[P-008]
ANTHRAQUINONE PRODUCTION IN CALLUS TISSUE CULTURES OF THE
PLANTS FROM Rhamnaceae FAMILY

Nada Kovačević1 and Dragoljub Grubišić2,3
1Faculty of Pharmacy, Vojvode Stepe 450, 11000 Belgrade, FR Yugoslavia
2Institute for Biological Research "Siniša Stanković", 29. Novembra 142, 11000 Belgrade, FR Yugoslavia
3Institute for Medicinal Plant Research "Dr Josif Pančić", Tadeuša Košćuška 1, 1100 Belgrade, FR Yugoslavia

Plant species from the family Rhamnaceae are known to contain substantial amounts of emodin type of anthraquinones (AQ). Some of these species are sources of laxative drugs (Frangulae cortex, Purshianae cortex, Rhamni cathartici fructus). In order to study the production of emodin type of AQ, root, shoot and callus cultures of Frangula alnus Mill., Frangula rupestris (Scop.) Schur and Rhamnus catharticus L. were established. Here will be presented the results of production of AQs in the callus tissue of these species.

The presence of free AQs and AQ O-glycosides in calluses of three investigated plant species was confirmed by Borntrager test reaction. Using the method of acid hydrolysis at the TLC plates it was confirmed the absence of C-glycosides of AQ. HPLC was applied for quantitative analysis of metabolites.

The best mass proliferation of callus tissue (27 ± 0.1) of Frangula rupestris was obtained on solid medium with combination of mineral salts and vitamins according to Murashige & Skoog (MS), with addition of sucrose (3%) and auxine NAA (0.5 mg/l). The best production of AQs (513 mg/100g) was observed in tissue grown on medium with addition of BAP (0.5 mg/l). Mainly, AQs were in free stage, and physcione and chrysophanol dominated.

The best production of AQs (2106 mg/100 g) was obtained in callus tissue of Frangula alnus which was grown on solid MS medium with addition of NAA (0.1 mg/l) and TDZ (1 mg/l). Mainly, AQs were in free stage (»60%); physcione and chrysophanol dominated.

The best production of AQs (2162 mg/100 g) was obtained in callus tissue of Rhamnus catharticus which was grown on solid MS medium with addition of NAA (0.1 mg/l) and BAP (0.5 mg/l), but the mass proliferation was very pure. Around 50% of total AQs were in free stage; chrysophanol and emodin dominated.

For the end in can be concluded that callus tissue of Frangula alnus and Rhamnus cathatricus were a good producers of AQs. This production was influenced by composition of mineral salts, plant growth regulators as well as with the other physical condition under which the cultures were grown.

[Full paper: NA]
[P-008]