Ecological geneticsEcological geneticsЭкологическая генетика1811-09322411-9202Eco-Vector70796510.17816/ecogen707965Methodology in ecological geneticsМетодология экологической генетикиUnknownPlant flow cytometry: current problems, aspects of application and data analysisПроточная цитометрия растений: современные проблемы, аспекты применения и анализа данныхhttps://orcid.org/0000-0002-4884-07684762-1760SkaptsovMikhailСкапцовМихаилmr.skaptsov@mail.ruhttps://orcid.org/0000-0002-8198-53604758-1180DegtyarevaGalinaДегтяреваГалинаdegavi@belozersky.msu.ruhttps://orcid.org/0000-0002-9657-39592706-6346SmirnovSergeyСмирновСергейserg_sm@mail.ruhttps://orcid.org/0000-0003-2284-68513424-9252KutsevMaximКуцевМаксимm_kucev@mail.ruhttps://orcid.org/0000-0002-1052-45752428-4653ShmakovAlexanderШмаковАлександрssbgbot@mail.ru290720262432905202629072026Copyright ©; , Eco-VectorCopyright ©; , Эко-ВекторEco-VectorЭко-Векторhttps://eco-vector.com/for_authors.php#07https://journals.eco-vector.com/ecolgenet/article/view/707965

Plant flow cytometry is a widely used method for analyzing and quantifying various cellular and nuclear characteristics, finding application in various fields of botany, plant physiology and biochemistry, evolutionary theory, genetics, and breeding. Flow cytometry is becoming an important tool for the certification and verification of biological collections, providing an objective criterion (genome size/ploidy) to confirm the taxonomic affiliation and genetic stability of collection specimens. Including cytometric data in collection metadata enhances their scientific value and facilitates interlaboratory reproducibility of results. While instrumentation advances have minimized the impact of hardware on results, the challenge of obtaining accurate and reproducible genome size data in plants remains pressing. In addition to the unique organization of plant genomes, which determines intraspecific variability, the sources of inaccuracies remain inherent in the flow cytometry method itself. This article covers key aspects of flow cytometry that influence the final result and affect all stages of sample preparation. Emphasis is placed on the quality of the source material, the issue of standardization, and key considerations when comparing previously obtained results. Illustrations are included to provide clarity and prevent common errors.

Проточная цитометрия растений – широко используемый метод для анализа и количественной оценки различных клеточных и ядерных характеристик, находящий применение в различных областях ботаники, физиологии и биохимии растений, эволюционном учении, генетике и селекции. Проточная цитометрия становится важным инструментом паспортизации и верификации биологических коллекций, обеспечивая объективный критерий (размер генома/плоидность) для подтверждения таксономической принадлежности и генетической стабильности коллекционных образцов. Включение цитометрических данных в метаданные коллекций повышает их научную ценность и способствует межлабораторной воспроизводимости результатов. Усовершенствование приборов сократило до минимума влияние аппаратной части на результат, однако проблема получения точных и воспроизводимых данных по размерам генома у растений продолжает оставаться актуальной. Помимо особенностей организации самих геномов растений, определяющих внутривидовую изменчивость, причины неточностей продолжают оставаться в самом методе проточной цитометрии. В статье затронуты ключевые моменты использования метода проточной цитометрии, влияющие на получаемый результат и затрагивающие все этапы пробоподготовки. Акцент сделан на качестве исходного материала, проблеме стандартизации и ключевых нюансах при сопоставлении ранее полученных результатов. Для наглядности и предотвращения типичных ошибок подобраны иллюстрации.

Doležel J., Bartoš J., Voglmayr H., et al. Nuclear DNA content and genome size of trout and human. Cytometry A. 2003;51:127–128. doi: 10.1002/cyto.a.10013Suda J., Krahulcová A., Trávníček P., et al. Genome size variation and species relationships in Hieracium sub-genus Pilosella (Asteraceae) as inferred by flow cytometry. Ann. Bot. 2007;100:1323–1335. doi: 10.1093/aob/mcm218Birstein V.J., Poletaev A.I., Goncharov B.F. DNA content in Eurasian sturgeon species determined by flow cytometry. Cytometry. 1993;14:377–383. doi: 10.1002/cyto.990140410MacCulloch R.D., Upton D.E., Murphy R.W. Trends in nuclear DNA content among amphibians and reptiles. Comp. Biochem. Physiol. B. 1996;113:601–605. doi: 10.1016/0305-0491(95)02076-4Hosny M., Gianinazzi-Pearson V., Dulieu H. Nuclear DNA contents of eleven fungal species in Glomales. Genome. 1998;41:422–428. doi: 10.1139/g98-038Brussaard C.P., Marie D., Bratbak G. Flow cytometric detection of viruses. J. Virol. Methods. 2000;85:175–182. doi: 10.1016/s0166-0934(99)00167-6Rowan B.A., Oldenburg D.J., Bendich A.J. A high-throughput method for detection of DNA in chloroplasts using flow cytometry. Plant Methods. 2007;3:5. doi: 10.1186/1746-4811-3-5Doležel J., Lucretti S., Molnár I., et al. Chromosome analysis and sorting. Cytometry A. 2021;99:328–342. doi: 10.1002/cyto.a.24331Kron P., Loureiro J., Castro S., et al. Flow cytometric analysis of pollen and spores: an overview of applications and methodology. Cytometry A. 2021;99:348–358. doi: 10.1002/cyto.a.24330Куцев М.Г., Скапцов М.В., Колтунова А.М., и др. ДНК-цитометрия изображений в исследовании растений: обзор. Turczaninowia. 2024;27:141–158. doi: 10.14258/turczaninowia.27.3.14 (In Russ.)Zonneveld B.J.M. The DNA weights per nucleus (genome size) of more than 2350 species of the flora of the Netherlands, of which 1370 are new to science, including the pattern of their DNA peaks. Forum Geobot. 2019;8:24–78. doi: 10.3264/FG.2019.1022Šmarda P., Knápek O., Březinová A., et al. Genome sizes and genomic guanine+cytosine (GC) contents of the Czech vascular flora with new estimates for 1700 species. Preslia. 2019;91:117–142. doi: 10.23855/preslia.2019.117Bennett M.D., Leitch I.J. Nuclear DNA amounts in angiosperms: targets, trends and tomorrow. Ann. Bot. 2011;107:467–590. doi: 10.1093/aob/mcq258Zonneveld B.J.M. Nuclear genome sizes of 343 accessions of wild collected Haworthia and Astroloba (Asphodelaceae, Alooideae), compared with the genome sizes of Chortolirion, Gasteria and 83 Aloe species. Plant Syst. Evol. 2015;301:931–953. doi: 10.1007/s00606-014-1123-8Kubešová M., Moravcová L., Suda J., et al. Naturalized plants have smaller genomes than their non-invading relatives: a flow cytometric analysis of the Czech alien flora. Preslia. 2010;82:81–96.Doležel J., Greilhuber J., Lucretti S., et al. Plant genome size estimation by flow cytometry: inter-laboratory comparison. Ann. Bot. 1998;82:17–26. doi: 10.1093/oxfordjournals.aob.a010312Greilhuber J. Intraspecific variation in genome size in angiosperms: identifying its existence. Ann. Bot. 2005;95:91–98. doi: 10.1093/aob/mci004Bourge M., Brown S.C., Siljak-Yakovlev S. Flow cytometry as tool in plant sciences, with emphasis on genome size and ploidy level assessment. Genet. Appl. 2018;2:1–12. doi: 10.31383/ga.vol2iss2pp1-12Skaptsov M.V., Kutsev M.G., Smirnov S.V., et al. Standards in plant flow cytometry: an overview, polymorphism and linearity issues. Turczaninowia. 2024;27:86–104. doi: 10.14258/turczaninowia.27.2.10Doležel J., Greilhuber J., Suda J. Estimation of nuclear DNA content in plants using flow cytometry. Nat. Protoc. 2007;2:2233–2244. doi: 10.1038/nprot.2007.310Galbraith D., Loureiro J., Antoniadi I., et al. Best practices in plant cytometry. Cytometry A. 2021;99:311–317. doi: 10.1002/cyto.a.24295Sliwinska E., Loureiro J., Leitch I.J., et al. Application-based guidelines for best practices in plant flow cytometry. Cytometry A. 2022;101:749–781. doi: 10.1002/cyto.a.24499Loureiro J., Čertner M., Lučanová M., et al. The use of flow cytometry for estimating genome sizes and DNA ploidy levels in plants. Methods Mol. Biol. 2023;2672:25–64. doi: 10.1007/978-1-0716-3226-0_2Čertner M., Lučanová M., Sliwinska E., et al. Plant material selection, collection, preservation, and storage for nuclear DNA content estimation. Cytometry A. 2022;101:737–748. doi: 10.1002/cyto.a.24482Doležel J., Greilhuber J., Suda J. Flow Cytometry with Plant Cells: Analysis of Genes, Chromosomes and Genomes. Weinheim: Wiley‐VCH Verlag GmbH & Co. KGaA; 2007.Doležel J., Sgorbati S., Lucretti S. Comparison of three DNA fluorochromes for flow cytometric estimation of nuclear DNA content in plants. Physiol. Plant. 1992;85:625–631. doi: 10.1111/j.1399-3054.1992.tb04764.xMichael T.P. Plant genome size variation: bloating and purging DNA. Brief. Funct. Genomics. 2014;13:308–317. doi: 10.1093/bfgp/elu005Fleischmann A., Michael T.P., Rivadavia F., et al. Evolution of genome size and chromosome number in the carnivorous plant genus Genlisea (Lentibulariaceae), with a new estimate of the minimum genome size in angiosperms. Ann. Bot. 2014;114:1651–1663. doi: 10.1093/aob/mcu189Pellicer J., Fay M.F., Leitch I.J. The largest eukaryotic genome of them all? Bot. J. Linn. Soc. 2010;164:10–15. doi: 10.1111/j.1095-8339.2010.01072.xFernández P., Amice R., Bruy D., et al. A 160 Gbp fork fern genome shatters size record for eukaryotes. iScience. 2024;27:109889. doi: 10.1016/j.isci.2024.109889Gordon S.P., Contreras-Moreira B., Woods D.P., et al. Extensive gene content variation in the Brachypodium distachyon pan-genome correlates with population structure. Nat. Commun. 2017;8:2184. doi: 10.1038/s41467-017-02292-8Lohaus R., Van de Peer Y. Of dups and dinos: evolution at the K/Pg boundary. Curr. Opin. Plant Biol. 2016;30:62–69. doi: 10.1016/j.pbi.2016.01.006Van de Peer Y., Mizrachi E., Marchal K. The evolutionary significance of polyploidy. Nat. Rev. Genet. 2017;18:411–424. doi: 10.1038/nrg.2017.26Шнеер В.С., Пунина Е.О., Родионов А.В. Внутривидовые различия в плоидности у покрытосеменных и их таксономическая интерпретация. Ботанический журнал. 2018;103:555–585. doi: 10.1134/S0006813618050010 (In Russ.)Husband B.C., Baldwin S.J., Suda J. The incidence of polyploidy in natural plant populations: major patterns and evolutionary processes. In: Greilhuber J., Doležel J., Wendel J.F., editors. Plant Genome Diversity. Vol. 2. Vienna: Springer; 2013. P. 255–276.Родионов А.В., Шнеер В.С., Гнутиков А.А., и др. Диалектика видов: от исходного единообразия, через максимальное возможное разнообразие к конечному единообразию. Ботанический журнал. 2020;105:835–853. doi: 10.31857/S0006813620070091 (In Russ.)Clark J.W. Genome evolution in plants and the origins of innovation. New Phytol. 2023;240:2204–2209. doi: 10.1111/nph.19242Carta A., Bedini G., Peruzzi L. A deep dive into the ancestral chromosome number and genome size of flowering plants. New Phytol. 2020;228:1097–1106. doi: 10.1111/nph.16668Chalhoub B., Denoeud F., Liu S., et al. Early allopolyploid evolution in the post-Neolithic Brassica napus oilseed genome. Science. 2014;345:950–953. doi: 10.1126/science.1253435Liang Z., Schnable J.C. Functional divergence between subgenomes and gene pairs after whole genome duplications. Mol. Plant. 2018;11:388–397. doi: 10.1016/j.molp.2017.12.010Rodionov A.V. Eupolyploidy as a mode in plant speciation. Russ. J. Genet. 2023;59:419–431. doi: 10.1134/S1022795423050113Leitch I.J., Bennett M.D. Genome downsizing in polyploid plants. Biol. J. Linn. Soc. 2004;82:651–663. doi: 10.1111/j.1095-8312.2004.00349.xGegas V.C., Wargent J.J., Pesquet E., et al. Endopolyploidy as a potential alternative adaptive strategy for Arabidopsis leaf size variation in response to UV-B. J. Exp. Bot. 2014;65:2757–2766. doi: 10.1093/jxb/ert473Scholes D.R., Siddappaji M.H., Paige K.N. The genetic basis of overcompensation in plants: a synthesis. Int. J. Mod. Bot. 2013;3:34–42. doi: 10.5923/s.ijmb.201310.05Greilhuber J., Doležel J., Lysák M.A., et al. The origin, evolution and proposed stabilization of the terms ‘genome size’ and ‘C-value’ to describe nuclear DNA contents. Ann. Bot. 2005;95:255–260. doi: 10.1093/aob/mci019Luo M.C., Deal K.R., Akhunov E.D., et al. Genome comparisons reveal a dominant mechanism of chromosome number reduction in grasses and accelerated genome evolution in Triticeae. Proc. Natl. Acad. Sci. U.S.A. 2009;106:15780–15785. doi: 10.1073/pnas.0908195106Panchy N., Lehti-Shiu M., Shiu S.H. Evolution of gene duplication in plants. Plant Physiol. 2016;171:2294–2316. doi: 10.1104/pp.16.00523Kron P., Suda J., Husband B.C. Applications of flow cytometry to evolutionary and population biology. Annu. Rev. Ecol. Evol. Syst. 2007;38:847–876. doi: 10.1146/annurev.ecolsys.38.091206.095504Smirnov S., Skaptsov M., Shmakov A., et al. Spontaneous hybridization among Allium tulipifolium and A. robustum (Allium subg. Melanocrommyum, Amaryllidaceae) under cultivation. Phytotaxa. 2017;303:155–164. doi: 10.11646/phytotaxa.303.2.5Urfus T., Chrtek J., Kaplan Z., et al. Flow cytometry in conservation: detecting hybridization risks in threatened plant species. Biodivers. Conserv. 2025;34:2337–2358. doi: 10.1007/s10531-025-03062-5Sokoloff D.D., Skaptsov M.V., Vislobokov N.A., et al. Morphological characterization of diploid and triploid Acorus calamus (Acoraceae) from southern Western Siberia, parthenocarpy in sterile plants and occurrence of aneuploidy. Bot. J. Linn. Soc. 2021;195:189–215. doi: 10.1093/botlinnean/boaa081Doležel J., Bartoš J. Plant DNA flow cytometry and estimation of nuclear genome size. Ann. Bot. 2005;95:99–110. doi: 10.1093/aob/mci005Parkhomenko A., Skaptsov M.V., Kondratieva A., et al. Nuclear DNA content in some Chondrilla taxa (Asteraceae) of European Russia and Western Kazakhstan. Acta Biol. Sibirica. 2025;11:49–69. doi: 10.5281/zenodo.14784067Avanzi S., Cionini P.G. A DNA cytophotometric investigation on the development of the female gametophyte of Ginkgo biloba. Caryologia. 1971;24:105–116. doi: 10.1080/00087114.1971.10796418Barlow P.W. Endopolyploidy: towards an understanding of its biological significance. Acta Biotheoretica. 1978;27:1–18. doi: 10.1007/BF00048400Barow M., Meister A. Endopolyploidy in seed plants is differently correlated to systematics, organ, life strategy and genome size. Plant Cell Environ. 2003;26:571–584. doi: 10.1046/j.1365-3040.2003.00988.xLeitch I.J., Dodsworth S. Endopolyploidy in plants. In: eLS. John Wiley & Sons, Ltd; 2017. doi: 10.1002/9780470015902.a0020097.pub2Kocová V., Mártonfi P. Endopolyploidy in Trifolium pratense L. Caryologia. 2011;64:419–426. doi: 10.1080/00087114.2011.10589809Barow M. Endopolyploidy in seed plants. BioEssays. 2006;28:271–281. doi: 10.1002/bies.20371Skaptsov M.V., Lomonosova M.N., Kutsev M.G., et al. The phenomenon of endopolyploidy in some species of the Chenopodioideae (Amaranthaceae). Bot. Lett. 2017;164:47–53. doi: 10.1080/23818107.2016.1276475Scholes D.R., Paige K.N. Plasticity in ploidy: a generalized response to stress. Trends Plant Sci. 2015;20:165–175. doi: 10.1016/j.tplants.2014.11.007Lim W.L., Loh C.S. Endopolyploidy in Vanda Miss Joaquim (Orchidaceae). New Phytol. 2003;159:279–287. doi: 10.1046/j.0028-646x.2003.00797.xLeite Montalvão A.P., Kersten B., Fladung M., et al. The diversity and dynamics of sex determination in dioecious plants. Front. Plant Sci. 2021;11:580488. doi: 10.3389/fpls.2020.580488Walas Ł., Mandryk W., Thomas P.A., et al. Sexual systems in gymnosperms: a review. Basic Appl. Ecol. 2018;31:1–9. doi: 10.1016/j.baae.2018.05.009Renner S.S. The relative and absolute frequencies of angiosperm sexual systems: dioecy, monoecy, gynodioecy, and an updated online database. Am. J. Bot. 2014;101:1588–1596. doi: 10.3732/ajb.1400196Ming R., Bendahmane A., Renner S.S. Sex chromosomes in land plants. Annu. Rev. Plant Biol. 2011;62:485–514. doi: 10.1146/annurev-arplant-042110-103914Doležel J., Göhde W. Sex determination in dioecious plants Melandrium album and M. rubrum using high-resolution flow cytometry. Cytometry. 1995;19:103–106. doi: 10.1002/cyto.990190203Koutecký P., Smith T., Loureiro J., et al. Best practices for instrument settings and raw data analysis in plant flow cytometry. Cytometry A. 2023;103:953–966. doi: 10.1002/cyto.a.24798Cires E., Cuesta C., Fernandez M.A., et al. Isolation of plant nuclei suitable for flow cytometry from species with extremely mucilaginous compounds: an example in the genus Viola L. (Violaceae). Anales Jard. Bot. Madrid. 2011;68:139–154. doi: 10.3989/ajbm.2293Suda J., Trávníček P. Reliable DNA ploidy determination in dehydrated tissues of vascular plants by DAPI flow cytometry – new prospects for plant research. Cytometry A. 2006;69:273–280. doi: 10.1002/cyto.a.20253Bainard J.D., Husband B.C., Baldwin S.J., et al. The effects of rapid desiccation on estimates of plant genome size. Chromosome Res. 2011;19:825–842. doi: 10.1007/s10577-011-9232-5Wang G., Yang Y. The effects of fresh and rapid desiccated tissue on estimates of Ophiopogoneae genome size. Plant Divers. 2016;38:190–193. doi: 10.1016/j.pld.2016.08.001Skaptsov M.V., Vaganov A.V., Kechaykin A.A., et al. The cytotypes variability of the complex Selaginella sanguinolenta s. l. Turczaninowia. 2020;23:5–14. doi: 10.14258/turczaninowia.23.2.1Overton W.R., McCoy J.P. Reversing the effect of formalin on the binding of propidium iodide to DNA. Cytometry. 1994;16:351–356. doi: 10.1002/cyto.990160410Sliwinska E., Zielinska E., Jedrzejczyk I. Are seeds suitable for flow cytometric estimation of plant genome size? Cytometry A. 2005;64:72–79. doi: 10.1002/cyto.a.20122Loureiro J., Kron P., Temsch E.M., et al. Isolation of plant nuclei for estimation of nuclear DNA content: overview and best practices. Cytometry A. 2021;99:318–327. doi: 10.1002/cyto.a.24237Sliwinska E. Nuclear DNA content analysis of plant seeds by flow cytometry. Curr. Protoc. Cytom. 2006;35:7.29.1–7.29.13. doi: 10.1002/0471142956.cy0729s35Matzk F., Meister A., Schubert I. An efficient screen for reproductive pathways using mature seeds of monocots and dicots. Plant J. 2000;21:97–108. doi: 10.1046/j.1365-313x.2000.00647.xОстроумова Т.А., Скапцов М.В., Гамова Н.С., и др. Таксономическое изучение двух видов рода Peucedanum (Umbelliferae) флоры Сибири и сопредельных территорий. Бюллетень Московского общества испытателей природы. Отдел биологический. 2026;131(3):40–52. doi: 10.55959/MSU0027-1403-BB-2026-131-3-40-52 (In Russ.)Дегтярева Г.В., Остроумова Т.А., Скапцов М.В. Использование плодов различного срока хранения в исследовании плоидности и содержания ДНК Peucedanum vaginatum Ledeb. (Umbelliferae Juss.) методом проточной цитометрии. Проблемы ботаники Южной Сибири и Монголии. 2025;(24-2):50–54. doi: 10.14258/pbssm.2025051 (In Russ.)Trávníček P., Ponert J., Urfus T., et al. Challenges of flow-cytometric estimation of nuclear genome size in orchids, a plant group with both whole-genome and progressively partial endoreplication. Cytometry A. 2015;87:958–966. doi: 10.1002/cyto.a.22681Brown S.C., Bourge M., Maunoury N., et al. DNA remodeling by strict partial endoreplication in orchids, an original process in the plant kingdom. Genome Biol. Evol. 2017;9:1051–1071. doi: 10.1093/gbe/evx063Schubert R., Hohlfeld H., Brugger M. Flow cytometry and cytological studies reveal consecutive rounds of strict partial genome duplications and endomitosis in cellular nuclei from vegetative organs of seeded Dactylorhiza majalis (subfamily Orchidoideae). Biologia. 2025;80:823–837. doi: 10.1007/s11756-025-01879-7Greilhuber J., Temsch E.M., Loureiro J.C.M. Nuclear DNA content measurement. In: Doležel J., Greilhuber J., Suda J., editors. Flow Cytometry with Plant Cells. Weinheim: Wiley-VCH Verlag GmbH & Co. KGaA; 2007. P. 67–101. doi: 10.1002/9783527610921.ch4Loureiro J., Rodriguez E., Doležel J., et al. Two new nuclear isolation buffers for plant DNA flow cytometry: a test with 37 species. Ann. Bot. 2007;100:875–888. doi: 10.1093/aob/mcm152Galbraith D.W., Harkins K.R., Maddox J.M., et al. Rapid flow cytometric analysis of the cell-cycle in intact plant tissues. Science. 1983;220:1049–1051. doi: 10.1126/science.220.4601.1049Doležel J., Binarová P., Lucretti S. Analysis of nuclear DNA content in plant cells by flow cytometry. Biol. Plant. 1989;31:113–120. doi: 10.1007/BF02907241Pfosser M., Amon A., Lelley T., et al. Evaluation of sensitivity of flow cytometry in detecting aneuploidy in wheat using disomic and ditelosomic wheat-rye addition lines. Cytometry. 1995;21:387–393. doi: 10.1002/cyto.990210412Otto F. DAPI staining of fixed cells for high-resolution flow cytometry of nuclear DNA. In: Radbruch A., editor. Flow Cytometry and Cell Sorting. Berlin: Springer-Verlag; 1992. P. 101–104.Darzynkiewicz Z., Traganos F., Kapuscinski J., et al. Accessibility of DNA in situ to various fluorochromes: relationship to chromatin changes during erythroid differentiation of Friend leukemia cells. Cytometry A. 1984;5:355–363. doi: 10.1002/cyto.990050411Merzlyak M.N., Chivkunova O.B., Solovchenko A.E., et al. Light absorption by anthocyanins in juvenile, stressed, and senescing leaves. J. Exp. Bot. 2008;59:3903–3911. doi: 10.1093/jxb/ern230Greilhuber J. “Self-tanning” — a new and important source of stoichiometric error in cytophotometric determination of nuclear DNA content in plants. Plant Syst. Evol. 1988;158:87–96. doi: 10.1007/BF00936335Labieniec M., Gabryelak T. Interactions of tannic acid and its derivatives (ellagic and gallic acid) with calf thymus DNA and bovine serum albumin using spectroscopic method. J. Photochem. Photobiol. B. 2006;82:72–78. doi: 10.1016/j.jphotobiol.2005.09.005Temsch E.M., Koutecký P., Urfus T., et al. Reference standards for flow cytometric estimation of absolute nuclear DNA content in plants. Cytometry A. 2022;101:710–724. doi: 10.1002/cyto.a.24495Barow M., Meister A. Lack of correlation between AT frequency and genome size in higher plants and the effect of nonrandomness of base sequences on dye binding. Cytometry. 2002;47:1–7. doi: 10.1002/cyto.10013Šmarda P., Bureš P., Horová L., et al. Genome size and GC content evolution of Festuca: ancestral expansion and subsequent reduction. Ann. Bot. 2008;101:421–433. doi: 10.1093/aob/mcm307Camunas-Soler J., Manosas M., Frutos S., et al. Single-molecule kinetics and footprinting of DNA bis-intercalation: the paradigmatic case of thiocoraline. Nucleic Acids Res. 2015;43:2767–2779. doi: 10.1093/nar/gkv087Greilhuber J., Temsch E.M., Loureiro J.C.M. Nuclear DNA content measurement. In: Doležel J., Greilhuber J., Suda J., editors. Flow Cytometry with Plant Cells. Weinheim: Wiley-VCH Verlag GmbH & Co. KGaA; 2007. P. 67–101. doi: 10.1002/9783527610921.ch4Veselý P., Bureš P., Šmarda P., et al. Genome size and DNA base composition of geophytes: the mirror of phenology and ecology? Ann. Bot. 2012;109:65–75. doi: 10.1093/aob/mcr267Marie D., Brown S.C. A cytometric exercise in plant DNA histograms, with 2C-values for 70 species. Biol. Cell. 1993;78:41–51. doi: 10.1016/0248-4900(93)90113-SBennett M., Johnston J., Hodnett G., et al. Allium cepa L. cultivars from four continents compared by flow cytometry show nuclear DNA constancy. Ann. Bot. 2000;85:351–357. doi: 10.1006/anbo.1999.1071Greilhuber J., Ebert I. Genome size variation in Pisum sativum. Genome. 1994;37:646–655. doi: 10.1139/g94-092Doležel J., Doleželová M., Novák F. Flow cytometric estimation of nuclear DNA amount in diploid bananas (Musa acuminata and M. balbisiana). Biol. Plant. 1994;36:351–357. doi: 10.1007/BF02920930Šmarda P., Bureš P., Horová L., et al. Ecological and evolutionary significance of genomic GC content diversity in monocots. Proc. Natl. Acad. Sci. U.S.A. 2014;111:4096–4102. doi: 10.1073/pnas.1321152111Veleba A., Šmarda P., Zedek F., et al. Evolution of genome size and genomic GC content in carnivorous holokinetics (Droseraceae). Ann. Bot. 2017;119:409–416. doi: 10.1093/aob/mcw229Tiersch T.R., Chandler R.W., Wachtel S.S., et al. Reference standards for flow cytometry and application in comparative studies of nuclear DNA content. Cytometry A. 1989;10:706–710. doi: 10.1002/cyto.990100606Nurk S., Koren S., Rhie A., et al. The complete sequence of a human genome. Science. 2022;376:44–53. doi: 10.1126/science.abj6987Yokoya K., Roberts A.V., Mottley J., et al. Nuclear DNA amounts in roses. Ann. Bot. 2000;85:557–561. doi: 10.1006/anbo.1999.1102Sokoloff D.D., Degtjareva G.V., Skaptsov M.V., et al. Diploids and tetraploids of Acorus (Acoraceae) in temperate Asia are pseudocryptic species with clear differences in micromorphology, DNA sequences and distribution patterns, but shared pollination biology. Taxon. 2024;73:718–761. doi: 10.1002/tax.13173Wang B., Yang X., Jia Y., et al. High-quality Arabidopsis thaliana genome assembly with Nanopore and HiFi long reads. Genomics Proteomics Bioinformatics. 2022;20:4–13. doi: 10.1016/j.gpb.2021.08.003Reiser L., Bakker E., Subramaniam S., et al. The arabidopsis information resource in 2024. Genetics. 2024;227(1):1–11. doi: 10.1093/genetics/iyae027Soni A., Henry R.J. Re-calibration of flow cytometry standards for plant genome size estimation. Front. Plant Sci. 2025;16:1548766. doi: 10.3389/fpls.2025.1548766Shang L., He W., Wang T., et al. A complete assembly of the rice Nipponbare reference genome. Mol. Plant. 2023;16(8):1232–1236. doi: 10.1016/j.molp.2023.08.003Abdullah M., Furtado A., Masouleh A.K., et al. An improved haplotype resolved genome reveals more rice genes. Tropical Plants. 2024;3(1):e007. doi: 10.48130/tp-0024-0007Tomato Genome Consortium. The tomato genome sequence provides insights into fleshy fruit evolution. Nature. 2012;485:635–641. doi: 10.1038/nature11119Su X., Wang B., Geng X., et al. A high-continuity and annotated tomato reference genome. BMC Genomics. 2021;22:898. doi: 10.1186/s12864-021-08212-xShen Y., Liu J., Geng H., et al. De novo assembly of a Chinese soybean genome. Sci. China Life Sci. 2018;61:871–884. doi: 10.1007/s11427-018-9360-0Yi X., Liu J., Chen S., et al. Genome assembly of the JD17 soybean provides a new reference genome for comparative genomics. G3 (Bethesda). 2022;12:jkac017. doi: 10.1093/g3journal/jkac017Hu Y., Resende Jr. M.F.R. Maize genome assembly with PacBio reads. *Bio-101*. 2022;e4456. doi: 10.21769/BioProtoc.4456Chen J., Wang Z., Tan K., et al. A complete telomere-to-telomere assembly of the maize genome. Nat. Genet. 2023;55:1221–1231. doi: 10.1038/s41588-023-01419-6Shirasawa K., Hirakawa H., Fukino N., et al. Genome sequence and analysis of a Japanese radish (Raphanus sativus) cultivar named ‘Sakurajima Daikon’ possessing giant root. DNA Res. 2020;27:dsaa010. doi: 10.1093/dnares/dsaa010Xu L., Wang Y., Dong J., et al. A chromosome-level genome assembly of radish (Raphanus sativus L.) reveals insights into genome adaptation and differential bolting regulation. Plant Biotechnol. J. 2023;21:990–1004. doi: 10.1111/pbi.14011Kreplak J., Madoui M.A., Cápal P., et al. A reference genome for pea provides insight into legume genome evolution. Nat. Genet. 2019;51:1411–1422. doi: 10.1038/s41588-019-0480-1Jayakodi M., Golicz A.A., Kreplak J., et al. The giant diploid faba genome unlocks variation in a global protein crop. Nature. 2023;615:652–659. doi: 10.1038/s41586-023-05791-5Finkers R., van Kaauwen M., Ament K., et al. Insights from the first genome assembly of onion (Allium cepa). G3 (Bethesda). 2021;11:jkab243. doi: 10.1093/g3journal/jkab243