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Published on June 11, 2008; 10.1104/pp.108.120493

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Received April 3, 2008
Accepted June 5, 2008

Quantitative 1H NMR metabolite profiling as a functional genomics platform to investigate alkaloid biosynthesis in opium poppy

Jillian M. Hagel , Aalim M. Weljie , Hans J. Vogel , and Peter J. Facchini *

Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada, T2N 1N4

* Corresponding author; email: pfacchin{at}ucalgary.ca.

Opium poppy (Papaver somniferum) produces a diverse array of bioactive benzylisoquinoline alkaloids and has emerged as a versatile model system to study plant alkaloid metabolism. The plant is widely cultivated as the only commercial source of the narcotic analgesics morphine and codeine. Variations in plant secondary metabolism as a result of genetic diversity are often associated with perturbations in other metabolic pathways. As part of a functional genomics platform, we have used 1H NMR metabolite profiling for the analysis of primary and secondary metabolism in opium poppy. Aqueous and chloroform extracts of six different opium poppy cultivars were subjected to chemometric analysis. Principle component analysis of the 1H NMR spectra for latex extracts clearly distinguished two varieties, including a low-alkaloid variety, and a high-thebaine, low-morphine cultivar. Distinction was also made between pharmaceutical-grade opium poppy cultivars and a condiment variety. Such phenotypic differences were not observed in root extracts. Loadings plots confirmed that morphinan alkaloids contributed predominantly to the variance in latex extracts. Quantification of 34 root and 21 latex metabolites, performed using Chenomx NMR Suite v. 4.6, showed major differences in the accumulation of specific alkaloids in the latex of the low-alkaloid and high-thebaine, low-morphine varieties. Relatively few differences were found in the levels of other metabolites indicating that the variation was specific for alkaloid metabolism. Exceptions in the low-alkaloid cultivar included an increased accumulation of the alkaloid precursor tyramine, and reduced levels of sucrose, some amino acids and malate. Real-time PCR analysis of 42 genes involved in primary and secondary metabolism showed differential gene expression mainly associated with alkaloid biosynthesis. Reduced alkaloid levels in the condiment variety were associated the reduced abundance of transcripts encoding several alkaloid biosynthetic enzymes.






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