| Autor: |
Frick, Susanne, Chitty, Julie, Kramell, Robert, Schmidt, Jürgen, Allen, Robert, Larkin, Philip, Kutchan, Toni |
| Zdroj: |
Transgenic Research; December 2004, Vol. 13 Issue: 6 p607-613, 7p |
| Abstrakt: |
The berberine bridge enzyme cDNA bbefrom Papaver somniferumL. was transformed in antisense orientation into seedling explants of the industrial elite line C048-6-14-64. In this way, 84 phenotypically normal T0plants derived from embryogenic callus cultures were produced. The selfed progeny of these 84 plants yielded several T1plants with an altered alkaloid profile. One of these plants T1-47, and its siblings T2-1.2 and T2-1.5 are the subject of the present work. The transformation of these plants was evaluated by PCR, and northern and Southern hybridisation. The transgenic plants contained one additional copy of the transgene. The alkaloid content in latex and roots was determined with HPLC and LC-MS. We observed an increased concentration of several pathway intermediates from all biosynthetic branches, e.g., reticuline, laudanine, laudanosine, dehydroreticuline, salutaridine and (S)-scoulerine. The transformation altered the ratio of morphinan and tetrahydrobenzylisoquinoline alkaloids in latex but not the benzophenanthridine alkaloids in roots. The altered alkaloid profile is heritable at least to the T2generation. These results are the first example of metabolic engineering of the alkaloid pathways in opium poppy and, to our knowledge, the first time that an alkaloid biosynthetic gene has been transformed into the native species, followed by regeneration into a mature plant to enable analyses of the effect of the transgene on metabolism over several generations. |
| Databáze: |
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