Evaluation of genetic stability of sugar beet (Beta vulgaris L.) plants obtained from unfertilized ovules using RAPD markers

Abstract

In order to improve the methods of embryo regeneration applied so far in in vitro cultures, a two-phased, innovative method was used. It involved using a liquid and solid media. Regeneration of ovules with the application of the gynogenesis technique took place mainly through the indirect organogenesis process during which callus got formed which was associated with the possibility of somaclonal variation occurrence. In order to identify the genetic variability taking plase during the culture, RAPD analyzes were performed. The DNA was isolated from the leaves of 10 regenerants, rooted and planted into the field. 5 of them were characterized by a haploid number of chromosomes whereas the others were of diploid nature. As a control, DNA originating from the leaves of a haploid plant growing in in vitro cultures on medium without growth regulators was used. Plants regenerated on media of various quantitative and qualitative composition in terms of phytohormonal characteristic. The results of the experiment showed that 9 out of 19 tested primers generate polymorphic bands while the remaining 10 amplify the same DNA fragments in all tested samples. The polymorphism effect was revealed in 4 plants with diploid genome and in 1 haploid plant. In total, 13 polymorphic bands were identified and 11 of them were detected in diploid DNA.

Keywords

• gynogenesis
• PCR
• RAPD
• somaclonal variation
• sugar beet