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| Naji, M. |
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| Motta, Antonella |
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| Aletan, Dirar |
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| Mohamed, Tarek |
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| Ertürk, Emre |
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| Taccardi, Nicola |
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| Petrov, R. H. | Madrid |
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| Kočí, Jan | Prague |
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| Kalteremidou, Kalliopi-Artemi | Brussels |
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| Azam, Siraj |
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| Ali, M. A. |
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| Popa, V. |
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| Rančić, M. |
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| Ollier, Nadège |
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| Azevedo, Nuno Monteiro |
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| Landes, Michael |
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Seppänen-Laakso, Tuulikki
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document
A functional genomics approach to unravel plant secondary metabolism by combining transcriptional profiling with targeted metabolome analysis
Abstract
Plant cell cultures are an option for producing secondary metabolitesuseful for diverse applications although only very few economically feasibleexamples exist so far. Engineering of plant cells with functionally testedgenes helps to understand the biosynthetic pathways and is the method ofchoice for creating high yielding strains for commercial production.On a genome wide scale genes involved in plant secondary metabolism aresimultaneously identified and isolated by an approach, in which a cDNA-AFLPbased transcript profiling technique in conjunction with metabolic pathwayprofiling is applied. Highly specific RNA fingerprints in function of time areobtained following elicitation of cell cultures. In parallel the quantitativeand qualitative changes of metabolites involved in selected pathways aredetermined by several hyphenated methods (e.g. GC-MS, HPLC-MS). The functionaltesting of promising genes obtained from full-length cloning is done byanalyzing the metabolic changes in overexpression/co-suppression experimentsof transformed cells.A well-defined correlation between the pathway specific metabolites and thetranscriptome was revealed using the model system of tobacco BY-2 cells. Insilico analysis of about 20000 visualised gene tags showed that about 600 weredifferentially regulated by the elicitor. The applied analytical methods weresensitive enough for the investigated secondary compounds but the need fordifferent extraction processes in order to analyze metabolites from differentbiosynthetic branches became obvious from comparing extraction efficiencies.First experiments with other plant species confirmed that the technology isprincipally applicable to any plant or plant cell culture of interest.