Abstract
Cryopreservation is a useful tool for the long-term storage of plant genetic resources, and different cryogenic procedures have recently been developed. The present study focused on the use of the Droplet-vitrification (DV) and V cryo-plate protocol for the cryopreservation of Stevia rebaudiana in vitro-derived apical shoot tips and axillary shoot tips. A preliminary test showed that 90 and 120 min PVS2 (Plant Vitrification Solution 2) treatment significantly reduced the regrowth of the explants before immersion in liquid nitrogen (LN). For both procedures tested, the best osmoprotective condition for obtaining a higher regrowth of cryopreserved explants occurred when explants were PVS2 treated for 60 min. After direct immersion in LN, thawing and plating, the highest regrowth recorded was 80% with DV and 93% with V cryo-plate. Moreover, shoot tips proved to be a more suitable material for Stevia cryopreservation. A satisfactory vegetative regrowth was observed in the subcultures following cryopreservation by DV and V cryo-plate cryogenic procedures.
Highlights
Stevia rebaudiana (Bertoni) is an herbaceous perennial plant of the Asteraceae family
The aim of this study was to assess the efficiency of the novel procedures, Droplet-vitrification
A protocol for in vitro conservation of S. rebaudiana under slow growth conditions and mass micropropagation after six months of storage without subculturing, was developed by Zayova et al [6], while cryopreservation to enable the long-term storage of shoot tips was carried out using a vitrification method [7]
Summary
Stevia rebaudiana (Bertoni) is an herbaceous perennial plant of the Asteraceae family. In addition to its sweetening properties, it has various medicinal properties and actions. For this reason, the Stevia plant is an extremely interesting crop for breeders who select varieties with high diterpene glycosides content as well as for propagators and the target market. S. rebaudiana is a self-incompatible plant and one of its limiting factors for large-scale cultivation is its poor seed germination [3]. Plants from seed propagation have a great variability in growth, maturity, and non-uniform plants, with considerable variations in the sweetening level and composition [4,5]. The recent results suggest that seed germination and stem cutting are not cost effective methods for higher biomass production, while the micropropagation can be a promising technique [3]
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