Abstract
Romanian tomato (Lycopersicon esculentum Mill.) cultivars have been cryopreserved by encapsulation-dehydration and successfully acclimatized to ex vitro growth conditions. Shoot tips were excised from in vitro grown plants then precultured for 24 h in various sucrose concentrations, dehydrated up to 6 h in laminar air flow prior to direct immersion in liquid nitrogen (−196°C) for 24 h. Different parameters have been studied: the effects of osmoprotection and desiccation duration on the regrowth of cryopreserved shoot tips, the effects of various IBA concentrations on rooting and the ex vitro cclimatization of plants recovered from liquid nitrogen. The highest frequency of regrowth (72% cv. ‘Pontica’) was obtained when encapsulated explants were precultured in 0.5 M sucrose and the moisture content (fresh weight basis) of alginate beads was 23%. The highest rooting rates (58% to 77%) for all cultivars were observed for shoots grown on MS medium supplemented with 1.0 mg/l IBA. The rooted plants could be easily acclimatized ex vitro with up to 100% survival.
Highlights
Lycopersicon esculentum (Mill.) is one of the most extensively studied species for their importance as crop species and as model system for genetic, molecular or physiological analyses (Gubiš et al, 2004)
Romanian tomato (Lycopersicon esculentum Mill.) cultivars have been cryopreserved by encapsulation-dehydration and successfully acclimatized to ex vitro growth conditions
Rooting and acclimatization For rooting, one node shoot segments (2-2.5 cm in length) from shoots regenerated following cryopreservation were transferred to MS medium supplemented with 1.5 mg/l N6benzyladenine (BA) and various concentrations (0, 0.5, 1.0, 1.5 mg/l) of indole-3-butyric acid (IBA)
Summary
Lycopersicon esculentum (Mill.) is one of the most extensively studied species for their importance as crop species and as model system for genetic, molecular or physiological analyses (Gubiš et al, 2004). Various aspects of in vitro grown tomato cultures have been studied including micropropagation, selection of cell lines resistant to biotic / abiotic stress and production of haploids (Bhatia et al, 2004) or cryopreservation (Zevallos et al, 2013). For the long-term conservation of genetic resources the availability or development of reliable and cost effective strategies and the subsequent regeneration of plants following storage are basic requirements (Kaviani, 2010, 2011). Methods involving alginate encapsulation followed by desiccation have been applied to numerous plant species (González-Arnáo and Engelmann, 2006; Harding et al, 2009; Engelmann, 2011; Cruz-Cruz et al, 2013) since their development by Fabre and Dereuddre (1990)
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