The Role of WOX Genes in Flower Development
The Role of WOX Genes in Flower Development
The Role of WOX Genes in Flower Development
Laboratory of Reproduction and Development of Plants, UMR5667 (ENS de Lyon, CNRS, INRA, UCBL), Ecole Normale Superieure
de Lyon, Lyon, France and 2Institute of Life Sciences, Scuola Superiore SantAnna, Pisa, Italy
* For correspondence. E-mail [email protected]
Received: 29 January 2014 Returned for revision: 12 March 2014 Accepted: 29 April 2014 Published electronically: 27 June 2014
Key words: WOX genes, HOMEOBOX, flower development, WUSCHEL, PRS/WOX3, MAW/WOX1,
EVERGREEN/WOX9, Petunia hybrida, Arabidopsis thaliana, dicots, monocots, plant evodevo.
algae, and many of them have been shown to play a role in plant
development (Mukherjee et al., 2009).
Plant HOMEOBOX genes sharing sequence identity with the
gene WUSCHEL (At-WUS) from arabidopsis are referred to as
WOX ((Wuschel-related homeobOX) genes. At-WUS was identified as a central player in stem cell maintenance in the shoot
apical meristem (SAM), although it is not required for SAM initiation (Laux et al., 1996; Liu et al., 2011). The name wuschel apparently derives from the bristled and bushy phenotype of the
mutants, in which ectopic meristems are repetitively produced
and prematurely terminated (Laux et al., 1996).
Since the discovery of At-WUS, several WOX genes have been
characterized in different species. Other members of the WOX
family are usually referred as WOX followed by an Arabic
numeral (with a few exceptions), and can be grouped in different
subfamilies or clades (van der Graaff et al., 2009; Vandenbussche
et al., 2009).
WOX G E N E S PL AY D I F F E R E N T ROL E S IN PL A N T
DEV E LOP M ENT
In arabidopsis, WOX genes have been shown to play a broad role
in plant development, from stem cell maintenance at the meristem level (WUSCHEL in shoot meristem, WOX4 in cambium,
WOX5 in root meristem) till embryo patterning (Laux et al.,
1996; Haecker et al., 2004; Sarkar et al., 2007; Ji et al., 2010).
We know from Picea abies that all the major WOX subfamilies,
with the exception of the MAW/WOX1 subfamily, probably
# The Author 2014. Published by Oxford University Press on behalf of the Annals of Botany Company. All rights reserved.
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Background WOX (Wuschel-like homeobOX) genes form a family of plant-specific HOMEODOMAIN transcription factors, the members of which play important developmental roles in a diverse range of processes. WOX genes
were first identified as determining cell fate during embryo development, as well as playing important roles in maintaining stem cell niches in the plant. In recent years, new roles have been identified in plant architecture and organ
development, particularly at the flower level.
Scope In this review, the role of WOX genes in flower development and flower architecture is highlighted, as evidenced from data obtained in the last few years. The roles played by WOX genes in different species and different
flower organs are compared, and differential functional recruitment of WOX genes during flower evolution is considered.
Conclusions This review compares available data concerning the role of WOX genes in flower and organ architecture among different species of angiosperms, including representatives of monocots and eudicots (rosids and asterids). These comparative data highlight the usefulness of the WOX gene family for evodevo studies of floral
development.
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AT- W US CH E L I S R E Q U I R E D FO R S T E M - C E L L
MAI NT E NA NC E I N T H E FLOW ER ( W US
S U B FA M ILY )
WUSCHEL (WUS) is the founding member of the WOX family
and is also representative of a clade, the WUS clade (Fig. 1).
WUS was initially isolated in arabidopsis (Laux et al. 1996) and
its function has been thoroughly investigated in this model species.
WUS promotes the identity and maintenance of stem cells, a
pool of undifferentiated and continuously dividing cells
located in the central zone of both the SAM and the flower meristem (FM) (Laux et al., 1996; Besnard et al., 2011). Thus, on a
wus genetic background, the SAM, instead of producing new
organs throughout the life of the plant, stops functioning prematurely in an aberrant flat morphology. However, wus plants
are still able to initiate a secondary meristem, but it fails to
self-maintain, resulting in plants with a highly disorganized,
bushy architecture. Similarly, wus flowers display many fewer
stamens (usually one or two) and no carpels, consistent with precocious FM termination (Laux et al., 1996). WUS is therefore
necessary for meristem maintenance, but is not required for
their initiation. Consistent with this function, WUS expression
is restricted to a small domain, the organizing centre, located
in the basal part of the central zone, beneath the L3 layer in the
SAM and beneath the L2 layer in the FM (Mayer et al., 1998).
Mechanistically, it is known now that WUS acts non cellautonomously to both promote stem cell identity and directly
activate CLAVATA3 (CLV3) expression within the central zone
(Schoof et al., 2000; Yadav et al., 2011). In turn, the CLE
peptide CLV3 diffuses outside of the central zone, binds to the
CLV1 and CLV2/CORYNE receptor kinases and thus triggers
the signalling pathway that eventually leads to the restriction of
WUS expression within the organizing centre (Brand et al.,
2000; Schoof et al., 2000; Lenhard and Laux 2003; Katsir
originated before the separation of angiosperms from gymnosperms (Hedman et al., 2013). A relationship between WOX
gene number and body pattern complexity among different
species, all from the green lineage (a grouping of land plants
and green algae), has also been proposed. In fact, WOX genes
can be divided into three different lineages that are supposed to
reflect their ancestry: an ancient lineage (comprising
At-WOX10, At-WOX13, At-WOX14 and their homologues), an
intermediate lineage (comprising At-WOX8, 9, 11 and 12 and
their homologues) and a new or WUS lineage (comprising
AtWOX17, including WUSCHEL, and their homologues)
(Haecker et al., 2004; Nardmann and Werr, 2012). Moreover,
WOX genes from the WUS lineage are absent from green algae,
bryophytes and ferns (with the exception of Leptosporangiatae).
Further diversification, sub-functionalization and recruitment in
different stem cell niches of these genes in angiosperms (but
also gymnosperms), has been considered as contributing to the
body plan diversity and evolutionary success of these groups
(Nardmann and Werr, 2012). At the molecular level, the acquisition of repressive activity by proteins from the modern lineage,
mainly due to an amino acid domain called the WUSCHEL
box (see red box on gene pictograms in Fig. 1), has been proposed
to play a major role in this process (Lin et al., 2013). In this review
we will further focus on the implication of WOX gene function
during floral development.
MAEWEST/WOX4 box
CLADE
OsWOX3A/Os11G01130
WOX3
99 Zm-NS2
98
KTLELFPIS
HD
KTLDLFPTK
HD
KTLDLFPTK
HD
KTLELFPTK
HD
OsWOX3B/Os05G02730
Mt-STF
WOX1
WUS
98 Ph-MAW
Ns-STF1
QTLELFPLR
HD
RKLR PLMPR
HD
QTLELFPLR
RKLR PLMPR
HD
QTLELFPLR
RKLR PLIPR
HD
QTLQLFPIR
YL PLS
HD
RTLNLFPVR
HD
At-WUS
RTLPLFPMH
77
98
Am-ROA
WUS
100 Ph-TER
OsWUS/Os04g56780
HD
97
72
WOX9
SI-Compound inflorescence
ETLPLFPVV
HD
Ph-SOE
Ph-EVG
ETLPLFPMH
HD
At-WOX9
9495
ETLPLFPMH
HD
At-WOX8
100
HD
HD
HD
Os01G47710
Os07G34880
Os05G48990
2.5 kb
At-WOX13
F I G . 1. Phylogenetic tree of WOX sequences. WOX sequences from different species (Am, Antirrhinum majus; At, Arabidopsis thaliana; Mt, Medicago truncatula;
Ns, Nicotiana sylvestris, Ph, Petunia hybrida; Os, Oryza sativa; Sl, Solanum lycopersicum; Zm, Zea mays) are clustered into different subfamilies (WOX3, WOX1,
WUS, WOX9) after phylogenetic analysis. The phylogenetic tree is based on the HOMEODOMAIN sequences. The WOX13 sequence (dark red) from arabidopsis is
used as outgroup. To support WOX relationships, 1000 bootstrap samples were obtained using the software TREECON (Van de Peer and De Wachter, 1994). Bootstrap
values ,65 % are not shown and corresponding branches are displayed as unresolved. On the right of the tree, gene structure is displayed for most of the WOX gene
sequences (solid blue bars are used for exons and thin black lines for introns), also depicting conserved amino acidic boxes (HD, HOMEODOMAIN; red rectangle,
WUSCHEL box; green rectangle, MAEWEST/WOX4 box, as in Vandenbussche et al., 2009). In addition, conserved amino acid residues are displayed for the
MAEWEST/WOX4 box and the WUSCHEL box. Note that WOX sequences also display characteristic C-terminal motifs with functional properties (not shown),
as illustrated in Vandenbussche et al. (2009). Where not specified, accession numbers are the same as in Vandenbussche et al. (2009); Mt-STF (AEL30892.1),
Nt-STF1 (AEL30893.1) and Sl-Compound Inflorescence (NP_001234072.1) are from GenBank.
termination is therefore closely linked to initiation of carpel development. However, these two processes are not coincident and
are uncoupled, although both are controlled by AG (Mizukami
and Ma, 1995; Ji et al., 2011). Very interestingly, the fact that organizing centre cells retain a molecular identity distinguishable
from that of surrounding cells even after the cessation of WUS expression further confirms the separation of the two processes but
also demonstrates that organizing centre cells persist after FM
termination and are not incorporated into carpels (Liu et al.,
2011). Recently, two different mechanisms of repression of
WUS by AG have been reported. They both explain why AG
does not repress WUS expression from stage 3. In the first mechanism, AG represses WUS expression indirectly by activating
At-WOX6
98
KTLPLFPIT
RKLR PLIPR
At-WOX1
98
92
KTLDLFP
HD
Ph-PRS
WUSCHEL box
HD
At-PRS
Zm-NS1
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T H E P R S / WO X 3 S U B FA M I LY
The second WOX gene that was found to play a role in flower
development is Arabidopsis PRESSED FLOWER (At-PRS, also
called WOX3). Mutants for this gene have flowers with a flattened
appearance (hence the name) because lateral sepal development
is affected: they are usually smaller, sometimes with a filamentous appearance, or can be completely absent (Matsumoto and
Okada, 2001). Although the size of the abaxial and adaxial
sepals is normal, marginal regions show defects. At-PRS was
shown to act independently of organ identity and meristem
size. The expression of At-PRS was detected at the lateral
regions of all lateral organs at very early stages, including
T HE M AW /WO X1 S U B FA M ILY
The evolutionary invention of petals, the usually brightly coloured organs of the flower, is generally believed to have played
a major role in the evolution of pollination syndromes. In
many taxa throughout the angiosperms, the petals fuse partly
or completely to form a tubular structure, thereby creating a protective barrier enclosing the reproductive organs and nectaries in
the centre of the flower. The maewest (maw) mutant in Petunia
was isolated in a genetic screen for mutants with defects in
petal fusion (Vandenbussche et al., 2009). Morphological analysis of maw flowers showed that petal fusion defects were
mainly due to reduced lateral outgrowth of the initially separate
petal primordia, which subsequently fail to fuse properly.
Similar defects were found in carpels, resulting in partly
unfused carpels, and sepals were narrower than wild-type. In
addition, leaf blade outgrowth was considerably reduced along
the lateral axis, as observed in floral organs, indicating that
MAW plays a general role in the lateral outgrowth of organs.
MAW was shown to encode a member of the WOX1 subfamily
of WOX transcription factors (Vandenbussche et al., 2009).
Similar phenotypes in leaf and flower development were found
for mutants of MAW/WOX1 homologues in Medicago truncatula
and Nicotiana sylvestris (McHale and Marcotrigiano, 1998; Lin
et al., 2013; Tadege et al., 2011a). In addition, mutants for MAW/
WOX1 homologues in two other species, narrow organs1 in
Lotus japonicus and lathyroides in Pisum sativum (garden
pea), have also been shown to be affected in lateral outgrowth
of organs such as leaves and petals (Zhuang et al., 2012),
further showing a broadly conserved role for MAW/WOX1
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Mutants
WT
Rosids
F I G . 2. Wox mutant flower phenotypes in petunia, Nicotiana, Medicago and arabidopsis (Vandenbussche et al., 2009; Tadege et al., 2011b; Lin et al., 2013). (A, C, D,
E) Wild-type (WT) flower phenotypes for Petunia hybrida, Nicotiana sylvestris, Medicago truncatula and Arabidopsis thaliana, respectively. (B) A Petunia wildtype pistil. (F, H, I, L) Mutant flowers: maw in Petunia (F), lam1 in Nicotiana (H), stf in Medicago (I) and wox1 prs in arabidopsis (L). (G) A strongly affected maw pistil
(carpels unfused). (C, D, H, I) Courtesy of M. Tadege.
Asterids
for the latter, the developmental defects found in the flower are
part of a more general phenotype, which also includes defects
in leaf blade expansion. Goethe (Goethe, 1790; Coen, 2001) proposed a long time ago that floral organs are in fact modified
leaves, so it is perhaps not very surprising to find that WOX
mutants are affected in both vegetative and floral development.
Yet it is clear that nature has exploited WOX gene function
during evolution for shaping floral architecture. Therefore,
while the homeotic function of animal HOX genes is fulfilled in
plant floral development by MADS box transcription factors,
WOX genes contribute to general aspects of floral architecture
and morphology. Classically, plant developmental biology has
focused mainly on Arabidopsis thaliana as a model organism.
Nevertheless, much of the progress made in our understanding
of the function of different WOX genes comes from studies in different species. We consider this to be a strong argument in favour
of the idea that plant developmental biology in general would
benefit from reorientation towards a more multi-model approach.
ACK NOW LED GE MENTS
M.V. is funded by an ATIP-AVENIR grant (Centre National de la
Recherche Scientifique). We are grateful to Million Tadege
(Oklahoma State University) and his team for providing pictures
of lam1 and stf mutants in Nicotiana and Medicago. We would
also like to thank the two anonymous reviewers, as well as the
editor, for very helpful suggestions to improve the manuscript.
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