Utjecaj antioksidansa u predkulturi mikroreznica in vitro na regeneraciju vinove loze nakon krioprezervacije

Abstract

Protokole krioprezervacije potrebno je testirati za autohtone sorte na zaraženom i zdravom sadnom materijalu, jer zaraženost može predstavljati problem u kulturi tkiva. Antioksidansi mogu potaknuti rast biljaka. Cilj rada bio je uspostaviti regeneraciju biljaka inficiranih genotipova u in vitro uvjetima, te na njima testirati standardni protokol krioprezervacije za vinovu lozu. Pupovi inficiranih genotipova veličine 1,5 mm postavljeni su na različite MS (Murashige i Skoog) medije uz dodatak 0,5 mg/L 6-benzilaminopurina (BAP) i 0,05 mg/L indol-3-octene kiseline (IAA) sa antioksidansima (M1: 0,1 mM salicilne kiseline, M2: 1 mM askorbinske kiseline, M3: 1 mM glutationa i M4:kombinacija tri antioksidansa navedenih koncentracija), te je praćen njihov rast in vitro. Iz potjeralih biljaka izolirani su vršni meristemi, te je proveden pokus krioprezervacije (LS-20´, ½ PVS2-30´i PVS2-50´ kontrola i smrzavanje). Najveći rast in vitro postignut je na mediju sa dodatkom glutationa (M3). Zaraženost je imala utjecaj u regeneraciji biljaka in vitro, ali nije utjecala na regeneraciju nakon krioprezervacije.

Cryopreservation protocols need to be tested for autochthonous cultivars on an infected and healthy plant material cause virus infection can be difficult in tissue culture. Antioxidants can improve the plant growth. The aim of this work was to establish the plant regeneration of infected plants in tissue culture conditions and testing a standard cryopreservation protocol for grapevine on them. Buds 1.5.mm-size of infected genotypes were inoculated on different MS (Murashige i Skoog) media with addition of 0.5 mg/L 6-benzilaminopurina (BAP) and 0.05 mg/L indol-3-acetic acid (IAA) with antioxidants (M1: 0.1 mM salycilic acid, M2: 1 mM ascorbic acid, M3: 1 mM glutathione and M4: combination of three antioxidants with mentioned concentrations) and in vitro growth was observed. From shooted plants apical meristems were excised and cryopreservation protocol was performed (LS-20´, ½ PVS2-30´and PVS2-50´ control and freezing). The highest in vitro growth was achieved on medium with addition of glutathione (M3). Infection has an effect on regeneration of plants, but has not effect on regeneration after cryopreservation.