Induction of Pyridine alkaloid formation in transformed root cultures of Nicotiana tabacum

Callus of Nicotiana tabacum was established from its explant and investigated. It has been found that the alkaloid yield was very little. Therefore hairy root cultures were set up by infection of root tips with Agrobacterium rhizogenes. The successful transformation was confirmed by analysing rolC and virC genes using polymerase chain reaction (PCR). Hairy root cultures were employed to study the formation of pyridine alkaloids, such as nicotine. The transformed cultures were incubated with potential elicitors, such as methyljasmonate, quercetin and salicylic acid in order to stimulate the biosynthesis of pyridine alkaloids. Profile and amounts of pyridine alkaloids were analysed using capillary GLC-MS. Treatment of the cultures with methyljasmonate (50 µm) increased the alkaloid accumulation ca 7-fold up to a level of 0.5mg/g fresh weight as compared to untreated controls. Quercetin (200 µm) enhanced the alkaloid production ˜ 4 fold (0.4mg/g fresh weight) within 24h. In contrast salicylic acid with all tested concentration decreased alkaloids to a level of 1 µg/g fresh weight. Also the inhibition effect of SA on the elicitation of MEJA and Q was investigated.