| Autor: |
R W J, Kortbeek, J, Xu, A, Ramirez, E, Spyropoulou, P, Diergaarde, I, Otten-Bruggeman, M, de Both, R, Nagel, A, Schmidt, R C, Schuurink, P M, Bleeker |
| Rok vydání: |
2016 |
| Předmět: |
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| Zdroj: |
Methods in enzymology. 576 |
| ISSN: |
1557-7988 |
| Popis: |
Glandular trichomes are specialized tissues on the epidermis of many plant species. On tomato they synthesize, store, and emit a variety of metabolites such as terpenoids, which play a role in the interaction with insects. Glandular trichomes are excellent tissues for studying the biosynthesis of specialized plant metabolites and are especially suitable targets for metabolic engineering. Here we describe the strategy for engineering tomato glandular trichomes, first with a transient expression system to provide proof of trichome specificity of selected promoters. Using microparticle bombardment, the trichome specificity of a terpene-synthase promoter could be validated in a relatively fast way. Second, we describe a method for stable expression of genes of interest in trichomes. Trichome-specific expression of another terpene-synthase promoter driving the yellow-fluorescence protein-gene is presented. Finally, we describe a case of the overexpression of farnesyl diphosphate synthase (FPS), specifically in tomato glandular trichomes, providing an important precursor in the biosynthetic pathway of sesquiterpenoids. FPS was targeted to the plastid aiming to engineer sesquiterpenoid production, but interestingly leading to a loss of monoterpenoid production in the transgenic tomato trichomes. With this example we show that trichomes are amenable to engineering though, even with knowledge of a biochemical pathway, the result of such engineering can be unexpected. |
| Databáze: |
OpenAIRE |
| Externí odkaz: |
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