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Efficiency of the use for doubled haploids in onion breeding (<i>Allium cepa</i> L.)
2022
E. A. Cherednichenko | V. F. Pivovarov | S. F. Gavrish | A. F. Pershin | M. V. Budylin
Relevance. In onion breeding, quickly obtain aligned material is an urgent target for create parent forms of heterosis hybrids. Using classical methods with helping of backcrosses, this is achieved in 10-12 years. Using the technology of doubled haploids, it is possible to reduce these terms several times, and also to avoid the manifestation of inbred depression when obtaining lines by self-pollination. At the same time, the most effective in the production of haploids is the use of whole flower buds as an explant, unlike ovules and ovaries, the production of which is more time-consuming and labor-intensive.Methods. The doubled onion haploids were obtained by the method of ovule culture on the basis of the biotechnology laboratory of the Gavrish Breeding Center using the technology that based on the methodological recommendations of Monakhos S.G. et al., 2014. DH–onion plants with a developed root system and leaf apparatus were planted in the open ground and grown according to the technology generally accepted for the zone at the sites of the Gavrish breeding center, Krymsk, Krasnodar Territory. The obtained commercial bulbs were evaluated according to the RTG/46/2 method.Results. It was obtain doubled haploids, which were tested for ploidness by flow cytometry. 40 digaploid onion plants have been created. As a result of further development after transplantation into the open ground, storage, springization and selection, 3 uterine bulbs were obtained for further reproduction and inclusion in the breeding process.
Show more [+] Less [-]Production of doubled haplois in Brassica purpuraria
2019
E. V. Kozar | K. S. Korottseva | O. V. Romanova | O. A. Chichvarina | L. Yu. Kan | V. A. Ahramenko | E. A. Domblides
Relevance. In recent years vegetable crop Brassica rapa ssp. chinensis var. purpuraria (synonyms: Brassica campestris L. var.purpurea Bailey; Brassica rapa L. ssp. chinensis var. purpurea) is gaining popularity as an object of genetic and molecular researches, and as an economically valuable vegetable plant due to the high content of biologically active compounds and distinctive economically valuable traits. Effective technology for development DH-plants to accelerate the breeding process for this culture has not been developed yet, so research in this area is relevant.Materials and methods. The study included two varieties from the collection of Vavilov AllRussian Research Institute of Plant Industry (VIR): No. 1301 (China) and No. 1357(Netherlands). Both protocols standard unmodified and with addition of silver nitrate (AgNO3) in the medium for embryogenesis induction were used in experiments for production of DH plants from isolated microspore in vitro. Direct chromosome counting in meristem cells and flow cytometry were used to determine the ploidy of regenerating plants.Results. As a result of the study embryogenesis in B. purpuraria culture can develop with the use of a standard protocol as well as with the addition of silver nitrate that showed a positive effect on the induction of embryogenesis. The yield of embryoids varied depending on the genotype of the individual plant within the variety accession. The highest yield of embryoids was 40 embryoids/petri dish. The main problem at the stage of regeneration is that about half of the regenerating plants occurred to be albinos and were not viable. We show a high degree of spontaneous chromosome doubling in regenerated plants (all analyzed plants were doubled haploids). In total 38 regenerated plants were obtained from accession No. 1301. It was shown that four DH-plants had self-incompatibility after self-pollination, but seed progeny from other plants was obtained. The created material was taken for genetics study and breeding work.
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