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RAPD and phytochemical analysis of Thymus moroderi plantlets after cryopreservation.

Cryo letters (2013-04-30)
Ana Marco-Medina, José Luis Casas
ABSTRACT

Cryopreservation is at present the most reliable strategy to preserve plant germplasm. When aromatic plants are the object of conservation it is necessary to assess not only the genetic but also the phytochemical stability to ensure that plant material maintains its qualities after storage. In this work we present molecular and phytochemical stability data related to a previously described vitrification-based cryopreservation protocol for Thymus moroderi Pau ex Martínez. RAPD markers have been used to assess the genetic stability of T. moroderi explants and revealed 0.34 percent of variation in the cryopreserved material studied. Phytochemical data collected from GC-MS analysis of dichloromethane extracts from cryopreserved plantlets rendered a profile in which 1,8-cineole (14.5 percent), camphor (5.9 percent) and borneol (5.2 percent) were the major components. Both data confirmed the suitability of the cryopreservation protocol applied.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Eucalyptol, 99%
Sigma-Aldrich
Camphor, 96%
Supelco
Eucalyptol, analytical standard
Supelco
D-Camphor, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
(1R)-(+)-Camphor, 98%
Sigma-Aldrich
Eucalyptol, natural, ≥99%, FCC, FG
Sigma-Aldrich
(+)-Borneol, 97%
Supelco
1,8-Cineole, primary reference standard
Supelco
(−)-Borneol, analytical standard
Sigma-Aldrich
Isoborneol, 95%
Sigma-Aldrich
(±)-Camphor, purum, synthetic, ≥95.0% (GC)
Sigma-Aldrich
(−)-Borneol, predominantly endo, 97%
Sigma-Aldrich
D-Camphor, ≥97%, FG
Sigma-Aldrich
(±)-Camphor, meets analytical specification of Ph. Eur., BP, ≥95% (GC), racemic
Sigma-Aldrich
(1S)-(−)-Camphor, 99%
Supelco
(−)-Camphor, analytical standard
Sigma-Aldrich
Isoborneol, ≥95%, FG
Cineole, European Pharmacopoeia (EP) Reference Standard
Camphor (racemic), European Pharmacopoeia (EP) Reference Standard
Camphor (dl), primary reference standard
Sigma-Aldrich
(±)-Camphor, ≥95.5%
Supelco
Eucalyptol, certified reference material, TraceCERT®
Sigma-Aldrich
(1S)-(−)-Camphor, 95%
Supelco
(+)-Borneol, analytical standard