Branching Hormone is Busy Both Underground and
Overground
Shinjiro Yamaguchi1 and Junko Kyozuka2
1RIKEN Plant Science Center, Japan
2Graduate School of Agriculture and Life Sciences, University of Tokyo, Japan
Abbreviations: AM, arbuscular mycorrhizal; CCD, CAROTENOID
CLEAVAGE DIOXYGENASE; SL, strigolactone.
The discovery of strigolactone (SL) or its derivatives as a new
hormone that inhibits shoot branching was a recent significant
breakthrough in plant biology (Gomez-Roldan et al. 2008,
Umehara et al. 2008). It was discovered independently by two
groups, who submitted their papers for publication only a few
weeks apart; it was clearly the moment for SL to be revealed.
Shoot branching is controlled by two distinct mechanisms,
namely formation and subsequent outgrowth of axillary buds
produced in the axils of leaves (Bennett and Leyser 2006). The
latter aspect, the control of bud outgrowth, is generally more
important in the determination of plant architecture. After
initiation, axillary buds make the decision to continue to grow
or to stay dormant, depending on a number of internal and
environmental cues. The presence or absence of an apical bud
is one such cue. Auxin supplied by the apical bud inhibits
axillary bud outgrowth, while auxin depletion caused by
decapitation triggers bud outgrowth. Physiological studies of
this phenomenon, called apical dominance, have predicted
the involvement of a second messenger of auxin; however, the
messenger has remained unknown for decades.
In the 1990s, mutants with an excess branching phenotype
were collected in garden pea, Arabidopsis and petunia. They
were named ramosus (rms), more axillary growth (max) and
decreased apical dominance (dad), respectively. Grafting experi-
ments using these mutants indicated that a long-distance,
graft-transmissible signal plays a critical role in the inhibition of
bud outgrowth (Foo et al. 2001). It was also suggested that,
among the branching mutants, rms1 and rms5 of pea and
max1, max3 and max4 of Arabidopsis are involved in the bio-
synthesis of the graft-transmissible signal, while rms5 and max2
probably have defects in perception or signalling.
Molecular cloning of the RMS and MAX genes stimulated
the progress of studies on shoot branching. First, the finding
that RMS5/MAX3 and RMS1/MAX4 encode CAROTENOID
CLEAVAGE DIOXYGENASE 7 (CCD7) and CCD8 led research-
ers to anticipate that the hormone might be derived from caro-
tenoids (Sorefan et al., 2003, Booker et al. 2004). In addition, the
Plant Cell Physiol. 51(7): 1091–1094 (2010) doi:10.1093/pcp/pcq088, available online at www.pcp.oxfordjournals.org
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Plant Cell Physiol. 51(7): 1091–1094 (2010) doi:10.1093/pcp/pcq088 © The Author 2010.
conservation of the mechanism controlling shoot branching
became apparent from studies in other plant species, in partic-
ular rice (Ishikawa et al. 2005).
SL was already known as an inducer of hyphal branching
in arbuscular mycorrhizal (AM) fungi, as well as a stimulator
of germination of root parasitic plants such as witchweeds
(Striga spp.) and broomrapes (Orobanche and Phelipanche spp.)
(Yoneyama et al. 2010). However, the link had never been made
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between SL and shoot branching in plants. In 2005, it was
reported that SL might be synthesized from carotenoids
(Matusova et al. 2005). Combining this information with the
finding that CCD action is required to synthesize the branch-
inhibiting hormone, two groups suspected that the hormone
might be SL or its derivatives. They tested this idea and found
that this is indeed the case (Gomez-Roldan et al. 2008,
Umehara et al. 2008). SLs were below the detectable level in
putative hormone-deficient mutants, whose defects were
rescued by application of GR24, an SL analog. The fact that
a level of GR24 as low as 10 nM was sufficient to inhibit bud
outgrowth was consistent with the idea that SL works as a plant
hormone. This was in fact the third discovery of SL. It was first
identified in the 1960s through its ability to stimulate seed
germination in root parasitic plants (Cook et al. 1966). Then,
in 2005 it was rediscovered as an inducer of AM fungi hyphal
branching (Akiyama et al. 2005). Because symbiosis between
AM fungi and plants is an ancient phenomenon, beginning
when plants landed 4.8 million years ago, parasitic weeds
probably imitated AM fungi to utilize the SL exudated from
plants roots. AM fungi aid plants in taking up phosphate (Pi)
from the soil. Interestingly, in planta SL synthesis is dramatically
induced under low Pi conditions (Yoneyama et al. 2007a,
Yoneyama et al. 2007b). This may imply that SL works as an
integrator of the nutrient signal and plant development
(Umehara et al. 2008).
So far, four genes that act in the SL biosynthesis pathway
have been identified (for a review, see Beveridge and Kyozuka
2010). Currently it is proposed that CCD7 and CCD8 function
in the chloroplast to cleave a carotenoid substrate. Then,
MAX1, a cytochrome P450, works on the substrate that is
mobile. D27 of rice, encoding an iron-containing protein, also
1091
 functions in the SL biosynthesis pathway; however, its molecu-
lar function remains to be determined. As observed for other
plant hormones, feedback regulation seems to be important in
the control of SL action. Expression of CCD7 and CCD8 genes is
up-regulated in pea, rice and Arabidopsis branching mutants.
Induction of CCD8 expression by auxin is also commonly
observed in all three species. According to recent studies, the
most plausible scenario is that the SL works downstream
of auxin as a second messenger to inhibit bud outgrowth
(Hayward et al. 2009). The auxin action in shoot branching is
mediated in part through regulation of the SL level, as well as
the control of cytokinin levels, as previously demonstrated
(Tanaka et al. 2006).
Genetic approaches using SL-insensitive branching mutants
are expected to be powerful in elucidating the SL signaling
pathway. Grafting experiments have shown that MAX2/RMS4 is
a putative component of SL signaling. MAX2/RMS4/D3 encodes
an F-box protein, suggesting that degradation of negative regu-
lators through ubiquitin-mediated protein degradation is likely
to be a key step in SL signaling (Stirnberg et al. 2002, Ishikawa
et al. 2005). Based on its sequence similarity to TIR1 and COI1,
receptors for auxin and jasmonic acids, respectively, MAX2/
RMS4/D3 has been proposed as a candidate for the SL receptor.
Recently, however, the discovery of the D14 gene in rice, encod-
ing a protein of the α/β-fold hydrolase superfamily, has changed
this view (Arite et al. 2009). d14 is an SL-insensitive increased
branching mutant. Considering that GID1, a receptor for gib-
berellins, is also a member of this family, and that protein deg-
radation is a crucial step in gibberellin signaling, it is reasonable
to predict that D14 may be an SL receptor (Ueguchi-Tanaka
et al. 2005). Nevertheless, the possibility that D14 acts as an
enzyme to catalyze the metabolic conversion(s) of SLs to
a bioactive form of the branching hormone still cannot be
ruled out.
This special issue covers a variety of topics related to SLs,
including their roles as rhizosphere signals and endogenous
hormones. Root parasitic plants have long been devastating
weeds of important crops in many parts of the world, especially
in Africa (Parker 2009). In this special issue, Yoneyama et al.
(pp. 1095–1103) provide a Mini-review article on the role of
SLs as seed germination stimulants in root parasitic plants in
their unique life cycle. In addition, they summarize recent
updates on the chemical diversity of SLs produced by plants
and their germination-stimulating activity on root parasite
seeds. The authors also discuss how quantitative and qualita-
tive differences in SL exudation between different sorghum
cultivars affect their susceptibility to Striga.
In contrast to root parasites, AM fungi are beneficial part-
ners in the rhizosphere of the host plant. To pursue the poten-
tial usefulness of SLs in agriculture, a key question is whether
any SL derivative can activate AM fungi without stimulating
root parasite seeds. Akiyama et al. (pp. 1104–1117) have carried
out extensive structure–activity relationship studies in the
induction of hyphal branching in AM fungi, using numerous SLs
and SL analogs. They find that the structural requirements for
1092 Plant Cell Physiol. 51(7): 1091–1094 (2010) doi:10.1093/pcp/pcq088 © The Author 2010.
SL to promote hyphal branching in AM fungi are largely similar
to those for stimulating the germination of root parasite seeds,
but are not identical, especially with respect to the enol ether
bridge. These findings suggest a unique feature of the recogni-
tion of SLs by AM fungi.
It has previously been established that Pi deficiency results in
elevated production/exudation of SLs from roots, in particular
in hosts of AM fungi (Yoneyama et al. 2007a, Yoneyama et al.
2007b). In this special issue, Umehara et al. (pp. 1118–1126)
address how endogenous SL levels elevated by Pi starvation
contribute to its hormonal action in rice. The authors show
that tiller growth is inhibited under low Pi conditions and that
this inhibition is dependent on SL biosynthesis and signaling.
They speculate that SL plays two roles in the adaptation to Pi
deficiency: as a rhizosphere signal that promotes AM fungi
symbiosis for efficient Pi acquisition; and as an endogenous
hormone (or biosynthetic precursor of a hormone) that opti-
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mizes shoot branching for efficient Pi utilization. This may
explain why plants use the same chemical signal to regulate
both shoot architecture and AM fungi symbiosis.
It has been well established that SL participates in the inhibi-
tion of axillary bud outgrowth, but little is known about its
mode of action. One potential downstream component in
the SL pathway in rice is FINE CULM1 (FC1), an ortholog of
maize TEOSINTE BRANCHED1. In this issue, Minakuchi et al.
(pp. 1127–1135) address this hypothesis. FC1 encodes a member
of the TCP family of transcription factors, and fc1 mutant plants
exhibit increased tillering (Takeda et al. 2003). The authors
show that the fc1 mutant is insensitive to a high dose of GR24
treatment, and that overexpression of FC1 partially rescues the
d3 mutant phenotype. They also found that FC1 expression is
negatively regulated by cytokinin treatment. The results sup-
port the idea that FC1 partly functions downstream of SL
action, and acts as an integrator of multiple hormone signaling
pathways that regulate shoot branching in rice.
Initial studies on the hormonal effects of SL focused on
its role as a shoot branching inhibitor (Gomez-Roldan et al.
1998, Umehara et al. 2008). However, previous work on the
rms/max/dad/d mutants had suggested that these mutations
affect other developmental processes as well, including leaf
morphology in the Arabidopsis max mutants and delayed
senescence and reduced root growth in the petunia dad1/ccd8
mutants (Stirnberg et al. 2002, Snowden et al. 2005). These
results suggest that SL regulates diverse developmental pro-
cesses as do other plant hormones. In this special issue, Hu et al.
(pp. 1136–1142) report another new role for SL in rice. Using
dark-grown SL-deficient and -insensitive mutant seedlings, the
authors demonstrate that SL inhibits mesocotyl elongation
by negatively regulating cell division. This discovery further
supports the notion that SL may have diverse roles in plant
growth and development.
Besides genetic mutations, an alternative way to study the
biological roles, biosynthesis and mechanisms of action of
a hormone is to employ chemical inhibitors of hormone bio-
synthesis or action. In this issue, Ito et al. (pp. 1143–1150) report
their unique approach toward discovering an SL biosynthesis
inhibitor. At least one cytochrome P450, CYP711A1/MAX1, is
involved in the SL biosynthesis pathway. The authors therefore
screened a chemical library consisting of triazole derivatives,
some of which have been known as effective inhibitors of
plant cytochrome P450s (Asami et al. 2000, Rademacher 2000).
They identified a chemical that induced tiller bud outgrowth
of rice seedlings, exactly as observed in SL-deficient mutant
seedlings. The discovery of a promising lead chemical in this
study will allow the authors to develop more specific SL bio-
synthesis inhibitors through structure–activity relationship
studies, an approach that has been successful in the develop-
ment of ABA and brassinosteroid inhibitors (Min et al. 1999,
Han et al. 2004).
SL research has entered a new phase, and there is still plenty
to learn about the biological roles, biosynthesis, transport
and signaling of SLs as endogenous hormones, as well as their
exudation from the root and recognition by symbiotic AM
fungi and root parasites. We believe that this special issue will
be a significant contribution to this research area, and hope
that it will attract the broader interest of PCP readers and
encourage more researchers to join this exciting and emerging
research field.
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1094 Plant Cell Physiol. 51(7): 1091–1094 (2010) doi:10.1093/pcp/pcq088 © The Author 2010.