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1 s2.0 S0031942212004876 Main
1 s2.0 S0031942212004876 Main
Phytochemistry
journal homepage: www.elsevier.com/locate/phytochem
Weed Science Center, Utsunomiya University, 350 Mine-machi, Utsunomiya 321-8505, Japan
Graduate School of Life and Environmental Sciences, Osaka Prefecture University, 1-1 Gakuen-cho, Naka-ku, Sakai, Osaka 599-8531, Japan
Department of Biosciences, Teikyo University, 1-1 Toyosatodai, Utsunomiya 320-8551, Japan
a r t i c l e
i n f o
Article history:
Received 5 October 2012
Received in revised form 8 November 2012
Available online 3 January 2013
Keywords:
Houttuynia cordata
Saururaceae
Strigol
Strigolactone
Strigone
Sorgomol
5-Deoxystrigol
Orobanche minor
Phelipanche ramosa
Striga hermonthica
Orobanchaceae
a b s t r a c t
(+)-Strigone was described earlier in a paper on isolation of strigol and then recently examined for hyphal
branching activity in arbuscular mycorrhizal fungi as a strigolactone. Herein, it was isolated from root
exudates of Houttuynia cordata, and its structure was conrmed by direct comparison with synthetic
standards in LCMS/MS, GCMS, and 1H and 13C NMR analyses. The stereochemistry of strigone was
determined by comparing the CD spectra and RRt in chiral LCMS/MS with those of synthetic (+)-strigone
and ()-strigone. Four stereoisomers of strigone exhibited clearly different levels of stimulation activity
on the seeds of three root parasitic plants, Orobanche minor, Phelipanche ramosa, and Striga hermonthica.
(+)-Strigone was a highly potent germination stimulant on S. hermonthica and also on P. ramosa, but less
active than ent-20 -epi-strigone on O. minor. In addition to strigone, H. cordata was found to produce strigol, sorgomol, and 5-deoxystrigol, indicating that this plant produces mainly strigol-type strigolactones
derived from 5-deoxystrigol.
2012 Elsevier Ltd. All rights reserved.
1. Introduction
Strigolactones (SLs) are carotenoid-derived plant secondary
metabolites which in the rhizosphere induce seed germination of
root parasitic plants, witchweeds (Striga spp.) and broomrapes
(Orobanche and Phelipanche spp.), and promote root colonization
by arbuscular mycorrhizal (AM) fungi (Akiyama et al., 2005; Bouwmeester et al., 2007; Xie et al., 2010). In planta, SLs function as
a novel class of hormones which are involved in the regulation of
shoot (Gomez-Roldan et al., 2008; Umehara et al., 2008) and root
architecture (Koltai, 2011; Ruyter-Spira et al., 2011), and various
developmental processes through cross-talk with other hormones.
SLs are distributed widely in the plant kingdom (Xie et al., 2010),
and even mosses and Charales produce and utilize them presumably in regulation and promotion of rhizoid elongation (Delaux
et al., 2012; Proust et al., 2011).
More than 15 SLs have been puried and characterized from
root exudates of various plant species (Xie et al., 2010) since strigol
was rst isolated from cotton root exudates as a germination stim Corresponding author. Tel.: +81 286495152.
E-mail address: yoneyama@cc.utsunomiya-u.ac.jp (K. Yoneyama).
0031-9422/$ - see front matter 2012 Elsevier Ltd. All rights reserved.
http://dx.doi.org/10.1016/j.phytochem.2012.11.013
ulant for S. lutea (Cook et al., 1966, 1972). All natural SLs characterized so far have a tricyclic lactone (ABC part) connected to a
butenolide group (D-ring) via an enol ether bridge (Xie et al.,
2010). They have different substituents on the A/B ring but the
same CD moiety which has been suggested to be essential for biological activity (Zwanenburg et al., 2009). Natural SLs can be divided into two groups, strigol-type and orobanchol-type SLs
carrying the a-oriented and b-oriented C ring, respectively (Yoneyama et al., 2012). The two simplest SLs, 5-deoxystrigol and its isomer ent-20 -epi-5-deoxystrigol appear to be the common precursors
for the other SLs, the former for strigol-type SLs and the latter for
orobanchol-type SLs. The biosynthesis of these two types of SLs
seems to be regulated somewhat independently in plant species
like Chinese milk vetch (Yoneyama et al., 2012).
In addition to these known SLs, detection of novel SLs including
methoxy-5-deoxystrigol isomers from rice (Jamil et al., 2011), SL1
and SL2 from maize (Jamil et al., 2012), and didehyro-orobanchol
(or strigol) isomers from red clover (Sato et al., 2003), tobacco
(Xie et al., 2007), and tomato (Lpez-Rez et al., 2008) have been
reported. Working with some of these novel SLs in our laboratory
was limited due to lack of enough pure samples for structural elucidation. In earlier studies, Ma et al. (2004) reported germination
61
stimulating activities of aqueous and methanol extracts from various Chinese medicinal herbs on the seeds of witchweed, Striga
hermonthica. In fact, the root culture of one of these medicinal
herbs, Menispermum dauricum, produces strigol (Yasuda et al.,
2003). By further analysis of the aqueous and methanol extracts
of Houttuynia cordata, that has been reported to be active on S. hermonthica seed germination (Ma et al., 2004, 2005), it was established that root exudates from this plant grown hydroponically
elicited the seed germination of Orobanche minor. Herein, are described the isolation and identication of (+)-strigone (1) (Cook
et al., 1972; Akiyama et al., 2010) as a natural SL from root exudates of H. cordata, and the signicantly different germination
stimulation activities of its stereoisomers on the seeds of three root
parasites, O. minor, Phelipanche ramosa, and S. hermonthica (Fig. 1).
10
8a
8b
O
3a
4a
6'
2'
5'
OH
4'
strigone (1)
HO
7'
strigol (2)
O
O
sorgomol (3)
ment with those of authentic ()-strigone and its epimer (Fig. 2a).
The RRt of 1 was identical to those of ()-strigone in both GCMS
and LCMS/MS analyses (Fig. 2b). Thus, 1 was determined to be
strigone.
To elucidate the stereochemistry of 1, (+)- and ()-strigones
were synthesized from optically pure strigol enantiomers (see Section 4) and the CD spectra of each were compared with that of 1.
The spectral pattern of (+)-strigone was identical to that of 1, and
chiral LCMS/MS analyses also supported this assignment. Consequently, 1 was determined to be (+)-strigone (1).
C
O
3'
Fig. 2. GCMS analysis of 1. (a) EIMS spectrum of 1. (b) Total ion chromatograms
of authentic strigones (upper) and 1 (lower).
5-deoxystrigol (4)
62
4. Experimental
O. minor
Germination (%)
100
80
60
ent-1
2'-epi-1
ent-2'-epi-1
40
20
0
0.01
0.1
10
100
10
100
Concentration (nM)
P. ramosa
Germination (%)
100
80
60
40
20
0
0.01
0.1
Concentration (nM)
S. hermonthica
Germination (%)
100
80
60
40
20
0
0.01
0.1
10
100
Concentration (nM)
Fig. 3. Germination stimulating activities of strigone stereoisomers toward O.
minor, P. ramosa and S. hermonthica seeds. Data are presented as means se (n = 3).
3. Concluding remarks
In the present study, strigone (1) was isolated as a natural SL.
Although 1 has the molecular formula same to those of putative
didehydro-orobanchol (strigol) isomers, 1 appeared to be different
from those detected in tobacco, red clover, and tomato by LCMS/
MS and GCMS analyses (data not shown). The observation that
highly potent germination stimulation activity of 1 on S. hermonthica while it is a rather weak stimulant on O. minor implies that different root parasitic plants may respond to an SL with different
sensitivities. Therefore, it would be expected to nd species-specic SLs (or specic mixtures of SLs) for root parasites with very
narrow host ranges such as O. hederae which parasitizes only ivy
(Fernndez-Aparicio et al., 2009).
63
4.8.2. ()-ent-Strigone
CD (c 0.0050, CH3CN) kmax (De) nm: 269 (1.17), 246 (24.4), 227
(4.06), 216 (2.85). [a]D25 222 (c 0.35, CHCl3). 1H and 13C NMR
and mass data were the same as for (+)-strigone.
64
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