Influences of different types of sugar and cytokinin on micropropagation of blackberry cultivar ‘Reuben’

Cilj istraživanja bio je utvrđivanje protokola za mikropropagaciju kupine sorte ‘Reuben’. Vegetacijski vršci izolirani su aseptično pod stereomikroskopom iz aksilarnih pupova i postavljeni na medij za uspostavljanje kulture (EM). Na mediju za proliferaciju (PM) ispitivan je utjecaj šećera u dvije varijante: s dodanih 30 g L-1 saharoze ili 20 g L-1 glukoze + 10 g L-1 saharoze. Da bi se utvrdila optimalna vrsta i koncentracija citokinina za mikropropagaciju, u drugom pokusu ispitano je 5 tretmana: 6-benzilaminopurin (BAP) u koncentracijama 1 mg L-1 i 0,3 mg L-1, meta-topolin (mT) u jednakim koncentracijama te kao kontrola, medij bez hormona. Zakorjenjivanje in vitro provedeno je u dvije varijante, na mediju s dodanom indolil-3-octenom kiselinom (IAA) u koncentraciji 1 mg L-1 i na mediju bez hormona. Najveći broj izdanaka po eksplantatu (2,6) dobiven je na mediju s 1 mg L-1 BAP-a, ali su oni ujedno bili najkraći. Najdulji izdanci dobiveni su na mediju s 0,3 mg L-1 BAP-a ili mT kao i na mediju bez hormona. Postotak zakorjenjivanja bio je viši (61%) na mediju za zakorjenjivanje bez hormona. Aklimatizacija je bila uspješna i iznosila 92%-95%, ovisno o mediju za zakorjenjivanje.The aim of the present study was to establish the protocol for micropropagation of blackberry cultivar ‘Reuben’. Using stereomicroscope, shoot tips were isolated from axillary buds and placed on medium (EM) to establish aseptic culture. The effect of different sugars in proliferation medium (PM) in the two variants: 30 g L-1 sucrose or 20 g L-1 glucose + 10 g L-1 sucrose was tested. In order to establish optimal constitution of cytokinins, in second experiment 5 treatments were tested: 6- benzylaminopurine (BAP) in concentration of 1 mg L-1 and 0.3 mg L-1 and meta-Topoline (mT) in the same concentrations, as well as a control hormone free medium. In vitro rooting was carried out in two experimental variants, on IAA added rooting medium and on hormone free medium. The greatest number of shoots/explant (2.6), but in the same time the shortest shoots, were produced on medium containing 1 mg L-1 BAP. Maximum shoots length was produced on medium containing 0.3 mg L-1 BAP or mT as well as on medium without hormones. The rooting percentage was higher (61%) on hormone free rooting medium. Acclimatization was very successful reaching 92% -95%, depending on rooting medium