HARNESSING PLANT BIOMASS FOR BIOFUELS AND BIOMATERIALS

Cell-wall carbohydrates and their modication as a resource

for biofuels
Markus Pauly
*
and Kenneth Keegstra
Department of Energy Plant Research Laboratory and Department of Biochemistry and Molecular Biology, Michigan State
University, East Lansing, MI 48824, USA
Received 1 December 2007; revised 5 February 2008; accepted 8 February 2008.
*
For correspondence (fax +1 517 353 9168; e-mail paulymar@msu.edu).
Summary
Plant cell walls represent the most abundant renewable resource on this planet. Despite their great abundance,
only 2% of this resource is currently used by humans. Hence, research into the feasibility of using plant cell
walls in the production of cost-effective biofuels is desirable. The main bottleneck for using wall materials is
the recalcitrance of walls to efcient degradation into fermentable sugars. Manipulation of the wall
polysaccharide biosynthetic machinery or addition of wall structure-altering agents should make it possible
to tailor wall composition and architecture to enhance sugar yields upon wall digestion for biofuel
fermentation. Study of the biosynthetic machinery and its regulation is still in its infancy and represents a
major scientic and technical research challenge. Of course, any change in wall structure to accommodate
cost-efcient biofuel production may have detrimental effects on plant growth and development due to the
diverse roles of walls in the life of a plant. However, the diversity and abundance of wall structures present in
the plant kingdom gives hope that this challenge can be met.
Keywords: cell walls, polysaccharide biosynthesis, hemicellulose, biofuel.
Plant cell walls are the most abundant renewable resource
on this planet
It has been estimated that the net CO
2
xation by land plants
per year is approximately 56 10
9
tonnes (Table 1) (Field
et al., 1998) and that the worldwide biomass production by
land plants is 170200 10
9
tonnes (Lieth, 1975). Of this
amount, 70% is estimated to represent plant cell walls
(Duchesne and Larson, 1989; Poorter and Villar, 1997).
Humans use these wall materials mainly in the formof wood
for heat production (Table 2), and as a building material
(timber), in the pulp and paper industry (Fenning and
Gershenzon, 2002), and as raw material in the textile
industry, e.g. cotton bers (http://faostat.fao.org/site/567/
default.aspx; parameter settings: production quantity, cot-
ton lint, world, 2006). Taken together, only 2% of the plant
cell-wall-based biomass is currently utilized by humans
(Table 2). It is thus not surprising that interest in using this
resource as a material for biofuels has increased in recent
years (Schubert, 2006). Important advantages of wall
materials as feedstocks for biofuel production are their great
abundance and the fact that they do not serve as food for
animals and humans as starch does, for example.
All cell walls of higher plants contain cellulose, a homo-
polymer of b-1,4-linked glucose units, mainly in the form of
crystalline microbrils as well as in an amorphous form
(Carpita and McCann, 2000). Walls also contain hemicellu-
loses (Figure 1), such as substituted glucans, xylans and/or
mannans, and anionic components such as the galacturonic
acid-containing pectic polysaccharides. Walls may also
contain polyphenols such as lignins, and, to a minor extent,
structural proteins. The prevalence of polysaccharides in the
wall is particularly advantageous for plants, as they are
generated directly from the products of photosynthesis
without the utilization of large amounts of nitrogen or phos-
phorus, two macronutrients that frequently limit plant
growth.
Not all cell walls have the same polysaccharide composition
When considering wall materials for the production of bio-
fuels, one should be aware that walls from higher plants
2008 The Authors 559
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The Plant Journal (2008) 54, 559568 doi: 10.1111/j.1365-313X.2008.03463.x
differ quite substantially in content, both qualitatively and
quantitatively. While the cell is still elongating, a primary
wall is formed. Primary walls contain cellulose and a
hydrated (65% water) matrix consisting of hemicelluloses
and pectins, with some structural proteins (Brett and
Waldron, 1996). Based on their polysaccharide composition,
primary cell walls are usually classied as type I or type II.
Type I walls are present in dicots and non-commelinoid
monocots; in addition to cellulose, they generally contain
xyloglucan as the main hemicellulose and abundant
amounts of pectic polysaccharides (Carpita and Gibeaut,
1993). In type II walls, the walls of the Poales, such as the
grasses, arabinoxylan is the major hemicellulose. In
addition, type II walls contain a higher percentage of
cellulose and only negligible amounts of pectins and
proteins (Carpita, 1996). Secondary walls, deposited once
cell elongation ceases, are usually thicker than primary
walls and may be deposited in a number of layers. Sec-
ondary walls contain cellulose and arabinoxylan and/or
glucomannans as the major hemicellulose (Brett and
Waldron, 1996). More importantly, in secondary walls,
water is largely replaced by lignin, making them nearly
impenetrable to solutes and enzymes. At the single-plant
level, nearly all of the approximately 35 different cell
types can be distinguished based on their varying wall
structures, as observed by microscopy (Carpita and
McCann, 2000), chemical composition analysis (Richmond
and Somerville, 2001) and labeling of wall polymers with
specic antibodies (Willats et al., 2000). Hence, it is not
Table 2 Annual human utilization of plant cell walls
Material Product Tonnes per year
Wood Energy 1.05 10
9
Wood Timber, pulp and paper 0.95 10
9
Figure 1. Examples of hemicellulose structures present in cell walls, including monosaccharides, and their linkage and ester constituents (modied fromSomerville
et al., 2004). Easily fermentable hexoses are distinguished by their square shape. Other sugars are shown in different shapes.
Table 1 Annual production of plant cell walls
Production Source Tonnes
Assimilated CO
2
Land plants, net primary
production
56 10
9
Plant biomass Land, worldwide 170200 10
9
Cell walls Land, worldwide 150170 10
9
560 Markus Pauly and Kenneth Keegstra
2008 The Authors
Journal compilation 2008 Blackwell Publishing Ltd, The Plant Journal, (2008), 54, 559568
surprising that plant-wall feedstocks that could be used
for biofuel production can differ quite signicantly in their
composition, even though the materials derive mainly
from secondary walls (Figure 2). In Table 3, some of the
prominent plant materials that could be used for biofuel
production are listed. Although the numbers cannot be
compared with each other because different methods
were used to establish them, some general features
become evident. The most dominant polysaccharide in
these walls is cellulose, making up 40.651.2% of the wall
material. The next largest fraction comprises the hemi-
celluloses, representing 28.537.2% of the walls. Lignin
occurs in these walls at a lower percentage (13.628.1%),
with a more than twofold difference between walls of
switchgrass, for example, and those of softwood. Viewed
in greater detail, composition is seen to vary widely with
regard to the hemicelluloses (Table 3, and references cited
therein; for structures, see Figure 1). The grasses contain
mainly arabinoxylan, but the degree of arabinosylation
can vary greatly. Wheat straw contains the lowest degree
of substitution, whereas sorghum xylans have an excep-
tionally high degree of substitution (Verbruggen et al.,
1995). Hardwood (from angiosperm trees) also contains
mainly xylan, but with a negligible degree of arabinosy-
lation. The xylans here are mainly substituted with
glucuronic acid or 4-O-methyl-glucuronic acid residues
(Ebringerova and Heinze, 2000). In contrast, softwood
(from conifers) contains mannans such as O-acetylated
galactoglucomannans (Capek et al., 2002) as their main
hemicellulose, although ample amounts of xylans are also
present. Most importantly, the differences and ranges
of wall components and their ne structures are also a
result of the differences in tissues from which the
feedstocks are derived (e.g. corn stover; Chundawat et al.,
2007).
Bottlenecks in utilizing cell-wall materials for biofuels
Plant cell-wall materials can be converted in a number of
ways. One way is the combustion and gasication of plant
material. The resulting CO and hydrogen gas (also called
syngas) can be converted to hydrocarbons of various
lengths via a catalyzed chemical reaction (FischerTropsch
process). Hydrocracking of the large hydrocarbons can be
used to produce diesel fuels (Tijmensen et al., 2002). Here,
the main objective from a cell-wall perspective is simply an
Figure 2. Cell-wall polymer composition (cellulose, hemicellulose and lignin)
and hemicellulose composition for a variety of plant materials that are
currently under discussion for use as biofuel feedstocks (based on the data
shown in Table 3 and references therein).
Table 3 Comparison of biomass feedstocks
Feedstock
a
Cellulose Hemi cellulose Lignin Ash Protein Solubles Reference
Corn stover
b
39.4 33.1 14.9 ND 3.7 8.9 Chundawat et al. (2007)
Wheat straw 34.9 22.5 21.3 9.4 ND 11.9 Lynd et al. (1999)
Rice straw 41.6 31.5 12.5 14.4
a
ND ND Jin and Chen (2007)
Miscanthus 41.9 26.6 13.3 3.2 ND 15.0 Magid et al. (2004)
Sorghum (whole sorghum
pith and bark)
15.0 12.3 5.8 0.4 ND 66.5
b
Billa et al. (1997)
Switchgrass (late cut) 46.1 32.2 12.3 4.7 4.6 ND Lynd et al. (1999)
Sugar cane 48.6 31.1 19.1 1.2 ND ND Sanjuan et al. (2001)
Hardwood (beech; Fagus sylvatica) 43.3 31.8 24.4 0.5 ND ND Fengel and Wegener (1989)
Softwood (spruce; Picia abies) 40.4 31.1 28.0 0.5 ND ND Fengel and Wegener (1989)
Values have been adjusted to a percentage basis (dry weight).
a
Mainly silicate.
b
Mainly sucrose.
ND, not determined in these studies.
Cell-wall carbohydrates 561
2008 The Authors
Journal compilation 2008 Blackwell Publishing Ltd, The Plant Journal, (2008), 54, 559568
increase in the production of biomass (cell walls) per
hectare, irrespective of its wall composition, although a
low water and ash content is desirable (Tijmensen et al.,
2002). A more sophisticated approach in biofuel produc-
tion involves the degradation of wall materials to mono-
saccharides and subsequent fermentation to liquid fuels
such as bioethanol (Schubert, 2006). However, plants have
evolved wall structures to accommodate their needs in
completing their lifecycle, not to suit mankinds desire to
exploit this resource for the production of biofuels. As a
result, cell walls are naturally resistant to breakdown by
mechanical and microbial forces, which are precisely the
processes needed for the cost-effective and efcient pro-
duction of monosaccharides. Hence, one major objective is
to make walls more accessible to degradation (Himmel
et al., 2007; Houghton et al., 2006). We could achieve this
goal by increasing water solubility and hence access of
enzymes to polysaccharides. One way to do this would be
to add de novo synthesized, water-soluble polysaccharides
to existing cells, leading to a greater abundance of these
polysaccharides in the wall. One alternative would be a
shift in the ratio of less soluble polysaccharides to soluble
ones. This objective would require: (1) for cellulose, an
increase in the abundance of amorphous glucan chains
rather than crystalline microbrils; (2) for hemicelluloses,
addition of side chains to decrease hydrogen bonding with
cellulose microbrils; and (3) for lignins, a general reduc-
tion in their amount or amendment to a more easily
degradable form (Akin, 2007), for example by introduction
of specic monolignols (Boerjan et al., 2003; Chen and
Dixon, 2007) and/or decrease of the existing ligninhemi-
cellulose linkages (Grabber et al., 2002). Another consid-
eration is the fermentability of the wall degradation
products, the resulting monosaccharides. Currently, the
sugars most easily fermentable by yeasts are the hexoses,
such as glucose and mannose, rather than the pentoses,
although yeast and bacterial strains have been developed
that can efciently ferment pentoses (Chu and Lee, 2007).
Hence, an increased production of hexose-containing
polymers such as cellulose, glucomannans and to some
extent xyloglucan is more desirable than an increase in
arabinoxylans, for example (see hexose and pentose
annotation in Figure 1). Depending on the fermentation- or
catalyst-based chemical process (Huber et al., 2005) used
to produce fuels, monosaccharide fermentation-inhibiting
components, such as aliphatic acids (e.g. acetic acid) or
phenolic compounds (Larsson et al., 1999) are present to
varying degrees in the degraded biomass. One goal should
be to reduce the abundance of such compounds to a
minimum.
All of the above-mentioned changes could be accom-
plished by either manipulation of the plant biosynthetic
pathways for the respective polymers and/or post-deposi-
tion metabolism alterations in planta.
Manipulation of the biosynthetic pathways
The natural variability in wall compositional quantity and
quality (Figure 2) suggests that there is an opportunity for
altering the abundance of specic wall components without
compromising the life cycle of a plant. Such a feat could
be accomplished by manipulation of the biosynthesis of
specic wall polysaccharides.
The two most abundant polysaccharides of plant cell
walls, cellulose and hemicellulose, are synthesized in
different compartments by signicantly different processes.
Cellulose, generally the most abundant component in
secondary cell walls (see Figure 1), is synthesized at the
plasma membrane by a complex machinery that we are just
beginning to understand (Somerville, 2006). Whereas the
glucosyl residues come from UDP-glucose molecules that
are present in the cytosol, the cellulose microbrils are
deposited into the extracellular wall matrix at a location
adjacent to the plasma membrane (Somerville, 2006). On the
other hand, the hemicellulosic polysaccharides present are
synthesized in the Golgi and packaged into secretory
vesicles before delivery to the cell surface and incorporation
into the wall matrix. The assembly events that combine
these components into the composite that exists in the wall
matrix are still poorly understood.
The CesA proteins are thought to be the catalytic subunits
of the cellulose synthase complexes (Somerville, 2006).
These proteins are encoded by a family of CesA genes that
are found throughout the plant kingdom (Hazen et al., 2002;
Richmond and Somerville, 2000). Genetic studies have led to
the conclusion that three CesA genes are needed for
cellulose biosynthesis in primary cell walls (Persson et al.,
2007b), and another set of three CesA genes is required for
cellulose synthesis in secondary cell walls (Somerville,
2006). The three different CesA proteins are thought to
cluster into a higher-order structure, which forms the rosette
structure observed in the plasma membrane (Somerville,
2006).
The rosettes containing multiple CesA proteins are
thought to move in the plasma membrane in a direction
that is dened by cortical microtubules, thereby producing
cellulose microbrils outside the plasma membrane. These
microbrils are deposited in a pattern that reects the
orientation of the cortical microtubules present on the
cytosolic side of the plasma membrane (Paredez et al.,
2006). Despite this emerging outline of how cellulose is
deposited, many important issues remain unresolved.
Genetic experiments provide evidence that additional pro-
teins are involved in cellulose deposition (Lane et al., 2001;
Pagant et al., 2002). One of these proteins has been shown to
be a membrane-bound endoglucanase/cellulase and is
thought to act as an editing/repairing protein during
cellulose biosynthesis (Mlhoj et al., 2002). However, the
precise roles of this and the other proteins are still not clear.
562 Markus Pauly and Kenneth Keegstra
2008 The Authors
Journal compilation 2008 Blackwell Publishing Ltd, The Plant Journal, (2008), 54, 559568
Whether the encoded proteins are part of the rosette
structure or whether they have other roles in cellulose
deposition remain to be determined.
As more is learned about the details of cellulose biosyn-
thesis, it may be possible to alter these processes in ways
that would render the walls more easily digestible during
processing to biofuels. For example, if one understood the
details of how the glucan chains come together to form
crystalline cellulose, it might be possible to modify this
process such that cellulose microbrils would have larger
amorphous regions.
The biosynthesis of hemicellulosic polysaccharides in the
Golgi apparatus differs signicantly from cellulose biosyn-
thesis. In the case of the mannans, the backbone is synthe-
sized by CslA proteins (Liepman et al., 2007) that have been
identied in a number of species (Dhugga et al., 2004;
Liepman et al., 2005, 2007; Suzuki et al., 2006). Each plant
species for which the complete genome is available has a
small family of CslA genes that are part of the CesA
superfamily. The CslA proteins produced in heterologous
systems not only have the ability to synthesize mannan
when GDP-mannose is present, they also have the ability to
synthesize glucomannan when a mixture of GDP-mannose
and GDP-glucose is present (Liepman et al., 2005, 2007;
Suzuki et al., 2006). Thus, the same protein is able to
incorporate both sugars into the backbone in vitro, and it
is likely that the same proteins produce both mannans and
glucomannans in vivo.
The degree of galactosylation of the mannan backbone
has implications for mannan solubility. Mannans with a low
degree of galactosyl substitution have limited solubility in
water, whereas polymers with a high degree of substitution
have important properties as emulsiers (Reid et al., 1988).
The galactosyltransferase enzymes that add side chains to
the mannan and glucomannan backbones have been iden-
tied and characterized (Edwards et al., 1999, 2002). The
levels of these enzymes appear to control the degree of
substitution of the backbone, with the side chains being
added in patterns that are described by hidden Markov
models (Edwards et al., 2004). Altering the degree of side-
chain substitutions will be vital in engineering more soluble
mannans.
The mannans are attractive candidates for enhancing wall
composition with the aimof creating improved biofuel crops
for several reasons. First, the genes and proteins needed for
mannan biosynthesis have been identied. Second, the
genes needed for mannan biosynthesis appear to be present
in all land plants, although their expression levels are such
that few mannans are present in the walls of most angio-
sperms. However, mannans accumulate to high levels in the
seeds of many plants, where they serve as storage carbo-
hydrates (Meier and Reid, 1982). During germination, seed-
lings have the ability to rapidly degrade the mannans and
use the resulting sugars as a source of carbon for early
seedling development. Because the released sugars are all
hexoses, they can easily speed up the central metabolism of
the developing seedling. Given these circumstances, it may
be possible to enhance mannan levels in vegetative tissues
such that the polymers could be easily degraded after
harvest to yield hexoses, which could be converted to
biofuels more efciently than the pentoses released fromthe
more abundant xylans.
Although xyloglucan is found mainly in the primary cell
walls of many plants (Hayashi, 1989), its biosynthesis is
relevant to our general understanding of hemicellulose
biosynthesis and therefore will be briey summarized here.
All the glycosyltransferases involved in synthesis of its side
chains have been tentatively identied (see reviews by
Scheible and Pauly, 2004; Lerouxel et al., 2006). However, it
remains unclear how the enzymes achieve the structural
side-chain diversity found in this polymer.
Cocuron et al. (2007) recently presented evidence that the
glucan synthase required for making the backbone of
xyloglucan is encoded by a CslC gene. When CslC genes
were expressed in Pichia pastoris cells, the cells accumu-
lated signicant quantities of oligosaccharides containing b-
1,4-linked glucosyl residues. When one of the xyloglucan
xylosyltransferase genes, which is responsible for substitut-
ing the glucan backbone with xylosyl residues (Faik et al.,
2002), was co-expressed with the CslC gene, the cells
produced large quantities of unsubstituted b-1,4-glucan
(Cocuron et al., 2007). These observations provide evidence
that these xylosyltransferase and glucan synthase enzymes
interact to form a complex that has an impact on the nature
of the resulting product, even though one of them does not
exhibit any activity. Further work is needed to conrm this
interesting hypothesis, as protein complexes involved in
hemicellulose biosynthesis have yet to be discovered.
Given that xylans are the most abundant hemicellulose
present in the secondary walls of plants being considered for
use in biofuel production (see Figure 1), it is unfortunate that
we know so little about their biosynthesis. Recently, several
groups have begun to make progress in this difcult area.
One of the most interesting observations comes from the
work of Pen a et al. (2007), who examined the xylan polysac-
charides present in two mutant lines of Arabidopsis that
have irregular xylem phenotypes. First, these authors redis-
covered an older, but little noticed, observation that xylan
polysaccharides often have an unusual oligosaccharide at
the reducing end. This oligosaccharide contains the glycosyl
sequence 4-b-D-Xyl-(1,4)-b-D-Xyl-(1,3)-a-L-Rha-(1,2)-a-D-GalA-
(1,4)-D-Xyl. Because it is at the reducing end of the polysac-
charide, it is possible that this oligosaccharide serves as the
primer for chain initiation, if chain elongation occurs from
the reducing end toward the non-reducing end, as is
commonly hypothesized. Both of the mutants have reduced
levels of xylan in the secondary walls of xylem elements,
leading to the irregular xylem phenotype.
Cell-wall carbohydrates 563
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Journal compilation 2008 Blackwell Publishing Ltd, The Plant Journal, (2008), 54, 559568
One of the mutants, irx9, has increased levels of the
unusual oligosaccharide, but the chains containing it are
shorter than in wild-type plants, suggesting that the IRX9
gene is involved in elongating the xylan chains. On the other
hand, the other mutant, irx8, has little of the unusual
oligosaccharide and lower quantities of xylan (Persson
et al., 2007a); the xylan that is present is longer and more
heterodisperse in size. These observations suggest that IRX8
may be involved in synthesizing the unusual oligosaccha-
ride, which may serve as a primer in wild-type plants. These
observations highlight the complexity of xylan biosynthesis,
but offer some hope that these new observations can lead to
an improved understanding of this important cell-wall
polymer.
As illustrated above, much is to be learned about the
biosynthetic machinery of polysaccharides. We are just
beginning to understand the carbon ux into the specic
wall polysaccharides (Sharples and Fry, 2007) and regulation
of polysaccharide biosynthesis.
Post-deposition wall changes
Another way to make wall structures more enzyme-acces-
sible is to add loosening agents through transgenic
approaches. Such agents include the expansins, plant pro-
teins that have been shown to induce the extensibility of
plant tissues under stress (Cosgrove, 2000). The precise
mechanism of expansin action is unknown, but it is thought
that they act at the interface of hemicellulose polymers and
cellulose microbrils (Cosgrove, 2000). Proteins with a
similar mode of action are the fungal swollenins, proteins
that consist of a cellulose-binding domain and an expansin-
like domain. They are thought to disrupt cellulose micro-
brils without hydrolytic activity, i.e. the release of reducing
sugars (Saloheimo et al., 2002). Adding expansins to wall
materials can double the yield of sugars released by fungal
cellulases (Cosgrove, 2001a,b), and it is expected that
swollenin might have a similar effect.
Another class of proteins that could be used to make
the wall more accessible is glycanases, in particular
endoglucanases. Expression of a poplar endoglucanase
in Arabidopsis leads to increased cell elongation and
subsequent plant growth (Park et al., 2003). A similar
effect was found when a fungal xyloglucanase (a xylo-
glucan-specic endoglucanase; Pauly et al., 1999) was
expressed in poplar (Park et al., 2004). This is not
surprising, as this enzyme is the only protein other than
expansin that is known to induce wall extension (Yuan
et al., 2001). One effect in the transgenic poplar material
is an observed increase in cellulose, which leads to
material with higher hexose content. A side-effect of the
enhanced growth of these transgenic plants is an increase
in the photosynthetic canopy, potentially allowing more
biomass to accumulate.
Other agents that work on the hemicellulosecellulose
network are xyloglucan transglycosylases/hydrolases. This
class of enzymes is thought to be involved in either
incorporating newly synthesized hemicelluloses and/or
remodeling existing hemicelluloses present in the wall by
loosening/re-ligating xyloglucan (Fry et al., 1992; Nishitani,
1997). It has been demonstrated that xyloglucan transgly-
cosylases/hydrolases are active in cell elongation and act at
the cellulose/xyloglucan interface (Vissenberg et al., 2000,
2005). Manipulating the levels of this agent thus has the
potential to loosen cell-wall structures.
Other examples of glycanases that have been expressed
in plants are pectin-degrading enzymes. Expression of a
galactanase in potato tubers led to signicant wall alteration
(i.e. reduction of galactans), but had no effect on plant or
tuber development (Srensen et al., 2000). Interestingly, the
tubers exhibited a marked change in physical tissue prop-
erties (Ulvskov et al., 2005). In particular, the water-binding
capacity was changed, indicating that removing pectin side
chains would probably render such wall material less
degradable.
Agents that work on the hemicelluloselignin interface,
i.e. that break the covalent bonds between the polymers, can
lead to more easily degradable wall materials. For example,
expression of phenolic esterases improves the release of
fermentable sugar (Akin, 2007).
Modication of plant cell walls will challenge
their biological function
Any strategy to improve wall materials in planta as feed-
stocks for biofuels needs to take into account the possibility
of functional failure of the cell wall, which could be detri-
mental to plant growth, leading to a concomitant reduction
in wall biomass and ultimately threatening the very survival
of the plant.
Cell walls are important for structural integrity of the
cell and indeed the whole plant. Through the evolutionary
introduction of polyphenol incorporation into the wall,
land plants were able to increase their sunlight-harvesting
capacity by increasing the plant canopy, not only in terms
of width, but also in terms of height, enabling the plant to
compete with other plants for sunlight. A number of
examples have demonstrated that altering walls can lead
to structural concessions such as dwarsm (Desprez et al.,
2007) and even to lethality (Goubet et al., 2003). Cell walls
determine the shape of the cell, so altering them can lead
to changes in morphology, such as an irregular xylem
(Turner et al., 2007), that may be disadvantageous to
water transport. Walls, in particular the pectinaceous
middle lamellae, ensure attachment of the cells. A num-
ber of mutants with altered pectic polysaccharides have
been shown to have reduced cell adhesion (Bouton et al.,
2002; Iwai et al., 2002; Krupkova et al., 2007), probably
564 Markus Pauly and Kenneth Keegstra
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Journal compilation 2008 Blackwell Publishing Ltd, The Plant Journal, (2008), 54, 559568
limiting the plants ability to withstand certain mechanical
stresses (such as wind). On the other hand, wall materials
from such plants might be more accessible to wall-
degrading enzymes, making processing of these plants
more rapid, easier, and thus more cost-effective. Another
function of the wall is keeping plant pathogens such as
bacteria and fungi away from the nutritious cytosolic
content of the cells. In addition to a simple mechanical
line of defense, walls contain signaling molecules that
allow the plant cell to recognize a pathogen attack and to
respond with various lines of defense (Cote and Hahn,
1994; Vorwerk et al., 2004). Alterations in wall composi-
tion and architecture thus also introduce the possibility of
increased susceptibility to pathogens, or endogenous
release of wall-derived oligosaccharides might lead to
disease symptoms. For example, expression of a fungal
arabinanase in potato tubers led to a severely stressed
plant morphology (Skjot et al., 2002), presumably through
the release of apoplastic arabinan-oligosaccharides. This
morphology was overcome when the enzyme was tar-
geted to the Golgi apparatus instead of the apoplast.
Targeting and retaining the arabinanase in the Golgi led
to plants and tubers with unaltered appearance but with a
signicant decrease in pectic arabinans.
In a few cases where inhibition of polysaccharide
biosynthesis through genetic engineering of the glycan
synthases was achieved, these alterations did not result in
diminishment of pathogen resistance (Jacobs et al., 2003),
and in some cases even increased resistance (Hernandez-
Blanco et al., 2007). The plant cell wall is a dynamic entity
that undergoes delicate metabolic changes during cell
elongation and differentiation. Throughout the elongation
process, the cell must balance loosening the wall with
maintaining turgor pressure and cohesiveness of the wall
structure. It is thought that metabolism of the hemicellu-
loses interlacing cellulose microbrils with wall-loosening
enzymes such as endoglucanases, xyloglucantransglycos-
ylases and expansins allows slippage of cellulose micro-
brils and thus controlled cell elongation (Cosgrove,
2001a,b). As cells elongate, new cell-wall material is
deposited (Refregier et al., 2004), probably leading to
strengthening of the wall. Consequently, changing the
abundance or structure of wall polymers may stiffen the
wall to the extent that the cell cannot enlarge effectively,
or may lead to mechanical failure and hence bursting of
the cell during the elongation process. It has become
clear that the plant cell has a hitherto unknown mecha-
nism for monitoring wall integrity and compensating for
change (Humphrey et al., 2007; Pilling and Ho fte, 2003).
Candidates for such a monitoring activity are plasma
membrane-localized, wall-associated kinases (Wagner and
Kohorn, 2001), which that have been shown to bind to
the pectin matrix in the apoplast (Kohorn et al., 2006).
Recently, another plasma membrane-localized receptor
has been identied that may also act as such a wall
sensor (Hematy et al., 2007). Manipulating putative sens-
ing mechanisms has the potential to overcome unex-
pected wall structural changes, even though they might
be benecial, such as decreasing lignin content but
increasing the relative content of cellulose (Hu et al.,
1999).
Concluding remarks
Owing to the abundance of cell-wall material generated
by plants, cell walls could play a prominent role in our
quest for the reduced utilization of carbon dioxide-emit-
ting fossil fuels. The scientic and technical challenges
inherent in realizing this goal are enormous. The pro-
duction of walls with tailored polysaccharide composition
and structures is still in its infancy due to our lack of
knowledge of polysaccharide biosynthesis and its regula-
tion. Despite the above-mentioned difculty of making the
wall polysaccharides more degradable, the current recal-
citrance of wall materials brings with it the advantage that
harvested wall materials, unlike grains and fruits, can be
stored relatively easily for extended periods without loss
of yield prior to factory processing. Also, identication of
specic bioenergy crop species with high biomass yields
grown in various climatic regions has just begun, as
have breeding programs for the increased production of
biomass.
References
Akin, D.E. (2007) Grass lignocellulose strategies to overcome
recalcitrance. Appl. Biochem. Biotechnol. 137, 315.
Billa, E., Koullas, D.P., Monties, B. and Koukios, E.G. (1997) Struc-
ture and composition of sweet sorghum stalk components.
Indust. Crops Prod. 6, 297302.
Boerjan, W., Ralph, J. and Baucher, M. (2003) Lignin biosynthesis.
Annu. Rev. Plant Biol. 54, 519546.
Bouton, S., Leboeuf, E., Mouille, G., Leydecker, M.T., Talbotec, J.,
Granier, F., Lahaye, M., Ho fte, H. and Truong, H.N. (2002) Quasi-
modo1 encodes a putative membrane-bound glycosyltransferase
required for normal pectin synthesis and cell adhesion in Ara-
bidopsis. Plant Cell, 14, 25772590.
Brett, C. and Waldron, K. (1996) Physiology and Biochemistry of
Plant Cell Walls. Topics in Plant Functional Biology. London:
Chapman and Hall.
Capek, P., Alfoldi, J. and Liskova, D. (2002) An acetylated galacto-
glucomannan from Picea abies L. Karst. Carbohydr. Res. 337,
10331037.
Carpita, N.C. (1996) Structure and biogenesis of the cell walls
of grasses. Annu. Rev. Plant Physiol. Plant Mol. Biol. 47, 445
476.
Carpita, N.C. and Gibeaut, D.M. (1993) Structural models of primary
cell walls in owering plants: consistency of molecular structure
with the physical properties of the walls during growth. Plant J. 3,
130.
Carpita, N. and McCann, M. (2000) The cell wall. In Biochemistry and
Molecular Biology of Plants (Buchanan, B.B., Gruissem, W. and
Cell-wall carbohydrates 565
2008 The Authors
Journal compilation 2008 Blackwell Publishing Ltd, The Plant Journal, (2008), 54, 559568
Jones, R.L., eds). Rockville, MD: American Society of Plant
Physiologists, pp. 52108.
Chen, F. and Dixon, R.A. (2007) Lignin modication improves
fermentable sugar yields for biofuel production. Nat. Biotechnol.
25, 759761.
Chu, B.C.H. and Lee, H. (2007) Genetic improvement of Saccharo-
myces cerevisiae for xylose fermentation. Biotechnol. Adv. 25,
425441.
Chundawat, S.P.S., Venkatesh, B. and Dale, B.E. (2007) Effect of
particle size based separation of milled corn stover on AFEX
pretreatment and enzymatic digestibility. Biotechnol. Bioeng. 96,
219231.
Cocuron, J.C., Lerouxel, O., Drakakaki, G., Alonso, A.P., Liepman,
A.H., Keegstra, K., Raikhel, N.V. and Wilkerson, C.G. (2007) A
gene from the cellulose synthase-like C family encodes a b1,4
glucan synthase. Proc. Natl Acad. Sci. USA, 104, 85508555.
Cosgrove, D.J. (2000) Loosening of plant cell walls by expansins.
Nature 407, 321326.
Cosgrove, D.J. (2001a) Wall structure and wall loosening. A look
backwards and forwards. Plant Physiol. 125, 131134.
Cosgrove, D.J. (2001b) Enhancement of Accessibility of Cellulose by
Expansins. US Patent No. 6326470.
Cote, F. and Hahn, M.G. (1994) Oligosaccharins: structures and
signal-transduction. Plant Mol. Biol. 26, 13791411.
Desprez, T., Juraniec, M., Crowell, E.F., Jouy, H., Pochylova, Z.,
Parcy, F., Ho fte, H., Gonneau, M. and Vernhettes, S. (2007)
Organization of cellulose synthase complexes involved in pri-
mary cell wall synthesis in Arabidopsis thaliana. Proc. Natl Acad.
Sci. USA, 104, 1557215577.
Dhugga, K.S., Barreiro, R., Whitten, B. et al. (2004) Guar seed
b-mannan synthase is a member of the cellulose synthase super
gene family. Science, 303, 363366.
Duchesne, L.C. and Larson, D.W. (1989) Cellulose and the evolution
of plant life. Bioscience, 39, 238241.
Ebringerova, A. and Heinze, T. (2000) Xylan and xylan derivatives
biopolymers with valuable properties. 1 Naturally occurring
xylans: structures, isolation procedures and properties. Macro-
mol. Rapid Commun. 21, 542556.
Edwards, M.E., Dickson, C.A., Chengappa, S., Sidebottom, C.,
Gidley, M.J. and Reid, J.S.G. (1999) Molecular characterization of
a membrane-bound galactosyltransferase of plant cell wall matrix
polysaccharide biosynthesis. Plant J. 19, 691697.
Edwards, M.E., Marshall, E., Gidley, M.J. and Reid, J.S.G. (2002)
Transfer specicity of detergent-solubilized fenugreek
galactomannan galactosyltransferase. Plant Physiol. 129,
13911397.
Edwards, M.E., Choo, T.S., Dickson, C.A., Scott, C., Gidley, M.J. and
Reid, J.S.G. (2004) The seeds of Lotus japonicus lines
transformed with sense, antisense, and sense/antisense galacto-
mannan galactosyltransferase constructs have structurally
altered galactomannans in their endosperm cell walls. Plant
Physiol. 134, 11531162.
Faik, A., Price, N.J., Raikhel, N.V. and Keegstra, K. (2002) An
Arabidopsis gene encoding an a-xylosyltransferase involved in
xyloglucan biosynthesis. Proc. Natl Acad. Sci. USA, 99,
77977802.
Fengel, D. and Wegener, G. (1989) Wood-Chemistry, Ultrastructure,
Reactions. Berlin: Walter de Gryter.
Fenning, T.M. and Gershenzon, J. (2002) Where will the wood come
from? Plantation forests and the role of biotechnology. Trends
Biotechnol. 20, 291296.
Field, C.B., Behrenfeld, M.J., Randerson, J.T. and Falkowski, P.
(1998) Primary production of the biosphere: integrating terrestrial
and oceanic components. Science, 281, 237240.
Fry, S.C., Smith, R.C., Renwick, K.F., Martin, D.J., Hodge, S.K. and
Matthews, K.J. (1992) Xyloglucan endotransglycosylase, a new
wall-loosening enzyme activity from plants. Biochem. J. 282,
821828.
Goubet, F., Misrahi, A., Park, S.K., Zhang, Z.N., Twell, D. and
Dupree, P. (2003) AtCSLA7, a cellulose synthase-like putative
glycosyltransferase, is important for pollen tube growth and
embryogenesis in Arabidopsis. Plant Physiol. 131, 547557.
Grabber, J.H., Ralph, J. and Hateld, R.D. (2002) Model studies of
ferulate-coniferyl alcohol cross-product formation in primary
maize walls: implications for lignication in grasses. J. Agric.
Food Chem. 50, 60086016.
Hayashi, T. (1989) Xyloglucans in the primary cell wall. Annu. Rev.
Plant Physiol. Plant Mol. Biol. 40, 139168.
Hazen, S.P., Scott-Craig, J.S. and Walton, J.D. (2002) Cellulose
synthase-like genes of rice. Plant Physiol. 128, 336340.
Hematy, K., Sado, P.E., Van Tuinen, A., Rochange, S., Desnos, T.,
Balzergue, S., Pelletier, S., Renou, J.P. and Ho fte, H. (2007) A
receptor-like kinase mediates the response of Arabidopsis cells to
the inhibition of cellulose synthesis. Curr. Biol. 17, 922931.
Hernandez-Blanco, C., Feng, D.X., Hu, J. et al. (2007) Impairment of
cellulose synthases required for Arabidopsis secondary cell wall
formation enhances disease resistance. Plant Cell, 19, 890903.
Himmel, M.E., Ding, S.Y., Johnson, D.K., Adney, W.S., Nimlos, M.R.,
Brady, J.W. and Foust, T.D. (2007) Biomass recalcitrance: engi-
neering plants and enzymes for biofuels production. Science, 315,
804807.
Houghton, J., Weatherwax, S. and Ferrell, J. (2006) Breaking
the Biological Barriers to Cellulosic Ethanol: A Joint Research
Agenda. Washington: US Department of Energy Ofce of Science
and Ofce of Energy Efciency and Renewable Energy, http://
www.doegenomestolife.org/biofuels.
Hu, W.J., Harding, S.A., Lung, J., Popko, J.L., Ralph, J., Stokke, D.D.,
Tsai, C.J. and Chiang, V.L. (1999) Repression of lignin biosyn-
thesis promotes cellulose accumulation and growth in transgenic
trees. Nat. Biotechnol. 17, 808812.
Huber, G.W., Chheda, J.N., Barrett, C.J. and Dumesic, J.A. (2005)
Production of liquid alkanes by aqueous-phase processing of
biomass-derived carbohydrates. Science, 308, 14461450.
Humphrey, T.V., Bonetta, D.T. and Goring, D.R. (2007) Sentinels
at the wall: cell wall receptors and sensors. New Phytol. 176,
721.
Iwai, H., Masaoka, N., Ishii, T. and Satoh, S. (2002) A pectin glu-
curonyltransferase gene is essential for intercellular attachment
in the plant meristem. Proc. Natl Acad. Sci. USA, 99, 16319
16324.
Jacobs, A.K., Lipka, V., Burton, R.A., Panstruga, R., Strizhov, N.,
Schulze-Lefert, P. and Fincher, G.B. (2003) An Arabidopsis callose
synthase, GSL5, is required for wound and papillary callose for-
mation. Plant Cell, 15, 25032513.
Jin, S.Y. and Chen, H.Z. (2007) Near-infrared analysis of the chem-
ical composition of rice straw. Indust. Crops Prod. 26, 207211.
Kohorn, B.D., Kobayashi, M., Johansen, S., Riese, J., Huang, L.F.,
Koch, K., Fu, S., Dotson, A. and Byers, N. (2006) An Arabidopsis
cell wall-associated kinase required for invertase activity and cell
growth. Plant J. 46, 307316.
Krupkova, E., Immerzeel, P., Pauly, M. and Schmulling, T. (2007) The
tumorous shoot development2 gene of Arabidopsis encoding a
putative methyltransferase is required for cell adhesion and
co-ordinated plant development. Plant J. 50, 735750.
Lane, D.R., Wiedemeier, A., Peng, L.C. et al. (2001) Temperature-
sensitive alleles of RSW2 link the KORRIGAN endo-1,4-beta-glu-
canase to cellulose synthesis and cytokinesis in Arabidopsis.
Plant Physiol. 126, 278288.
566 Markus Pauly and Kenneth Keegstra
2008 The Authors
Journal compilation 2008 Blackwell Publishing Ltd, The Plant Journal, (2008), 54, 559568
Larsson, S., Reimann, A., Nilvebrant, N.O. and Jonsson, L.J. (1999)
Comparison of different methods for the detoxication of ligno-
cellulose hydrolyzates of spruce. Appl. Biochem. Biotechnol. 77,
91103.
Lerouxel, O., Cavalier, D.M., Liepman, A.H. and Keegstra, K. (2006)
Biosynthesis of plant cell wall polysaccharides a complex
process. Curr. Opin. Plant Biol. 9, 621630.
Liepman, A.H., Wilkerson, C.G. and Keegstra, K. (2005) Expression
of cellulose synthase-like (Csl) genes in insect cells reveals that
CslA family members encode mannan synthases. Proc. Natl
Acad. Sci. USA, 102, 22212226.
Liepman, A.H., Nairn, C.J., Willats, W.G.T., Srensen, I., Roberts,
A.W. and Keegstra, K. (2007) Functional genomic analysis sup-
ports conservation of function among cellulose synthase-like
A gene family members and suggests diverse roles of mannans
in plants. Plant Physiol. 143, 18811893.
Lieth, H. (1975) Primary production of the major vegetation units of
the world. In Primary Production of the Biosphere (Lieth, H. and
Whittaker, R.H., eds). Berlin: Springer-Verlag, pp. 203215.
Lynd, L.R., Wyman, C.E. and Gerngross, T.U. (1999) Biocommodity
engineering. Biotechnol. Prog. 15, 777793.
Magid, J., Luxhoi, J. and Lyshede, O.B. (2004) Decomposition of
plant residues at lowtemperatures separates turnover of nitrogen
and energy rich tissue components in time. Plant Soil, 258, 351
365.
Meier, H. and Reid, J.S.G. (1982) Reserve polysaccharides other
than starch in higher plants. In Encyclopedia of Plant Physiology,
Vol. 13A (Loewus, F.A. and Tanner, W., eds). Berlin: Springer,
pp. 418471.
Mlhoj, M., Pagant, S.R. and Hofte, H. (2002) Towards under-
standing the role of membrane-bound endo-beta1,4-glucanases
in cellulose biosynthesis. Plant Cell Physiol. 43, 13991406.
Nishitani, K. (1997) The role of endoxyloglucan transferase in the
organization of plant cell walls. Int. Rev. Cytol. 173, 157206.
Pagant, S., Bichet, A., Sugimoto, K., Lerouxel, O., Desprez, T.,
McCann, M., Lerouge, P., Vernhettes, S. and Ho fte, H. (2002)
KOBITO1 encodes a novel plasma membrane protein necessary
for normal synthesis of cellulose during cell expansion in Ara-
bidopsis. Plant Cell, 14, 20012013.
Paredez, A.R., Somerville, C.R. and Ehrhardt, D.W. (2006) Visuali-
zation of cellulose synthase demonstrates functional association
with microtubules. Science, 312, 14911495.
Park, Y.W., Tominaga, R., Sugiyama, J., Furuta, Y., Tanimoto, E.,
Samejima, M., Sakai, F. and Hayashi, T. (2003) Enhancement of
growth by expression of poplar cellulase in Arabidopsis thaliana.
Plant J. 33, 10991106.
Park, Y.W., Baba, K., Furuta, Y., Iida, I., Sameshima, K., Arai, M. and
Hayashi, T. (2004) Enhancement of growth and cellulose accu-
mulation by overexpression of xyloglucanase in poplar. FEBS
Lett. 564, 183187.
Pauly, M., Andersen, L.N., Kauppinen, S., Kofod, L.V., York, W.S.,
Albersheim, P. and Darvill, A. (1999) A xyloglucan-specic endo-
beta1,4-glucanase from Aspergillus aculeatus: expression clon-
ing in yeast, purication and characterization of the recombinant
enzyme. Glycobiology, 9, 93100.
Pen a, M.J., Zhong, R., Zhou, G.-K., Richardson, E.A., ONeill, M.A.,
Darvill, A.G., York, W.S. and Ye, Z.-H. (2007) Arabidopsis irregular
xylem8 and irregular xylem9: implications for the complexity of
glucuronoxylan biosynthesis. Plant Cell, 19, 549563.
Persson, S., Caffall, K.H., Freshour, G., Hilley, M.T., Bauer, S.,
Poindexter, P., Hahn, M.G., Mohnen, D. and Somerville, C.
(2007a) The Arabidopsis irregular xylem8 mutant is decient in
glucuronoxylan and homogalacturonan, which are essential for
secondary cell wall integrity. Plant Cell, 19, 237255.
Persson, S., Paredez, A., Carroll, A., Palsdottir, H., Doblin, M.,
Poindexter, P., Khitrov, N., Auer, M. and Somerville, C.R. (2007b)
Genetic evidence for three unique components in primary
cell-wall cellulose synthase complexes in Arabidopsis. Proc. Natl
Acad. Sci. USA, 104, 1556615571.
Pilling, E. and Ho fte, H. (2003) Feedback from the wall. Curr. Opin.
Plant Biol. 6, 611616.
Poorter, H. and Villar, R. (1997) The fate of acquired carbon in plants:
chemical composition and construction costs. In Plant Resource
Allocation (Bazzaz, F.A. and Grace, J., eds). San Diego, CA: Aca-
demic Press, pp. 3972.
Refregier, G., Pelletier, S., Jaillard, D. and Ho fte, H. (2004) Interac-
tion between wall deposition and cell elongation in dark-grown
hypocotyl cells in Arabidopsis. Plant Physiol. 135, 959968.
Reid, J.S.G., Edwards, M. and Dea, I.C.M. (1988) Enzymatic modi-
cation of natural seed gums. In Gums and Stabilizers for the Food
Industry (Phillips, G.O., Wedlock, D.J. and Williams, P.A., eds).
Washington DC: IRL Press, pp. 391398.
Richmond, T.A. and Somerville, C.R. (2000) The cellulose synthase
superfamily. Plant Physiol. 124, 495498.
Richmond, T.A. and Somerville, C.R. (2001) Integrative approaches
to determining Csl function. Plant Mol. Biol. 47, 131143.
Saloheimo, M., Paloheimo, M., Hakola, S., Pere, J., Swanson, B.,
Nyyssonen, E., Bhatia, A., Ward, M. and Penttila, M. (2002)
Swollenin, a Trichoderma reesei protein with sequence similarity
to the plant expansins, exhibits disruption activity on cellulosic
materials. Eur. J. Biochem. 269, 42024211.
Sanjuan, R., Anzaldo, J., Vargas, J., Turrado, J. and Patt, R. (2001)
Morphological and chemical composition of pith and bers from
Mexican sugarcane bagasse. Holz als Roh- und Werkstoff. 59,
447450.
Scheible, W.-R. and Pauly, M. (2004) Glycosyltransferases and cell
wall biosynthesis: novel players and insights. Curr. Opin. Plant
Biol. 7, 285295.
Schubert, C. (2006) Can biofuels nally take center stage? Nat.
Biotechnol. 24, 777784.
Sharples, S.C. and Fry, S.C. (2007) Radioisotope ratios discriminate
between competing pathways of cell wall polysaccharide and
RNA biosynthesis in living plant cells. Plant J. 52, 252262.
Skjot, M., Pauly, M., Bush, M.S., Borkhardt, B., McCann, M.C. and
Ulvskov, P. (2002) Direct interference with rhamnogalacturonan I
biosynthesis in Golgi vesicles. Plant Physiol. 129, 95102.
Somerville, C. (2006) Cellulose synthesis in higher plants. Annu.
Rev. Cell Dev. Biol. 22, 5378.
Somerville, C., Bauer, S., Brininstool, G. et al. (2004) Toward a
systems approach to understanding plant-cell walls. Science,
306, 22062211.
Srensen, S.O., Pauly, M., Bush, M., Skjot, M., McCann, M.C.,
Borkhardt, B. and Ulvskov, P. (2000) Pectin engineering: modi-
cation of potato pectin by in vivo expression of an endo-1,4beta-
D-galactanase. Proc. Natl Acad. Sci. USA, 97, 76397644.
Suzuki, S., Li, L.G., Sun, Y.H. and Chiang, V.L. (2006) The cellulose
synthase gene superfamily and biochemical functions of xylem-
specic cellulose synthase-like genes in Populus trichocarpa.
Plant Physiol. 142, 12331245.
Tijmensen, M.J.A., Faaij, A.P.C., Hamelinck, C.N. and van
Hardeveld, M.R.M. (2002) Exploration of the possibilities for
production of Fischer Tropsch liquids and power via biomass
gasication. Biomass Bioenerg. 23, 129152.
Turner, S., Gallois, P. and Brown, D. (2007) Tracheary element
differentiation. Annu. Rev. Plant Biol. 58, 407433.
Ulvskov, P., Wium, H., Bruce, D., Jorgensen, B., Qvist, K.B., Skjot,
M., Hepworth, D., Borkhardt, B. and Sorensen, S.O. (2005)
Biophysical consequences of remodeling the neutral side chains
Cell-wall carbohydrates 567
2008 The Authors
Journal compilation 2008 Blackwell Publishing Ltd, The Plant Journal, (2008), 54, 559568
of rhamnogalacturonan I in tubers of transgenic potatoes. Planta,
220, 609620.
Verbruggen, M.A., Beldman, G. and Voragen, A.G.J. (1995) The
selective extraction of glucuronoarabinoxylans from sorghum
endosperm cell-walls using barium and potassium hydroxide
solutions. J. Cereal Sci. 21, 271282.
Vissenberg, K., Martinez-Vilchez, I.M., Verbelen, J.P., Miller, J.G.
and Fry, S.C. (2000) In vivo colocalization of xyloglucan endo-
transglycosylase activity and its donor substrate in the elongation
zone of Arabidopsis roots. Plant Cell, 12, 12291237.
Vissenberg, K., Fry, S.C., Pauly, M., Ho fte, H. and Verbelen, J.P.
(2005) XTH acts at the microbrilmatrix interface during cell
elongation. J. Exp. Bot. 56, 673683.
Vorwerk, S., Somerville, S. and Somerville, C. (2004) The role of
plant cell wall polysaccharide composition in disease resistance.
Trends Plant Sci. 9, 203209.
Wagner, T.A. and Kohorn, B.D. (2001) Wall-associated kinases are
expressed throughout plant development and are required for
cell expansion. Plant Cell, 13, 303318.
Willats, W.G.T., Steele-King, C.G., McCartney, L., Orla, C.,
Marcus, S.E. and Knox, J.P. (2000) Making and using antibody
probes to study plant cell walls. Plant Physiol. Biochem. 38,
2736.
Yuan, S., Wu, Y.J. and Cosgrove, D.J. (2001) A fungal endoglucan-
ase with plant cell wall extension activity. Plant Physiol. 127, 324
333.
568 Markus Pauly and Kenneth Keegstra
2008 The Authors
Journal compilation 2008 Blackwell Publishing Ltd, The Plant Journal, (2008), 54, 559568