Cryopreservation of Limonium serotinum apical meristems from in vitro plantlets using droplet-vitrification

Abstract In this study in vitro shoot tips of a Sicilian genotype of Limonium serotinum were successfully cryopreserved using the droplet-vitrification technique. Growth recovery of cryopreserved shoot tips was possible only when samples were pretreated for 16 h in liquid medium with 0.3 M sucrose, then for 5 h in liquid medium with 0.7 M sucrose before performing the cryopreservation protocol. Optimal conditions included treatment for 20 min in a loading solution containing 1.9 M glycerol + 0.5 M sucrose, treatment with vitrification solution B5 (glycerol 40.0%, sucrose 40.0%, w/v) for 60 and 90 min or vitrification solution A9 (glycerol 30.0%, dimethylsulfoxide 20.0%, ethylene glycol 20.0…

THE EFFECTS OF GROWTH REGULATORS ON IN VITRO AXILLARY SHOOT PROLIFERATION AND ROOTING OF ERICA MULTIFLORA

Multiple shoots were induced on nodal segments of a mature plant of Erica multiflora L. Axillary shoots produced on uncontaminated explants were excised, segmented, and recultured on the same medium. The effect of Anderson medium, augmented with different concentrations of 2iP either singly or in combination with NAA, as potential medium for shoot multiplication by nodal segments was investigated. In the following experiment equal molar concentrations of four cytokinins (2iP, kinetin, zeatin, and BA) were studied for ability to induce axillary shoot development from single node stem segments. The highest rate of axillary shoot proliferation was induced on Anderson medium supplemented with 1…

Investigating the cryopreservation of nodal explants of Lithodora rosmarinifolia (Ten.) Johnst., a rare, endemic Mediterranean species

In this study, we investigated the possibility of using the droplet-vitrification technique for cryopreserving nodal segments of in vitro plantlets of the endangered plant species Lithodora rosmarinifolia. Among the three vitrification solutions tested, only solutions B1, containing (w/v) 50 % glycerol and 50 % sucrose, and B3, containing 40 % glycerol and 40 % sucrose, were able to induce cryotolerance in nodal explants, resulting in intermediate survival and recovery after cryopreservation. A three-step vitrification protocol, including an additional dehydration treatment with half-strength vitrification solution for 30 min before the treatment with full-strength vitrification solution, d…

La coltura in vitro per la conservazione del germoplasma vegetale.

Comparing encapsulation-dehydration and droplet-vitrification for cryopreservation of sugarcane (Saccharum spp.) shoot tips

Abstract In this study, in vitro shoot tips of two sugarcane clones were successfully cryopreserved using encapsulation-dehydration and droplet-vitrification with two vitrification solutions, PVS2 and PVS3. For both clones, encapsulation-dehydration induced significantly higher recovery, reaching 60% for clone H70-144 and 53% for clone CP68-1026, compared with droplet-vitrification in which recovery was 33–37% for clone H70-144 and 20–27% for clone CP68-1026. Optimal conditions included preculture of encapsulated shoot apices for 24 h in liquid medium with 0.75 M sucrose and dehydration with silica gel to 20% moisture content (fresh weight basis) before direct immersion in liquid nitrogen. …