To establish plant culture systems for product synthesis, a multi-scale engineering approach is necessary. At the intracellular level, the influx of 'omics' data has necessitated development of new methods to properly annotate and establish useful metabolic models that can be applied to elucidate unknown steps in specialized metabolite biosynthesis, define effective metabolic engineering strategies and increase enzyme diversity available for synthetic biology platforms. On an intercellular level, the presence of aggregates in culture leads to distinct metabolic sub-populations. Recent advances in flow cytometric analyses and mass spectrometry imaging allow for resolution of metabolites on the single cell level, providing an increased understanding of culture heterogeneity. Finally, extracellular engineering can be used to enhance culture performance through media manipulation, co-culture with bacteria, the use of exogenous elicitors or modulation of shear stress.
