CRYOPRESERVATION OF ADVENTITIOUS ROOTS OF Cleome rosea Vahl (CLEOMACEAE) USING A VITRIFICATION TECHNIQUE AND ASSESSMENT OF GENETIC STABILITY.

ABSTRACT

BACKGROUND: Cleome rosea, a Brazilian native species, has medicinal potential. Previously a cryopreservation protocol for in vitro roots using the vitrification solution PVS2 has been developed. However, the genetic stability of the cryopreserved material is yet to be assessed. OBJECTIVES: To evaluate the effects of loading and vitrification solutions (PVS2 and PVS3) on post-cryopreservation recovery of C. rosea roots, and to assess their genetic stability using Random Amplified Polymorphic DNA (RAPD) markers. MATERIALS AND METHODS: Root segments were pretreated with increasing concentrations of sucrose (0.2 to 0.4 M), followed by osmoprotection with loading solution and treatment with one of the vitrification solutions tested. RESULTS: The highest recoveries using PVS2 and PVS3 were obtained when root segments were exposed to these solutions for 15 min, reaching 77% and 100% respectively. The RAPD band profiles were monomorphic with most of the primers used. This molecular analysis revealed high genetic similarity (similarity coefficients among 0.98 and 1.00) between the cryopreserved roots and their mother plants. CONCLUSION: Roots from in vitro-propagated plants of C. rosea, were successfully cryopreserved using the vitrification technique. No major variations were observed on the genetic stability of cryopreserved roots, validating the use of this protocol as an efficient long-term conservation option for this species.