Gorislavets S. Peculiarities of morphogenesis induction and anthocyanin synthesis in callus culture of the grapes identified using microsatellite markers

The Thesis presents results of research into in vitro plant stability and hormonal regulation of morphogenetic responses and anthocyanin synthesis in callus culture of four closely related grape varieties with colored berries: Antaeus magarachskii, Rubinovyi Magaracha, Granatovyi Magaracha and Rubin Golodrigi, and their putative parental forms Saperavi and Сabernet Sauvignon. Results of studies concerned with effects of NAA, BA and 2,4D on the processes of the callus formation, rhizogenesis and anthocyanin synthesis in the callus culture of six grape cultivars are presented. Selection of an optimum parity of phytohormones in a nutrient medium with the 2 mg/l NAA and 3 mg/l BAP has allowed to differentiate in vitro grapes on capacity of root system. An optimum phytohormone ratio with the 2 mg/l NAA and 2 mg/l BAP of nutrient medium for induction of anthocyanin synthesis in grape callus cultures was adjusted. Anthocyanin spectra of extracts of berries and callus cultures were studied by high performance liquid chromatography (HPLC). Up to eight anthocyanin peaks were detected in chromatograms of the extracts of callus cultures of six study varieties. Malvidin-3-O-glucoside was the major component of berry extracts while cyanidin-3-O-glucoside and peonidin-3-O- glucoside dominanated in the extracts of callus cultures of all study varieties. The variety Granatovyi Magaracha preved to be the most promising from a standpoint of induction of anthocyanin synthesis in callus culture. The varieties were genotyped and identified based on 10 nuclear and 3 chloroplast microsatellite loci. Despite their close kinship, the microsatellite profiles obtained were unique for each study variety. Analysis on a comparative basis of the allelic composition of the 6 microsatellite loci revealed that plants cultivated in vitro for long periods of time were identical to the original in situ genotypes.