4130654600

Phytochemistry 77 (2012) 79-88

F.l „SFYTF.R

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Phytochemistry

journal homepage: www.elsevier.com/locate/phytochem

Transcript and metabolite profiling in celi cultures of 18 plant species that produce benzylisoquinoline alkaloids

Scott C. Fanów¹, Jillian M. Hagel Peter J. Facchini*

Department of Biological Sciences. University of Calgary. Calgary. Alberta. Canada T2N IN4

ABSTRACT

ART1CLE 1NF0

Artide bisiory:

Received 14 Sepiember 2011

Receiwd in revised form 29 November 2011

Available Online 14 March 2012

Keynwrds:

Ranunculales

Benzylisoquinoline alkaloids Transcriptomics Targeted metabolite profiling Plant celi culture Gene discovery

Benzylisoquinoline alkaloids (BIAs) are a large and diverse group ol'^2500 specialized metabolites found predominanlly in planls of the order Ranunculales. Research focused on BIA melabolism in a restricled number of plant species bas identified many enzymes and cognate genes involved in the biosynthesis of compounds such as morphine, sanguinarine and berberine. However. the formation of most BIAs remains uncharacterized at the molecular biochemical level. Herein a compendium of sequence- and melabolite-profiling resources from 18 species of BlA-accumulating celi cultures was established, repre-senting four related plant families. Our integrated approach consisled of the construction of EST libraries each containing approximately 3500 unigenes per species for a total of 58,787 unigenes. The EST libraries were manually triaged usingknown BlA-biosynthetic genes as queries to identify putative homologs with similaror potentially different functions. Sequence resources were analyzed in the context of the targeted metabolite profilesobtained for each celi culture using electrospray-ionization andcollision-induced dis-sociation mass spectrometry. Fragmentation analysis was used for the identification or structural charac-terization coupled with the relative quantification of 72 BIAs, which establishes a key resource for futurę work on alkaloid biosynthesis. The metabolite profile obtained for each species provides a rational basis for the prediclion of enzyme function in BIA melabolism. The metabolic frameworks assembled through the integralion of transcript and metabolite profiles allowa comparison ol BIA metabolism across several plant species and families. Taken together, thesedata representan imporlant tool for the discovery ofBIA biosynthetic genes.

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