Biochemical Systematics and Ecology 32 (2004) 631635

www.elsevier.com/locate/biochemsyseco

Eects of quercetin and its seven derivatives

on the growth of Arabidopsis thaliana
and Neurospora crassa
Mohammad Masud Parvez a, Kaori Tomita-Yokotani b,
,
Yoshiharu Fujii a, Tatsuo Konishi c, Tsukasa Iwashina c
a

Chemical Ecology Unit, National Institute for Agro-Environmental Sciences, Tsukuba,

Ibaraki 305-8604, Japan
b
Institute of Applied Biochemistry, University of Tsukuba, Tsukuba, Ibaraki 305-8572, Japan
c
Tsukuba Botanical Garden, National Science Museum, Tsukuba, Ibaraki 305-0005, Japan
Received 20 January 2003; accepted 20 December 2003

Abstract
Structureactivity-relationships of quercetin and its seven derivatives were investigated
using a shoot growth test with Arabidopsis thaliana seedlings and conidial germination test
with Neurospora crassa. All the tested substances inhibited the shoot growth of A. thaliana.
On the other hand, in conidial germination test of N. crassa, some avonoids did not show
any inhibitory activity. Quercetin 3-methyl ether and its glycosides especially showed the
highest inhibitory eect among them in the conidial germination test of Neurospora. These
results indicate that the presence of methyl group in avonoid nucleus have some important
roles as inhibiting eect to A. thaliana and N. crassa.
# 2004 Elsevier Ltd. All rights reserved.
Keywords: Allelopathy; Arabidopsis thaliana; Flavonoid; Neurospora crassa; Quercetin; Quercetin
3-methyl ether

1. Introduction
There have been many kinds of compounds in nature that have allelopathic
properties. It is well known that some avonoids, e.g. hesperetin 7-O-rutinoside,
taxifolin 7-O-arabinoside and formononetin 7-O-glucoside, have allelopathic

Corresponding author. Tel.: +81-298-53-4617; fax: +81-298-53-7229.

E-mail address: kaboka@sakura.cc.tsukuba.ac.jp (K. Tomita-Yokotani).

0305-1978/$ - see front matter # 2004 Elsevier Ltd. All rights reserved.
doi:10.1016/j.bse.2003.12.002

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M.M. Parvez et al. / Biochemical Systematics and Ecology 32 (2004) 631635

properties in plant such as Pluchea lanceolata (DC) C.B. Clarke (Compositae)

(Inderjit and Dakshini, 1991, 1992, 1994; Rice-Evans and Packet, 1998). Recently,
two avonoids, isorhamnetin 3-O-rutinoside and isorhamnetin 3-O-robinobioside
were isolated from the pericarps of the cactus, Hylocereus undatus Britton Rose
and their allelopathic activity against Celosia cristata L. (Amaranthaceae) and a
fungus, Neurospora crassa Sheat B.D. Dodge, were elucidated (Tomita-Yokotani
et al., 2004).
In this study, we study the eects of a common avonoid, quercetin and its derivatives against plant and fungal growth, and discuss the eect of avonoids as allelopathic substances in the environment. Arabidopsis thaliana (L.) Heynh. and
Neurospora crassa were used as materials for assay. These organisms are very useful for allelopathy studies, because A. thaliana and N. crassa are used as plant and
fungus models in molecular studies and much is known about dierent aspects of
their biology (Tomita-Yokotani et al., 2003).
2. Materials and methods
2.1. Flavonoid materials
Quercetin and its seven derivatives were tested in the seedling and conidial germination assays (Fig. 1). The origins of the avonoids are as follows: Quercetin
from the owers of Astrophytum ornatum Britton Rose (Iwashina et al., 1988).
Quercetin 3- O-glucoside (isoquercitrin) and quercetin 3-O-rhamnoside (quercitrin)
(Synthese, Lab Sarget, Merignac), quercetin 3-O-galactoside (hyperin) from the

Fig. 1. Structures of quercetin and its derivatives. 1 Quercetin 3-O-galactoside, 2 Quercetin 3-O-glucoside, 3 Quercetin 3-O-arabinofuranoside, 4 Quercetin 3-O-rhamnoside, 5 Quercetin, 6 Quercetin 3methyl ether 4-O-glucoside, 7 Quercetin 3-methyl ether 7-O-glucoside, 8 Quercetin 3-methyl ether.

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633

ower of Notocactus ottonis (Lehm.) Berger (Iwashina and Ootani, 1986). Quercetin 3-O-arabinofuranoside (avicularin) (Extrasythese, Genay France). Quercetin
3-methyl ether, quercetin 3-methyl ether 40 -O-glucoside (neochilenin) and quercetin
3-methyl ether 7-O-glucoside (transilin) from the owers of Neochilenia spp. (Iwashina et al., 1984).
2.2. Arabidopsis seedling growth test
Arabidopsis thaliana ecotype Columbia was used for the seedling growth test.
The seeds were surface sterilized with 0.1% sodium hypochlorite for 10 min,
washed with distilled water three times, and then sown in a 9 cm Petri-dish with a
v
moistened lter paper (Toyo no. 1) and incubated at 5 C in the dark for 4 days.
Five seeds were incubated in a 100 ml ask with 1 ml of Murashige-Skoog medium
v
(MS medium) at 23 C for 4 days in the light (8 lmol m2 s1) with 30 shakings
per min. Test compounds were dissolved into ethyl alcohol and diluted with incubation medium. The suspended solutions were added into the medium at the
initiation of incubation. The same volume of ethyl alcohol in the medium was
added into the medium as control. The nal percentage of alcohol in the medium
was lower than 0.05%. Growth of seedlings was recorded by taking photographs
and then measuring the growth with a ruler on enlarged copies of the photographs.
2.3. Neurospora germination test
Wild type strain of Neurospora crassa 74A was used as the test fungus. The conv
idia were inoculated on the complete agar medium and incubated at 26 C for 6
days in the dark, then exposed to normal laboratory uorescent light for 1 day, as
reported previously (Tomita et al., 1996). The conidia of the cultures were suspended in cold sterile water, and then ltered through two layers of gauze to
remove the contamination of mycelia fragments. The suspension was centrifuged at
v
1780  g for 10 min at 4 C and the pellet was washed with sterile water. The conidia were re-suspended and incubated in a 8 ml tube with a 1 ml volume of Fries
minimal medium, containing 1.5% sucrose with test solution or without test solv
ution as control at an optimum concentration of 2  106 conidia ml1 at 26 C in
the dark with 120 shakings per min. The method for testing the solutions was the
same as that used in the Arabidopsis seedling growth test. Conidial germination
was counted 3 h after incubations. These conidial germination tests were carried
out as described previously (Tomita et al., 1996, Tomita-Yokotani et al., 2003).
3. Results and discussion
Allelopathy plays an important role in the agro-ecosystems and a wide range of
interaction in biotic communities. Extensive information on allelopathy phenomenon is available, however, very few substances with allelopathy activity have been
identied.

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M.M. Parvez et al. / Biochemical Systematics and Ecology 32 (2004) 631635

Fig. 2. Eects of ve quercetin derivatives and three methoxyquercetin derivatives on the growth of
Arabidopsis thaliana (A) and conidial germination of Neurospora crassa (B) (see Fig. 1 for key to
compounds).

Flavonoids are universally distributed in dierent tissues like owers, roots,

leaves and stems of higher plants. Some of the avonoids like quercetin and
kaempferol have already been isolated as allelopathic substances (Williams, 1960;
Arntzen et al., 1974), however, the details of their functions and mechanisms as
allelopathic substances have not yet been investigated.
Quercetin 3-methyl ether (8) and its 40 -O-glucoside (6) and 7-O-glucoside (7)
completely inhibited the growth of A. thaliana at 100 ppm (Fig. 2A). At lower concentrations (30 ppm), they completely inhibited growth. These three compounds
also inhibited conidial germination and they acted dierently (Fig. 2B). Quercetin
3-O-glucoside was the only non-methylated avonoids to inhibit conidial germination (Fig. 2B).
These results indicate that the presence of a methyl group in the avonoid
nucleus has an important role in inhibiting A. thaliana and N. crassa. Further studies are being undertaken on a range of other avonoids to investigate structure
allelopathy relationships.

Acknowledgements
The authors are grateful to Prof. Dr. Nobuharu Goto, Miyagi University of
Education for providing seeds of Arabidopsis thaliana and to Prof. Dr. Koji Hasegawa,
University of Tsukuba for cooperation. A part of this work is supported by the
fund from the Cooperative System for Supporting Priority Research (CSSPR),
Japan Science and Technology Corporation, Ministry of Education, Culture,
Science, Technology and Sports, Japan to M.M.P. and Y.F.

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