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Nat Biotechnol. 2004 Dec;22(12):1559-66. Epub 2004 Nov 14.

RNAi-mediated replacement of morphine with the nonnarcotic alkaloid reticuline in opium poppy.

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  • 1CSIRO Plant Industry, PO Box 1600, Canberra, Australian Capital Territory 2601, Australia.

Abstract

We report on the silencing of codeinone reductase (COR) in the opium poppy, Papaver somniferum, using a chimeric hairpin RNA construct designed to silence all members of the multigene COR family through RNA interference (RNAi). After gene silencing, the precursor alkaloid (S)-reticuline-seven enzymatic steps upstream of codeinone-accumulated in transgenic plants at the expense of morphine, codeine, oripavine and thebaine. Methylated derivatives of reticuline also accumulated. Analysis verified loss of Cor gene transcript, appearance of 22-mer degradation products and reduction of enzyme activity. The surprising accumulation of (S)-reticuline suggests a feedback mechanism preventing intermediates from general benzylisoquinoline synthesis entering the morphine-specific branch. However transcript levels for seven other enzymes in the pathway, both before and after (S)-reticuline, were unaffected. This is the first report of gene silencing in transgenic opium poppy and of metabolic engineering to cause the high-yield accumulation of the nonnarcotic alkaloid reticuline.

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PMID:
15543134
[PubMed - indexed for MEDLINE]

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