Genetic variation and field evaluation of buckwheat plants (Fagopyrum esculentum Moench) obtained in vitro with the use of mutagenic media
Abstract
Background. Obtaining starting material for buckwheat breeding is an important task of the breeding process. Heavy metals used in vitro as mutagenic factors induce genetic variation in buckwheat germplasm sources and expand the pool of promising genotypes with important agronomic traits.
Materials and methods. Regenerated buckwheat plants were grown in vitro on MS mutagenic media with zinc ions (184–299 mg/L) and under mineral starvation in the Laboratory of Breeding and Genetic Research on Field Crops at the Federal Scientific Center of Agricultural Biotechnology of the Far East named after A.K. Chaika. Genetic variation of the accessions was studied with four ISSR-markers (M1, M2, M7, and M11). Cv. ʽIzumrudʼ served as the control. Field evaluation of buckwheat plants was performed to assess their main agronomic characters.
Results. Regenerated buckwheat accessions R 1069, R 1070 and R 1071 possessed valuable agronomic traits and were genetically different from the original form. Under extreme meteorological conditions, associated with waterlogging of the soil, they showed a statistically significant and valid increase in productivity per plant (1.8–2.2 times) and higher values of morphological indicators compared to the control. Accession R 1069 combined high protein content (11.98%) with high productivity.
Conclusion. Mutagenic media with zinc ions and mineral starvation in vitro induced genetic variation in the source cultivar of buckwheat, leading to the emergence of genotypes with improved morphological and agronomic traits. Genetic differences from the original form were confirmed with ISSR markers M1, M2, M7, and M11. The regenerated buckwheat accessions (R 1069, R 1070, and R 1071) may be recommended as promising starting material for breeding.
About the Authors
S. A. BorovayaRussian Federation
Svetlana A. Borovaya, Researcher
30 Volozhenina St., Timiryazevsky Settlem., Ussuriysk 692539, Russia
A. G. Klykov
Russian Federation
Alexey G. Klykov, Dr. Sci. (Biology), Full Member (Academician) of the RAS, Head of a Department
30 Volozhenina St., Timiryazevsky Settlem., Ussuriysk 692539, Russia
References
1. Aljanabi S.M., Martinez I. Universal and rapid salt-extraction of high quality genomic DNA for PCR-based techniques. Nucleic Acids Research. 1997;25(22):4692-4693. DOI: 10.1093/nar/25.22.4692
2. Barsukova E.N., Klykov A.G., Chikina E.L. Usage of the tissue culture method for the development of new forms of Fagopyrum esculentum Moench. Russian Agricultural Science. 2019;(5):3-6. [in Russian]. DOI: 10.31857/S2500-2627201953-6
3. Borovaya S.А., Klykov A.G., Barsukova E.N. Influence of the toxic effect of zinc and mineral starvation on the growth and development of buckwheat plantlets in vitro. Proceedings on Applied Botany, Genetics and Breeding. 2023;184(2):9-18. [in Russian]. DOI: 10.30901/2227-8834-2023-2-9-18
4. Butovets E.S., Lukyanchuk L.M., Kodirova G.A., Kubankova G.V., Efremova O.S. Studying regenerated soybean lines for their useful agronomic and biochemical characteristics. Proceedings on Applied Botany, Genetics and Breeding. 2024;185(2):38-49. [in Russian]. DOI: 10.30901/2227-8834-2024-2-38-49
5. Dospekhov B.A. Methodology of field trial (with fundamentals of statistical processing of research results) (Metodika polevogo opyta [s osnovami statisticheskoy obrabotki rezultatov issledovaniy]). 4th ed. Moscow: Kolos; 1979. [in Russian]
6. Erturk F.A., Agar G., Arslan Е., Nardemir G. Analysis of genetic and epigenetic effects of maize seeds in response to heavy metal (Zn) stress. Environmental Science and Pollution Research International. 2015;22(13):10291-10297. DOI: 10.1007/s11356-014-3886-4
7. GOST 10843-76. Interstate standard. Grain. Method for determination of filmness. Moscow: Standartinform; 2009. [in Russian]. URL: http://gost.gtsever.ru/Data2/1/4294840/4294840035.pdf [дата обращения: 15.05.2025].
8. GOST 10846-91. Interstate standard. Grain and products of its processing. Method for determination of protein. Moscow: Standartinform; 2009. [in Russian]. URL: https://normativ.kontur.ru/document?moduleId=9&documentId=468933 [дата обращения: 15.05.2025].
9. GOST 12042-80. Interstate standard. Seeds of farm crops. Methods of determination of 1000 seed weight. Moscow: Standartinform; 2011. [in Russian]. URL: https://fsvps.gov.ru/files/gost-12042-80-mezhgosudarstvennyj-standart-s [дата обращения: 15.05.2025].
10. GOST 29033-91. Interstate standard. Grain and derived products. Determination of fat content. Moscow: Publishers of the Standards; 2004. [in Russian]. URL: https://internet-law.ru/gosts/gost/10417 [дата обращения: 15.05.2025].
11. Harbison S.T., Chang S., Kamdar K.P., Масkay T.F.C. Quantitative genomics of starvation stress resistance in Drosophila. Genome Biology. 2005;6(4):R36. DOI: 10.1186/gb-2005-6-4-r36
12. Kadyrova G.D., Kadyrova F.Z., Martirosyan E.V., Ryzhova N.N. Analysis of genomic diversity of samples and cultivars in common buckwheat (Fagopyrum esculentum Moench) by the ISSR-method. Agricultural Biology. 2010;45(5):42-48. [in Russian]
13. Khalafyan A.A. STATISTICA 6. Statistical data analysis (STATISTICA 6. Statisticheskiy analiz dannykh). Moscow: Binom-Press; 2008. [in Russian]
14. Klykov A.G., Barsukova E.N. Biotechnology and buckwheat breeding in the Russian Far East. Vladivostok: PSP95; 2021. [in Russian]
15. Kolyasnikova N.L. Problems of genetic safety (Problemy geneticheskoy bezopasnosti). Perm: Prokrost; 2019. [in Russian]
16. Rakhmikhudoev G. Buckwheat crop in the Pamir Mountains (Kultura grechikhi na Pamire) [dissertation]. Dushanbe; 2000. [in Russian]
17. Sabreena, Nazir М., Mahajan R., Hashim M.J., Iqbal J., Alyemeni M.N. et al. Deciphering allelic variability and population structure in buckwheat: An analogy between the efficiency of ISSR and SSR markers. Saudi Journal of Biological Sciences. 2021;28(11):6050-6056. DOI: 10.1016/j.sjbs.2021.07.061
18. Shmaraev G.E. (ed.). Guidelines for studying collection accessions of maize, sorghum and groat crops (millet, buckwheat, and rice) (Metodicheskiye ukazaniya po izucheniyu kollektsionnykh obraztsov kukuruzy, sorgo i krupyanykh kultur [proso, grechikha, ris]). Leningrad: VIR; 1968. [in Russian]
19. Singh S., Parihar P., Singh R., Singh V.P., Prasad S.M. Heavy metal tolerance in plants: Role of transcriptomics, proteomics, metabolomics, and ionomics. Frontiers in Plant Science. 2016;6:1143. DOI: 10.3389/fpls.2015.01143
20. Suvorova G. Buckwheat tissue cultures and genetic transformation. In: M. Zhou, I. Kreft, S.H. Woo, N. Chrungoo, G. Wieslander (eds). Molecular Breeding and Nutritional Aspects of Buckwheat. London: Academic Press; 2016. р.365-375. DOI: 10.1016/B978-0-12-803692-1.00029-8
21. Ul’yanenko L.N., Reva E.V., Synzynys B.I. Cytogenetic effects in Allium cepa L. resulted from separate and combined exposure to Cu, Zn and Ni. Agricultural Biology. 2017;52(1):183-190. [in Russian]. DOI: 10.15389/agrobiology.2017.1.183rus
22. Üstündağ Ü., Çavuşoğlu K., Yalçın E. Comparative analysis of cyto-genotoxicity of zinc using the comet assay and chromosomal abnormality test. Environmental Science and Pollution Research International. 2024;31(44):56140-56152. DOI: 10.1007/s11356-024-34940-0
23. Viana V.E., Pegoraro C., Busanello C., Costa de Oliveira A. Mutagenesis in rice: The basis for breeding a new super plant. Frontiers in Plant Science. 2019;10:1326. DOI: 10.3389/fpls.2019.01326
24. Zhang J., Jiang F., Shen Y., Zhan Q., Bai B., Chen W. et al. Transcriptome analysis reveals candidate genes related to phosphorus starvation tolerance in sorghum. BMC Plant Biology. 2019;19(1):306. DOI: 10.1186/s12870-019-1914-8
25. Zinchenko M., Dubrovnaya O., Bavol A. In vitro selection of wheat for complex resistance and analysis of obtained form. Izvestia of Samara Scientific Center of the Russian Academy of Sciences. 2013;15(3-5):1610-1614. [in Russian]
26. Zlobin I.E. Early stressor responses of rapeseed plants to an increase in the levels of copper and zinc (Ranniye stressornye otvety rasteniy rapsa na povyshennye urovni medi i tsinka v srede) [dissertation]. Moscow: Timiryazev Institute of Plant Physiology; 2015. [in Russian]
Review
For citations:
Borovaya S.A., Klykov A.G. Genetic variation and field evaluation of buckwheat plants (Fagopyrum esculentum Moench) obtained in vitro with the use of mutagenic media. Proceedings on applied botany, genetics and breeding.
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