Agrobacterium-mediated transformation of Artemisia absinthium L. (wormwood) and production of secondary metabolites |
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Authors: | S Nin A Bennici G Roselli D Mariotti S Schiff R Magherini |
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Institution: | (1) Dipartimento di Ortoflorofrutticoltura, Università di Firenze, Via Donizetti 6, I-50144 Firenze, Italy, IT;(2) Dipartimento di Biologia Vegetale, Università di Firenze, Piazzale delle Cascine 28, I-50144 Firenze, Italy, IT;(3) Istituto sulla Propagazione delle Specie Legnose, Consiglio Nazionale delle Ricerche, Via Ponte di Formicola 76, I-50018 Scandicci, Firenze, Italy,, IT;(4) Istituto di Radiochimica ed Ecofisiologia Vegetali, Consiglio Nazionale delle Ricerche, Via Salaria km 29300, I-00016 Monterotondo Scalo, Roma, Italy, IT |
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Abstract: | Hairy roots were obtained after infection of Artemisia absinthium shoots with Agrobacterium rhizogenes strains 1855 and LBA 9402. The susceptibility to hairy root transformation varied between plant genotypes and bacterial strains.
Hairy roots showed macroscopic differences from control root cultures. Southern blot hybridization confirmed the integration
of T-DNA from both p1855 and pBin19, while polymerase chain reaction analysis indicated the presence of the neomycin phosphotransferase
gene in the hairy root genome. Subcultured transformed root lines grew well in selective B5 agar-solidified medium containing
kanamycin or rifampicin and without hormones. Shake-flask experiments with fast-growing root lines showed that 40 g l–1 was the best sucrose concentration for biomass production, yielding a 463-fold increase in dry weight after 28 days of culture.
Great differences were found in the profiles of the essential oils isolated from normal and hairy roots. Gas chromatography/mass
spectrometry analysis showed the oil produced by transformed cultures to be a mixture of 50 compounds with only one major
component representing 37% of the oil content.
Received: 19 March 1996 / Revision received: 15 July 1996 / Accepted: 13 December 1996 |
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Keywords: | Wormwood Hairy roots Genetic transformation Essential oil Secondary metabolites |
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