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Ewelina Piątczak, Aleksandra Królicka and Halina Wysokińska

Morphology, Secoiridoid Content and RAPD Analysis of Plants Regenerated from Callus of Centaurium erythraea RAFN

Centaurium erythraea plants obtained by indirect organogenesis are described in the paper. The plants were initiated from a single adventitious shoot regenerated from callus derived from the cotyledon of a 30-day-old seedling. The shoot was multiplied on MS medium supplemented with IAA (0.1 mg·L-1) and BAP (1.0 mg·L-1). The multiplication rate (28 shoots per culture within 4 weeks) was highest at the first subculture and decreased in further subcultures. The shoots were rooted on MS medium. The effect of IBA (0.1 mg·L-1) on the number of shoots forming roots differed depending on the composition of the basal medium (MS). The rooted shoots were transplanted to soil and grown in a greenhouse with 90% effectiveness. RAPD analysis was done with adventitious shoots of C. erythraea from in vitro culture. In shoots and whole plants regenerated from the callus tissue, secoiridoid content was determined by the HPLC method. We showed significant differences in morphology (leaf size, fresh and dry weight and height of plants) and changes in the DNA profiles as compared to earlier reports for shoot tip-derived shoots and plants of C. erythraea, but the two groups of plants biosynthesized the same qualitative pattern and similar levels of secoiridoids, up to 150 mg·g-1 dry weight; the increased biomass of plants regenerated from callus tissue makes them a better source of secondary metabolites.

Open access

Katarzyna Bergier, Elżbieta Kuźniak, Marzena Wielanek, Aleksandra Królicka and Maria Skłodowska

We compared the biochemical profiles of Physalis ixocarpa hairy roots transformed with Agrobacterium rhizogenes ATCC and A4 strains with non-transformed root cultures. The studied clones of A4- and ATCC-induced hairy roots differed significantly; the latter showed greater growth potential and greater ability to produce secondary metabolites (tropane alkaloids) and to biotransform hydroquinone to arbutin. We compared glucose content, alanine and aspartate aminotransferase activity, and L-phenylalanine ammonia-lyase activity. We analyzed markers of prooxidant/antioxidant homeostasis: catalase, ascorbate peroxidase, oxidase, glutathione peroxidase and transferase activity, and the levels of ascorbate, glutathione, tocopherol and lipid peroxidation. We found that transformation induced strain-specific regulation, including regulation based on redox signals, determining the rate of allocation of carbon and nitrogen resources to secondary metabolism pathways. Our results provide evidence that A. rhizogenes strain-specific modification of primary metabolites contributed to regulation of secondary metabolism and could determine the ability of P. ixocarpa hairy root clones to produce tropane alkaloids and to convert exogenously applied hydroquinone to pharmaceutically valuable arbutin. Of the studied parameters, glucose content, L-phenylalanine ammonia-lyase activity and alanine aminotransferases activity may be indicators of the secondary metabolite-producing potential of different P. ixocarpa hairy root clones.