Update on Xylan Biosynthesis in Grasses
Xylan Biosynthesis: News from the Grass1
Ahmed Faik*
Department of Environmental and Plant Biology, Molecular and Cellular Biology Program, Ohio University,
Athens, Ohio 45701
WHY XYLANS? WHY NOW?
Plant cell wall polysaccharides are critical compo-
nents of numerous products in our everyday life.
Because of this, modification of their constituent com-
ponents offers unique opportunities for product im-
provement and economic advancement. Xylans are
one of these polysaccharides that have a variety of
applications that affect our well-being. For example,
xylans are important functional ingredients in baked
products. They affect the quality of cereal flours for
bread making and the mechanical properties of dough.
They also impact brewing properties of grains (Vinkx
and Delcour, 1996). Xyl, the main constituent of xylans,
can be converted into important value-added products
such as xylitol, used as a natural food sweetener, a
dental cavities reducer, and a sugar substitute for
diabetics; in 1997, its market was estimated to approx-
imately $125 million (Saha and Bothast, 1997). Xylans
are also important for the livestock industry, as they
are critical factors for silage digestibility (Vinkx and
Delcour, 1996). Xylans are major constituents in the
nonnutritional constituent of feed (mostly cereal grain
by-products) in monogastric animals (poultry; Choct
and Annison, 1990). Thus, any small change in the
xylan content of grains that go into poultry and swine
feed could reflect billions of dollars of savings through
an improvement of the food conversion ratio (Donohue
and Cunningham, 2009). Given the functional and
economical importance of these polymers, their bio-
synthesis has attracted and puzzled plant cell wall
biologists for decades. Only in the last 5 years has
significant progress been made in identifying candi-
dates for the glycosyltransferase (GT) genes involved
in the biosynthetic process in dicots and monocots.
The focus of this Update, which should complement
the extensive review on xylan biosynthesis in dicots by
York and O’Neill (2008), is the recent progress made on
the biosynthesis of xylans in grasses.
Xylans are one of the major hemicelluloses in sec-
ondary cell walls of dicots and all walls of grasses.
Grasses have typical type II walls that are rich in
1
This work was supported by the National Science Foundation
(grant no. IOS–0724135).
* E-mail faik@ohio.edu.
The author responsible for distribution of materials integral to the
findings presented in this article in accordance with the policy
described in the Instructions for Authors (www.plantphysiol.org) is:
Ahmed Faik (faik@ohio.edu).
www.plantphysiol.org/cgi/doi/10.1104/pp.110.154237
396 Plant PhysiologyÒ, June 2010, Vol. 153, pp. 396–402, www.plantphysiol.org Ó 2010 American Society of Plant Biologists
glucurono(arabino)xylans (GAXs) and b-(1,3/4)glucan
(also called mixed-linkage glucan [MLG]; Bacic et al.,
1988; Ebringerova et al., 2005). Therefore, grasses have
been increasingly used as models to investigate xylans
and MLG biosynthesis. Progress has been made in
understanding the biosynthesis of many hemicellu-
loses (including MLG); however, investigating the
biosynthetic mechanism of xylans at the biochemical
and molecular levels has proven to be more challenging.
Now, with the genome sequences of rice (Oryza sativa)
and Brachypodium distachyon available, it is an exciting
time to work on xylan biosynthesis. Only recently,
through an extensive bioinformatics approach (Mitchell
et al., 2007), the identity of some GTs has been postu-
lated as candidates for xylan biosynthetic enzymes in
grasses. Despite this major advance, the biosynthetic
mechanism has yet to be fully characterized. Cur-
rently, we cannot delve deeper into the bioinformatics
classification and predict the exact biochemical func-
tion of putative GTs without isolation of the proteins
involved and functional enzymatic assays.
XYLANS FROM GRASSES: REGULAR/REPETITIVE
STRUCTURE OR NOT?
Xylans from grasses have the general structure of all
xylans (i.e. substitution with a-Araf, GlcA, or MeGlcA
residues) but also some unique features (for review, see
Ebringerova et al., 2005). The presence of Xyl-arabino-
furanosyl side chains (Fig. 1) is considered one of their
unique features (Wende and Fry, 1997; Höije et al., 2006;
Pastell et al., 2009). Another unique structural feature
of grass xylans is the presence of feruloyl groups on the
C-5 position of Araf residues (Fig. 1; Bacic et al., 1988;
Wende and Fry, 1997). Although the number of sub-
stituents and side chains may vary among xylans, two
main groups of xylans in grasses can be distinguished:
GAX, which makes up 35% of cell walls of vegetative
tissues and usually contains both Araf and GlcA/
MeGlcA residues; and AX, which lacks GlcA residues
and is found mainly in the starchy endosperm cell
walls in cereals (up to 70%; Ebringerova et al., 2005).
The substitution pattern of the xylan backbone is not
well known; one outstanding question is whether
GAXs from grasses have a regular/repetitive struc-
ture. Extensive postsynthesis modifications of these
polymers (loss of uronic acid and arabinosyl residues)
make a regular structure, originally present in the
newly synthesized GAX, difficult to detect. The first
suggestion of a repeating structure for GAX came from
 Xylan Biosynthesis in Grasses

saccharides, the authors concluded that the GAX

polymer had a regular structure. Two of the three
oligosaccharides released coeluted with authentic AX
oligosaccharides having six Xyl and two Ara or five
Xyl and two Ara (Zeng et al., 2008). This regular
structure was recently confirmed using an affinity-
purified GAX synthase activity (A. Faik, unpublished
data). Figure 2 illustrates the regular/repetitive struc-
ture of the nascent GAX polymer and the possible
degradation pattern to explain the formation of the
GAX fragments, which are expected to have two
unbranched Xyl residues at the reducing end and
one or two unbranched Xyl residues at the nonreduc-
ing end (due to the mechanism of degradation by
endoxylanases from the GH11 family; Gruppen et al.,

Figure 1. Structural features of the main side chains found in xylans

from grasses. The asterisks indicate the position of the attachment of
feruloyl groups on the a-(1,3)-linked Araf residues. Ac, O-acetyl
groups; Me, O-methyl groups.

early work carried out by Carpita and Whittern (1986).

Using Smith degradation and methylation analysis,
they identified a structure for GAX from developing
maize (Zea mays) coleoptiles and found that the aver-
age length of the released oligosaccharides was ap-
proximately six residues, but the exact distribution of
GlcA and Araf residues was not determined (Carpita
and Whittern, 1986). Other structural studies of AXs
from barley (Hordeum vulgare) and malt showed that
substituted Xyl residues were not randomly distrib-
uted in the polysaccharide but have a pattern in which Figure 2. Proposed structures of the repeating unit in the newly
up to four isolated unsubstituted residues are sepa- synthesized GAX polymer and the oligosaccharides generated by
rated by one or two substituted residues (Hoffmann digestion with a purified endoxylanase III from Aspergillus niger
et al., 1992; Vietor et al., 1994; Trogh et al., 2004). More (GH11 family). These structures do not take into account the GlcUA
residues, as their positions on the xylan backbone are not known. To
recently, Zeng et al. (2008) used a purified endoxyla-
cleave the xylan backbone, the endoxylanase III requires at least three
nase from the GH11 family (endoxylanase III) and adjacent and unsubstituted Xyl residues between substituted ones
high-pH anion-exchange chromatography (Dionex) (Davies et al., 1997). The sequence of degradation reactions steps by
methods to elucidate the structure of the newly syn- the endoxylanase is proposed to explain the formation of the final
thesized GAX polymer produced by microsomal oligosaccharides and monosaccharides detected by high-pH anion-
membranes of wheat (Triticum aestivum). Because the exchange chromatography separation (Zeng et al., 2008). Araf, Arabi-
endoxylanase III treatment released only three oligo- nofuranose residues; Xylp, xylopyranose residues.

Plant Physiol. Vol. 153, 2010 397

Faik
1993; Davies et al., 1997). This type of regular structure
is usually the result of a cooperative mechanism be-
tween enzymes, which lends further support to the
coordinated action between xylan synthase (XylT),
arabinosyltransferase (AraT), and glucuronosyltrans-
ferase (GlcAT) activities during GAX biosynthesis, as
proposed by Zeng et al. (2008).
COOPERATIVE ACTION: A COMMON MECHANISM
IN XYLAN BIOSYNTHESIS?
Progress in the polysaccharide biosynthesis field has
been limited by the lack of enzymological studies,
especially considering that most polysaccharides are
likely to be the result of well-organized, multienzyme
complexes. Xylan biosynthesis is not an exception and
appears to be a surprisingly difficult process to study.
Although several biochemical studies have investi-
gated xylan biosynthesis in vitro using microsomal
membranes from many grass species (Porchia and
Scheller, 2000; Kuroyama and Tsumuraya, 2001;
Porchia et al., 2002; Urahara et al., 2004), these efforts
were not conclusive regarding the biosynthetic mech-
anism. More recently Zeng et al. (2008) reported that
wheat Golgi-enriched microsomes contained a GAX-
GlcAT that was stimulated by the presence of UDP-Xyl
in the reaction mixture. There is a striking similarity
between this GAX-GlcAT activity in etiolated wheat
hypocotyls and the glucuronoxylan (GX)-GlcAT activ-
ity in pea (Pisum sativum) epicotyls (Waldron and
Brett, 1983; Baydoun et al., 1989). As in wheat, the
presence of UDP-Xyl enhanced the capacity of micro-
somes from pea epicotyls to transfer GlcA from UDP-
GlcA into the ethanol-insoluble material. This similarity
between wheat and pea GlcAT activities may be the
first hint of a similar mechanism for xylan biosynthesis
in primary cell walls of both dicots and monocots. Lee
et al. (2007a) demonstrated that crude microsomes
isolated from Arabidopsis (Arabidopsis thaliana) stems
contained GlcAT activity that appeared to slightly
stimulate the XylT activity. Furthermore, Zeng et al.
(2008) showed that a coordinated action existed be-
tween the wheat XylT and AraT; as a result of this
cooperative mechanism, the newly synthesized GAX
polymer had a regular structure, as represented in
Figure 2. The cooperative biosynthetic mechanism and
regular structure have also been associated with
xyloglucans (XyGs; Ray, 1980; Gordon and Maclachlan,
1989; White et al., 1993) and galactomannans in dicots
(Edwards et al., 1999). Thus, plants may have evolved
this biosynthetic mechanism to build many of their
hemicellulosic polymers, including xylans.
GLUCURONOARABINOXYLAN BIOSYNTHETIC
ENZYMES: DO GRASSES AND DICOTS USE
MEMBERS OF THE SAME GT FAMILIES?
According to the structure of xylans from grasses
(Fig. 1), at least five GT activities may be involved in the
398
biosynthetic process in addition to ester-forming trans-
ferases that add hydroxycinammic acids (i.e. ferulic and
p-coumaric acids). These GT activities include four
inverting enzyme activities, a-(1,2)arabinofuranosyl-
transferase [a-(1,2)AraT], a-(1,3)arabinofuranosyltrans-
ferase [a-(1,3)AraT], b-(1,2)xylosyltransferase [b-(1,2)
XylT], and b-(1,4)xylan synthase (XylT), and one retain-
ing GT activity, a-(1,2)glucuronyltransferase (GlcAT). In
addition, a mutase activity is required for the conver-
sion of UDP-L-arabinopyranose (UDP-L-Arap) to UDP-
L-arabinofuranose (UDP-L-Araf). Of particular interest
is whether grasses use the same mechanism and mem-
bers of the same GT families to synthesis AX and GAX
as dicot plants (i.e. Arabidopsis) use to synthesize GX
polymers. To identify candidate GT genes, Mitchell
et al. (2007) did an extensive bioinformatics analysis
that was based on differential expression of cereal
orthologs of the dicot GT genes that might be involved
in AX and GAX. Their hypothesis was that the expres-
sion of the GT genes involved in these polymers (the
most abundant polymers in grass walls) would be
higher in cereals as compared with dicots. This analysis
revealed that members from the GT43, GT47, and GT61
families (http://afmb.cnrs-mrs.fr/CAZY) were poten-
tial candidates for xylan biosynthesis in cereals. The rice
genes identified by Mitchell et al. (2007) clustered with
IRREGULAR XYLEM14 (IRX14; for the GT43 family)
and with IRX10 and IRX10-L (for the GT47 family; Fig.
3); therefore, it is reasonable to assume that the rice
genes have functions similar to their Arabidopsis ho-
mologs. Thus, bioinformatics analysis in conjunction
with the genetic studies carried out in Arabidopsis
indicate that members of the GT43 and GT47 families
are shared in the biosynthetic process of xylans in both
dicots and monocots. The question, then, is what these
functions would be.
Compelling data indicate that Arabidopsis members
of the GT43 family are XylTs responsible for the
synthesis of the xylan backbone: (1) the Arabidopsis
mutants irx9 and irx14 showed a drastic reduction in
xylan chain length in GX (Brown et al., 2007; Lee et al.,
2007a; Pena et al., 2007); and (2) microsomes from these
two mutant plants have reduced capacity to transfer
Xyl from UDP-Xyl onto xylooligosaccharide acceptors
(Brown et al., 2007; Lee et al., 2007a). Of course, the
inverting mechanism of the GT43 family is also pre-
dicted as a requirement for XylT. The fact that irx9 and
irx14 mutants show a similar phenotype may suggest
that they perform similar functions and/or comple-
ment each other in the same machinery (complex).
Actually, a complex of two XylTs (IRX9/IRX14) would
work better, because it would eliminate the need for a
180° rotation during Xyl transfer. Confirmation of the
XylT activity is still awaiting experimental evidence
using isolated proteins. If this function is confirmed, it
would completely eliminate the need for any other
proteins, such as members of the cellulose synthase-
like family, which includes several b-glycan synthases
(Dhugga et al., 2004; Liepman et al., 2005; Burton et al.,
2006; Cocuron et al., 2007; Doblin et al., 2009).
Plant Physiol. Vol. 153, 2010

Prediction of the biochemical function for GT47
members (IRX7/IRX10/IRX10-L) is even more com-
plicated. Possible functions include GlcAT and AraT.
Since xylans from both dicots and monocots have
GlcA substituents in common and differ largely in the
amount of Araf residues, it would be tempting to
attribute the GlcAT function to GT47 members. Be-
cause the mechanism of this GT47 family is not con-
sistent with the retaining mechanism of GlcAT activity,
an intermediate donor substrate containing b-linked
GlcA was proposed by Zhong et al. (2005). However, if
we assume that the inverting mechanism is valid and
no intermediate donor is involved, then some GT47
members could possibly be AraTs. This function could
connect the high content in Araf in GAX (compared
with GX in dicots) and the high representation of
IRX10 and IRX10-L homologs in grasses. Mitchell et al.
(2007) reached the same conclusion in their bioinfor-
matics work. The question now is how to explain the
involvement of some GT47 members (such as AraTs)
in the biosynthesis of a xylan polymer (i.e. GX from
Arabidopsis) that presumably does not contain any
Araf residues. One possible explanation would be that
the nascent GX polymers in dicots contain certain
levels of Araf residues that are removed during the
secretion/incorporation of GX into cell walls. In other
words, Arabidopsis GX is synthesized as GAX during
secondary cell wall deposition and then converted into
GX by the action of an a-arabinosidase. This biosyn-
thetic pathway is supported by the observation that
the expression of an Arabidopsis a-arabinosidase
ARAF1 gene (At3g10740) is well correlated with ex-
pression of the FRA8 gene (Chávez Montes et al., 2008;
see the ExpressionProfiling tool at http://genecat.
mpg.de/cgi-bin/Ainitiator.py) and that GXs from
some woody plants have small amounts of Araf res-
idues (Kormelink and Voragen, 1993). The absence of
Plant Physiol. Vol. 153, 2010
Xylan Biosynthesis in Grasses
Figure 3. Phylogenetic trees re-
flecting the relationships between
rice and Arabidopsis proteins from
the GT43 (A) and GT47 (B) families.
IRX7, IRX10, IRX10-L, and IRX14
along with XGD1 (At5g33290; a
xylogalacturonan xylosyltransfer-
ase), ARAD1 (At2g35100; a pu-
tative arabinosyltransferase), and
MUR3 (At2g20370; a xyloglucan
galactosyltransferase) proteins
were included in the analysis. The
IRX proteins are known to be in-
volved in GX biosynthesis during
secondary cell wall deposition in
Arabidopsis (York and O’Neill,
2008; Brown et al., 2009; Wu
et al., 2009). Trees were constructed
under the maximum-likelihood
principle using the PhyML program
(Guindon et al., 2005).
Araf in GXs from the Arabidopsis stem cell wall may
explain why Brown et al. (2005) could not detect a de-
crease in Araf content in stem cell walls from the Arab-
idopsis irx10 mutant. The removal of Araf residues
in GAX during coleoptile growth was also reported
in barley (Gibeaut et al., 2005) and maize (Carpita,
1984), presumably due to an a-arabinofurnosidase
activity.
The feruloylation of the a-(1,3)Araf side chains of
AX/GAX polymers (Fig. 1) is unique to grasses and
plays an important role in the integrity of type II cell
walls (de O Buanafina, 2009). Members of the Pfam
family PF02458 (12 members) were suggested as pu-
tative AX feruloyltransferases by EST-based differen-
tial expression analysis (Mitchell et al., 2007). This
conclusion was experimentally supported by recent
work by Piston et al. (2010), who showed that down-
regulation of four rice genes (Os05g08640, Os06g39470,
Os01g09010, and Os06g39390) from this Pfam family
resulted in reduced ferulate content in the cell walls of
transgenic rice plants.
Despite this progress in xylan biosynthesis in
grasses, several questions remain unanswered and
would require proof of function for certain genes. For
example, if some members of the GT47 family are
AraTs, what would be the identity of the GlcATs? The
GT8 family members could be candidates for GlcATs,
as this GT family has a retaining mechanism that fits
the GlcAT requirement. But this hypothesis has not
been supported by genetic studies, as both irx8 and
parvus mutations did not decrease GlcAT activity in
their microsomal fractions (Lee et al., 2007a, 2007b). In
addition, if members of the GT8 family are GlcAT, why
they are not highly expressed in grasses is a puzzle.
York and O’Neill (2008) proposed that members of this
GT family (IRX8, PARVUS) have a role in generating a
unique sequence (called sequence 1) identified at the
399
Faik
reducing ends of GXs from dicots and gymnosperms.
This sequence 1 seems to be absent in grass xylans
(Fincher, 2009).
Another question is what function the GT61 mem-
bers would have. Some members of this GT family
are highly represented in the cereal EST databases
(Mitchell et al., 2007) and were proposed to catalyze
the transfer of a b-(1,2)XylT onto the a-(1,3)Araf sub-
stituents (Fig. 1). This enzyme activity is still in need of
experimental evidence from isolated proteins and the
use of well-characterized acceptors. Since the disac-
charide Xyl-arabinofuranosyl side chain was found in
large amounts in AXs from corn cobs and barley husks
(Pastell et al., 2009), it would be possible to develop an
enzymatic assay for b-(1,2)XylT activity using micro-
somes from these plants and to use this assay to screen
candidates from the GT61 family.
Despite the fact that Konishi et al. (2007, 2010)
provided strong biochemical evidence that members
of the GT75 family (also called reversibly glycosylated
polypeptides [RGPs]) have UDP-Arap mutase activity
(presumably necessary for AraT), bioinformatics anal-
ysis (Mitchell et al., 2007) did not reveal members of
this family as candidates for AX and GAX biosynthe-
sis. Therefore, direct biochemical evidence that links
these mutases to AX and GAX biosynthesis is needed.
However, in their study of AX biosynthesis in wheat,
Porchia et al. (2002) have reported reversible arabino-
sylation of a 41-kD protein (possibly an RGP).
CONTROL OF THE XYLAN LENGTH IN GRASSES
York and O’Neill (2008) proposed a mechanistic
model for GX elongation in Arabidopsis in which the
xylan backbone could be elongated from its reducing
end, but then a mechanism to terminate the elongation
of the backbone is needed (Bodevin-Authelet et al.,
2005; Mukerjea and Robyt, 2005; Tlapak-Simmons
et al., 2005). The identification of sequence 1 at the
reducing ends of GXs from dicots and gymnosperms
was proposed to act as a signal to terminate the xylan
backbone (Pena et al., 2007; York and O’Neill, 2008)
and may suggest an elongation mechanism from the
reducing end. The question of interest is whether
grasses use a similar mechanism to control the length
of GAX polymers during biosynthesis. Zeng et al.
(2008) reported that in vitro, wheat microsomes gen-
erated only short GAX polymers that eluted on CL-6B
columns as a sharp peak around 10 to 15 kD, corre-
sponding to a degree of polymerization (DP) of 50 to
80. Longer reaction incubations and changes in reac-
tion conditions did not produce longer GAX polymers
(Zeng et al., 2008). A DP of approximately 50 to 80 is
slightly shorter as compared with the size (DP of
approximately 80–90) of GX from Arabidopsis (Pena
et al., 2007). Thus, it is conceivable that both dicots and
grasses evolved mechanisms that terminate xylan
backbone elongation after a number of Xyl residues.
However, experimental evidence is required to verify
400
this hypothesis using a purified GAX synthase activity
and the identification of the sequence terminator. The
fact that wheat microsomes could also produce xylan
polymers with an apparent molecular mass larger than
500 kD (Porchia and Scheller, 2000) may suggest the
existence of several xylan synthase activities in wheat.
One of the most challenging questions concerns the
initiation of xylan chain elongation in grasses. For
example, several works showed that microsomal
membranes or partially purified enzymes make in
vitro xylan polymers without the addition of exoge-
nous primers, but they did not eliminate the possibil-
ity of the presence of endogenous primers in their
enzyme preparations (Baydoun et al., 1989; Porchia
and Scheller, 2000; Porchia et al., 2002; Zeng et al.,
2008). On the other hand, other research groups
demonstrated that xylooligosaccharides added ex-
ogenously to permeabilized Golgi membranes act as
acceptors (possibly primers), but only a few Xyl resi-
dues were added to these oligosaccharides (Kuroyama
and Tsumuraya, 2001; Urahara et al., 2004; Brown
et al., 2007; Lee et al., 2007a). To accommodate these
findings, York and O’Neill (2008) proposed that xylan
backbone elongation occurred from its reducing end.
In this biosynthetic mechanism, the elongating xylan
chain would form a covalent intermediate with XylT or
with other proteins associated with XylT. Whether
some RGPs could fulfill this function is another chal-
lenge for cell wall biology.
CONCLUSION
The recent progress with respect to the biochemistry
of GAX biosynthesis in wheat has enhanced our un-
derstanding of the biosynthetic process of xylans. The
picture that emerges here is that plant cells appear to
use a cooperative mechanism (through multienzyme
complexes) to synthesize many hemicellulosic poly-
mers found in primary cell walls, namely GAXs, GXs,
and XyGs. Thus, the “arabinosyl-Xyl” backbone of GAX
is synthesized in a manner similar to the “xylosyl-
Glc” backbone of XyGs in dicots. Furthermore, as a
result of this cooperative mechanism, the newly syn-
thesized polymers have regular structures. However,
many challenges are still before us. For example, direct
proof of the biochemical functions of the GT43, GT47,
and GT61 members is still lacking. Whether or not
primary and secondary cell walls require xylan syn-
thases with different protein compositions, as was
demonstrated for cellulose synthesis (for review, see
Somerville, 2006), is not known. Clearly, progress in
the xylan biosynthesis field is limited by the lack of
enzymological studies.
A thorough understanding of any regulating mech-
anism, including the identification of transcription
factors (Zhong and Ye, 2007; Aohara et al., 2009)
would also certainly increase our understanding of
xylan biosynthesis in the context of primary and
secondary cell wall elaboration and would offer op-
Plant Physiol. Vol. 153, 2010

portunities to genetically manipulate cell wall compo-
sition and structure to better fit human needs.
ACKNOWLEDGMENTS
My thanks go to Dr. Sarah Wyatt for critical reading of the manuscript and
comments. Thank you also to all students in my laboratory for their hard
work to make this difficult project succeed.
Received February 1, 2010; accepted March 31, 2010; published April 7, 2010.
LITERATURE CITED
Aohara T, Kotake T, Kaneko Y, Takatsuji H, Tsumuraya Y, Kawasaki S
(2009) Rice BRITTLE CUM 5 (BRITTLE NODE) is involved in secondary
cell wall formation in the sclerenchyma tissue of nodes. Plant Cell
Physiol 50: 1886–1897
Bacic A, Harris PJ, Stone BA (1988) Structure and function of plant cell
walls. In J Preiss, ed, The Biochemistry of Plants, Vol 14. Academic Press,
San Diego, pp 297–371
Baydoun EAH, Waldron KW, Brett CT (1989) The interaction of xylosyl-
transferase and glucuronyltransferase involved in glucuronoxylan syn-
thesis in pea (Pisum sativum) epicotyls. Biochem J 257: 853–858
Bodevin-Authelet S, Kusche-Gullberg M, Pummill PE, DeAngelis PL,
Lindahl U (2005) Biosynthesis of hyaluronan: direction of chain elon-
gation. J Biol Chem 280: 8813–8818
Brown DM, Goubet F, Wong VW, Goodacre R, Stephans E, Dupree P,
Turner S (2007) Comparison of five xylan synthesis mutants reveals new
insight into the mechanisms of xylan synthesis. Plant J 52: 1154–1168
Brown DM, Zeef LAH, Ellis J, Goodacre R, Turner SR (2005) Identification
of novel genes in Arabidopsis involved in secondary cell wall formation
using expression profiling and reverse genetics. Plant Cell 17: 2281–2295
Brown DM, Zhang ZN, Stephens E, Dupree P, Turner SR (2009) Charac-
terization of IRX10 and IRX10-like reveals an essential role in glucur-
onoxylan biosynthesis in Arabidopsis. Plant J 57: 732–746
Burton RA, Wilson SM, Hrmova M, Harvey AJ, Shirley NJ, Medhurst A,
Stone BA, Newbigin EJ, Bacic A, Fincher GB (2006) Cellulose synthase-
like CslF genes mediate the synthesis of cell wall (1,3;1,4)-beta-D-glucans.
Science 311: 1940–1942
Carpita NC (1984) Cell wall development in maize coleoptiles. Plant
Physiol 76: 205–212
Carpita NC, Whittern D (1986) A highly substituted glucuronoarabinox-
ylan from developing maize coleoptiles. Carbohydr Res 146: 129–140
Chávez Montes RA, Ranocha P, Martinez Y, Minic Z, Jouanin L, Marquis
M, Saulnier L, Fulton LM, Cobbett CS, Bitton F, et al (2008) Cell wall
modifications in Arabidopsis plants with altered a-L-arabinofuranosi-
dase activity. Plant Physiol 147: 63–77
Choct M, Annison G (1990) Anti-nutritive activity of wheat pentosans in
broiler diets. Br Poult Sci 30: 811–821
Cocuron JC, Lerouxel O, Drakakaki G, Alonso AP, Liepman AH, Keegstra
K, Raikhel N, Wilkerson CG (2007) A gene from the cellulose synthase-
like C family encodes a b-1,4 glucan synthase. Proc Natl Acad Sci USA
140: 8550–8555
de O Buanafina MM (2009) Feruloylation in grasses: current and future
perspectives. Molecular Plant 2: 861–872
Dhugga KS, Barreiro R, Whitten B, Stecca K, Hazebroek J, Randhawa GS,
Dolan M, Kinney AJ, Tomes D, Nichols S, et al (2004) Guar seed beta-
mannan synthase is a member of the cellulose synthase super gene
family. Science 303: 363–366
Doblin MS, Pettolino FA, Wilson SM, Campbell R, Burton RA, Fincher
GB, Newbigin E, Bacic A (2009) A barley cellulose synthase-like CSLH
gene mediates (1,3;1,4)-beta-D-glucan synthesis in transgenic Arabi-
dopsis. Proc Natl Acad Sci USA 106: 5996–6001
Donohue M, Cunningham DL (2009) Effects of grain and oilseed prices on
the costs of US poultry production. J Appl Poult Res 18: 325–337
Edwards ME, Dickson CA, Chengappa S, Sidebottom C, Gidley MJ, Reid
JSG (1999) Molecular characterization of a membrane-bound galacto-
syltransferase of plant cell wall matrix polysaccharide biosynthesis.
Plant J 19: 691–697
Plant Physiol. Vol. 153, 2010
Xylan Biosynthesis in Grasses
Davies GJ, Wilson KS, Henrissat B (1997) Nomenclature for sugar-binding
subsites in glycosyl hydrolases. Biochem J 321: 557–559
Ebringerova A, Hromadkova Z, Heinze T (2005) Hemicellulose. Adv
Polym Sci 186: 1–67
Fincher GB (2009) Revolutionary times in our understanding of cell wall
biosynthesis and remodeling in the grasses. Plant Physiol 149: 27–37
Gibeaut DM, Pauly M, Bacic A, Fincher GB (2005) Changes in cell wall
polysaccharides in developing barley (Hordeum vulgare) coleoptiles.
Planta 221: 729–738
Gordon R, Maclachlan G (1989) Incorporation of UDP-[14C]glucose into
xyloglucan by pea membranes. Plant Physiol 91: 373–378
Gruppen H, Kormelink FJM, Voragen AGJ (1993) Differences in efficacy of
xylanases in the breakdown of wheat flour arabinoxylans due to their
mode of action. In C Wenk, M Boessinger, eds, Enzymes in Animal
Nutrition. Kartause Ittingen, Thurgau, Switzerland, pp 276–280
Guindon S, Lethiec F, Duroux P, Gascuel O (2005) PHYML Online: a Web
server for fast maximum likelihood-based phylogenetic inference.
Nucleic Acids Res 33: W557–W559
Hoffmann RA, Geijtenbeek T, Kamerling JP, Vliegenthart JFG (1992)
1H-NMR study of enzymatically generated wheat-endosperm arabinox-
ylan oligosaccharides: structures of hepta- to tetradeca-saccharides con-
taining two or three branched xylose residues. Carbohydr Res 223: 19–44
Höije A, Sandström C, Roubroeks JP, Andersson R, Gohil S, Gatenholm P
(2006) Evidence of the presence of 2-O-b-xylopyranosyl-a-L-arabino-
furanose side chains in barley husk arabinoxylan. Carbohydr Res 341:
2959–2966
Konishi T, Ohnishi-Kameyama M, Funane K, Miyazaki Y, Konishi T,
Ishii T (2010) An arginyl residue in rice UDP-arabinopyranose mutase
is required for catalytic activity and autoglycosylation. Carbohydr Res
345: 787–791
Konishi T, Takeda T, Miyazaki Y, Ohnishi-Kameyama M, Hayashi T,
O’Neill MA, Ishii T (2007) A plant mutase that interconverts UDP-
arabinofuranose and UDP-arabinopyranose. Glycobiology 17: 345–354
Kormelink FJM, Voragen AGJ (1993) Degradation of different [(glucurono)
arabino]xylan by a combination of purified xylan-degrading enzymes. Appl
Microbiol Biotechnol 38: 688–695
Kuroyama H, Tsumuraya Y (2001) A xylosyltransferase that synthesizes
b-(1,4)-xylans in wheat (Triticum aestivum L.) seedlings. Planta 213: 231–240
Lee C, O’Neill MA, Tsumuraya Y, Darvill AG, Ye ZH (2007a) The irregular
xylem9 mutant is deficient in xylan xylosyltransferase activity. Plant Cell
Physiol 48: 1624–1634
Lee C, Zhong R, Richardson EA, Himmelsbach D, McPhail BT, Ye ZH
(2007b) The PARVUS gene is expressed in cells undergoing secondary
wall thickening and is essential for glucuronoxylan biosynthesis. Plant
Cell Physiol 48: 1659–1672
Liepman AH, Wilkerson CG, Keegstra K (2005) Expression of cellulose
synthase-like (Csl) genes in insect cells reveals that CslA family mem-
bers encode mannan synthases. Proc Natl Acad Sci USA 102: 2221–2226
Mitchell RAC, Dupree P, Shewry PR (2007) A novel bioinformatics
approach identifies candidate genes for the synthesis and feruloylation
of arabinoxylan. Plant Physiol 144: 43–53
Mukerjea R, Robyt JF (2005) Starch biosynthesis: the primer nonreducing-
end mechanism versus the nonprimer reducing-end two-site insertion.
Carbohydr Res 340: 245–255
Pastell H, Virkki L, Harju E, Tuomainen P, Tenkanen M (2009) Presence of
1,3-linked 2-O-b-D-xylopyranosesyl-a-L-arabinofuranosyl side chains
in cereal arabinoxylans. Carbohydr Res 344: 2480–2488
Pena MJ, Zhong R, Zhou GK, Richardson EA, O’Neill MA, Darvill AG,
York WS, Ye ZH (2007) Arabidopsis irregular xylem8 and irregular xylem9:
implications for the complexity of glucuronoxylan biosynthesis. Plant
Cell 19: 549–563
Piston F, Uauy C, Fu L, Langston J, Labavitch J, Dubcovsky J (2010) Down-
regulation of four putative arabinoxylan feruloyl transferase genes from
family PF02458 reduces ester-linked ferulate content in rice cell walls.
Planta 231: 677–691
Porchia AC, Scheller HV (2000) Arabinoxylan biosynthesis: identification
and partial characterization of a b-1,4-xylosyltransferase from wheat.
Physiol Plant 110: 350–356
Porchia AC, Sørensen SO, Scheller HV (2002) Arabinoxylan biosynthesis
in wheat: characterization of arabinosyltransferase activity in Golgi
membranes. Plant Physiol 130: 432–441
Ray P (1980) Cooperative action of b-glucan synthase and UDP-xylose
401
Faik
xylosyltransferase of Golgi membranes in the synthesis of xyloglucan-
like polysaccharide. Biochim Biophys Acta 629: 431–444
Saha BC, Bothast RJ (1997) Microbial production of xylitol. In BC Saha, J
Woodward, eds, Fuels and Chemicals from Biomass. American Chem-
ical Society, Washington, DC, pp 307–319
Somerville C (2006) Cellulose synthesis in higher plants. Annu Rev Cell
Dev Biol 22: 53–78
Tlapak-Simmons VL, Baron CA, Gostschall R, Haque D, Canfield WM,
Weigel PH (2005) Hyaluronan biosynthesis by class I streptococcal
hyaluronan synthase occurs at the reducing end. J Biol Chem 280:
13012–13018
Trogh I, Courtin CM, Delcour JA (2004) Isolation and characterization of
water-extractable arabinoxylan from hull-less barley flours. Cereal
Chem 81: 555–680
Urahara T, Tsuchiya K, Kotake T, Tohno-oka T, Komae K, Kawada N,
Tsumuraya Y (2004) A b-(1,4)xylosyltransferase involved in the synthe-
sis of arabinoxylans in developing barley endosperms. Physiol Plant
122: 169–180
Vietor RJ, Kormelink FJM, Angelino SAGF, Voragen AGJ (1994) Substi-
tution patterns of water-unsoluble arabinogalactans from barley and
malt. Carbohydr Polym 24: 113–118
Vinkx CJA, Delcour JA (1996) Rye (Secale cereale L.) arabinoxylans: a
critical review. J Cereal Sci 24: 1–14
402
Waldron KW, Brett CT (1983) A glucuronyltransferase involved in glu-
curonoxylan synthesis in pea (Pisum sativum) epicotyls. Biochem J 213:
115–122
Wende G, Fry SC (1997) 2-O-Beta-D-xylopyranosyl-(5-O-feruloyl)-L-arabinose,
a widespread component of grass cell walls. Phytochemistry 44: 1019–1030
White AR, Xin Y, Pezeshk V (1993) Xyloglucan synthase in Golgi mem-
branes from Pisum sativum (pea). Biochem J 294: 1–8
Wu AM, Rihouey C, Seveno M, Hornblad E, Singh SK, Matsunaga T, Ishii
T, Lerouge P, Marchant A (2009) The Arabidopsis IRX10 and IRX10-like
glycosyltransferases are critical for glucuronoxylan biosynthesis during
secondary cell wall formation. Plant J 57: 718–731
York WS, O’Neill MA (2008) Biochemical control of xylan biosynthesis:
Which end is up? Curr Opin Plant Biol 11: 258–265
Zeng W, Chatterjee M, Faik A (2008) UDP-xylose stimulated glucuronyltrans-
ferase activity in wheat (Triticum aestivum L.) microsomal membranes:
characterization and role in glucurono(arabino)xylan biosynthesis. Plant
Physiol 147: 78–91
Zhong R, Pena MJ, Zhou GK, Naim J, Wood-Jones A, Richardson EA,
Morrison WH, Darvill AG, York WS, Ye ZH (2005) Arabidopsis Fragile
Fiber8, which encodes a putative glucuronyltransferase, is essential for
normal secondary wall synthesis. Plant Cell 17: 3390–3408
Zhong R, Ye ZH (2007) Regulation of cell wall biosynthesis. Curr Opin
Plant Biol 10: 564–572
Plant Physiol. Vol. 153, 2010