Cytological methods of analysis of haploid plants-regenerants of cabbage (<i>Brassica oleracea</i> L.) obtained <i>in vitro</i>
2019
Rima N. Kirakosyan | Elena A. Kalashnikovа
Relevance Currently, in genetic studies and selection of cabbage cultures, biotechnological methods for creating clean lines — doubled haploids in the culture of anthers and in the culture of isolated microspores are widely used. A common feature of these technologies is that the plants obtained in vitro have different levels of ploidy and along with doubled haploids there are haploid, tetraploid and mixoploid forms. Therefore, the use of new cytological methods of analysis of haploid plants remains an urgent problem. Material and method The aim of this work is to establish the genetic nature of regenerated plants of Brassica oleracea L., obtained from reproductive organs in vitro. Isolated anthers and ovaries of white cabbage were cultivated on solid nutrient media containing mineral salts according to the recipe Murashige and Skoog (MS). The obtained regenerated plants were used to calculate the number of chromosomes in the root meristem, as well as the number of chloroplasts in the cells of the closing stomata of leaves using the new universal method of preparing preparations of plant chromosomes – “SteamDrop”. Results As a result of the research, the dependence of the level of ploidy on the cultivation conditions was studied. It has been shown that plants-regenerants of white cabbage, obtained in vitro from reproductive organs, had a different set of chromosomes (n, 3n, 5n). It was established that the number of chloroplasts in the stomatal cells of regenerated plants was from 9 to 45, while the original donor plants had 18–20.
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