INTRODUCTION TO PROCESS OF in vitro PROPAGATION AND ROOTS CLUSTER FORMATION OF THE Tabernanthe iboga (L.) Nutt. AND ANALYSIS OF IBOGAIN LEVELS

Proučevali smo možnost in vitro razmnoževanja iboge (Tabernanthe iboga (L.) Nutt.) z različnimi metodami rastlinskih tkivnih kultur. Semena rastline, ki smo jih pridobili iz štirih plodov, so kalila na treh različnih gojiščih. Od tega je bilo eno brez hormona, drugi dve pa sta vsebovali avksina IBA ali NAA. Za razmnoževanje poganjkov smo testirali B5, WPM, MS in DKW gojišča, ki so vsebovala različne mešance makro- in mikroelementov, dodali pa smo citokinin BAP. Na gojišču RP1 (gojišče B5) smo namnožili največ poganjkov. Na gojiščih za koreninjenje poganjkov ni prišlo do nastanka korenin na odrezanem delu. Na njem je nastal samo kalus. Problematično je bilo rumenenje poganjkov tako na gojiščih za namnoževanje poganjkov kot na gojiščih za koreninjenje poganjkov. Zamenjava kelatne oblike železa s FeEDDH je kasneje izboljšala pojav rumenenja. Proučevali smo tudi možnost indukcije rasti koreninskega sistema na trdnih gojiščih in v tekočih gojiščih. Na trdnih gojiščih nam je uspelo doseči izraščanje novih korenin iz izsečkov korenin, listov in stebla, vendar je bila rast zelo počasna. V tekočih gojiščih so se sprva tvorile nove korenine, kasneje pa smo poleg tvorbe novih korenin opazili tudi tvorbo somatskih embrijev. Embriji so se tvorili v več subkulturah, posamične od teh pa smo uspeli tudi vzgojiti do faze sadik ter jih aklimatizirati. Rezultate meritev vsebnosti ibogaina smo prejeli od dr. Gordane Koželj z Inštituta za sodno medicino Medicinske fakultete Univerze v Ljubljani, in sicer je »zeleni vzorec« (poganjki) vseboval 0,010 ut. % ibogaina, »rjavi vzorec« (korenine rastlin) pa 0,012 ut. % ibogaina.Protocols for micropropagation of iboga Tabernanthe iboga (L.) Nutt. was studied by various methods of plant tissue culture. One them was hormone-free, the other contained auxins IBA or NAA. Seeds germinates on all germination media and seedlings developed into plants. Multiplication of shoots was tested on various compositions of macro and micro elements - B5, WPM, MS, DKW. Medium RP1 (B5 medium) was the best for shoots multiplication. On rooting media no rooting occurred but formation of callus. Leaf yellowing was later reduced in media containing iron chelated with FeEDDH. Unexpectedly in liquid media supplemented with auxins somatic formation of embryos occurred. Induction of root growth from shoot and root explants was tested on solid and liquid media. On solid media growth was slow but root explants formed a limited quantity of new roots and shoots. Roots cultured in liquid media formed additional roots but after third subculture somatic embryos were formed. Somatic embryos were formed in several subcultures and were grown to plantlet stage being successfully acclimatized. The results of measurements of ibogaine was received from Dr. Gordane Koželj from the Institute of Forensic Medicine Faculty of Medicine, University of Ljubljana, the "green pattern" (shoots) containing 0.010 wt. % Ibogaine, "brown sample" (the roots of the plant) 0.012 wt. % ibogaine