title: Biochemical genomics for gene discovery in benzylisoquinoline alkaloid biosynthesis in opium poppy and related species.
abstract: Benzylisoquinoline alkaloids (BIAs) are a large, diverse group of ∼2500 specialized plant metabolites. Many BIAs display potent pharmacological activities, including the narcotic analgesics codeine and morphine, the vasodilator papaverine, the cough suppressant and potential anticancer drug noscapine, the antimicrobial agents sanguinarine and berberine, and the muscle relaxant (+)-tubocurarine. Opium poppy remains the sole commercial source for codeine, morphine, and a variety of semisynthetic drugs, including oxycodone and buprenorphine, derived primarily from the biosynthetic pathway intermediate thebaine. Recent advances in transcriptomics, proteomics, and metabolomics have created unprecedented opportunities for isolating and characterizing novel BIA biosynthetic genes. Here, we describe the application of next-generation sequencing and cDNA microarrays for selecting gene candidates based on comparative transcriptome analysis. We outline the basic mass spectrometric techniques to perform deep proteome and targeted metabolite analyses on BIA-producing plant tissues and provide methodologies for functionally characterizing biosynthetic gene candidates through in vitro enzyme assays and transient gene silencing in planta.
authors.affiliation: Department of Biological Sciences, University of Calgary, Calgary, Alberta, Canada.
authors.forename: Thu Thuy T
authors.initials: TT
authors.lastname: Dang
authors.forename: Akpevwe
authors.initials: A
authors.lastname: Onoyovwi
authors.forename: Scott C
authors.initials: SC
authors.lastname: Farrow
authors.forename: Peter J
authors.initials: PJ
authors.lastname: Facchini
mesh_terms: D044182:Benzylisoquinolines / Q000737:chemistry*; D053898:Biosynthetic Pathways; D004591:Electrophoresis, Polyacrylamide Gel; D057075:Enzyme Assays; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D020868:Gene Silencing; D017343:Genes, Plant; D023281:Genomics / Q000379:methods*; D013058:Mass Spectrometry; D055432:Metabolomics / Q000379:methods; D020411:Oligonucleotide Array Sequence Analysis; D010207:Papaver / Q000737:chemistry* / Q000235:genetics*; D010940:Plant Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification; D053719:Tandem Mass Spectrometry
chemical_list: D044182:Benzylisoquinolines; D010940:Plant Proteins