Registration Number
0225U004657,
(0123U100680)
, R & D reports
Title
Development of methods of in vitro culture introduction of seed germs of buckwheat in order to obtain haploid lines
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Розроблення методів введення в культуру in vitro насіннєвих зачатків гречки з метою отримання гаплоїдних ліній
Head
Roik Mykola V., Доктор сільськогосподарських наук
Registration Date
16-12-2025
Organization
Institute of Bioenergy Crops and Sugar Beet of National Academy of Agrarian Sciences of Ukraine
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Develop and select conditions for introduction of embryos into sterile culture in vitro and induction of haploids. Establish the optimal conditions for the cultivation of explants in vitro and develop the parameters of light and temperature regimes and nutrient media for the regeneration of clones.
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During the research, it was established that varietal characteristics significantly influenced the formation of macrostructures: the highest yield was recorded in the varieties Krupnozelena and Ukrainka, while the lowest was observed in Yulianna. The plant material was collected from field donor plants of the varieties Ukrainka, Syn 3/02, Yulianna, and Krupnozelena. A 24-hour cold pretreatment was found to be optimal before introduction into culture. Sterilization with the “Bilyzna” solution ensured 84–96% viable and sterile structures, whereas Sublimate (HgCl₂) provided only 10–23%. Callus induction occurred on a medium supplemented with 2,4-D; the optimal concentration of 0.2 mg/L resulted in 49–66% callus formation, while a lower concentration of 0.1 mg/L produced only 11–22%. A dark period of 12–15 days, followed by cultivation at 24±2 °C, illumination of 3500–4500 lx, and a 16-hour photoperiod, enabled the formation of callus globules sized 5.0–6.8 mm. The highest level of rhizogenesis (65–91%) was obtained on a modified Murashige–Skoog medium with the addition of NAA and IAA at 0.3 mg/L. The root system length of haploids ranged from 3.5 to 5.7 cm. Plant survival in varieties compared to lines varied between 84–92%, while in haploid lines it reached 70–87%; the adaptation coefficient increased with the development of a taproot system (r = 0.82). The substrate had a substantial effect on the adaptive capacity of regenerants; the best results were achieved with mixtures of peat, sod soil, and vermiculite (survival 91–93%). The optimal medium pH was 6.7–6.9, combined with high air permeability. Heteroauxin at 1.0 mg/L and stimulants (heteroauxin 1.0–1.5 mL/L, Epin 0.1 mL/L) ensured 100% survival of plants within 7–10 days. Succinic acid (1.0 mmol/L) increased seedling survival after transfer from in vitro conditions to 94–97%.
Product Description
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Mykola V. Roik
Olesia А. Zinchenko
Victoria I. Voitovska
Olha A. Potapovych
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2025-12-16
