Influences of various factors on hairy root induction in Agastache foeniculum (Pursh) Kuntze | Vpliv različnih dejavnikov na indukcijo lasastih korenin pri janežnem ožepu (agastache foeniculum (pursh) kuntze)

Inglés. Agrobacterium rhizogenes is known as a natural tool of genetic engineering in many plant species. For the first time, hairy root induction in Agastache foeniculum using A. rhizogenes, rosmarinic acid content and the effect of different culture media and inoculation methods on hairy root growth rate were investigated. Hairy root culture of A. foeniculum was established by inoculation of the 1-month-old leaf explant with A4 strain of A. rhizogenes and the effectiveness of light – dark conditions and two inoculation methods (immersion and injection) were tested. Furthermore, in immersion method, the effects of inoculation time (3, 5 and 7 min) on root induction were investigated. In the second part of the study, the hairy root culture of A. foeniculum was studied using different basal culture media (MS, 1/2 MS and B5). Rosmarinic acid content in hairy roots and non- transformed roots was analyzed using high-performance liquid chromatography (HPLC). There was no significant difference between various inoculation methods in the ability of hairy roots induction. Observations showed that percentage of hairy root induction was higher when the explants were immersed for 5 min in bacterial suspension. Light conditions displayed the highest hairy root induction rates compared with dark condition. Various culture media are different in terms of types and amounts of nutrients and have influence on growth rate. The maximum growth rate (1.61 g fr wt/50 ml) of hairy roots were obtained in 1/2 MS medium. Rosmarinic acid content in transformed roots (213.42 µg/g dry wt) was significantly higher than non-transformed roots (52.28 µg/ g dry wt).

Esloveno. Bakterija Agrobacterium rhizogenes je znana kot naravno orodje genskega inženiringa pri mnogih rastlinskih vrstah. V tej raziskavi so prvič preučevali indukcijo lasastih korenin pri janežnem ožepu (Agastache foeniculum (Pursh.) Kuntze) z uporabo bakterije A. rhizogenes, vsebnost rožmarinske kisline in vpliv različnih gojišč ter inokulacijskih metod na rast lasastih korenin. Kulturo lasastih korenin janežnega ožepa so vzgojili iz izsečkov enomesečnih listov inokuliranih z bakterijo A. rhizogenes sev A4 z uporabo dveh inokulacijskih metod – potapljanje in vbrizgavanje, v razmerah na svetlem in v temi. Pri metodi potapljanja izsečkov v bakterijsko suspenzijo so preučevali tudi vpliv časa (3, 5 in 7 min) na indukcijo korenin. V drugem delu raziskave so preučevali vpliv različnih osnovnih gojišč (MS, 1/2 MS in B5) na kulturo lasastih korenin. Vsebnost rožmarinske kisline v transformiranih lasastih koreninah in netransformiranih koreninah so analizirali s HPLC metodo. Značilna razlika v indukciji lasastih korenin v odvisnosti od metod inokulacije ni bila ugotovljena. Opazovanja so pokazala, da je bil odstotek indukcije lasastih korenin večji, če so izsečke listov potopili v bakterijsko suspenzijo za pet minut. Indukcija lasastih korenin je bila največja v razmerah gojitve na svetlobi v primerjavi z gojenjem v temi. Različna gojišča so se razlikovala v vrsti in količini hranil in so vplivala na rast korenin. Največja rast lasastih korenin (1.61 g sveže mase/50 ml) je bila dobljena pri polovični koncentraciji osnovnega gojišča (1/2 MS). Vsebnost rožmarinske kisline je bila v transformiranih koreninah značilno večja (213.42 µg/g suhe mase) kot v netransformiranih (52.28 µg/ g suhe mase).