Evaluation of genetic fidelity of in vitro-propagated blackberry plants using RAPD and SRAP molecular markers

Authors: Orsolya BORSAI, Monica HÂRȚA, Katalin SZABO, Cristina-Daniela KELEMEN, Flavia Andreea ANDRECAN, Maria-Maruni CODREA, Doina CLAPA.

Abstract:

The aim of this research was to evaluate the genetic uniformity of blackberry plants (Rubus fruticosus L. ‘Loch Ness’ and ‘Chester Thornless’) obtained by micropropagation. Genetic uniformity was analysed by using RAPD and SRAP markers. For the in vitro multiplication, the slightly modified Murashige and Skoog (1962) basal medium was used, supplemented with 0.5 mg/L 6-Benzyladenine (BA), prepared with tap water and 50 g/L wheat starch as the gelling agent. This culture medium ensured the regeneration of well-developed plantlets, with multiplication rates of more than 42 for both cultivars. The in vitro multiplication was carried out in 30 months including 12 subcultures. The plants obtained from the 3rd and 11th subcultures were compared with the mother plants using 64 SRAP primer combinations (eight forward and eight reverse primers) and 20 RAPD primers to check their genetic fidelity. The amplification products were monomorphic in the micropropagated plants and the mother plant. No polymorphism was detected, thus proving the genetic fidelity and uniformity of the micropropagated plants.

Published: Horticultural Science (Prague), 47, 2020 (1): 21–27