Towards isolated microspores culture of red beet

UDC 635.112:576

Grigolava T.R., Vishnyakova A.V., Monakhos S.G.

Doubled haploids (DH) production in vitro is an accelerated way to create pure lines for breeding commercial F1 hybrids. The most common method for the production of doubled haploids in plants of the genus Beta, mainly sugar beet, is the culture of isolated ovules (gynogenesis). The disadvantages of the gynogenic method for the DH production are high labor intensity due to manual isolation and inoculation of ovules on a nutrient medium and a high probability of development of somatic clones from the tissues surrounding the embryo sac. To eliminate the described disadvantages, the culture of isolated microspores is a significant alternative. However, protocols for the routine DH production of red beet in culture of isolated microspores have not yet been developed. The aim of this study was to study the factors affecting callusogenesis in the culture of isolated red beet microspores and to identify the optimal conditions for their cultivation in vitro. Microspores were isolated from buds 1,2–2,7 mm with a step of 0,3 mm to study the relationship between the bud size and the stage of microspores development with the frequency of callusogenesis and 1,2–1,5 mm to study of the nutrient medium effect, cultivation temperature and the effect of exposure to heat treatment at 32,5 °C on callusogenesis of isolated red beet microspores. Microspores were cultivated on NLN nutrient medium supplemented with 2,4-D 0,1 mg/l, NAA 0,1 mg/l, 130 g/l sucrose and on NLN medium without the growth regulators addition containing 130 g/l sucrose. As a result of the study, the sizes of flower buds were found to be 1,2–1,5 mm, containing the most responsive microspores of the mononuclear stage of development to IMT, and the nutrient medium NLN with the addition of 130 g/l of sucrose, 0,1 mg/l of 2,4-D and 0,1 mg/l NAA, providing the highest yield of microsporogenic callus. It has been shown that under the applied conditions, thermal treatment of isolated microspores at different exposures at 32,5 °C does not induce microspore embryogenesis.

Key words: Beta vulgaris, DH-technologies, androgenesis, isolated microspore, callogenesis, callus, doubled haploids, red beet

Grigolava T.R., postgraduate student of the Department of Botany, Breeding and Seed Production of Garden Plants of the Federal State Budgetary Educational Institution of Higher Education «Russian State Agrarian University – Moscow Agricultural Academy after K.A. Timiryazev» (RSAU –MTAA after K.A. Timiryazev). E-mail: ORCHID 0000-0002-1594-8430

Vishnyakova A.V., Cand. Sci. (Agr.), associate professor of the Department of Botany, Breeding and Seed Production of Garden Plants of the RSAU – MTAA after K.A. Timiryazev. E-mail: ORCHID 0000-0002-9160-1164

Monakhos S.G., D.Sci. (Agr.), Head of the Department of Botany, Breeding and Seed Production of Garden Plants of the RSAU – MTAA after K.A. Timiryazev. E-mail: ORCHID 0000-0001-9404-8862

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For citing: Grigolava T.R., Vishnyakova A.V., Monakhos S.G. Towards isolated microspores culture of red beet. Potato and vegetables. 2022. No5. Pp. 37-40. (In Russ.).

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