Ocimum basilicum L. is one of the most famous plant used in the Mediterranean foods. It is rich in terpenoidic compounds like estragol, linalool, camphor, eugenol and in the well studied antioxidant molecule Rosmarinic Acid. Protocols for tissue culture and genetic transformation were set up with the aim to obtain in vitro biomass cell culture and/or hairy roots (HR), for secondary metabolite extraction in controlled conditions. In vitro transformation of O. basilicum was performed by leaf co-cultivation (for 15 or 30 minutes) with A. rhizogenes strains LBA 9402 and ATCC 15834. Then the explants were transferred onto basal medium MS for 48 hours in light. Afterword they were transplanted onto Petri dishes with MS medium added with cefotaxime 100 mg/L. After 20 days 100% of leaves treated with LBA 9402 strain developed new roots; these roots showed the characteristic hairy root behaviour and grew fast. Only 10% of leaves treated with 15834 strain developed putative HR roots. More than 15 HR clonal lines showed the characteristic transgenic morphology and had a high and fast growth rate. The presence of a fragment of rolC gene, was confirmed in a HR putative transformed line.
Utilizzo di due ceppi di Agrobacterium rhizogenes per la trasformazione di Ocimum basilicum L.: risultati preliminari
D'ANGIOLILLO, FRANCESCA;BARBERINI, SARA;PISTELLI, LUISA;PISTELLI, LAURA;
2012-01-01
Abstract
Ocimum basilicum L. is one of the most famous plant used in the Mediterranean foods. It is rich in terpenoidic compounds like estragol, linalool, camphor, eugenol and in the well studied antioxidant molecule Rosmarinic Acid. Protocols for tissue culture and genetic transformation were set up with the aim to obtain in vitro biomass cell culture and/or hairy roots (HR), for secondary metabolite extraction in controlled conditions. In vitro transformation of O. basilicum was performed by leaf co-cultivation (for 15 or 30 minutes) with A. rhizogenes strains LBA 9402 and ATCC 15834. Then the explants were transferred onto basal medium MS for 48 hours in light. Afterword they were transplanted onto Petri dishes with MS medium added with cefotaxime 100 mg/L. After 20 days 100% of leaves treated with LBA 9402 strain developed new roots; these roots showed the characteristic hairy root behaviour and grew fast. Only 10% of leaves treated with 15834 strain developed putative HR roots. More than 15 HR clonal lines showed the characteristic transgenic morphology and had a high and fast growth rate. The presence of a fragment of rolC gene, was confirmed in a HR putative transformed line.I documenti in IRIS sono protetti da copyright e tutti i diritti sono riservati, salvo diversa indicazione.
