| Abstract Detail
Recent Topics Posters Ricigliano, Vincent [1], Brooks, Christopher [2], Howarth, Dianella [1]. Agrobacterium rhizogenes-mediated transformation of Valeriana officinalis for the study of sesquiterpenoid biosynthesis. Medicinal plants have a long history of human use across many cultures. Two-thirds of the world’s population still relies on plants as their primary pharmaceutical care. Root extracts of plants in the genus Valeriana are principally used as an herbal sedative, anxiolytic and spasmolytic agent. We have developed a cell culture system for the study of secondary metabolism in Valeriana officinalis. An agrobacterium-mediated transformation system was developed for functional characterization of genes implicated in terpenoid biosynthesis. Agrobacterium tumefaciens strain EHA105 harboring a binary vector was used for all transformation experiments. Neomycin phosphotransferase (nptII) was used as a selectable marker. Explants growing on selection were confirmed by PCR and a ~20% transformation efficieny was echieved. An intron-containing beta-glucuronidase driven by the constitutive CaMV35s promoter was employed as a reporter gene. Beta-glucuronidase activity was detected in 87% of PCR-positive lines by GUS histochemical assay. Farnesyldiphosphate synthase (FDS), a rate-limiting step in sesquiterpenoid metabolism, was characterized in yeast and subsequently in-planta. A dose dependent up-regulation of FDS was detected in response to biotic and abiotic elicitation. Cells treated with methyl jasmonate, a known regulator of secondary metabolism, displayed a 3-fold increase in FDS transcript over control. GC-MS analysis of MeJA treated cells quantified significant increases in sesquiterpenoid contents of the cultures. An improved understanding of terpenoid biosynthesis will enable metabolic engineering for the production of pharmaceutically important molecules in medicinal plants, cell cultures, and heterologous hosts. Broader Impacts:
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1 - St. John's University, Department of Biological Sciences, 8000 Utopia Parkway, Queens, NY, 11439, USA
2 -
Keywords:
agrobacterium
secondary metabolites
sesquiterpenoid.
Presentation Type: Recent Topics Poster
Session: P
Location: Grand Salon A - D/Riverside Hilton
Date: Monday, July 29th, 2013
Time: 5:30 PM
Number: PRT029
Abstract ID:1324
Candidate for Awards:None |
