Introduction
Although many plants with interesting phenotypes have been generated by
genetic manipulation, the central metabolic objective of being able to make predictable
changes to specified fluxes generally remains elusive. The numerous
reports of engineered plants with metabolic phenotypes that are not usefully
different from the wild type, for example, in starch metabolism (Fernie
et al., 2002), show that the rational manipulation of plant metabolism is far from
straightforward, and that in many instances our understanding of plant metabolic
networks is insufficient to permit accurate predictions about the metabolic consequences
of genetic manipulation. Unexpected metabolic phenotypes are interesting
in their own right since they often provide information about the structure
and regulatory properties of the network, but from an engineering perspective,
they are undesirable since they consume resources and reduce the efficiency of
the process.
If the production of unwanted metabolic phenotypes is to be avoided, then
metabolic engineering has to be based on a detailed quantitative understanding of
the capabilities of the metabolic network. Essentially this requires: (1) definition of
the network of reactions, (2) definition of all the molecular interactions in the
system that have an impact on the functioning of the network, and (3) specification
of the intracellular and external environments in which the network is
functioning. Unfortunately, each of these requirements is potentially very
demanding: the plant metabolic network is of necessity complex, reflecting the
demands placed on sessile organisms that live in a fluctuating environment;
this complexity increases the scope for regulation of the network through
changes in enzyme level (via changes in gene expression and protein turnover)
and enzyme activity (via covalent modification, effector binding, and changes in
substrate and product concentrations); and for most purposes, plants have to be
grown under non–steady-state conditions, thus complicating any prediction
of metabolic performance. The net result of these complications is that models of
plant metabolism (Giersch, 2000; Morgan and Rhodes, 2002) tend to be relatively
limited in scope and to fall some way short of the virtual cell that is required if
accurate predictions are to be made of the impact of genetic manipulation on
metabolic fluxes.
Three topics central to the development of a quantitative understanding of the
metabolic capabilities of plant cells are discussed in this chapter. First, the complexity
of the plant metabolic network is described and the prospects for obtaining
a complete description of the network are assessed. Second, a review is provided
of some of the tools that are now available for understanding the structure and
performance of the network. Finally, to emphasize the level of sophistication that
is required for models with real predictive value, we review some landmark
studies that highlight the complexity of the system-wide mechanisms that permit
the integration of plant metabolism. The emphasis is on the primary pathways of
carbon metabolism since these pathways are fundamentally important for the
functioning and manipulation of the network.