Transgenic hairy root cultures of Catharanthus roseus were established with a glucocorticoid-inducible promoter controlling the expression of green fluorescent protein (GFP), and GFP expression was characterized. The inducible system shows a tightly controlled, reversible, and dosage-dependent response to the glucocorticoid dexamethasone in C. roseus hairy roots. Full induction was noted after 12-18 h in the mature regions of the root tips and after 6 h in the meristem tissue. Upon removal of the inducing agent, GFP expression declined to undetectable levels in the mature tissues after 24 h and in the meristem after 48 h. Although no dosage-dependent response was noted in the meristem region, such a response was apparent in the mature region of the tip and verified by quantitative GFP analysis. The inducible promoter system allowed quantitative control of GFP expression between 0.01 and 10 microM dexamethasone with saturation occurring at higher levels. Using GFP as a model system allowed demonstration of the ability to control temporal and quantitative gene expression with the glucocorticoid-inducible promoter in transgenic C. roseus hairy roots.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1021/bp025603o | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Development of an efficient -mediated transformation method and its application in tryptophan pathway modification in .
Plant Biotechnol (Tokyo)
December 2023
Biosolutions Development Section, Biosolutions Labs, Research Institute for Bioscience Products & Fine Chemicals, Ajinomoto Co., Inc., 1-1 Suzuki-cho, Kawasaki-ku, Kawasaki-shi, Kanagawa 210-8681, Japan.
The biosynthetic pathway of vinca alkaloids has a long research history, including not only identification of metabolic intermediates but also the mechanisms of inter-cellular transport and accumulation of biosynthesized components. Vinca alkaloids pathway begins with strictosidine, which is biosynthesized by condensing tryptamine from the tryptophan pathway and secologanin from the isoprenoid pathway. Therefore, increasing the supply of precursor tryptophan may enhance vinca alkaloid content or their metabolic intermediates.
View Article and Find Full Text PDFHairy roots: An untapped potential for production of plant products.
Front Plant Sci
August 2022
Department of Chemical and Biological Engineering, Colorado State University, Fort Collins, CO, United States.
While plants are an abundant source of valuable natural products, it is often challenging to produce those products for commercial application. Often organic synthesis is too expensive for a viable commercial product and the biosynthetic pathways are often so complex that transferring them to a microorganism is not trivial or feasible. For plants not suited to agricultural production of natural products, hairy root cultures offer an attractive option for a production platform which offers genetic and biochemical stability, fast growth, and a hormone free culture media.
View Article and Find Full Text PDFIdentification and Characterization of Transcription Factors Regulating Terpenoid Indole Alkaloid Biosynthesis in Catharanthus roseus.
Methods Mol Biol
June 2022
Department of Plant and Soil Sciences, and the Kentucky Tobacco Research and Development Center, University of Kentucky, Lexington, KY, USA.
Biosynthesis of the therapeutically valuable terpenoid indole alkaloids (TIAs), in the medicinal plant Catharanthus roseus, is one of the most elaborate and complex metabolic processes. Although genomic and transcriptomic resources have significantly accelerated gene discovery in the TIA pathway, relatively few genes of transcription factors (TFs) have been identified and characterized thus far. Systematic identification of TFs and elucidation of their functions are crucial for understanding TIA pathway regulation.
View Article and Find Full Text PDFGeneration of Stable Catharanthus roseus Hairy Root Lines with Agrobacterium rhizogenes.
Methods Mol Biol
May 2022
Department of Chemical Engineering, Northeastern University, Boston, MA, USA.
Agrobacterium rhizogenes is the bacterial agent that causes hairy root disease in dicots and is purposefully engineered for the development of transgenic hairy root cultures. Due to their genetic and metabolic stability, hairy root cultures offer advantages as a tissue culture system for investigating the function of transgenes and as a production platform for specialized metabolites or proteins. The process for generating hairy root cultures involves first infecting the explant with A.
View Article and Find Full Text PDFRepression of , and affects expression of terpenoid indole alkaloid biosynthetic and regulatory genes.
PeerJ
July 2021
Plant and Microbial Biology, University of Minnesota-Twin Cities, Saint Paul, MN, United States.
Terpenoid indole alkaloids (TIAs) include several valuable pharmaceuticals. As remains the primary source of these TIA pharmaceuticals, several research groups have devoted substantial efforts to increase production of these compounds by . Efforts to increase TIA production by overexpressing positive regulators of TIA biosynthetic genes have met with limited success.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!