We have chosen to focus on lignification as an important
aspect of wood formation. Lignin is a complex polymer of
three phenylpropanoid subunits deposited in the cell walls of
vascular elements after cell growth has ceased. Sucrose supplied
by the phloem is transformed into phenylalanine via the
shikimic acid pathway, and lignin monomers are produced
from phenylalanine by a branch of phenylpropanoid metabolism.
The pathway of lignin precursor biosynthesis is well
established (see ref. 7 for review). The first step of the phenylpropanoid
pathway is deamination of phenylalanine by
' Supported by the North Carolina State University Forest Biotechnology
Industrial Associates Group.
PAL2 (EC 4.3.1.5) to yield trans-cinnamic acid. Hydroxylation
of cinnamic acid produces p-coumaric acid; additional
hydroxylation and methylation of p-coumarate produce first
ferulic acid, then sinapic acid. These three acids are coupled
to CoA and reduced to the corresponding alcohols. The end
products of this pathway are p-coumaryl, coniferyl, and sinapyl
alcohols. The relative proportions of the three precursors
in lignin varies from species to species and even between cell
types within a single individual (14). In general, lignin in
angiosperms contains more p-coumaryl and sinapyl alcohol
residues than does gymnosperm lignin; coniferyl alcohol is
the primary lignin monomer in most gymnosperms.
The regulation of synthesis and polymerization of lignin
monomers is still poorly understood. Synthesis of lignin monomers
could be regulated at multiple points; the entry of
sugars into the shikimic acid pathway or the entry of phenylalanine
into the phenylpropanoid pathway are obvious possibilities.
Feedback regulation of many or all of the enzymes
in the pathway is another alternative. None of the intermediates
in the phenylpropanoid pathway, from phenylalanine
to coniferyl alcohol, accumulates to significant levels in differentiating
xylem of lodgepole pine (24). The only ligninrelated
compound that does accumulate is coniferin, the 4-0-
f3-D-glucoside of coniferyl alcohol (25). This observation supports
the suggestion (26) that the enzymes involved in synthesizing
lignin monomers may be coordinately regulated and
arranged in multi-enzyme complexes, so that intermediates
are not allowed to accumulate.