Influence of Initial Media and Duration of Cold Pretreatment on the Efficiency of Androgenesis in the Culture of Isolated Anthers of Winter Wheat (Triticum aestivum L.) Variety Irkutskaya

Abstract

Haploid technologies are based on the regeneration of gamete cells to form whole plants. The main advantage of haploid technologies is the ability to quickly in one generation obtain homozygous plants with the required parameters. Nevertheless, the use of these technologies today is limited, what is associated with insufficient knowledge of the physiological and biochemical features of the application of this method. In this regard, this work is devoted to the study of the features of androgenesis in vitro in the culture of isolated anthers of winter wheat cultivar Irkutskaya and the determination of the methodological features of obtaining intact plants. In the autumn-winter period, donor plants were grown at the artificial climate station SIPPB SB RAS. In the spring-summer period, donor plants were grown in the field at the experimental plots of the Zalarinsky agroecological station (Tungui village). The plant material was selected at the booting stage. Anthers were isolated from 6-10 spikelets of the middle third of the spike. The features of androgenesis were assessed by the frequency of formation of calli and embryo-like structures to the total number of anthers and the frequency of formation of the total number of seedlings and the number of green seedlings to the number of embryo-like structures and calli. It was found that the optimal medium for the induction of androgenesis in the anther culture of this common wheat cultivar is the modification of medium N6 containing 9% sucrose, with the addition of glycine (2 mg/L), 2.4-D (1 mg/L) and NAA (1 mg/L). The maximum yield of embryoid-like structures was observed in the case of low-temperature pretreatment of donor plants at + 4 °C for 6-12 days. At the same time, the most intensive calli formation in the culture of isolated anthers occurred after 3-6 days of donor plant cold pretreatment. Regeneration of green plants took place on 190-2Cu medium with the addition of growth regulators (2.4-D, 0.5 mg/L; kinetin, 0.5 mg/L) with obligatory cold pretreatment of androclinic structures at +4 °C for 3-7 days. Obtained results indicate that low-temperature exposure at different stages, from pretreatment of donor ears to exposure to androclinic structures, is a key factor for winter wheat that promotes androgenesis in the culture of isolated anthers and the successful formation of intact plants.