Plant cell cultures are an option for producing secondary metabolites useful for diverse applications although only very few economically feasible examples exist so far. Engineering of plant cells with functionally tested genes helps to understand the biosynthetic pathways and is the method of choice for creating high yielding strains for commercial production. On a genome wide scale genes involved in plant secondary metabolism are simultaneously identified and isolated by an approach, in which a cDNA-AFLP based transcript profiling technique in conjunction with metabolic pathway profiling is applied. Highly specific RNA fingerprints in function of time are obtained following elicitation of cell cultures. In parallel the quantitative and qualitative changes of metabolites involved in selected pathways are determined by several hyphenated methods (e.g. GC-MS, HPLC-MS). The functional testing of promising genes obtained from full-length cloning is done by analyzing the metabolic changes in overexpression/co-suppression experiments of transformed cells. A well-defined correlation between the pathway specific metabolites and the transcriptome was revealed using the model system of tobacco BY-2 cells. In silico analysis of about 20000 visualised gene tags showed that about 600 were differentially regulated by the elicitor. The applied analytical methods were sensitive enough for the investigated secondary compounds but the need for different extraction processes in order to analyze metabolites from different biosynthetic branches became obvious from comparing extraction efficiencies. First experiments with other plant species confirmed that the technology is principally applicable to any plant or plant cell culture of interest.
|Publication status||Published - 2003|
|MoE publication type||Not Eligible|
|Event||2nd International Conference Plant Metabolomics - Potsdam, Germany|
Duration: 25 Apr 2003 → 28 Apr 2003
|Conference||2nd International Conference Plant Metabolomics|
|Period||25/04/03 → 28/04/03|