Micropropagation of blackberry cultivar "Reuben"

Cilj ovog istraživanja bio je utvrđivanje protokola za uspješnu mikropropagaciju kupine sorte “Reuben”. Vegetacijski vršci izolirani su aseptično pod stereomikroskopom iz aksilarnih pupova i postavljeni na medij za uspostavljanje kulture (EM). Ispitivan je utjecaj različitih šećera u PM2 mediju u dvije varijante 30 g/L saharoze i 20 g/L glukoze + 10 g/L saharoze. Kako bi se utvrdila optimalna koncentracija regultora rasta za mikropropagaciju, istraživane su 4 varijante: 6-benzilaminopurin (BAP) u koncentracijama 0,3 mg/L i 1 mg/L te meta-Topolin (mT) (u jednakim koncentracijama). Zakorjenjivanje in vitro provedeno je u dvije varijante, na IAA mediju za zakorjenjivanje i na mediju bez hormona. Najveći broj izdanaka po eksplantatu (2,6) dobiven je na mediju s koncentracijom 1 mg/L BAP-a, a najveća duljina najdužeg izdanka (8,9 mm) na mediju s koncentracijom 0,3 BAP-a. Postotak zakorjenjivanja je bio veći (61%) na mediju za zakorjenjivanje bez hormona, ali postotak aklimatizacije je bio sličan u obadvije varijante (92% i 95%).The aim of the present study was to establish the protocol for micropropagation of blackberry cultivar “Reuben”. In aseptic conditions using stereomicroscope, shoot tips were isolated from axillary buds and placed on medium (EM) to establish aseptic culture. The effect of different sugars on PM2 medium in the two variants of 30 g / L sucrose and 20 g / L glucose + 10 g / L sucrose was tested. In order to establish optimal variant of growth regulators 4 variants were experimented, 6- benzylaminopurine (BAP) in concentration of 0.3 mg/L and 1 mg/L and meta-Topolin (mT) in same concentrations. In vitro rooting was carried out in two experimental variants, on IAA rooting medium and on hormone free rooting medium. The greatest number of shoots with average of 2.6 were produced in medium containing 1 mg/L BAP and the maximum shoot length of 8.9 was produced in medium containing 0.3 mg/L BAP. The rooting percentage was higher (61%) on hormone free rooting medium but the acclimatization percentages for both variants were very similar (92% and 95%)