An optimized method for total DNA extraction from genetically modified sugar beet plants has been presented. Uses of high toxic organic compounds and high cost extraction kits for nucleic acid isolation are excluded. A comparative analysis of polymerase chain reaction products was performed with amplifica tion of DNA obtained by two methods.

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Here we are testing the specific primers NEM06FWD2/NEM06REV2 and nem06FWD1/ nem06REV1 for the R6m-1 resistance gene to root-knot nematodes Meloidogyne spp. in breeding samples of sugar beet. Sugar beet plants of domestic and foreign breeding lines were the object of the study. To identify the relationship between R6m-1 gene, which is localized on the chromosome 1 and controls the stable level of the kinase activity signal, with sugar beet resistance to phytopathogens, PCR-analysis of 10 sugar beet samples were carried out using 2 pairs of molecular genetic markers. DNA amplification revealed a fragments ~500 bp and ~100 bp in length and as a result of sequencing of nucleotide sequences of R6m-1 gene region with subsequent alignment by Geneious Prime program, 3 single nucleotide substitutions (A/G, G/C, and G/A) in the resistant MS11018 genotype and one nucleotide substitution (A/G) and 3 deletions in a foreign hybrid Humber were identified. It can be assumed that these SNPs can form resistance by amino acid substitutions in the polypeptide chain. Finally, possibility to differentiate homozygous and heterozygous genotypes for this allele was shown.

Изучение гена кислой хитиназы SE2 в генотипах сахарной свеклы РЕЗЮМЕЦель работы -апробация специфического праймера FusA1F/FusA1R для изучения гена SE2, ответственного за экспрессию кислой хитиназы при стрессовых ситуациях. Материалом для исследования служили растения сахарной свеклы отечественной и зарубежной селекции. Для подтверждения связи гена SE2, локализованного на хромосоме 3 и контролирующего стабильный уровень работы кислой хитинизы, с устойчивостью сахарной свеклы к корневой гнили проведено генотипирование 10 образцов сахарной свеклы с использованием молекулярного-генетического маркера FusA1. Были выявлены по 2 ДНК-фрагмента длиной 600 п.н. и 400 п.н. во всех исследуемых генотипах, кроме растений дикой свеклы (Beta corolliflora Zoss.). В результате проведенных исследований гена SE2 идентифицированы восемь однонуклеотидных замен (3 T/C, 2 C/G, A/G, G/A, C/T) и 3 однонуклеотидные вставки (нуклеотид А) в растениях селекционного номера 9 (Sh.1) по сравнению с устойчивым генотипом. Растения данного гибрида и в полевых условиях проявляли признаки зараженности фузариозом. Можно предположить, что данные SNPs могут приводить к преодолению устойчивости (путем замены аминокислотной единицы в полипептиде) и, как следствие, снижению адаптационной способности растений. Studying of the acid chitinase SE2 gene in sugar beet genotypes ABSTRACTAim of the work was specific primers (FusA1F/FusA1R) testing to study the SE2 gene controlling acid chitinase effect under stress conditions. Plants of domestic and foreign sugar beet were the material for the investigation. To confirm relationship between the SE2 gene (localized to the chromosome 3) controlling steady effect of acid chitinase and sugar beet resistance to root rot, 10 sugar beet samples were genotyped using the FusA1 molecular marker. DNA-fragments of 600 and 400 b.p.in length were revealed in each of the investigated genotypes except plants of wild beet (Beta corolliflora Zoss.). As a result of molecular-genetic studies of the SE2 gene, eight single nucleotide polymorphism (3 T/C, 2 C/G, A/G, G/A, C/T) and 3 single nucleotide inserts (nucleotide A) were identified in plants of the breeding sample No. 9 (Sh.1). Plants of this genotype showed symptoms of fusariose infection under field conditions as well. It can be concluded that the SNPs data lead to overcoming of resistance (by changing an amino-acid unit in a polypeptide), and, as consequence, to decrease of plant adaptability.

The purpose of this study was to isolate the Saccharomyces cerevisiae present on different fruits and performing RAPD and ISSR analyses to know the genetic interrelationship between different S. cerevisiae isolates. Some fruits namely apple, plum, dates, and peach were used as natural sources for S. cerevisiae isolation. The isolated S. cerevisiae was designated as SUC1, SUC2, SUC3, SUC4, SUC5 respectively. Amplicon fingerprints for the isolated species were obtained by RAPD assay using six different primers and ISSR assay using six different primers. RAPD assay showed the lowest genetic distance (0.1559) between SUC2 and SUC3 isolates whereas ISSR assay showed the lowest genetic distance (0.06899) between SUC4 and SUC5 isolates.  Both genetic markers showed the highest genetic distance for SUC1 when compared to the other isolates.