vavilovВавиловский журнал генетики и селекцииVavilov Journal of Genetics and Breeding2500-3259Institute of Cytology and Genetics of Siberian Branch of the RAS10.18699/VJGB-23-38vavilov-3769Research ArticleГЕНОМИКА И ТРАНСКРИПТОМИКАPLANT GENETICS AND BREEDINGГенетическая структура популяции дикорастущих форм винограда заповедника «Утриш»Genetic structure of the population of wild-growing vines of the Utrish Nature Reservehttps://orcid.org/0000-0002-2446-0971ИльницкаяЕ. Т.IlnitskayaE. T.

Краснодар

Krasnodar

ilnitskaya79@mail.ruhttps://orcid.org/0000-0002-3397-0666МакаркинаМ. В.MakarkinaM. V.

Краснодар

Krasnodar

https://orcid.org/0000-0002-4702-9148ГорбуновИ. В.GorbunovI. V.

Анапа

Anapa

https://orcid.org/0000-0002-6251-300XСтепановИ. В.StepanovI. V.

Краснодар

Krasnodar

https://orcid.org/0000-0003-2908-6461КозинаТ. Д.KozinaT. D.

Краснодар

Krasnodar

https://orcid.org/0000-0003-1305-3614КожевниковЕ. А.KozhevnikovE. A.

Краснодар

Krasnodar

https://orcid.org/0000-0002-4865-0323КотлярВ. К.KotlyarV. K.

Краснодар

Krasnodar

Северо-Кавказский федеральный научный центр садоводства, виноградарства, виноделияРоссияNorth Caucasian Federal Scientific Center of Horticulture, Viticulture, Wine-makingRussian FederationАнапская зональная опытная станция виноградарства и виноделия – филиал Северо-Кавказского федерального научного центра садоводства, виноградарства, виноделияРоссияAnapa Zonal Experimental Station of Viticulture and Wine-making – Branch of North Caucasian Federal Scientific Center of Horticulture, Viticulture, Wine-makingRussian Federation202313072023274316322Copyright © Ильницкая Е.Т., Макаркина М.В., Горбунов И.В., Степанов И.В., Козина Т.Д., Кожевников Е.А., Котляр В.К., 20232023Ильницкая Е.Т., Макаркина М.В., Горбунов И.В., Степанов И.В., Козина Т.Д., Кожевников Е.А., Котляр В.К.Ilnitskaya E.T., Makarkina M.V., Gorbunov I.V., Stepanov I.V., Kozina T.D., Kozhevnikov E.A., Kotlyar V.K.This work is licensed under a Creative Commons Attribution 4.0 License.https://vavilov.elpub.ru/jour/article/view/3769

Виноград – одна из самых распространенных сельскохозяйственных культур в мире. В настоящее время наиболее точным методом для изучения генофонда растений считается анализ генотипов на уровне ДНК. Дикорастущие формы и древние сорта различных регионов виноградарства – актуальные объекты исследований в данной области. Целью работы было изучение популяции дикорастущего винограда, произрастающей на территории заповедника «Утриш» на Черноморском побережье Краснодарского края. Территория заповедника представляет интерес для подобного рода исследований, так как является местом древних поселений, а экологические условия пригодны для произрастания диких форм винограда. В процессе обследования территории обнаружено 24 образца дикорастущего винограда, которые были описаны по основным морфологическим характеристикам и проанализированы молекулярно-генетическим методом. Найденные формы генотипированы с помощью 15 ДНК-маркеров, в том числе девяти общепринятых для ДНК-паспортизации сортов винограда (VVS2, VVMD5, VVMD7, VVMD25, VVMD27, VVMD28, VVMD32, VrZAG62, VrZAG79) и маркера VVIb23, позволяющего определять обоеполые и двудомные формы. Статистическая обработка данных полиморфизма микросателлитных локусов выполнена в программе GenAlEx 6.5. Генетические взаимосвязи исследуемых форм винограда оценивали в программе PAST 2.17c. В результате обнаружено, что образцы характеризуются морфологическим и генетическим полиморфизмом. Количество идентифицированных аллелей в выборке варьировало от 5 до 18 и составило в среднем 8 аллелей на локус. Проведение статистической обработки данных ДНК-анализа позволило выявить две генетически различные популяции среди обнаруженных дикорастущих форм. Оценено генетическое сходство найденных форм с некоторыми аборигенными сортами географически близких регионов виноградарства, подвойными сортами и представителями Vitis sylvestris с других территорий. Одна из обнаруженных в заповеднике «Утриш» популяций близка к ряду генотипов V. sylvestris, ДНК-профили которых представлены в международной базе данных Vitis International Variety Catalogue.

Grapes are one of the most common agricultural crops in the world. Currently, the analysis of genotypes directly at the DNA level is considered to be the most accurate method for studying the plant gene pool. The study of wild vines and ancient varieties in various regions of viticulture is an important direction of research in this field. The purpose of this work was to study the population of wild grapes growing on the territory of the Utrish Nature Reserve on the Black Sea coast of Krasnodar Region. The territory of the reserve is of interest as it is a site of ancient settlements, and the environmental conditions are suitable for the growth of wild grapes. During the survey of the territory, 24 samples of wild grapes were found, which were described according to the main morphological characteristics and analyzed by the molecular genetic method. The found vines were genotyped using 15 DNA markers, including nine commonly used for DNA fingerprinting (VVS2, VVMD5, VVMD7, VVMD25, VVMD27, VVMD28, VVMD32, VrZAG62, VrZAG79) and VVIb23, which allows determining hermaphrodite and dioecious vines. Statistical processing of microsatellite loci polymorphism data was carried out using the GenAlEx 6.5 program. The genetic relationships of the studied vines were evaluated using the PAST 2.17c program. The samples were found to be morphologically and genetically polymorphic. The number of alleles identified in the sample varied from 5 to 18 and averaged 8 alleles per locus. Statistical processing of DNA analysis data made it possible to identify two genetically different populations among the wild discovered vines. An assessment of genetic similarity of the found vines with some local varieties of geographically close viticulture regions, rootstocks and representatives of Vitis sylvestris from other territories was made. One of the populations found in the Utrish Nature Reserve is close to a number of V. sylvestris genotypes, the DNA profiles of which are presented in the Vitis International Variety Catalogue.

дикорастущие формы виноградаVitis sylvestrisДНК-профилированиегенетический полиморфизмSSR-локусыwild-growing vinesVitis sylvestrisDNA profilinggenetic polymorphismSSR lociThis study was supported by the Russian Foundation for Basic Research and the Administration of the Krasnodar Territory (grant No. 19-416-230025 r_а).ReferencesChernodubov A.I., Rudenok Y. Phytocenoses biodiversity in “Utrish” Re serve. Lesotehnicheskiy Zhurnal = Forestry Engineering Jour nal. 2015;5(1):120-127. DOI 10.12737/11269. (in Russian)Chernodubov A.I., Rudenok Y. Phytocenoses biodiversity in “Utrish” Re serve. Lesotehnicheskiy Zhurnal = Forestry Engineering Jour nal. 2015;5(1):120-127. DOI 10.12737/11269. (in Russian)Cunha J., Ibáñez J., Teixeira-Santos M., Brazão J., Fevereiro P., Martínez-Zapater J.M., Eiras-Dias J.E. Genetic relationships among portuguese cultivated and wild Vitis vinifera L. germplasm. Front. Plant Sci. 2020;11:127. DOI 10.3389/fpls.2020.00127.Cunha J., Ibáñez J., Teixeira-Santos M., Brazão J., Fevereiro P., Martínez-Zapater J.M., Eiras-Dias J.E. Genetic relationships among portuguese cultivated and wild Vitis vinifera L. germplasm. Front. Plant Sci. 2020;11:127. DOI 10.3389/fpls.2020.00127.De Michele R., La Bella F., Gristina A.S., Fontana I., Pacifico D., Garfi G., Motisi A., Crucitti D., Abbate L., Carimi F. Phylogenetic relationship among wild and cultivated grapevine in Sicily: a hotspot in the middle of the Mediterranean basin. Front. Plant Sci. 2019;10:1506. DOI 10.3389/fpls.2019.01506.De Michele R., La Bella F., Gristina A.S., Fontana I., Pacifico D., Garfi G., Motisi A., Crucitti D., Abbate L., Carimi F. Phylogenetic relationship among wild and cultivated grapevine in Sicily: a hotspot in the middle of the Mediterranean basin. Front. Plant Sci. 2019;10:1506. DOI 10.3389/fpls.2019.01506.Di Gaspero G., Copetti D., Coleman C., Castellarin S.D., Eibach R., Kozma P., Lacombe T., Gambetta G., Zvyagin A., Cindrić P., Kovács L., Morgante M., Testolin R. Selective sweep at the Rpv3 locus during grapevine breeding for downy mildew resistance. Theor. Appl. Genet. 2012;124(2):227-286. DOI 10.1007/s00122-011-1703-8.Di Gaspero G., Copetti D., Coleman C., Castellarin S.D., Eibach R., Kozma P., Lacombe T., Gambetta G., Zvyagin A., Cindrić P., Kovács L., Morgante M., Testolin R. Selective sweep at the Rpv3 locus during grapevine breeding for downy mildew resistance. Theor. Appl. Genet. 2012;124(2):227-286. DOI 10.1007/s00122-011-1703-8.Doulati-Baneh H., Mohammadi S., Labra M., De Mattia F., Bruni I., Mezzasalma V., Abdollahi R. Genetic characterization of some wild grape populations (Vitis vinifera subsp. sylvestris) of Zagros mountains (Iran) to indentify a conservation strategy. Plant Genet. Resour. 2015;13(1):27-35. DOI 10.1017/S1479262114000598.Doulati-Baneh H., Mohammadi S., Labra M., De Mattia F., Bruni I., Mezzasalma V., Abdollahi R. Genetic characterization of some wild grape populations (Vitis vinifera subsp. sylvestris) of Zagros mountains (Iran) to indentify a conservation strategy. Plant Genet. Resour. 2015;13(1):27-35. DOI 10.1017/S1479262114000598.Ellstrand N.C., Heredia S.M., Leak-Garcia J.A., Heraty J.M., Burger J.C., Li Y., Nohzadeh-Malakshah S., Ridley C.E. Crops gone wild: evolution of weeds and invasive from domesticated ancestors. Evol. Appl. 2010;3(5-6):494-504. DOI 10.1111/j.1752-4571.2010.00140.x.Ellstrand N.C., Heredia S.M., Leak-Garcia J.A., Heraty J.M., Burger J.C., Li Y., Nohzadeh-Malakshah S., Ridley C.E. Crops gone wild: evolution of weeds and invasive from domesticated ancestors. Evol. Appl. 2010;3(5-6):494-504. DOI 10.1111/j.1752-4571.2010.00140.x.Gorbunov I.V., Lukyanov A.A. Kuban wild-growing grapes and their morphological features. Vinogradarstvo i Vinodelie = Viticulture and Winemaking. 2020;49:30-33. (in Russian)Gorbunov I.V., Lukyanov A.A. Kuban wild-growing grapes and their morphological features. Vinogradarstvo i Vinodelie = Viticulture and Winemaking. 2020;49:30-33. (in Russian)Gorbunov I.V., Lukyanov A.A., Bykhalova O.N. Morphological peculiarities of the Kuban wild-growing forms of grapes. Plodovodstvo i Vinogradarstvo Yuga Rossii = Fruit Growing and Viticulture of South Russia. 2020;65(5):70-82. DOI 10.30679/2219-5335-2020-5-65-70-82. (in Russian)Gorbunov I.V., Lukyanov A.A., Bykhalova O.N. Morphological peculiarities of the Kuban wild-growing forms of grapes. Plodovodstvo i Vinogradarstvo Yuga Rossii = Fruit Growing and Viticulture of South Russia. 2020;65(5):70-82. DOI 10.30679/2219-5335-2020-5-65-70-82. (in Russian)Gorislavec S.M., Risovannaya V.I., Volkov Y.A., Kolosova A.A., Volodin V.A. Identification and evaluation of wild growing vines on the territory of Yalta mountain-forest nature reserve using molecular markers. “Magarach”. Vinogradarstvo i Vinodelie = “Magarach”. Viti culture and Winemaking. 2017;1:19-21. (in Russian)Gorislavec S.M., Risovannaya V.I., Volkov Y.A., Kolosova A.A., Volodin V.A. Identification and evaluation of wild growing vines on the territory of Yalta mountain-forest nature reserve using molecular markers. “Magarach”. Vinogradarstvo i Vinodelie = “Magarach”. Viti culture and Winemaking. 2017;1:19-21. (in Russian)Hammer Ø., Harper D.A.T., Ryan P.D. PAST: Paleontological statistics software package for education and data analysis. Palaeontol. Electron. 2001;4(1):1-9.Hammer Ø., Harper D.A.T., Ryan P.D. PAST: Paleontological statistics software package for education and data analysis. Palaeontol. Electron. 2001;4(1):1-9.Kupe M., Ercisli S., Jovanović-Cvetković T., Eyduran S.P., Ayed R.B. Molecular characterization of wild grapes from northeastern part of Turkey. Genetika. 2021;53(1):93-102. DOI 10.2298/GENSR2101093K.Kupe M., Ercisli S., Jovanović-Cvetković T., Eyduran S.P., Ayed R.B. Molecular characterization of wild grapes from northeastern part of Turkey. Genetika. 2021;53(1):93-102. DOI 10.2298/GENSR2101093K.Lukšić K., Zdunić G., Hančević K., Mihaljević M.Ž., Mucalo A., Maul E., Riaz S., Pejić I. Identification of powdery mildew resistance in wild grapevine (Vitis vinifera subsp. sylvestris Gmel Hegi) from Croatia and Bosnia and Herzegovina. Sci. Rep. 2022;12(1):2128. DOI 10.1038/s41598-022-06037-6.Lukšić K., Zdunić G., Hančević K., Mihaljević M.Ž., Mucalo A., Maul E., Riaz S., Pejić I. Identification of powdery mildew resistance in wild grapevine (Vitis vinifera subsp. sylvestris Gmel Hegi) from Croatia and Bosnia and Herzegovina. Sci. Rep. 2022;12(1):2128. DOI 10.1038/s41598-022-06037-6.Margaryan K., Maul E., Muradyan Z., Hovhannisyan A., Melyan G., Aroutiounian R. Evaluation of breeding potential of wild grape originating from Armenia. BIO Web Conf. 2019;15:01006. DOI 10.1051/bioconf/20191501006.Margaryan K., Maul E., Muradyan Z., Hovhannisyan A., Melyan G., Aroutiounian R. Evaluation of breeding potential of wild grape originating from Armenia. BIO Web Conf. 2019;15:01006. DOI 10.1051/bioconf/20191501006.McGovern P.E. Ancient Wine: The Search for the Origins of Viniculture. Princeton: Princeton Univ. Press, 2004. DOI 10.2307/j.ctvfjd0bk.McGovern P.E. Ancient Wine: The Search for the Origins of Viniculture. Princeton: Princeton Univ. Press, 2004. DOI 10.2307/j.ctvfjd0bk.Merdinoglu D., Butterlin G., Bevilacqua L., Chiquet V., Adam-Blondon A.F., Decroocq S. Development and characterization of a large set of microsatellite markers in grapevine (Vitis vinifera L.) suitable for multiplex PCR. Mol. Breed. 2005;15:349-366. DOI 10.1007/s11032-004-7651-0.Merdinoglu D., Butterlin G., Bevilacqua L., Chiquet V., Adam-Blondon A.F., Decroocq S. Development and characterization of a large set of microsatellite markers in grapevine (Vitis vinifera L.) suitable for multiplex PCR. Mol. Breed. 2005;15:349-366. DOI 10.1007/s11032-004-7651-0.Peakall R., Smouse P.E. GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research – an update. Bioinformatics. 2012;28(19):2537-2539. DOI 10.1093/bioinformatics/bts460.Peakall R., Smouse P.E. GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research – an update. Bioinformatics. 2012;28(19):2537-2539. DOI 10.1093/bioinformatics/bts460.Pritchard J.K., Stephens M., Donnelly P. Inference of population structure using multilocus genotype data. Genetics. 2000;155(2):945-959. DOI 10.1093/genetics/155.2.945.Pritchard J.K., Stephens M., Donnelly P. Inference of population structure using multilocus genotype data. Genetics. 2000;155(2):945-959. DOI 10.1093/genetics/155.2.945.Riaz S., Boursiquot J.M., Dangl G.S., Lacombe T., Laucou V., Tenscher A.C., Walker M.A. Identification of mildew resistance in wild and cultivated Central Asian grape germplasm. BMC Plant Biol. 2013;13(1):149. DOI 10.1186/1471-2229-13-149.Riaz S., Boursiquot J.M., Dangl G.S., Lacombe T., Laucou V., Tenscher A.C., Walker M.A. Identification of mildew resistance in wild and cultivated Central Asian grape germplasm. BMC Plant Biol. 2013;13(1):149. DOI 10.1186/1471-2229-13-149.Rogers S.O., Bendich A.J. Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues. Plant Mol. Biol. 1985;5:69-76. DOI 10.1007/BF00020088.Rogers S.O., Bendich A.J. Extraction of DNA from milligram amounts of fresh, herbarium and mummified plant tissues. Plant Mol. Biol. 1985;5:69-76. DOI 10.1007/BF00020088.Schwander F., Eibach R., Fechter I., Hausmann L., Zyprian E., Töpfer R. Rpv10: a new locus from the Asian Vitis gene pool for pyra miding downy mildew resistance loci in grapevine. Theor. Appl. Genet. 2012;124(1):163-176. DOI 10.1007/s00122-011-1695-4.Schwander F., Eibach R., Fechter I., Hausmann L., Zyprian E., Töpfer R. Rpv10: a new locus from the Asian Vitis gene pool for pyra miding downy mildew resistance loci in grapevine. Theor. Appl. Genet. 2012;124(1):163-176. DOI 10.1007/s00122-011-1695-4.This P. Microsatellite markers analysis. In: Minutes of the First Grape Gen06 Work-shop, March 22nd and 23rd. INRA, Versailles (France). Paris: INRA, 2007;3-42.This P. Microsatellite markers analysis. In: Minutes of the First Grape Gen06 Work-shop, March 22nd and 23rd. INRA, Versailles (France). Paris: INRA, 2007;3-42.This P., Jung A., Boccacci P., Borrego J., Botta R., Costantini L., Crespan M., Dangl G.S., Eisenheld C., Ferreira-Monteiro F., Grando S., Ibáñez J., Lacombe T., Laucou V., Magalhães R., Meredith C.P., Milani N., Peterlunger E., Regner F., Zulini L., Maul E. Development of a standard set of microsatellite reference alleles for identification of grape cultivars. Theor. Appl. Genet. 2004;109(7):1448-1458. DOI 10.1007/s00122-004-1760-3.This P., Jung A., Boccacci P., Borrego J., Botta R., Costantini L., Crespan M., Dangl G.S., Eisenheld C., Ferreira-Monteiro F., Grando S., Ibáñez J., Lacombe T., Laucou V., Magalhães R., Meredith C.P., Milani N., Peterlunger E., Regner F., Zulini L., Maul E. Development of a standard set of microsatellite reference alleles for identification of grape cultivars. Theor. Appl. Genet. 2004;109(7):1448-1458. DOI 10.1007/s00122-004-1760-3.This P., Lacombe T., Thomas M.R. Historical origins and genetic diversity of wine grapes. Trends Genet. 2006;22(9):511-519. DOI 10.1016/j.tig.2006.07.008.This P., Lacombe T., Thomas M.R. Historical origins and genetic diversity of wine grapes. Trends Genet. 2006;22(9):511-519. DOI 10.1016/j.tig.2006.07.008.van Heerden C.J., Burger P., Vermeulen A., Prins R. Detection of downy and powdery mildew resistance QTL in a ‘Regent’ × ‘RedGlobe’ population. Euphytica. 2014;200:281-295. DOI 10.1007/s10681-014-1167-4.van Heerden C.J., Burger P., Vermeulen A., Prins R. Detection of downy and powdery mildew resistance QTL in a ‘Regent’ × ‘RedGlobe’ population. Euphytica. 2014;200:281-295. DOI 10.1007/s10681-014-1167-4.VIVC. Vitis International Variety Catalogue. Microsatellites by profile. Last modified: 2022-08-10. Julius Kühn-Institut, 2022. Available at: https://www.vivc.de/index.php?r=eva-analysis-mikrosatelliten-vivc%2FindexVIVC. Vitis International Variety Catalogue. Microsatellites by profile. Last modified: 2022-08-10. Julius Kühn-Institut, 2022. Available at: https://www.vivc.de/index.php?r=eva-analysis-mikrosatelliten-vivc%2FindexZdunić G., Lukšić K., Nagy Z.A., Mucalo A., Hančević K., Radić T., Butorac L., Jahnke G.G., Kiss E., Ledesma-Krist G., Regvar M., Likar M., Piltaver A., Žulj Mihaljević M., Maletić E., Pejić I., Werling M., Maul E. Genetic structure and relationships among wild and cultivated grapevines from Central Europe and part of the Western Balkan Peninsula. Genes. 2020;11(9):962. DOI 10.3390/genes11090962.Zdunić G., Lukšić K., Nagy Z.A., Mucalo A., Hančević K., Radić T., Butorac L., Jahnke G.G., Kiss E., Ledesma-Krist G., Regvar M., Likar M., Piltaver A., Žulj Mihaljević M., Maletić E., Pejić I., Werling M., Maul E. Genetic structure and relationships among wild and cultivated grapevines from Central Europe and part of the Western Balkan Peninsula. Genes. 2020;11(9):962. DOI 10.3390/genes11090962.Zendler D., Schneide P., Töpfe R., Zyprian E. Fine mapping of Ren3 reveals two loci mediating hypersensitive response against Erysiphe necator in grapevine. Euphytica. 2017;213(68):1029. DOI 10.1007/s10681-017-1857-9.Zendler D., Schneide P., Töpfe R., Zyprian E. Fine mapping of Ren3 reveals two loci mediating hypersensitive response against Erysiphe necator in grapevine. Euphytica. 2017;213(68):1029. DOI 10.1007/s10681-017-1857-9.

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