ISHS Acta Horticulturae 502: II WOCMAP Congress Medicinal and Aromatic Plants, Part 3: Agricultural Production, Post Harvest Techniques, Biotechnology
IN VITRO CULTURE OF FOUR MEDICINAL ASTERACEAE SPECIES FOR AGROBACTERIUM RHIZOGENES TRANSFORMATION |
| Authors: | A.P. Pellegrino, D.R.G. Joaquim, S.L. Kirszenzaft Shepherd |
| Keywords: | Artemisia annua, Calendula officinalis, hairy-roots, Helianthus annuus, in vitro cultivation, Mikania glomerata |
| Abstract:
Hairy roots produced by infection with Agrobacterium rhizogenes are phenotypically altered and show an overproduction of secondary compounds. Four Asteraceae species were tested: Artemisia annua (with proven antimalarial activity), Calendula officinalis (antiseptic), Mikania glomerata (a Brazilian plant used for respiratory disease treatment) and Helianthuus annuus (sunflower with tonic and anti-neuralgic activities). Seeds of Artemisia annua, Calendula officinalis and Helianthus annuus were surface disinfected with 25% NaOCl solution for 15 min., and Mikania glomerata stems were surface disinfected with 25% NaOCl solution for 30 min. Sterile and viable explants were mantained in complete Murashige & Skoog's culture medium and subcultured after 28 days. Once established in culture, the explants of all species were inoculated with Agrobacterium rhizogenes strains 8196 and 15834. Only Artemisia annua showed a positive hairy root response. These roots were excised and cultured “in vitro”. In order to confirm the transgenic character of the hairy roots a Southern Blot hybridization was carried out. The culture protocols established for the other three species were used to obtain explants for further transformation tests. | |
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