Abstract

Relevance: One of the most important ways of classical breeding is creation of highly productive hybrids and varieties of agricultural plants. To speed up considerately the solution of this problem it is necessary to use in vitro technologies, in particular, production of haploid and diploid plants. Such technologies are of particular interest for growing important agricultural crops, for instance, broccoli. However, the frequency of plant regeneration from reproductive organs remains low. Therefore, nowadays it is necessary to search for high-performance and replicable technologies to produce DH plants.

Materials and methods: The objective of the work is to develop a procedure for producing plantsregenerants from the reproductive organs of broccoli (Brassica oleracea L. convar. Botrytis (L.) Alef. var cymosa Duch.) using the method of vacuum infiltration. The objects of the study were isolated anthers and ovaries of broccoli. The sort that was used is called “Dan` Nikolayu Vavilovu”. Vacuum infiltration of broccoli buds was carried out with a 15% solution of Aminoven at the concentration of 4ml/l and with a solution of Dropp at the concentration of 0.1mg/l. After the infiltration anthers and ovaries were isolated from the buds, which had been cultivated on MC nutrient medium containing the Dropp specimen at the concentration of 0.01mg/l and NAA 0.1mg/l.

Result: The use of vacuum infiltration of broccoli buds with Aminoven for 10 minutes along with pressure of 0.8 atm. was found to have a significant impact on embryogenesis process. Besides, this process took place directly on isolated anthers. In this case the frequency of embryoid formation averaged at about 3.7%. In the case of isolated ovaries usage, the frequency of embryogenesis was 2%. The proposed technology makes it possible to increase the frequency of the formation of haploid plants from isolated anthers and ovaries by 25-30% in comparison with the existing technologies of other authors

Keywords: Embryogenesis; Antheres; Ovaires; Broccoli sprout; Vacuum infiltration; In vitro