центр Всероссийский институт генетических ресурсов растений имени Н.И. ВавиловаРоссияN.I. Vavilov All-Russian Institute of Plant Genetic ResourcesRussian FederationСанкт-Петербургский государственный университетРоссияSt. Petersburg State UniversityRussian Federation202414022025745667Copyright © Анисимова И.Н., Алпатьева Н.В., Рязанова М.К., Бердиган Р.Д., Радченко Е.Е., Гаврилова В.А., 20242024Анисимова И.Н., Алпатьева Н.В., Рязанова М.К., Бердиган Р.Д., Радченко Е.Е., Гаврилова В.А.Anisimova I.N., Alpatieva N.V., Ryazanova M.K., Berdigan R.D., Radchenko E.E., Gavrilova V.A.This work is licensed under a Creative Commons Attribution 4.0 License.https://biosel.elpub.ru/jour/article/view/259

Актуальность. При создании промышленных гибридов подсолнечника (Helianthus annuus L.) преимущественно используется генетическая система ЦМС-Rf на основе цитоплазматической мужской стерильности РЕТ1-типа. Ключевым геном в селекции гибридов является Rf1, необходимый для восстановления фертильности пыльцы растений F1. Эффективным инструментом для идентификации генотипов родительских линий по локусу Rf1, контроля однородности, а также определения генетической чистоты партий гибридных семян являются молекулярно-генетические маркеры, апробированные на различном генетическом материале. В настоящем исследовании для идентификации генотипов линий генетической коллекции подсолнечника ВИР и межлинейных гибридов F2 использованы доступные из литературных источников аллель-специфичные маркеры генов-кандидатов локуса Rf1. Материал и методы. Изучены две выборки генотипов: 46 линий генетической коллекции подсолнечника ВИР, ранее охарактеризованных в полевых опытах по способности к восстановлению фертильности пыльцы, и 80 фенотипированных по признакам фертильности/стерильности растений, из расщепляющихся популяций F2 от скрещиваний линии ЦМС ВИР 116А с восстановителями фертильности ВИР 740 и RIL 130. Линии различались по типу цитоплазмы и наличию SCAR-маркера HRG02, тесно сцепленного с локусом Rf1. Линии генотипированы с использованием маркеров, специфичных для доминантного (PPR621.5R, SRF833, 67N04_P_170) и рецессивного (PPR621.5M, 67N04_P_155) аллелей генов-кандидатов Rf1. Маркерные фрагменты PPR621.5M и PPR621.5 R, амплифицированные у шести генотипов, были выделены и секвенированы. Результаты. Нуклеотидные последовательности маркеров PPR621.5М и PPR621.5R отличались по четырём SNP и были полностью идентичными с опубликованными в литературе. У линий ЦМС и большинства закрепителей стерильности идентифицированы маркеры PPR621.5М и 67N04_P_155, специфичные для аллеля rf1. Девятнадцать из 21 линии, характеризовавшейся стерильной цитоплазмой и наличием маркера HRG02, имели по три маркера, специфичных для доминантного аллеля; у двух линий обнаружены по два аллель-специфичных маркера. У четырёх из семи восстановителей фертильности (стерильная цитоплазма, без маркера HRG02) выявлено по два или три маркера, специфичных для доминантного аллеля, у трёх линий идентифицированы лишь маркеры рецессивного аллеля. Обнаружены генотипы F2, возникшие в результате рекомбинации между SCAR-маркером HRG02 и аллель-специфичными маркерами. Заключение. Подтверждены эффективность аллель-специфичных маркеров генов-кандидатов локуса Rf1 для генотипирования линий подсолнечника и их диагностическая ценность для отбора целевых генотипов из расщепляющихся гибридных популяций.

Background. The CMS-Rf genetic system based on the PET1-type cytoplasmic male sterility (CMS) is commonly used to create commercial sunflower (Helianthus annuus) hybrids. The Rf1 gene, of key importance for hybrid breeding, is necessary for restoring pollen fertility in F1 plants. The molecular genetic markers tested on various genetic materials are an effective tool for identifying parental line genotypes at the Rf1 locus, controlling homogeneity, and determining the genetic purity of hybrid seed lots. In the present study, the allele-specific markers of the Rf1 candidate genes available from literature were used to genotype lines from the VIR sunflower genetic collection and F2 hybrids. Material and methods. The study concentrated on two sample sets of genotypes, one of which contained 46 lines from the VIR sunflower genetic collection, previously characterized in field experiments for the pollen restoration ability, and the other 80 plants from segregating F2 populations from crosses of the CMS VIR 116A line with fertility restorers VIR 740 and RIL 130, phenotyped for fertility/sterility. The lines differed in respect of the cytoplasm type and the presence of the SCAR marker HRG02 closely linked to the Rf1 locus. The lines have been genotyped using markers specific for the dominant (PPR621.5R, SRF833, 67N04_P_170) and recessive (PPR621.5M, 67N04_P_155) alleles of the Rf1 candidate genes. The PPR621.5M and PPR621.5 R, marker fragments amplified in six genotypes, have been isolated and sequenced. Results. The nucleotide sequences of PPR621.5M and PPR621.5R turned out to be different in four SNPs and completely identical to those presented in the published literature. The PPR621.5M and 67N04_P_155 markers specific for the rf1 allele were identified in CMS lines and the majority of sterility maintainers. Nineteen out of 21 lines characterized by sterile cytoplasm and the presence of the HRG02 marker had three markers specific for the dominant allele; two lines had two allele-specific markers. Four out of seven fertility restorers (sterile cytoplasm, without the HRG02 marker) were found to contain two or three markers specific for the dominant allele, while three lines had only markers for the recessive allele. The F2 genotypes resulting from recombination between the SCAR marker HRG02 and allele specific markers were detected. Conclusion. The study confirmed efficiency of allele-specific markers of the Rf1 locus candidate genes for genotyping sunflower lines, as well as their diagnostic value for selecting target genotypes from segregating hybrid populations.

Helianthus annuusсистема ЦМС-Rfгибридывосстановление фертильности пыльцыгенотипSNPгаплотипHelianthus annuusCMS-Rf systemhybridspollen fertility restorationgenotypeSNPhaplotypeработа выполнена в рамках Государственного задания согласно тематическому плану ВИР по проекту № FGEM –2022-0005 «Растительные ресурсы масличных и прядильных культур ВИР как основа теоретических исследований и их практического использования».The research was performed within the framework of the State Assignment according to the Theme Plan of VIR, Project No. FGEM-2022-0005 “Plant resources of oil and fiber crops at VIR as the basis for theoretical research and their practical utilization”.ReferencesAbratti G., Bazzalo M.E., Leon A. Mapping a novel fertility restoration gene in sunflower. In: Proceedings of the 17th International Sunflower Conference; 2008 June 8-12; Cordoba, Spain. Consejería de Agricultura y Pesca; 2008. Vol. 2. P. 617-621. Available from https://www.isasunflower.org/fileadmin/documents/aaProceedings/17thISC_CordobaVol2/617sonia.pdf [accessed Nov. 15, 2024]Abratti G., Bazzalo M.E., Leon A. Mapping a novel fertility restoration gene in sunflower. In: Proceedings of the 17th International Sunflower Conference; 2008 June 8-12; Cordoba, Spain. Consejería de Agricultura y Pesca; 2008. Vol. 2. P. 617-621. Available from https://www.isasunflower.org/fileadmin/documents/aaProceedings/17thISC_CordobaVol2/617sonia.pdf [accessed Nov. 15, 2024]Анащенко А.В., Дука М.В. Изучение генетической системы ЦМС-Rf у подсолнечника (Helianthus annuus L.). Сообщение II. Восстановление мужской фертильности у гибридов на основе ЦМС. Генетика. 1985;21(12):1999-2004.Anashchenko A.V., Duka M.V. Study of the genetic system of CMS-Rf in sunflower (Helianthus annuus L.). Communication II. Restoration of male fertility in hybrids based on CMS. Genetika. 1985;21(12):1999-2004. [in Russian]Анисимова И.Н., Карабицина Ю.И., Алпатьева Н.В., Кузнецова Е.Б., Титов Н.В., Лютко А.Ю., Гаврилова В.А. Диагностическая ценность молекулярных маркеров гена Rf1 подсолнечника. Биотехнология и селекция растений. 2021;4(2):28-37. DOI: 10.30901/2658-6266-2021-2-o3Anisimova I.N., Karabitsina Yu.I., Alpatieva N.V., Kuznecova E.B., Titov N.V., Lyutko A.Yu., Gavrilova V.A. Diagnostic value of Rf1 gene molecular markers in sunflower. Biotechnology and Plant Breeding. 2021;4(2):28-37. [in Russian]. DOI: 10.30901/2658-6266-2021-2-o3Badouin H., Gouzy J., Grassa C.J., Murat F., Staton S.E., Cottret L., Lelandais-Brière C., Owens G.L., Carrère S., Mayjonade B., Legrand L., Gill N., Kane N.C., Bowers J.E., Hubner S., Bellec A., Bérard A., Bergès H., Blanchet N., Boniface M-C., Brunel D., Catrice O., Chaidir N., Claudel C., Donnadieu C., Faraut T., Fievet G., Helmstetter N., King M., Knapp S.J., Lai Z., Le Paslier M-C., Lippi Y., Lorenzon L., Mandel J.R., Marage G., Marchand G., Marquand E., Bret-Mestries E., Morien E., Nambeesan S., Nguyen T., Pegot-Espagnet P., Pouilly N., Raftis F., Sallet E., Schiex T., Thomas J., Vandecasteele C., Varès D., Vear F., Vautrin S., Crespi M., Mangin B., Burke J.M., Salse J., Muños S., Vincourt P., Loren H. Rieseberg L.H., Langlade N.B. The sunflower genome provides insights into oil metabolism, flowering and Asterid evolution. Nature. 2017;546:148-152. DOI: 10.1038/nature22380Badouin H., Gouzy J., Grassa C.J., Murat F., Staton S.E., Cottret L., Lelandais-Brière C., Owens G.L., Carrère S., Mayjonade B., Legrand L., Gill N., Kane N.C., Bowers J.E., Hubner S., Bellec A., Bérard A., Bergès H., Blanchet N., Boniface M-C., Brunel D., Catrice O., Chaidir N., Claudel C., Donnadieu C., Faraut T., Fievet G., Helmstetter N., King M., Knapp S.J., Lai Z., Le Paslier M-C., Lippi Y., Lorenzon L., Mandel J.R., Marage G., Marchand G., Marquand E., Bret-Mestries E., Morien E., Nambeesan S., Nguyen T., Pegot-Espagnet P., Pouilly N., Raftis F., Sallet E., Schiex T., Thomas J., Vandecasteele C., Varès D., Vear F., Vautrin S., Crespi M., Mangin B., Burke J.M., Salse J., Muños S., Vincourt P., Loren H. Rieseberg L.H., Langlade N.B. The sunflower genome provides insights into oil metabolism, flowering and Asterid evolution. Nature. 2017;546:148-152. DOI: 10.1038/nature22380Baute G.J., Kane N.C., Grassa C.J., Lai Z., Rieseberg L.H. Genome scans reveal candidate domestication and improvement genes in cultivated sunflower, as well as post-domestication introgression with wild relatives. New Phytologist. 2015;206(2):830-838. DOI: 10.1111/nph.13255Baute G.J., Kane N.C., Grassa C.J., Lai Z., Rieseberg L.H. Genome scans reveal candidate domestication and improvement genes in cultivated sunflower, as well as post-domestication introgression with wild relatives. New Phytologist. 2015;206(2):830-838. DOI: 10.1111/nph.13255Choudhari A.K., Bagade A.B. Diverse cytosteriles in sunflower: a review. International Journal of Current Microbiology and Applied Sciences. 2019;8(11):1641-1644. DOI: 10.20546/ijcmas.2019.811.189Choudhari A.K., Bagade A.B. Diverse cytosteriles in sunflower: a review. International Journal of Current Microbiology and Applied Sciences. 2019;8(11):1641-1644. DOI: 10.20546/ijcmas.2019.811.189Feng J., Jan C.C. Introgression and molecular tagging of Rf4, a new male fertility restoration gene from wild sunflower Helianthus maximiliani L. Theoretical and Applied Genetics. 2008;117(2):241. DOI: 10.1007/s00122-008-0769-4Feng J., Jan C.C. Introgression and molecular tagging of Rf4, a new male fertility restoration gene from wild sunflower Helianthus maximiliani L. Theoretical and Applied Genetics. 2008;117(2):241. DOI: 10.1007/s00122-008-0769-4Гаврилова В.А., Анисимова И.Н., Алпатьева Н.В., Рожкова В.Т., Ступникова Т.Г., Карабицина Ю.И., Кузнецова Е.Б. Каталог мировой коллекции ВИР. Вып. 853. Генетическая коллекция подсолнечника. Санкт-Петербург: ВИР; 2017.Gavrilova V.A., Anisimova I.N., Alpatyeva N.V., Rozhkova V.T, Stupnikova T.G., Karabitsina Yu.I., Kuznetsova E.B. Catalogue of the VIR global collection. Iss. 853. Genetic collection of sunflower (Geneticheskaya kollektsiya podsolnechnika). St. Petersburg: VIR; 2017. [in Russian]Goryunov D.V., Anisimova I.N., Gavrilova V.A., Chernova A.I., Sotnikova E.A., Martynova E.U., Boldyrev S.V., Ayupova A.F., Gubaev R.F., Mazin P.V., Gurchenko E.A., Shumskiy A.A., Petrova D.A., Garkusha S.V., Mukhina Z.M., Benko N.I., Demurin Y.N., Khaitovich P.E., Goryunova S.V. Association mapping of fertility restorer gene for CMS PET1 in sunflower. Agronomy. 2019;9(2):49. DOI: 10.3390/agronomy9020049Goryunov D.V., Anisimova I.N., Gavrilova V.A., Chernova A.I., Sotnikova E.A., Martynova E.U., Boldyrev S.V., Ayupova A.F., Gubaev R.F., Mazin P.V., Gurchenko E.A., Shumskiy A.A., Petrova D.A., Garkusha S.V., Mukhina Z.M., Benko N.I., Demurin Y.N., Khaitovich P.E., Goryunova S.V. Association mapping of fertility restorer gene for CMS PET1 in sunflower. Agronomy. 2019;9(2):49. DOI: 10.3390/agronomy9020049Heiser C.B. Hybridization in the annual sunflowers: Helianthus annuus × H. debilis var. cucumerifolius. Evolution. 1951;5(1):42-51. DOI: 10.2307/2405429Heiser C.B. Hybridization in the annual sunflowers: Helianthus annuus × H. debilis var. cucumerifolius. Evolution. 1951;5(1):42-51. DOI: 10.2307/2405429Horn R., Köhler R.H., Zetsche K. A mitochondrial 16 kDa protein is associated with cytoplasmic male sterility in sunflower. Plant Molecular Biology. 1991;17(1):29-36. DOI: 10.1007/BF00036803Horn R., Köhler R.H., Zetsche K. A mitochondrial 16 kDa protein is associated with cytoplasmic male sterility in sunflower. Plant Molecular Biology. 1991;17(1):29-36. DOI: 10.1007/BF00036803Horn R., Kusterer B., Lazarescu E., Prüfe M., Friedt W. Molecular mapping of the Rf1 gene restoring fertility in PET1-based F1 hybrids in sunflower (Helianthus annuus L.). Theoretical and Applied Genetics. 2003;106:599-606. DOI: 10.1007/s00122-002-1078-yHorn R., Kusterer B., Lazarescu E., Prüfe M., Friedt W. Molecular mapping of the Rf1 gene restoring fertility in PET1-based F1 hybrids in sunflower (Helianthus annuus L.). Theoretical and Applied Genetics. 2003;106:599-606. DOI: 10.1007/s00122-002-1078-yHorn R., Radanovic A., Fuhrmann L., Sprycha Y., Hamrit S., Jockovic M., Miladinovic D., Jansen C. Development and validation of markers for the fertility restorer gene Rf1 in sunflower. International Journal of Molecular Sciences. 2019:20(6):1260. DOI: 10.3390/ijms20061260Horn R., Radanovic A., Fuhrmann L., Sprycha Y., Hamrit S., Jockovic M., Miladinovic D., Jansen C. Development and validation of markers for the fertility restorer gene Rf1 in sunflower. International Journal of Molecular Sciences. 2019:20(6):1260. DOI: 10.3390/ijms20061260Jan C.C., Vick B.A., Miller J.F., Kahler A.L., Butler E.T. Construction of an RFLP linkage map for cultivated sunflower. Theoretical and Applied Genetics. 1998;96(1):15-22. DOI: 10.1007/s001220050703Jan C.C., Vick B.A., Miller J.F., Kahler A.L., Butler E.T. Construction of an RFLP linkage map for cultivated sunflower. Theoretical and Applied Genetics. 1998;96(1):15-22. DOI: 10.1007/s001220050703Jan C.C., Vick B.A. Inheritance and allelic relationships of fertility restoration genes for seven new sources of male-sterile cytoplasm in sunflower. Plant Breeding. 2007;126(2):213-217. DOI: 10.1111/j.1439-0523.2007.01350.xJan C.C., Vick B.A. Inheritance and allelic relationships of fertility restoration genes for seven new sources of male-sterile cytoplasm in sunflower. Plant Breeding. 2007;126(2):213-217. DOI: 10.1111/j.1439-0523.2007.01350.xKarabitsina Y.I., Gavrilova V.A., Alpatieva N.V., Kuznetsova E.B., Anisimova I.N. Peculiarities of inheritance of pollen fertility restoration trait in sunflower with cytoplasmic male sterility. Russian Journal of Genetics. 2019;55:1375-1382. DOI: 10.1134/S1022795419110073Karabitsina Y.I., Gavrilova V.A., Alpatieva N.V., Kuznetsova E.B., Anisimova I.N. Peculiarities of inheritance of pollen fertility restoration trait in sunflower with cytoplasmic male sterility. Russian Journal of Genetics. 2019;55:1375-1382. DOI: 10.1134/S1022795419110073Kinman M.L. New developments in the USDA and state experiment station sunflower breeding programs. In: Proceedings of the 4th International Sunflower Conference; 1970 June 23-25; Memphis, Tennessee, USA. 1970. P. 181-183. Available at: https://www.isasunflower.org/ fileadmin/documents/Proceedings/4thISC1970/T1970BRE08.pdf [accessed Nov. 15, 2024].Kinman M.L. New developments in the USDA and state experiment station sunflower breeding programs. In: Proceedings of the 4th International Sunflower Conference; 1970 June 23-25; Memphis, Tennessee, USA. 1970. P. 181-183. Available at: https://www.isasunflower.org/ fileadmin/documents/Proceedings/4thISC1970/T1970BRE08.pdf [accessed Nov. 15, 2024].Leclerq P. Une sterilite cytoplasmique chez le tournesol. Annales de l'Amélioration des Plantes. 1969;19(2):99-106. [In French]Leclerq P. Une sterilite cytoplasmique chez le tournesol. Annales de l'Amélioration des Plantes. 1969;19(2):99-106. [In French]Li J.T., Yang J., Chen D.C., Zhang X.I., Tang Z.S. An optimized mini-preparation method to obtain high-quality genomic DNA from mature leaves of sunflower. Genetics and Molecular Research. 2007;6(4):1064-1071.Li J.T., Yang J., Chen D.C., Zhang X.I., Tang Z.S. An optimized mini-preparation method to obtain high-quality genomic DNA from mature leaves of sunflower. Genetics and Molecular Research. 2007;6(4):1064-1071.Liu Z., Mulpuri S., Feng J., Vick B.A., Jan C.-C. Molecular mapping of the Rf3 fertility restoration gene to facilitate its utilization in breeding confection sunflower. Molecular Breeding. 2012;29:275-284. DOI: 10.1007/s11032-011-9563-0Liu Z., Mulpuri S., Feng J., Vick B.A., Jan C.-C. Molecular mapping of the Rf3 fertility restoration gene to facilitate its utilization in breeding confection sunflower. Molecular Breeding. 2012;29:275-284. DOI: 10.1007/s11032-011-9563-0Liu Z., Wang D., Feng J., Seiler G.J., Cai X., Jan C.-C. Diversifying sunflower germplasm by integration and mapping of a novel male fertility restoration gene. Genetics. 2013;193(3):727-37. DOI: 10.1534/genetics.112.146092Liu Z., Wang D., Feng J., Seiler G.J., Cai X., Jan C.-C. Diversifying sunflower germplasm by integration and mapping of a novel male fertility restoration gene. Genetics. 2013;193(3):727-37. DOI: 10.1534/genetics.112.146092Liu Z., Zhang L., Seiler G.J., Jan C.-C. Molecular mapping of theRf9 gene from RCMG 1 for CMS ANN3 derived from wild sunflower (Helianthus annuus L.). Euphytica. 2023;219:46. DOI: 10.1007/s10681-023-03176-3Liu Z., Zhang L., Seiler G.J., Jan C.-C. Molecular mapping of theRf9 gene from RCMG 1 for CMS ANN3 derived from wild sunflower (Helianthus annuus L.). Euphytica. 2023;219:46. DOI: 10.1007/s10681-023-03176-3NCBI. National Center for Biotechnology Information. Available from: https://www.ncbi.nlm.nih.gov [accessed Nov. 15, 2024].NCBI. National Center for Biotechnology Information. Available from: https://www.ncbi.nlm.nih.gov [accessed Nov. 15, 2024].Oil seed sunflower description of released restorer line germplasm. Available from: https://www.ag.ndsu.edu/fss North Dakota Foundation Seedstocks [accessed Nov. 15, 2024].Oil seed sunflower description of released restorer line germplasm. Available from: https://www.ag.ndsu.edu/fss North Dakota Foundation Seedstocks [accessed Nov. 15, 2024].Polivanova O.B., Sivolapova A.B., Goryunov D.V., Fedorova A.V., Sotnikova E.A., Chebanova Y.V., Karabitsina Y.U.; Benko N.I., Demurin Y.N., Goryunova S.V. Structural diversity of sunflower (Helianthus annuus L.) candidate Rf1 loci based on gene-specific PCR. Research on Crops. 2021;22(1):40-46. DOI: 10.31830/2348-7542.2021.034Polivanova O.B., Sivolapova A.B., Goryunov D.V., Fedorova A.V., Sotnikova E.A., Chebanova Y.V., Karabitsina Y.U.; Benko N.I., Demurin Y.N., Goryunova S.V. Structural diversity of sunflower (Helianthus annuus L.) candidate Rf1 loci based on gene-specific PCR. Research on Crops. 2021;22(1):40-46. DOI: 10.31830/2348-7542.2021.034Qi L.L., Seiler G.J., Hulke B.S., Vick B.A., Gulya T.J. Genetics and mapping of the R11 gene conferring resistance to recently emerged rust races, tightly linked to male fertility restoration, in sunflower (Helianthus annuus L.). Theoretical and Applied Genetics. 2012;125:921-932. DOI: 10.1534/genetics.112.146092Qi L.L., Seiler G.J., Hulke B.S., Vick B.A., Gulya T.J. Genetics and mapping of the R11 gene conferring resistance to recently emerged rust races, tightly linked to male fertility restoration, in sunflower (Helianthus annuus L.). Theoretical and Applied Genetics. 2012;125:921-932. DOI: 10.1534/genetics.112.146092Radanovi´c A.; Sprycha Y.; Jockovi´c M.; Sundt M.; Miladinovi´c D.; Jansen C.; Horn R. KASP markers specific for the fertility restorer locus Rf1 and application for genetic purity testing in sunflowers (Helianthus annuus L.). Genes. 2022;13:465. DOI: 10.3390/genes13030465Radanovi´c A.; Sprycha Y.; Jockovi´c M.; Sundt M.; Miladinovi´c D.; Jansen C.; Horn R. KASP markers specific for the fertility restorer locus Rf1 and application for genetic purity testing in sunflowers (Helianthus annuus L.). Genes. 2022;13:465. DOI: 10.3390/genes13030465Sajer O., Schirmak U., Hamrit S., Horn R. Mapping of the new fertility restorer gene Rf-PET2 close to Rf1 on linkage group 13 in sunflower (Helianthus annuus L.). Genes. 2020;11:269. DOI: 10.3390/genes11030269Sajer O., Schirmak U., Hamrit S., Horn R. Mapping of the new fertility restorer gene Rf-PET2 close to Rf1 on linkage group 13 in sunflower (Helianthus annuus L.). Genes. 2020;11:269. DOI: 10.3390/genes11030269Schnabel U., Engelmann U., Horn R. Development of markers for the use of the PEF1 cytoplasm in sunflower hybrid breeding. Plant Breeding. 2008;127(6):587-591. DOI: 10.1111/j.1439-0523.2008.01516.xSchnabel U., Engelmann U., Horn R. Development of markers for the use of the PEF1 cytoplasm in sunflower hybrid breeding. Plant Breeding. 2008;127(6):587-591. DOI: 10.1111/j.1439-0523.2008.01516.xSerieys H. Identification, study, utilization in breeding programs of new CMS sources. In: Proceedings of the Sunflower Subnetwork Progress Report; 2005 July 17-20; Novi Sad, Serbia and Montenegro. Rome, Italy: FAO. 2005. P. 47-53.Serieys H. Identification, study, utilization in breeding programs of new CMS sources. In: Proceedings of the Sunflower Subnetwork Progress Report; 2005 July 17-20; Novi Sad, Serbia and Montenegro. Rome, Italy: FAO. 2005. P. 47-53.Sivolapova A.B., Polivanova O.B., Goryunov D.V., Chebanova Y.V., Fedоrova A.V., Sotnikova E.A., Karabitsina Y.I., Benko N.I., Mukhina Z.M., Anisimova I.N., Demurin Y.N., Goryunova S.V. Refinement of Rf1-gene localization and development of the new molecular markers for fertility restoration in sunflower. Molecular Biology Reports. 2023;50(9):7919-7926. DOI: 10.1007/s11033-023-08646-4Sivolapova A.B., Polivanova O.B., Goryunov D.V., Chebanova Y.V., Fedоrova A.V., Sotnikova E.A., Karabitsina Y.I., Benko N.I., Mukhina Z.M., Anisimova I.N., Demurin Y.N., Goryunova S.V. Refinement of Rf1-gene localization and development of the new molecular markers for fertility restoration in sunflower. Molecular Biology Reports. 2023;50(9):7919-7926. DOI: 10.1007/s11033-023-08646-4Talukder Z., Ma G., Hulke B., Jan C.-C., Qi L. Linkage mapping and genome-wide association studies of the Rf gene cluster in sunflower (Helianthus annuus L.) and their distribution in world sunflower collections. Frontiers in Genetics. 2019;10:216. DOI: 10.3389/fgene.2019.00216Talukder Z., Ma G., Hulke B., Jan C.-C., Qi L. Linkage mapping and genome-wide association studies of the Rf gene cluster in sunflower (Helianthus annuus L.) and their distribution in world sunflower collections. Frontiers in Genetics. 2019;10:216. DOI: 10.3389/fgene.2019.00216Трубачеева Н.В., Салина Е.А., Шумный В.К. Использование системы ЦМС-Rf в гибридной селекции подсолнечника. Письма в Вавиловский журнал генетики и селекции. 2024;10(2):119-131. DOI: 10.18699/letvjgb-2024-10-14Trubacheeva N.V., Salina E.A., Shumny V.K., The use of CMS/Rf system for sunflower hybrid breeding. Pisma v Vavilovskii Zhurnal Genetiki i Selektsii = Letters to Vavilov Journal of Genetics and Breeding. 2024;10(2):119-131. [in Russian]. DOI: 10.18699/letvjgb-2024-10-14Yue B., Vick B.A., Cai X., Hu J. Genetic mapping for the Rf1 (fertility restoration) gene in sunflower (Helianthus annuus L.) by SSR and TRAP markers. Plant Breeding. 2010;129:24-28. DOI: 10.1111/j.14390523.2009.01661.xYue B., Vick B.A., Cai X., Hu J. Genetic mapping for the Rf1 (fertility restoration) gene in sunflower (Helianthus annuus L.) by SSR and TRAP markers. Plant Breeding. 2010;129:24-28. DOI: 10.1111/j.14390523.2009.01661.x

The authors declare that there are no conflicts of interest present.