Engineering metabolic pathways in plants by multigene transformation

Metabolic engineering in plants can be used to increase the abundance of specific valuable metabolites, but single-point interventions generally do not improve the yields of target metabolites unless that product is immediately downstream of the intervention point and there is a plentiful supply of precursors. In many cases, an intervention is necessary at an early bottleneck, sometimes the first committed step in the pathway, but is often only successful in shifting the bottleneck downstream, sometimes also causing the accumulation of an undesirable metabolic intermediate. Occasionally it has been possible to induce multiple genes in a pathway by controlling the expression of a key regulator, such as a transcription factor, but this strategy is only possible if such master regulators exist and can be identified. A more robust approach is the simultaneous expression of multiple genes in the pathway, preferably representing every critical enzymatic step, therefore removing all bottlenecks and ensuring completely unrestricted metabolic flux. This approach requires the transfer of multiple enzyme-encoding genes to the recipient plant, which is achieved most efficiently if all genes are transferred at the same time. Here we review the state of the art in multigene transformation as applied to metabolic engineering in plants, highlighting some of the most significant recent advances in the field. Research at the Universitat de Lleida is supported by the Ministerio de Ciencia e Innovación (grants no. BFU2007-61413, BIO2011-23324, BIO02011-22525, PIM2010PKB-0074, Acciones complementarias BIO2007-30738-E and BIO2011-22525, and the Centre CONSOLIDER on Agrigenomics), the European Union Framework 7 Program (SmartCell Integrated Project 222716), the European Research Council IDEAS Advanced Grant Program (BIOFORCE) (to PC), the European Cooperation in Science and Technology (COST Action FA0804), and RecerCaixa.