Influence of genotype and culture conditions on in vitro gynogenesis in red beet (Beta vulgaris subsp. vulgaris)

The process was examined or the effect of culture conditions on in vitro gynogenesis in red beet was analyzed, conditions were modified or optimized. A significant influence of the genotype on the gynogenesis process was demonstrated. Of the eight genotypes, 58.3% planted ovules regenerated embryo-like structures in breeding line 411, 2.1% in RA-10, RA-11, RA-12 breeding lines and 0.9% embryo-like structures in Opolski. For the gynogenesis induction, B5 medium containing 0.1 mg L–1 2,4-dichlorophenoxyacetic acid was the most effective from all tested media. On this medium, the highest number of gynogenetic embryo-like structures was obtained. Most of the plants were regenerated on MS medium supplemented with 30 g L–1 sucrose, 0.2 mg L–1 6-benzylaminopurine and 1 mg L–1 indole-3-acetic acid. Thirty nine percent of regenerated plants acclimatized. Cytometric evaluation of gynogenetic plants of four tested genotypes revealed that in three genotypes, 100% of tested plants were haploid. Plants showed diploid ploidy level in one genotype. Isoenzymatic analysis of gynogenetic plants demonstrated that 95% and 70% of examined populations were homozygotic for the phosphohexose isomerase isoenzyme and the aspartato aminotransferase isoenzyme, respectively. During the next generation sequencing, 93% of reads were successfully mapped, from which 83% to 85% were mapped in pairs. For 15% of pairs it was clear that obtained sequence was fully homozygous, the rest of the readings were not unambiguous, but similar to the sequence of a homozygous base pair system.