Cryopreservation of hairy root cultures of Maesa lanceolata and Medicago truncatula

Abstract

To study the production of secondary metabolites of Maesa lanceolata and Medicago truncatula, hairy root cultures of both plant species were established. Because maintenance of large numbers of cultures is laborious and costly, we developed a cryopreservation protocol and stored different isolated lines over time. Using encapsulation-dehydration, high survival rates were observed for both Maesa and Medicago hairy roots. Root tips were isolated and encapsulated in calcium-alginate beads, containing 0.1 M sucrose. The encapsulated hairy roots were precultured for 3 days using basal medium containing high sucrose concentrations. Medicago root tip growth during the preculturing time lead to unwanted outgrowth which could be tempered by addition of plant growth inhibitors. After preculturing, the beads were dehydrated in the air flow of a laminar flow until 35-40% of the initial bead weight was reached. Dehydrated beads were plunged into liquid nitrogen and after different storage times thawed in a water bath at 40°C. The survival rates were 90% for Maesa and 53% for Medicago, which are sufficient to allow implementation in large storage experimental set-ups.

Hairy roots cultures from different Solanaceous species have varying capacities to produce E. coli B-subunit heat-labile toxin antigen

“…For species which do not readily regenerate into plants with otherwise potentially desirable bio-containment, growth and biomass productivity features, such as petunia as identified in the current study, long-term storage solutions may be the cryopreservation of tissues in liquid N 2 (Benson and Hamill 1991;Hirata et al 2002;Lambert et al 2009). This approach does of course require a moderate-high level of expertise and technological infrastructure.…”

Hairy root biotechnology of Rauwolfia serpentina: a potent approach for the production of pharmaceutically important terpenoid indole alkaloids

“…Cryopreservation and revival of hairy root clones of medicinal plants is a known practice (Lambert et al 2009). The survival potential of Rauwolfia hairy root cultures during and after preservation have indicted an easy adaptation to growth medium and restitution of normal growth.…”

Multiplication and cryopreservation of adventitious roots of Cleome rosea Vahl

“…Hairy root cultures of numerous medicinal plants have been cryopreserved (Teoh et al 1996;Hirata et al 2002;Xue et al 2008;Lambert et al 2009). However, few studies have reported on the application of cryopreservation techniques to untransformed or adventitious root cultures (Jung et al 2001;Oh et al 2009).…”