See discussions, stats, and author profiles for this publication at: https://www.researchgate.

net/publication/6131265

Brassinosteroid transport

Article  in  Journal of Experimental Botany · February 2008

DOI: 10.1093/jxb/erm098 · Source: PubMed

CITATIONS READS
88 235

4 authors, including:

John J. Ross James Reid

University of Tasmania University of Tasmania
153 PUBLICATIONS   5,849 CITATIONS    266 PUBLICATIONS   9,671 CITATIONS   

SEE PROFILE SEE PROFILE

Some of the authors of this publication are also working on these related projects:

Auxin Biosynthesis in Pisum sativum View project

All content following this page was uploaded by John J. Ross on 27 January 2016.

The user has requested enhancement of the downloaded file.

Journal of Experimental Botany, Vol. 59, No. 1, pp. 17–24, 2008
Transport of Plant Growth Regulators Special Issue
doi:10.1093/jxb/erm098 Advance Access publication 19 August, 2007
SPECIAL ISSUE REVIEW PAPER

Brassinosteroid transport

Gregory M. Symons*, John J. Ross, Corinne E. Jager and James B. Reid

School of Plant Science, University of Tasmania, Private Bag 55, Hobart, Tasmania, Australia 7001

Received 18 February 2007; Revised 13 March 2007; Accepted 16 April 2007

Abstract elongation, xylem differentiation, and fruit ripening. These

compounds appear to be ubiquitous across the plant
Brassinosteroids (BRs) are steroidal plant hormones
kingdom, having been detected in a large number of
that are important regulators of plant growth. These
species from a broad range of taxonomic groups (Bajguz

Downloaded from http://jxb.oxfordjournals.org/ by guest on November 6, 2015

compounds are widely distributed throughout repro-
and Tretyn, 2003). The last 10 years have seen significant
ductive and vegetative plant tissues. This raises the
progress in our understanding of BR biology, including
question of whether or not BRs are transported over
their distribution in the plant, the role of BRs in plant
long distances between these tissues. Several lines of
growth, BR biosynthesis, BR perception and signal trans-
evidence indicate that this is not the case. Exogenous
duction, and the interactions between BRs and other plant
BRs move only slowly, if at all, after application to
hormones (for reviews see Fujioka and Yokota, 2003;
leaves; grafting BR-deficient mutants to wild-type
Belkhadir and Chory, 2006; Nomura and Bishop, 2006; Li
plants has no phenotypic effect; removal of the apical
and Jin, 2007). However, until recently there had been
bud or mature leaves does not reduce BR levels in the
little focus on the question of whether BRs are transported
remaining internodes; and, in tomato, wild-type sectors
around the plant and whether BR transport could play
do not substantially alter the growth of BR-deficient
a role in regulating BR levels and BR action. The aim of
sectors when the two types are together in a variegated
this review is to provide an update on recent develop-
leaf. Although BRs do not undergo long-distance trans-
ments in our understanding of BR transport, and on the
port they may influence long-distance signalling by
implications of these new findings for the regulation of
altering auxin transport. At the cellular level, BRs do
BR levels.
appear to be transported. The enzymes for BR bio-
synthesis appear to be located within the cell, and to be
associated with the endoplasmic reticulum, in particu-
lar. BR reception, on the other hand, is thought to occur Distribution of BRs in plants
on the exterior cell surface. Therefore, BRs must move
Before the question of whether BRs undergo transport in
from the interior of the cell to the exterior, where they
the plant is addressed, it is essential to understand where
are perceived by the same cell or by neighbouring cells.
they are located. Generally speaking, the distribution of
The existence of a feedback system, whereby bioactive
BRs is widespread, with these compounds being present
BRs negatively regulate their own biosynthesis, pro-
in most plant tissues (Bajguz and Tretyn, 2003). However,
vides further evidence that individual cells are able to
levels of the bioactive BRs, castasterone (CS) and
both perceive and synthesize BRs.
brassinolide (BL), differ greatly in different parts of the
Key words: Brassinosteroids, perception, plant hormones,
plant, in a pattern that is similar in a wide range of plant
synthesis, transport.
species (Bancos et al., 2002; Bajguz and Tretyn, 2003;
Shimada et al., 2003; Symons and Reid, 2004). For
instance, the highest levels of bioactive BRs generally
occur in reproductive organs such as pollen [5–1000 ng
g
1 fresh weight (FW)], from which BRs were originally
Introduction
isolated by Mitchell et al. (1970), seeds (0.3–1600 ng g
1
Brassinosteroids (BRs) are a group of plant steroidal FW), and fruits (0.2–3.5 ng g
1 FW) (Bajguz and
hormones, which regulate processes as diverse as cell Tretyn, 2003; Montoya et al., 2005; Symons et al., 2006;

* To whom correspondence should be addressed. E-mail: g_symons@utas.edu.au

ª The Author [2007]. Published by Oxford University Press [on behalf of the Society for Experimental Biology]. All rights reserved.
For Permissions, please e-mail: journals.permissions@oxfordjournals.org
18 Symons et al.
Nomura et al., 2007). This is consistent with the important
roles proposed for BRs in processes such as fruit
development and ripening (Montoya et al., 2005;
Symons et al., 2006). In contrast, bioactive BR levels in
vegetative (shoot) tissues are generally much lower (0.12–
2 ng g
1 FW), whilst the lowest levels (<0.05 ng g
1 FW)
of bioactive BRs are found in the roots (Bancos et al.,
2002; Shimada et al., 2003; Symons and Reid, 2004).
Within shoot tissues, the greatest accumulation of bio-
active BRs usually occurs in the young actively growing
regions such as the shoot tip (apex) and the young
internodes, which is again consistent with the proposed
roles for BRs in the promotion of cell division and
elongation (Shimada et al., 2003; Symons and Reid,
2004).
Such a widespread distribution of endogenous BRs
throughout the plant immediately raises the question of
whether BRs are synthesized in a localized region of the
plant and then transported to distant sites of action.
Do BRs undergo long-distance transport?
Experiments with exogenous BRs
The possible transport of endogenous BRs has been
studied by monitoring the movement of radiolabelled
BRs after application to various parts of the plant
(previously reviewed by Arteca, 1995; Adam and
Schneider, 1999; Sasse, 1999, 2003; Bishop and Yokota,
2001; Ross et al., 2006). However, caution is required
when interpreting such studies, because the observed
movement of exogenous radiolabelled substances may not
reflect the transport of endogenous hormones (Hoad,
1995). For example, even if it can be shown that
exogenous BRs move within the plant, this does not prove
that endogenous BRs are normally transported in a similar
manner (Ross et al., 2006). However, keeping this in
mind, it is interesting to note that there is some evidence
for root-to-shoot transfer of exogenous BRs. For example,
after application of [3H]BL or [3H]CS to the roots of rice
plants, approximately 10% of the radioactivity taken up
by the plant was found in the shoot, after only 6 h
(Yokota et al., 1992). Of this 10%, the large majority was
in the form of unmetabolized BRs, suggesting that
exogenous BRs taken up through the roots were trans-
located, unchanged, to the shoot (Yokota et al., 1992).
Similarly, in both cucumber and wheat seedlings, there
appeared to be a rapid transfer of radioactivity to the shoot
after [14C]epi-brassinolide ([14C]epiBL) was applied to the
roots (Nishikawa et al., 1994). However, in this study it
was not determined whether the radioactivity found in the
shoot was in the form of unmetabolized BRs or more
polar BR metabolites. As a consequence, it is not possible
to be confident that the observed movement of radioactiv-
ity accurately represents the movement of a BR in its
bioactive form. Consistent with results showing some
root-to-shoot movement of exogenous BRs is the obser-
vation that the aberrant shoot phenotype of BR-deficient
Arabidopsis mutants can be restored to normal when these
plants are grown on media containing bioactive BRs
(Choe et al., 1998). Again this may suggest that root-to-
shoot transfer of exogenous BRs is possible, but does not
prove that endogenous BRs are transported in a similar
manner.
When applied directly to shoot tissues, exogenous BRs
appear to be relatively immobile. For instance, when
[3H]BL or [3H]CS was applied to leaves of pea, it entered
those leaves, but was not exported from them (Symons
and Reid, 2004). Similarly, the majority of exogenous,
radiolabelled BL or CS incorporated into the leaves of rice
remained in the treated leaves 24 h after it was applied
(Yokota et al., 1992). In this last study, some radioactivity
was detected in the roots after 72 h, but was shown to be
Downloaded from http://jxb.oxfordjournals.org/ by guest on November 6, 2015
largely in the form of water-soluble BR metabolites
(Yokota et al., 1992). In wheat, radioactivity did not
move from the treated leaf, even 7 d after exogenous
[14C]epiBL was applied, while in cucumber, some
radioactivity moved out of the leaf, but not until after 3
d (Nishikawa et al., 1994).
Grafting experiments
A more satisfactory method for studying the possible
transport of endogenous BRs involves grafting BR-
deficient dwarf mutants to WT plants. In pea, the usual
grafting technique results in a composite plant with the
young, apical portion of the shoot (or scion) being mutant,
and the basal portion WT. The basal portion (or stock) can
consist only of below-ground parts or can also include
several mature leaves. Reciprocal grafts (WT scion grafted
onto a mutant stock) are also informative.
The reasoning is that if BRs are normally transported
acropetally, from the roots and/or mature shoot tissue into
the apical portion of the shoot, then the BR deficiency and
dwarf stature of a mutant scion should be reversed by
grafting to a WT stock (assuming that BRs can move
across the graft union). For another group of plant growth-
promoting hormones, the gibberellins, spectacular growth
promotion can be demonstrated in this way (although it is
the precursors of bioactive gibberellins, rather than the
active compounds themselves, which are transported;
Reid, 1983; Ross et al., 2006).
In pea, lkb is the main BR-deficient mutant that has been
used for grafting studies. It is now known that the lkb mut-
ation blocks BR synthesis between 24-methylenecholesterol
and campesterol in both shoot and root tissue (Nomura
et al., 1999; Schultz et al., 2001), and that it is this
blockage that results in the dwarf phenotype. Grafting lkb
scions onto WT stocks did not restore internode elonga-
tion in the mutant, while WT scions grafted to lkb stocks
elongated normally (Reid and Ross, 1989; Symons and
 Brassinosteroid transport 19
Reid, 2004). These results indicate that BRs do not
undergo acropetal transport from the more basal parts of
the plant into the young shoot tissues.
The results discussed above do not, however, preclude
the possibility that BRs are transported basipetally from
the shoot to the root. This was addressed by grafting WT
shoots onto an lkb rootstock and then quantifying
endogenous BRs in the grafted plants (Symons and Reid,
2004). Interestingly, the presence of the WT shoot did not
increase BR levels in the lkb rootstock (Fig. 1), indicating
that the maintenance of bioactive BR levels in pea roots is
not dependent on basipetal BR transport from the shoots.
Tomato is a second species in which reciprocal grafting
between a BR-deficient mutant (dx) and WT plants has
been carried out (Fig. 2; Montoya et al., 2005). An
important difference between these studies and those in
pea was that the tomato stocks continued to produce new
internodes and leaves. Therefore, when used as a stock,

Downloaded from http://jxb.oxfordjournals.org/ by guest on November 6, 2015

the dx mutant had the potential phenotypically to reflect
whether or not BRs were transported basipetally from the
WT scion. When used as a scion, the dx mutant, as with
lkb in the pea grafts, had the potential phenotypically to
reflect acropetal transport of BRs from the WT stock.
However, neither graft combination had a visible effect on
the dx phenotype, suggesting that BRs are not transported
long distances, in either the acropetal or basipetal
direction, in tomato.
It should be noted that because the pea mutant lkb
blocks relatively early in the BR pathway, the evidence
discussed above pertains to the transport not only of
bioactive BRs, but of all BRs past the lkb block. In
tomato, in contrast, the dx mutant blocks immediately
before the bioactive BR, CS, and the conclusions apply
only to that compound and its derivatives.
Given the outcomes of these grafting studies in tomato
and pea, it is possible that the acropetal movement of
exogenously applied BRs from the roots (reported in
Yokota et al., 1992; Nishikawa et al., 1994) does not
accurately reflect the transport of endogenous BRs in
plants.

Tissue excision experiments

The possibility of BR transport within the shoot was
further investigated in pea by excising the apical bud
(decapitation) or the mature leaves of WT plants and then
quantifying endogenous BRs in the remaining tissues
(Symons and Reid, 2004). Interestingly, decapitation did
not reduce BR levels in the remaining internodes and
leaves. In this respect, the BRs clearly differ from the
other growth-promotory hormones, auxin (indole-3-acetic
acid, IAA) and the bioactive gibberellin, GA1, the levels
of which are dramatically reduced in internodes after
decapitation. These studies indicate that BRs are not
imported into the stem or mature leaves from the apical
Fig. 1. Grafting of BR-deficient lkb and WT pea plants. (A) Phenotype
of the shoot in 45-d-old grafted plants, indicating the lack of acropetal
BR transport from the roots. (B) Endogenous BR levels in shoot and
root tissues of 30-d-old grafted plants, indicating the lack of acropetal
BR transport from the roots or basipetal BR transport from the
shoot. Reprinted from Plant Physiology, Vol. 135, Symons GM, Reid
JB. Brassinosteroids do not undergo long-distance transport in pea.
Implications for the regulation of endogenous brassinosteroid levels,
2196–2206, Copyright (2004). With permission from the American
Society of Plant Biologists.
20 Symons et al.
Fig. 2. Grafting of brassinosteroid-deficient dx and WT tomato plants.
(a) dx stock with WT scion, indicating the lack of basipetal BR
transport. (b) WT stock with dx scion, indicating the lack of acropetal
BR transport. Reprinted from The Plant Journal, Vol. 42, Montoya T
et al. Patterns of Dwarf expression and brassinosteroid accumulation in
tomato reveal the importance of brassinosteroid synthesis during fruit
development. 262–269, Copyright (2005), with permission from Black-
well Publishing.
bud, and that the maintenance of BR levels in internodes
does not depend on normal levels of IAA or GA1.
Similarly, excising the mature leaves did not substan-
tially alter BR levels in the remaining internodes and
apical bud. This indicates a lack of BR transport from
those leaves, consistent with the results from grafting
studies. In contrast, a subsequent study (Jager et al.,
2007) showed that defoliation reduced both IAA and GA1
levels in the remaining internodes. This indicates that
mature leaves make some contribution to the basipetal
auxin transport stream, although there is little doubt that
entry into that stream occurs mainly in the apical bud.
Strong evidence indicates that the effect of decapitation or
defoliation on GA1 content is a secondary consequence of
the drop in IAA (Ross et al., 2000).
Two lines of evidence indicate that the failure of
defoliation or decapitation to reduce BR content results
from an inability of the leaves or apical bud to export
endogenous BRs, rather than an inability to synthesize
these compounds. First, both the apical bud and mature
leaves contained substantial amounts of BRs; and sec-
ondly, as discussed earlier, mature pea leaves failed to
export exogenous BRs (Symons and Reid, 2004).
Information from variegated plants
Other experiments with the BR-deficient dx mutant of
tomato have also provided evidence for the absence of
long-distance BR transport within plant shoots (Bishop
et al., 1996). In this research, several transposon-induced
dx mutant lines were shown to exhibit a variegated pheno-
Downloaded from http://jxb.oxfordjournals.org/ by guest on November 6, 2015
type, consisting of revertant WT sectors within shoots that
otherwise exhibited a dx mutant phenotype (Bishop et al.,
1996). The fact that these WT sectors (presumably with
normal BR levels) existed within a dx background,
without completely restoring the dx phenotype, indicates
that BRs are not freely transported, even within the leaf
(Bishop and Yokota, 2001; Montoya et al., 2005).
Recently, Savaldi-Goldstein et al. (2007) used tissue-
specific promoters to drive BR biosynthesis and reception
genes in Arabidopsis, and reported only limited move-
ment of BRs between different tissues within an organ.
Implications for the regulation of BR levels
Plant hormone levels in cells or tissues must be strictly
controlled in order to facilitate an appropriate growth
response. It is widely accepted that this kind of control
over hormone levels is achieved by balancing the relative
rates of hormone synthesis, catabolism, inactivation, and
transport within the plant. However, unlike some other
plant hormones, it appears that BR levels are not regulated
by the long-distance transport of these compounds around
the plant. This clearly implies that BRs in a given tissue
(such as the fruit, roots, internodes, or leaves) are
produced in situ, rather than being produced elsewhere
and transported to that location. Consistent with this idea
are results showing that BR biosynthesis genes are widely
expressed in plant tissues, and transcript levels are
generally higher in tissues with high BR levels; although
this cannot be used as a quantitative tool for determining
BR levels (Bancos et al., 2002; Shimada et al., 2003;
Montoya et al., 2005; Symons et al., 2006; reviewed by
Nomura and Bishop, 2006). There is also good evidence
that BR synthesis is tightly regulated because several steps
in the BR biosynthesis pathway undergo feedback
regulation in response to BR levels (reviewed by Fujioka
and Yokota, 2003; Nomura and Bishop, 2006). Similarly,

it is also clear that the regulation of BR deactivation plays
an important role in the regulation of endogenous BR
levels (reviewed by Nomura and Bishop, 2006). Thus, it is
likely that endogenous BR levels in plant tissues are
primarily regulated through the tissue-specific control of
BR synthesis, catabolism, and inactivation rather than
through long-distance transport (Symons and Reid, 2004).
Do BRs undergo short-distance transport?
Although the available evidence rules out long-distance
BR transport within the plant, it is possible, indeed likely,
that endogenous BRs do move over short distances both
within and between cells. This is because there appears to
be a spatial separation of BR synthesis and perception at
the cellular level. For instance, BRs are perceived at the
exterior cell surface by a leucine-rich receptor-like kinase
named BRI1 (Fig. 3; Li and Chory, 1997; Friedrichson
et al., 2000; He et al., 2000; Wang et al., 2001).
According to the current model of BR action, BRI1 is
incorporated in the plasma membrane, with the actual site
of BR binding on the outside of that membrane; sub-
sequent to binding, the BR signal is transferred into the
cell by a domain of BRI1 that is located in the cytoplasm
(Belkhadir and Chory, 2006; Li and Jin, 2007).
Unlike BR perception, the available evidence suggests
that BR synthesis is associated with internal cell mem-
branes. For example, the DIM/DWF1 protein, which
catalyses an early step in BR synthesis, is thought to be
Fig. 3. A proposed model of short-distance BR transport within and between neighbouring cells. The model outlines the movement of BRs from
their intracellular sites of synthesis, across the plasma membrane, to the extracellular space where they can bind to BR receptors that are located on
the exterior surface of the same, or neighbouring cells.
Brassinosteroid transport 21
an integral membrane protein associated with the endo-
plasmic reticulum (Klahre et al., 1998). Similarly, the
tomato DWARF and Arabidopsis DWF4 and CYP85A1
proteins, which also catalyse steps in BR biosynthesis,
each have repeated hydrophobic residues at the N-terminus
that are likely to function as membrane-anchor regions
(Bishop et al., 1996; Choe et al., 1998; Shimada et al.,
2001). Similar to DIM/DWF1, CYP85A1, which catalyses
a BR activation step, is thought to anchor specifically to
the endoplasmic reticulum membrane (Shimada et al.,
2001). Indeed, cytochrome P450 proteins (which include
many of the BR biosynthesis enzymes characterized to
date) are predominantly membrane-bound, and the syn-
thesis of BR on or near such membranes is thought to be
necessary because steroid hormones are relatively hydro-
phobic (Bishop et al., 1996; Klahre et al., 1998; Nomura
and Bishop, 2006). Based on this evidence, it can be
assumed that BRs are produced within cells, yet it is clear
Downloaded from http://jxb.oxfordjournals.org/ by guest on November 6, 2015
that they are perceived at the exterior cell surface.
Logically, this means that BRs must be transported, either
actively or passively, from their intracellular site(s) of
synthesis to the extracellular sites of perception (Fig. 3).
Interestingly, Yamamoto et al. (2001) detected BRs in the
medium in which Zinnia elegans callus tissue was
cultured, consistent with BR movement out of these cells.
Furthermore, they reported that 6-oxo BRs, which include
the bioactive CS, are preferentially secreted from the
Zinnia cells, compared with the less active 6-deoxo BRs
(Yamamoto et al., 2001).
22 Symons et al.
While one view could be that BRs are transported out of
the cell where they are produced, to be perceived only at
the surface of neighbouring ‘target’ cells, other evidence
suggests that individual cells both synthesize and perceive
BRs. For instance, BR biosynthesis and deactivation
undergo clear feedback regulation (reviewed by Fujioka
and Yokota, 2003; Nomura and Bishop, 2006), which is
facilitated by a direct interaction between a component of
the BR signal transduction pathway, BZR1, and the
regulatory sequences of BR biosynthesis genes in the
nucleus of the cell (He et al., 2005). This feedback system
provides compelling evidence that BR synthesis and
perception can occur in/on the same cell; it seems unlikely
that these processes take place exclusively in separate
cells because if this were the case, feedback regulation of
BR levels would require a second ‘signal’ to be trans-
ported from the cells where BRs are perceived to the cells
where they are synthesized.
Thus a situation is proposed where most, if not all, cells
in actively growing tissues can both produce and perceive
endogenous BRs, and where short-distance transport of
BRs occurs within and between similar cells in a given
plant tissue. Since BRs are synthesized within the cell, it
appears that they are transported, actively or passively, out
of the cell, to interact subsequently with the BR receptor at
the surface of the same, or of a neighbouring, cell (Fig. 3).
This would allow similar cells within that tissue effec-
tively to communicate with each other regarding their BR
status, to act together to regulate BR levels, and all to
respond in a co-ordinated manner to the BR signal.
Additional evidence for short-distance cell-to-cell BR
transport comes from the BR-deficient dx mutant of
tomato (Bishop et al., 1996). As discussed earlier, several
transposon-induced dx mutant lines exhibited a variegated
phenotype, consisting of revertant WT sectors within
shoots that otherwise showed a dx mutant phenotype
(Bishop et al., 1996). While the WT sectors were unable
to rescue the mutant portions substantially, the boundaries
between the two types were diffuse, and occurred mainly
at physical boundaries such as the veins (Bishop et al.,
1996). Thus, there may have been some short-distance,
cell-to-cell transport of BRs, resulting in the rescue of
some mutant cells.
Whilst short-distance intracellular or intercellular trans-
port of BRs appears likely, a key question relates to how
this transport occurs. At present, the mechanism(s) that
might facilitate such transport are largely unknown. How-
ever, it is unlikely that it occurs through the simple passive
diffusion of free bioactive BRs within and between cells.
Under the scenario proposed above, BRs are thought to
be produced on membranes within the cell (such as the
endoplasmic reticulum), then enter the cytosol and move
to the plasma membrane, cross the plasma membrane, and
enter the extracellular environment, where they are per-
ceived at the surface of surrounding cells. For the BRs,
which are relatively non-polar molecules, this series of
movements presents some significant challenges. As a
consequence, it is likely that short-distance BR transport
involves carrier mechanisms, which allows these mole-
cules to move through these diverse cellular environ-
ments. For instance, BR transport through the cytosol
could well be facilitated by binding of BRs to other
molecules. It has been suggested that BR conjugates, such
as those formed by the binding of BRs to fatty acids or
glucose (reviewed by Fujioka and Yokota, 2003), may be
candidates for transport (Sasse, 2003; Choe, 2006).
Alternatively, BR transport could also be facilitated by
the binding of these hormones to specific proteinaceous
transporters (Sasse, 2003). Indeed, a potential mechanism
for BR transport in plants has been proposed by
Markovic-Housley et al. (2003). This involves binding of
BRs to Bet v 1, a pathogenesis-related (PR-10) protein, to
form a complex that could allow the relatively water-
Downloaded from http://jxb.oxfordjournals.org/ by guest on November 6, 2015
insoluble BRs to be transported in the cytosol (Markovic-
Housley et al., 2003). Formation of these BR conjugates
or BR–protein complexes may also enable the transport of
BRs across the plasma membrane into the extracellular
environment. However, in this instance, it is possible that
as yet unknown, membrane-bound, transporters may
also be involved. Indeed, membrane-bound, ABC (ATP-
binding cassette) transporters facilitate the movement of
steroids out of animal cells, and it is possible that
homologous mechanisms may exist in plants (Young and
Fielding, 1999; Choe, 2006). There are also many other
questions about short-distance BR transport that remain to
be answered. For instance, is the transport of BR within
and between cells an active or a passive process? Do cell
polarity and the localization of cells within tissues and
organs affect the direction of BR transport? Is the rate of
transport regulated? And if so, how? There is clearly
considerable scope for future investigations on short-
distance BR transport, the mechanism(s) by which this
occurs, and its potential role in the regulation of BR levels
and plant development.
The effect of BRs on auxin (IAA) transport
The BRs themselves do not appear to undergo long-
distance transport, but evidence is emerging that they may
affect the long-distance transport of auxin (IAA). For
example, in BR-deficient mutants of Arabidopsis, the
expression of certain PIN genes is reduced (Nakamura
et al., 2004; Li et al., 2005). The PIN gene family plays
a key role in the regulation of auxin transport, by
encoding facilitators of auxin efflux (Paponov et al.,
2005; Wisniewska et al., 2006). Auxin transport and
distribution studies also supported a possible role for BRs
in auxin transport (Li et al., 2005).
Consistent with this hypothesis are previously unpub-
lished results from pea (Fig. 4), which show that a real or

Fig. 4. Endogenous IAA levels (as determined by GC–MS–SIM; see
Symons and Reid, 2004) in different tissue types of 21-d-old WT and
lkb pea plants. Data are shown as mean 6SE (n¼2).
perceived deficiency in endogenous BR levels leads to an
altered distribution of auxin (IAA) levels in the upper
region of the shoot. In both the lkb mutant (which is
defective in BR biosynthesis; Schultz et al., 2001), and
the lka mutant (which is defective in BR signalling;
Nomura et al., 2003), a significant accumulation of IAA
was found in the apical bud, compared with WT plants
(Fig. 4; unpublished data). IAA levels in the unexpanded
internode directly below the apical bud were similar to
those of WT plants (Fig. 4). However, IAA levels in the
more mature internodes, located further down the stem,
are consistently reduced in the lka and lkb mutants,
compared with WT plants, by about 50–70% (Fig. 4;
McKay et al., 1994; Jager et al., 2005). In addition, it was
found that, after application of [3H]IAA to the apical bud,
less label appears to be transported down the stem in the
lkb mutant, compared with the WT (data not shown).
These results strongly indicate that in pea, endogenous
BRs are required for normal auxin transport from the
apical bud into the internodes below that bud. It will
therefore be interesting to investigate the expression of
pea PIN homologues in the lkb mutant.
Conclusions
The available evidence clearly indicates that endogenous
BRs do not undergo long-distance transport between the
Brassinosteroid transport 23
shoot and root or between different organs within the
shoot. As such, the BRs cannot legitimately be considered
to act as a long-distance signals like other plant hormones,
such as IAA or abscisic acid, although it is conceivable
that BRs may have an indirect role in long-distance
signalling in plants, via their putative effect on IAA
transport.
In contrast to the absence of long-distance BR transport,
it appears that BRs are transported over short distances,
since they are synthesized inside the cell but perceived on
the cell’s exterior. It seems reasonable to suggest that the
transport of BRs out of the BR-producing cell does not
stop at the exterior surface of that cell, but continues on to
the surfaces of neighbouring cells (which themselves are
exporting BRs), thereby ensuring that a relatively large
number of cells within the tissue perceive the same
concentration of bioactive BR. This would result in a co-
ordinated BR response across the cells concerned. Pre-
Downloaded from http://jxb.oxfordjournals.org/ by guest on November 6, 2015
sumably, the presence of the receptor on the exterior of
the cell facilitates this co-ordinated response. If BRs were
both produced and perceived exclusively by the same cell,
with no intercellular BR movement, this might render
difficult a co-ordinated growth response across the tissue
concerned.
Acknowledgements
The authors would like to acknowledge the financial support of the
Australian Research Council (ARC).
References
Adam G, Schneider B. 1999. Uptake, transport and metabolism.
In: Sakurai A, Yokota T, Clouse SD, eds. Brassinosteroids:
steroidal plant hormones. Tokyo, Japan: Springer, 113–136.
Arteca RN. 1995. Brassinosteroids. In: Davies PJ, ed. Plant
hormones: physiology, biochemistry and molecular biology.
Dordrecht, The Netherlands: Kluwer Academic Press, 206–213.
Bajguz A, Tretyn A. 2003. The chemical characteristic and distri-
bution of brassinosteroids in plants. Phytochemistry 62, 1027–1046.
Bancos S, Nomura T, Sato T, Molnar G, Bishop GJ, Koncz C,
Yokota T, Nagy F, Szekeres M. 2002. Regulation of transcript
levels of the Arabidopsis cytochrome P450 genes involved in
brassinosteroid biosynthesis. Plant Physiology 130, 1–10.
Belkhadir Y, Chory J. 2006. Brassinosteroid signalling: a paradigm
for steroid hormone signalling from the cell surface. Science 314,
1410–1411.
Bishop GJ, Harrison K, Jones JDG. 1996. The tomato Dwarf
gene isolated by heterologous transposon tagging encodes the
first member of a new cytochrome p450 family. The Plant Cell 8,
959–969.
Bishop GJ, Yokota T. 2001. Plant steroid hormones, brassinoste-
roids: current highlights of molecular aspects on their synthesis/
metabolism, transport, perception and response. Plant Cell
Physiology 42, 114–120.
Choe S. 2006. Brassinosteroid biosynthesis and action. Physiologia
Plantarum 126, 539–548.
Choe S, Dilkes BP, Fujioka S, Takatsuto S, Sakurai A,
Feldmann KA. 1998. The DWF4 gene of Arabidopsis encodes
 24 Symons et al.
a cytochrome P450 that mediates multiple 22a-hydroxylation
steps in brassinosteroid biosynthesis. The Plant Cell 10, 231–243.
Friedrichsen DM, Joazeiro CP, Li J, Hunter T, Chory J. 2000.
Brassinosteroid insensitive-1 is a ubiquitously expressed leucine-
rich repeat receptor serine/threonine kinase. Plant Physiology
123, 1247–1256.
Fujioka S, Yokota T. 2003. Biosynthesis and metabolism of
brassinosteroids. Annual Review of Plant Biology 54, 137–164.
He JX, Gendron JM, Sun Y, Gampala SS, Gendron N, Sun CQ,
Wang ZY. 2005. BZR1 is a transcriptional repressor with dual
roles in brassinosteroid homeostasis and growth responses.
Science 307, 1634–1638.
He Z, Wang Z-Y, Li J, Zhu Q, Lamb C, Ronald P, Chory J.
2000. Perception of brassinosteroids by the extracellular domain
of the receptor kinase BRI1. Science 288, 2360–2363.
Hoad GV. 1995. Transport of hormones in the phloem of higher
plants. Journal of Plant Growth Regulation 16, 173–182.
Jager CE, Symons GM, Glancy NE, Reid JB, Ross JJ. 2007.
Evidence that the mature leaves contribute auxin to the immature
tissues of pea (Pisum sativum L.). Planta (in press).
Jager CE, Symons GM, Ross JJ, Smith JJ, Reid JB. 2005. The
brassinosteroid growth response in pea is not mediated by
changes in gibberellin content. Planta 221, 141–148.
Klahre U, Noguchi T, Fujioka S, Takatsuto S, Yokota T,
Yoshida S, Chua N-H. 1998. The Arabidopsis DIMINUTO/
DWARF1 gene encodes a protein involved in steroid synthesis.
The Plant Cell 10, 1677–1690.
Li J, Chory J. 1997. A putative leucine-rich repeat receptor
kinase involved in brassinosteroid signal transduction. Cell 90,
929–938.
Li J, Jin H. 2007. Regulation of brassinosteroid signalling. Trends
in Plant Science 12, 37–41.
Li L, Xu J, Xu Z-H, Xue H-W. 2005. Brassinosteroids stimulate
plant tropisms through modulation of polar auxin transport in
Brassica and Arabidopsis. The Plant Cell 17, 2738–2753.
McKay MJ, Ross JJ, Lawrence NL, Cramp RE, Beveridge CA,
Reid JB. 1994. Control of internode length in Pisum sativum.
Further evidence for the involvement of indole-3-acetic acid.
Plant Physiology 106, 1521–1526.
Markovic-Housley Z, Degano M, Lamba D, von Roepenack-
Lahaye E, Clemens S, Susani M, Ferreira F, Scheiner O,
Breiteneder H. 2003. Crystal structure of a hypoallergenic
isoform of the major birch pollen allergen Bet v 1 and its likely
biological function as a plant steroid carrier. Plant Molecular
Biology 325, 123–133.
Mitchell JW, Mandava N, Worley JF, Plimmer JR, Smith MV.
1970. Brassins: a new family of plant hormones from rape pollen.
Nature 225, 1065–1066.
Montoya T, Nomura T, Yokota T, Farrar K, Harrison K,
Jones JGD, Kaneta T, Kamiya Y, Szekeres M, Bishop GJ.
2005. Patterns of dwarf expression and brassinosteroid accumula-
tion in tomato reveal the importance of brassinosteroid synthesis
during fruit development. The Plant Journal 42, 262–269.
Nakamura A, Goda H, Shimada Y, Yoshida S. 2004. Brassinos-
teroid selectively regulates PIN gene expression in Arabidopsis.
Bioscience, Biotechnology and Biochemistry 68, 952–954.
Nishikawa N, Toyama S, Shida A, Fatatsuya F. 1994. The uptake
and transport of 14C-labeled epibrassinolide in intact seedlings
of cucumber and wheat. Journal of Plant Research 107, 125–130.
Nomura T, Bishop GJ. 2006. Cytochrome P450s in plant steroid
hormone synthesis and metabolism. Phytochemistry Review 5,
421–432.
Nomura T, Bishop GJ, Kaneta T, Reid JB, Chory J, Yokota T.
2003. The LKA gene is a BRASSINOSTEROID INSENSITIVE 1
homolog of pea. The Plant Journal 26, 291–300.
View publication stats
Nomura T, Kitasaka Y, Takatsuto S, Reid JB, Fukami M,
Yokota T. 1999. Brassinosteroid/sterol synthesis and plant growth
as affected by lka and lkb mutations of pea. Plant Physiology
119, 1517–1526.
Nomura T, Ueno M, Yamada Y, Takatsuto S, Takeuchi Y,
Yokota T. 2007. Roles of brassinosteroids and related mRNAs in
pea seed growth and germination. Plant Physiology (in press).
Paponov IA, Teale WD, Trebar M, Blilou I, Palme K. 2005. The
PIN auxin efflux facilitators: evolutionary and functional perspec-
tives. Trends in Plant Science 10, 170–177.
Reid JB. 1983. Internode length in Pisum. Plant Physiology 72,
759–763.
Reid JB, Ross JJ. 1989. Internode length in Pisum. Two further
gibberellin insensitivity genes lka and lkb. Physiologia Plantarum
75, 81–88.
Ross JJ, O’Neill DP, Smith JJ, Kerckhoffs HJ, Elliot RC. 2000.
Evidence that auxin promotes gibberellin A1 biosynthesis in pea.
The Plant Journal 21, 547–552.
Ross JJ, Symons GM, Abas L, Reid JB, Luschnig C. 2006.
Hormone distribution and transport. In: Hedden P, Thomas S,
eds. Plant hormone signalling, Vol. 24. Oxford, UK: Blackwell
Downloaded from http://jxb.oxfordjournals.org/ by guest on November 6, 2015
Publishing, 257–291.
Sasse J. 1999. Physiological actions of brassinosteroids. In: Sakurai A,
Yokota T, Clouse SD, eds. Brassinosteroids: steroidal plant
hormones. Tokyo: Springer-Verlag, 137–161.
Sasse J. 2003. Physiological action of brassinosteroids: an update.
Journal of Plant Growth Regulation 22, 276–288.
Savaldi-Goldstein S, Peto C, Chory J. 2007. The epidermis both
drives and restricts plant shoot growth. Nature 446, 199–202.
Schultz L, Kerckhoffs LHJ, Klahre U, Yokota T, Reid JB. 2001.
Molecular characterization of the brassinosteroid-deficient lkb
mutant in pea. Plant Molecular Biology 47, 491–498.
Shimada Y, Fujioka S, Miyauchi N, Kushiro M, Takatsuto S,
Nomura T, Yokota T, Kamiya Y, Bishop GJ, Yoshida S. 2001.
Brassinosteroid-6-oxidases from Arabidopsis and tomato catalyse
multiple C-6 oxidations in brassinosteroid biosynthesis. Plant
Physiology 126, 770–779.
Shimada Y, Goda H, Nakamura A, Takatsuto S, Fujioka S,
Yoshida S. 2003. Organ-specific expression of brassinosteroid-
biosynthetic genes and distribution of endogenous brassinoste-
roids in Arabidopsis. Plant Physiology 131, 287–297.
Symons GM, Reid JB. 2004. Brassinosteroids do not undergo
long-distance transport in pea. Implications for the regulation
of endogenous brassinosteroid levels. Plant Physiology 135,
2196–2206.
Symons GM, Davies C, Shavrukov Y, Dry IB, Reid JB,
Thomas MR. 2006. Grapes on steroids: brassinosteroids are
involved in grape berry ripening. Plant Physiology 140, 150–158.
Wang Z-Y, Seto H, Fujioka S, Yoshida S, Chory J. 2001. BRI1 is
a critical component of a plasma-membrane receptor for plant
steroids. Nature 410, 380–383.
Wisniewska J, Xu J, Seifertova D, Brewer PB, Ruzicka K,
Blilou I, Rouquie D, Benkova E, Scheres B, Friml J. 2006.
Polar PIN localization directs auxin flow in plants. Science 312,
883.
Yamamoto R, Fujioka S, Demura T, Takatsuto S, Yoshida S,
Fukuda H. 2001. Brassinosteroid levels increase drastically prior
to morphogenesis of tracheary elements. Plant Physiology 125,
556–563.
Yokota T, Higuchi K, Kosaka Y, Takahashi N. 1992. Transport and
metabolism of brassinosteroids in rice. In: Karssen CM, van Loon
LC, Vreugdenhil D, eds. Progress in plant growth regulation. The
Netherlands: Kluwer Academic Publishers, 298–305.
Young SG, Fielding CJ. 1999. The ABCs of cholesterol efflux.
Nature Genetics 22, 316–318.