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Early Phenylpropanoid Biosynthetic Steps in Cannabis sativa: Link between Genes and Metabolites
Institute of Agricultural Biology and Biotechnology, CNR, Via Bassini 15, Milan 20133, Italy
Institute for Macromolecular Studies, NMR Department, CNR, Via Bassini 15, Milan 20133, Italy
* Author to whom correspondence should be addressed.
Received: 13 May 2013; in revised form: 13 June 2013 / Accepted: 14 June 2013 / Published: 28 June 2013
Abstract: Phenylalanine ammonia-lyase (PAL), Cinnamic acid 4-hydroxylase (C4H) and 4-Coumarate: CoA ligase (4CL) catalyze the first three steps of the general phenylpropanoid pathway whereas chalcone synthase (CHS) catalyzes the first specific step towards flavonoids production. This class of specialized metabolites has a wide range of biological functions in plant development and defence and a broad spectrum of therapeutic activities for human health. In this study, we report the isolation of hemp PAL and 4CL cDNA and genomic clones. Through in silico analysis of their deduced amino acid sequences, more than an 80% identity with homologues genes of other plants was shown and phylogenetic relationships were highlighted. Quantitative expression analysis of the four above mentioned genes, PAL and 4CL enzymatic activities, lignin content and NMR metabolite fingerprinting in different Cannabis sativa tissues were evaluated. Furthermore, the use of different substrates to assay PAL and 4CL enzymatic activities indicated that different isoforms were active in different tissues. The diversity in secondary metabolites content observed in leaves (mainly flavonoids) and roots (mainly lignin) was discussed in relation to gene expression and enzymatic activities data.
Keywords: Cannabis sativa; phenylalanine ammonia lyase; 4-Coumarate: CoA ligase; cinnamic acid 4-hydroxylase; chalcone synthase; phenylpropanoid; secondary metabolism; expression analysis; NMR metabolic profiling
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Docimo, T.; Consonni, R.; Coraggio, I.; Mattana, M. Early Phenylpropanoid Biosynthetic Steps in Cannabis sativa: Link between Genes and Metabolites. Int. J. Mol. Sci. 2013, 14, 13626-13644.
Docimo T, Consonni R, Coraggio I, Mattana M. Early Phenylpropanoid Biosynthetic Steps in Cannabis sativa: Link between Genes and Metabolites. International Journal of Molecular Sciences. 2013; 14(7):13626-13644.
Docimo, Teresa; Consonni, Roberto; Coraggio, Immacolata; Mattana, Monica. 2013. "Early Phenylpropanoid Biosynthetic Steps in Cannabis sativa: Link between Genes and Metabolites." Int. J. Mol. Sci. 14, no. 7: 13626-13644.