Accelerated development of rice populations using anther culture in vitro method

Keywords: Oryza sativa L. rice, in vitro pollen culture, doubled haploid (DH) lines, economic and biological traits, productivity elements

Abstract

Background.  Inclusion of doubled haploids in the breeding process allows increasing the speed and reliability of selection of desired forms with smaller population volumes. To stabilize the genotype, anthers of hybrid combinations of F2, F3, F4 and F5 generations, developed in crossings of white-grained samples with traits of high nutritional value of grain, and anthers of combinations of F4 generation from crossings of varieties contrasting in pericarp color and amylose content, were introduced into the culture. The responsiveness to gamete technologies was studied in 21 hybrid combinations. New genetically stable material (DH - doubled haploids) was developed. Phenotyping was carried out in conditions of a vegetation experiment for economic and biological traits and elements of plant productivity in four populations, which included 45 DH lines. Variability was noted in a number of traits within the DH line populations. Based on the results of the biometric analysis of the regenerated populations, 6 sources with a "1000 grain mass" of more than 30 grams were identified.

The study was carried out with the financial support of the Kuban Science Foundation and the Russian Science Foundation within the framework of the scientific project No. 25-16-20103 "Application of the genomic approach in rice breeding for high technological grain quality"

Purpose. To study the response of hybrids obtained from crossing contrasting rice samples to in vitro pollen culture, to accelerate the creation of DH line populations based on the studied genotypes, and to phenotype them.

Materials and methods. The research was conducted at the Laboratory of Biotechnology and Molecular Biology at the Federal Research Center for Rice, using the culture of isolated anthers in vitro according to the generally accepted method of R.G. Butenko (1990).

Results. The genetic determinism of the "regeneration" trait in the donor plants used in the crossbreeding was noted. The genotypes of the crossbreeding involving the varieties Favorit and Azovsky proved to be the most productive in terms of the output of highly morphogenic calluses and androgenic lines. Phenotypic analysis revealed significant diversity among the plants in the individual DH populations in terms of the shape of the panicle, the angle of the flag leaf deviation, the length of the growing season, the weight of 1,000 grains, the height of the plants, and the length of the growing season.

Conclusion. Phenotypic analysis of the DH lines' traits showed that their genesis originates from microspores, thus these lines are a valuable genetic resource. The inclusion of doubled haploids in the breeding process will help to facilitate the assessment of recombinant genotypes arising from the cross, will allow to detect rare recessive alleles, will increase the speed and reliability of the selection of desired forms with smaller population volumes.

EDN: NOSNRM

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Author Biographies

Elena G. Savenko, Federal Scientific Rice Centre

Candidate of Biological Sciences, Head of the Laboratory of Biotechnology and Molecular Biology, Leading Researcher

Valentina А. Glazyrina, Federal Scientific Rice Centre

Senior Researcher of the Laboratory of Biotechnology and Molecular Biology

Ludmila A. Shundrina, Federal Scientific Rice Centre

Scientist of Laboratory of the Laboratory of Biotechnology and Molecular

Zhanna M. Mukhina, Federal Scientific Rice Centre

Doctor of Biological Sciences, Dr Chief Scientist of the Laboratory of Biotechnology and Molecular Biology

Lyubov V. Esaulova, Federal Scientific Rice Centre

Candidate of Biological Sciences, Deputy Director of Science

References

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Список литературы

Dwivedi, S. L. (2015). Haploids: Constraints and opportunities in plant breeding. Biotechnology Advances, 33, 812–829. https://doi.org/10.1016/j.biotechadv.2015.07.001

Wedzony, M., Foster, B. P., Zur, I., Golemiec, E., Szechynska Hebda, M., Dubas, E., & Gotebiowska, G. (2009). Chapter 1. Progress in doubled haploid technology in higher plants. In: Advances in haploid production in higher plants (pp. 1–35). Springer Science + Business Media B.V.

Weyen, J., Touraev, A., Foster, B. P., & Jain, E. M. (2009). Chapter 15. Barley and wheat doubled haploids in breeding. In: Advances in haploid production in higher plants (pp. 179–189). Springer Science + Business Media B.V.

Уразалиев, К. Р., Орсини, Х. М., Абекова, А. М., Базылова, Т. А., & Даниярова, А. К. (2013). Ускорение селекции пшеницы с использованием дигаплоидов, полученных методом культуры микроспор. Вестник КазНУ. Серия экологическая, 2/2(38), 369–374.

Mishra, R., Gundimeda, J., & Narashima, R. (2016). In vitro androgenesis in rice: Advantages, constraints and future prospects. Rice Science, 23, 57–68. https://doi.org/10.1016/j.rsci.2016.02.001

Bernardo, R. (2009). Should maize double haploids be induced among F1 or F2 plants? Theoretical and Applied Genetics, 119, 255–262. https://doi.org/10.1007/s00122-009-1034-1. EDN: https://elibrary.ru/EKWFIE

Zheng, M. Y. (2003). Microspore culture in wheat (Triticum aestivum): Doubled haploid production via induced embryogenesis. Plant Cell, Tissue and Organ Culture, 73, 213–230. EDN: https://elibrary.ru/EQLTAR

Maluszynski, M., Kasha, K. J., & Szarejko, I. (2003). Published doubled haploid protocols in plant species. In: Doubled haploid production in crop plants (pp. 309–335). Springer: Dordrecht, the Netherlands. https://doi.org/10.1007/978-94-017-1293-4_46. ISBN: 978 94 017 1293 4. EDN: https://elibrary.ru/XWCXXB

Li, W., Gang, L., Deming, Z., Feng, W., & Jiabin, C. (2011). Tissue culture system for different hybrid of indica rice. Northeast Agricultural University Journal, 18, 13–17.

Wren, J., Wu, P., Trumpe, B., Tian, H., Lubberstedt, T., & Chen, S. (2017). New technologies in the extraction of doubled haploid lines. Plant Biotechnology, 15, 1361–1370.

Forster, B. P., Heberle Bors, E., Kasha, K. J., & Turaev, A. (2007). Revival of haploids in higher plants. Trends in Plant Science, 12(8), 368–375. https://doi.org/10.1016/j.tplants.2007.06.007. EDN: https://elibrary.ru/MMXAAH

Bishnoy, U., Jain, R. K., Rohilla, J. S., Chowdhury, W. K., Gupta, K. R., & Chowdhury, J. B. (2000). Another culture of recalcitrant indica rice hybrids × Basmati. Euphytica, 114, 93–101. EDN: https://elibrary.ru/AGJSPB

Dwivedi, S. L., Britt, A. B., Tripathi, L., Sharma, S., Upadhyaya, H. D., & Ortiz, R. (2015). Haploids: Limitations and opportunities in plant breeding. Biotechnology Advances, 33, 812–829.

Belicuas, P. R., Guimarães, C. T., Paiva, L. V., Duarte, J. M., Maluf, W. R., & Paiva, E. (2007). Androgenetic haploids and SSR markers as tools for the development of tropical maize hybrids. Euphytica, 156, 95–102. https://doi.org/10.1007/s10681-007-9356-z. EDN: https://elibrary.ru/XPVIZH

Germana, M. A. (2011). Anther culture for haploid and doubled haploid production. Plant Cell, Tissue and Organ Culture, 104, 283–300. https://doi.org/10.1007/s11240-010-9852-z. EDN: https://elibrary.ru/VXVTHW

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Blaydes, D. F. (1966). Interaction of kinetin and various inhibitors in the growth of soybean tissue. Physiologia Plantarum, 19, 748–753.

Murashige, T. A., & Skoog, F. (1962). Revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiologia Plantarum, 15, 473–497.


Published
2025-12-30
How to Cite
Savenko, E., Glazyrina, V., Shundrina, L., Mukhina, Z., & Esaulova, L. (2025). Accelerated development of rice populations using anther culture in vitro method. Siberian Journal of Life Sciences and Agriculture, 17(6-2), 149-164. https://doi.org/10.12731/2658-6649-2025-17-6-2-1542
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Articles