Enhancement of Tropane Alkaloid Production among Several Clones and Explants Types of Hairy Root of Atropa belladonna L.

Abstract

Agrobacterium rhizogenes (pRi), a causative agent of hairy root disease, effectively induces hairy root formation in a variety of plant species. In our study four bacterial strains AR15834, A4, 9435 and C318 and three explants types leaf, stems and roots, were examined. Hairy roots were induced from roots, stems and leaf explants. The highest transformation efficiency of 77% was achieved by using strain AR15834. The transgenic status of hairy roots was confirmed by polymerase chain reaction using rolB specific primers for the presence of rolB gene in the genomes of transformed roots. Six clones of hairy roots were established that differed in their morphology. The results indicated that 4–11 fold increasing in root biomass after 28 days compared with non-transformed seedling roots. The concentration of tropane alkaloids hyoscyamine and scopolamine obtained in transformed roots (4 mg/g dry weight) was 3-11 folds more than in non-transformed cultured roots (0.36 mg/g dry weight).

Keywords

• hairy root
• Bacterial strain
• Explants type
• Tropane alkaloid