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J. Bio. & Env. Sci. 2014
82 | Shams et al
RESEARCH PAPER OPEN ACCESS
Allelopathic effects of weeds extract on seed germination of
Trachyspermum copticum L., Foeniculum vulgare and
Cuminum cyminum
Mostafakemal Shams1*
, Ehsan Zandi Esfahan2
, Mahdi Ramezani3
, Mahnaz
Ghandkanlou1
1
Department of agriculture, Payame Noor University, P.O. Box 19395- 3697, Tehran, Iran
2
Rangeland Research Division, Research Institute of Forests and Rangelands, P.O. Box 13185-116,
Tehran, Iran
3
Science and Research Branch, Islamic Azad University, P.O. Box 14778. Tehran, Iran
Article published on July 08, 2014
Key words: Allelopathic effects, weed species, Trachyspermum copticum L., Foeniculum vulgare, Cuminum
cyminum
Abstract
Seed germination stage is one of the most basic growth stages in medicinal plants that may be adversely affected
by the chemicals released from other plants in the soil environment. The present study was performed to
investigate the allelopathic effects of weeds, including (Xanthium strumarium, Amaranthus retroflexus and
Chenopodium album) as common weeds in medicinal plant's farms, on seed germination parameters, total dry
weight and allometry coefficient of Trachyspermum copticum L., Foeniculum vulgare and Cuminum cyminum
L. in laboratory conditions. Treatments included extracts of all organs of three weed species, applied on the seeds
of the medicinal plants at three concentrations of zero (distilled water), 50 and 100% by volume. The results
showed that the extracts of mentioned weed species had a significant inhibitory effect on germination
characteristics of the studied medicinal plants. The greatest negative impact on seed germination was recorded
for the extract of Amaranthus retroflexus, but Trachyspermum copticum L. showed a better resistance to the
extract of Amaranthus retroflexus as compared to Foeniculum vulgare and Cuminum cyminum.
*Corresponding Author: Mostafakemal Shams  shams100@inbox.com
Journal of Biodiversity and Environmental Sciences (JBES)
ISSN: 2220-6663 (Print) 2222-3045 (Online)
Vol. 5, No. 1, p. 82-90, 2014
http://www.innspub.net
J. Bio. & Env. Sci. 2014
83 | Shams et al
Introduction
The world's need for medicinal plants as medicines,
cosmetics and as a food supplement for improving
human welfare is rising day by day. Allelopathic is the
interaction of produced plant chemicals on each
other.
Allelopathy is one of the major problems in most
agricultural lands so that many weed species affect
the germination and growth of other plants by
producing secondary metabolites (Anowar Razvy et
al., 2011; Mizutani, 1999) and the Production of
secondary metabolites in plants depends on
environmental conditions (Shams et al., 2013).
The reduction of plant yield is not only due to weed
interference in field but allelopathic agents can play
an effective role in this regard (Iqbal and Wright,
1999).
In the farms, Xanthium strumarium (Rough
Cocklebur), Amaranthus retroflexus (Redroot
Amaranth) and Chenopodium album (Lamb's
Quarters) as weed species cause a sharp decline in the
yield of cultivated plants (Azizi and Fuji, 2006).
Allelopathic activity of Redroot Amaranth has been
fully proven. Saponins, phenolics, chlorogenicacid,
scopoline and benzoic acid are allelopathic
compounds of Redroot Amaranth (Inderjit and Duke,
2003).
According to the new researches, seven sesquiterpens
have been reported for Redroot Amaranth. The
inhibitory effects of these compounds on the
germination of Lettuce at minimum concentration
have been demonstrated by biochemical studies
(D'Abrosca et al., 2006).
Aldehydes, alkaloids, apo-carotenoids, flavonoids,
chlorogenic acid and saponins have been reported by
the studies conducted on Lamb's Quarters (Della
Greca et al., 2004).
The aqueous extract of the roots and shoots of Lamb's
Quarters reduced the growth of coleoptiles in Maize
(El-Khatib et al., 2004).
According to the results reported by (Mallik et al.,
1994), new compounds were identified in Lamb's
Quarters, including chlorogenic acid and seven
phenolic compounds, having a toxic effect on the
germination of Radish.
Rough Cocklebur is also one of the weeds, threatening
medicinal plants so that it severely competes in the
fields through changing the physiological
characteristics of medicinal plants (Wassom et al.,
2002).
(Cutler and Cole, 1983) reported that potassium
carboxyactractyloside and hypoglyaemic, extracted
from plant residues of Rough Cocklebur, strongly
inhibited the growth of hypocotyls of wheat. The
extracts of Rough Cocklebur also inhibited the
germination of pepper, pumpkin and white clover
(Kadioglu, 2004).
In general, Xanthium strumarium (Rough
Cocklebur), Amaranthus retroflexus (Redroot
Amaranth) and Chenopodium album (Lamb's
Quarters) are common weeds having high effect on
the growth and germination of crops (Azizi and Fuji,
2006). There is little information on the mechanism
between their allelopatic substance and medicinal
plants growth in farms. The fight against weeds
requires spending high costs but it can be done easily
through recognizing the allelopathic agents and its
mechanism.
Therefore, the current study was aimed to investigate
the harmful effects of secreted allelopathic substances
and the extracts of (Chenopodium album,
amaranthus retroflexus and Xanthium strumarium)
on seed germination and growth of Trachyspermum
copticum L., Foeniculum vulgare and Cuminum
cyminum.
J. Bio. & Env. Sci. 2014
84 | Shams et al
Materials and methods
The present research was conducted at the laboratory
of medicinal plants, Faculty of Agriculture of Arak
University in 2008. The experiment was performed
in a completely randomized design with four
replications.
Treatments were included extracts of Xanthium,
Amaranthus retroflexusand Chenopodium album at
three concentration levels of 0, 50, and 100%. The
medicinal species studied in this research were as
follows: Trachyspermum copticum L., Foeniculum
vulgare and Cuminum cyminum.
The aqueous extracts of weeds were prepared from all
plant parts (seeds, stems, leaves, and roots). The
plants were collected from the field and then were
dried in an electric oven at 60 °C and were powdered
by grinding. 100 ml distilled water was added to 100
grams of powder of each plant and then it was placed
on a shaker for 72 hours to prepare the main solution
(100%).
The treatments included the solutions of 100% (100%
main solution), 50% (50% main solution+50%
distilled water), and distilled water was considered as
control.
To prevent fungal infection, solution containers, the
petri dishes, and seeds were disinfected by soaking in
sodium hypochlorite 10% for 10 minutes and to
remove the effect of sodium hypochlorite they were
washed five times with distilled water. Then, 100
seeds of medicinal plants were placed on Whatman
filter paper No. 2, inside the Petri dishes and placed
into germinator with a temperature of 23+2 °C.
In this experiment, the traits presented in Table 2
were measured and germinated seeds were counted
daily. A 2-mm radicle was considered as seed
germination and evaluation ended when the number
of germinated seeds did not differ in two consecutive
counts. The length of radicle and plumule was
measured on the eleventh day.
Data analysis was performed by SPSS15 software and
mean comparisons were done by Duncan's Multiple
Range Test at a significance level of 5%.
Results and discussion
Germination percentage
According to the results of ANOVA, the effect of the
type of extract and different levels of extract was
significant on germination percentage.
Table 1. Definition and formula related to the variables related to seed germination used in this study. N (the
number of germinated seeds), D (the number of days after germination), SFW (seedling fresh weight), SDW
(seedling dry weight), RL (radicle length) and PL (plumule length).
Variable Formula References
UG Ultimate Germination The maximum number of seeding's that germinated
during the experiment.
GR Germination rate (Saxena et al., 1996)
MPUG Mean Period of
Ultimate Germination
(Saxena et al., 1996)
IP Inhibitory percent (Saxena et al., 1996)
SVI Seed vigour index SVI= UG × Mean (RL+PL) (Biradar et al., 2007)
TWC Total water capacity (Samad et al., 2008)
CA Coefficient of allometry Radicle length/Plumule length (Saxena et al., 1996)
J. Bio. & Env. Sci. 2014
85 | Shams et al
Mean comparison of the effects of Lamb's Quarters,
Redroot Amaranth and Rough Cocklebur extracts
showed that Lamb's Quarters at concentration level of
50 had the lowest effect on the reduction of seed
germination percentage compared to Redroot
Amaranth while at the same concentration Redroot
Amaranth and Rough Cockleburhad the highest
effect.
Seed germination was reduced to 100% by the extract
of Redroot Amaranth and Rough Cockleburat
concentration level of 100 but a reduction of 95% was
recorded for Lamb's Quarters extract, indicating a
lower inhibitory effect.
Therefore, according to the obtained results, the
increased extract concentration caused decreased
germination percentage of the studied medicinal
plants. Our results are in agreement with the findings
of (D'Abrosca et al., 2006; Mallik et al., 1994).
Previous researches indicate that weed extracts with a
negative impact on the induction of germination
hormones such as gibberellin and the activity of
specific enzymes including amylase and proteinase
which are essential for the germination process cause
reduced germination percentage (Kruse et al., 2000).
Reduced germination percentage caused by Rough
Cocklebur extract is due to the chlorogenic acid and
phenolic in the extract (Mallik et al., 1994). In
addition, the inhibitory effects of Rough Cocklebur
extract on germination percentage may be due to the
toxic effects of folic acid in the extract (Einhelling,
2008). Therefore, the cultivation of medicinal plants
is not recommended on the farms with the mentioned
weeds.
Table 2. Effect of Chenopodium album, Amaranthus retroflexus and Xanthium strumarium extracts at
concentration levels of 0, 50 and 100 on Carum copticum, Foeniculum vulgare andPortulaca oleraceae seed
germination. Data followed by a different letter were significantly different (P≤ 0.05) according to the Duncan
Multiple Range Test.
Plant Weed Substance level UG GR MPUG IP SVI CA TWC
T.copticum
Ch. album 0 71 D 11.9 F 7.5 BCD 30 B 2.6 EF 4.6 B 0.70 B
50 20 BC 1.7 C 13.2 EF 72 C 1.4 BC 0.8A 0.90 C
100 16 B 1.1 BC 9.8 CDE 77 C 1.39 B 0.82A 0.93 C
A. retroflexus 0 70 D 11.9 F 7.5 BCD 0.0 A 3.0 F 4.8 B 0.92 C
50 20 BC 1.3 C 16.5 F 72 C 1.5 BC 1.3 A 0.95 C
100 0.0 A 0.0 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A
X. strumarium 0 70 D 11.9 F 7.5 BCD 0.0 A 3 F 4.8 B 0.92 C
50 26 C 1.5 C 5.1 ABC 63 A 1.7 BCD 1.3 A 0.93 C
100 0 A 0.0 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A
F.vulgare
Ch. album 0 70 D 8.5 E 10.1 CDE 0.0 A 3.0 F 4.8 B 0.92 C
50 18 B 1.5 B 13.4 EF 75 C 2.2 DE 4.6 B 0.89 C
100 0 A 0.0 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A
A. retroflexus 0 70 D 8.5 E 10.1 CDE 0.0 A 3.0 F 4.8 B 0.92 C
50 0 A 0.0 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A
100 0 A 0.0 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A
X. strumarium 0 70 D 8.5 E 10.1 CDE 0.0 A 3.0 F 4.8 B 0.92 C
50 21 BC 2.2 B 3.4 AB 71 C 1.9 CD 7.4 C 0.88 A
100 0 A 0.0A 0.0 A 100 D 0.0 A 0.0 A 0.00 A
C.cyminum
Ch. album 0 70 D 5.5 D 12.6 DEF 0.0 A 3.0 F 4.8 B 0.92 C
50 0 A 0.0 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A
100 0 A 0.00 A 0.0A 100 D 0.0 A 0.0 A 0.00 A
A. retroflexus 0 70 D 5.5 D 12.6 DEF 0.0 A 3.0 F 4.8 B 0.92 C
50 0 A 0.00 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A
100 0 A 0.00 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A
X. strumarium 0 70 D 5.56 D 12.6 DEF 0.0 A 3.0 F 4.8 B 0.92 C
50 0 A 0.00 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A
100 0 A 0.00 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A
J. Bio. & Env. Sci. 2014
86 | Shams et al
In this research, the highest and the lowest
germination percentage caused by different
concentrations of weed extracts were related to
Ajwain and Cumin, respectively (table 3). This
indicates that the seeds of Ajwain have a better
resistance against the germination inhibiting factors
in the weed extracts.
Germination rate
The results of mean comparisons with Duncan’s
Multiple Range Test showed that the extracts of
Redroot Amaranth and Rough Cockleburat
concentration level of 100 dramatically reduced the
germination rate (GR).
Lamb's Quarters extract at low concentration had less
impact on reducing the GR compared to the control
group, indicating the lower allelopathic effects of
Lamb's Quarters extract compared to Redroot
Amaranth and Rough Cocklebur (table 2).
According to the results, the highest (maximum) GR
in control group (distilled water) was related to
Ajwain while the lowest GR was recorded for Cumin.
The same was found for different concentrations of
weed extract. Ajwain showed a better resistance
against high concentrations of weed extracts in
comparison with Fennel and Cumin.
(ZH and SG, 2012) showed that small seeds were
more exposed to allelochemicals due to having a large
contact surface and are more vulnerable to the
allelopathic effects of weeds. Our results showed that
seed size had no effective role in this regard since the
seeds of Ajwain were smaller than that of Fennel and
Cumin (Table 3).
However, the highest resistance against
allelochemicals was related to Ajwain and it is
probably due to its specific physiological properties.
As seen in Table 2, allelopathic effects not only reduce
the germination but also cause a delay in
germination, affecting the species competition (T.
Nilsen and M. Orcutt, 1996).
Allelochemichals in the extract lead to the slowing of
plant vital processes due to the reduction of seed
respiration which ultimately reduce the GR.
Mean Period of Ultimate Germination
According to the results of Table 2, significant
differences were found among the germination onset
of the seeds affected by different concentrations of
weed extracts.
Among the treatments, Rough Cocklebur and Redroot
Amaranth extracts at concentration level of 100%
increased the mean period of ultimate germination
(MPUG) (table 2), Our results are in consistence with
the findings of (Rezaei et al., 2008), who studied the
effect of Redroot Amaranth extract on the
germination of Rapeseed.
In other words, Rough Cocklebur had the most
impact on MPUG compared to Lamb's Quarters (table
4).
Therefore, MPUG increased by increase of weed
extract concentration. This may be due to the reduced
respiration caused by allelochemicals in the extract,
ultimately leading to reduced energy for vital plant
processes such as germination.
Among the studied species, the lowest MPUG was
recorded for Ajwain (Table 3) because Ajwain showed
higher resistance against allelochemicals and for this
reason, its high germination rate reduced the MPUG.
Inhibitory percent
Triterpens are one of the allelopathic compounds in
Redroot Amaranth. Evidence has shown that terpens
inhibit cell elongation and cell division and also are
considered as germination inhibiting factors.
Sesquiterpenes, with nerolidol skeleton, are one of the
germination inhibiting factors in lettuce (D'Abrosca et
al., 2006).
The Inhibitory percent (IP) of Redroot Amaranth
extract increased by increasing the concentration of
J. Bio. & Env. Sci. 2014
87 | Shams et al
the extract as compared to control group, germination
was inhibited to 100% in the seeds of Ajwain, Fennel,
and Cumin (Table 2). This is due to the presence of
sesquiterpens in Redroot Amaranth extract and this
compound is considered as the most important
reason for allelopathic effects of Redroot Amarant
(D'Abrosca et al., 2006).
Seed germination of the studied medicinal species
decreased by increasing the concentration of Rough
Cocklebur extract as Rough Cocklebur extract at
concentration level of 100 increased the IP up to
100% in Ajwain, Fennel, and Cumin compared to
control group (Table 2).
Table 3. Mean values of seed germination characteristics of T. copticum, Foeniculum vulgare and Cuminum
cyminum caused by weed extracts. Data followed by a different letter were significantly different (P≤ 0.05)
according to the Duncan Multiple Range Test.
Plant UG GR MPUG IP SVI CA TWC
T. copticum 32.56 A 4.59 A 7.46 A 57.11 A 1.62 A 2.05 A 0.69 A
Foeniculum vulgare 27.67 B 3.24 B 5.23 B 60.67 A 1.46 B 2.93 A 0.50 B
Cuminum cyminum 23.33 C 1.84 C 4.20 C 66.67 A 1.00 C 1.60 B 0.31 C
Also, Lamb's Quarters extract at concentration level
of 100 reduced the seed germination of Fennel and
Cumin up to 100% as compared to control group but
Ajwain was less affected (70%). The allelopathic effect
of Lamb's Quarters extract may be due to flavonoids
in its extract, reducing seed germination by inhibition
of energy transmission.
The studied medicinal species showed different
resistance against IP of different weed extracts and
the highest resistance was obtained for Ajwain
compared to Fennel and Cumin.
Seed vigor index
Seedling length and germination percentage of the
studied medicinal species decreased by increasing the
concentration of weed extracts. As a result, seed vigor
index (SVI) decreased, indicating that seedling
resistance depends on the toxicity of weed extract.
According to the obtained results, different
concentrations of weed extracts significantly affected
SVI so that weed extracts at high concentration level
had the most impact on the reduction of SVI
compared to control group. The extract of Redroot
Amaranth had the most impact on reducing the seed
viability of the study medicinal species. This result is
in agreement with the findings of (D'Abrosca et al.,
2006) who studied the effect of Redroot Amaranth
extract on the seed viability of lettuce.
Among the weed extracts at low concentration level,
the lowest impact on SVI of medicinal species was
recorded for Lamb's Quarters extract (Table 4).
The highest SVI was obtained for the seeds of Ajwain,
resulting from its genetic and physiological
characteristics (Table 3).
Seedling fresh weight
According to the results, different concentrations of
weed extract significantly affected seedling fresh
weight. The lowest seedling fresh weight was obtained
for Rough Cocklebur and Redroot Amaranth extracts,
caused by the concentration level of 100 (Table 2).
Reduced seedling fresh weight of Corn and Soya,
caused by Lamb's Quarters extract, also has been
reported (El-Khatib et al., 2004).
Reduced absorption of storage nutrients required for
growth is of the factors reducing seedling fresh
weight.
Allelopathic compounds are effective on cell division
and the synthesis of proteins and hormones, resulting
in the decrease of cell growth and ultimately the
J. Bio. & Env. Sci. 2014
88 | Shams et al
reduction of seedling fresh weight (Shahrokhi et al.,
2011).
Phenolic compounds in the weed extracts cause
reduced ATP production by decreasing mitochondrial
respiration. Also, phenolics have the ability to change
the mitochondrial membrane and prevent the
transmission of required energy for vital growth
processes. This irregularity followed by a series of
physiological effects leads to reduced growth and
ultimately seedling fresh weight (Yang et al., 2002).
Coefficient of allometry (Radicle length/Plumule
length)
Previous results indicate the negative effect of
Redroot Amaranth extract on Coefficient of allometry
(CA). In the current experiment, CA decreased by
increasing the concentration of Lamb's Quarters,
Rough Cocklebur, and Redroot Amaranth extracts
(table 2).
Table 4. Mean values effect of Chenopodium album, Amaranthus retroflexus and Xanthium strumarium
extracts on seed germination. Data followed by a different letter were significantly different (P≤ 0.05) according
to the Duncan Multiple Range Test.
Weed UG GR MPUG IP SVI CA TWC
Chenopodium album 29.44 B 3.36 B 7.40 C 61.56 C 1.51 B 2.27 B 0.58 C
Amaranthus retroflexus 25.56 A 3.02 A 5.19 B 63.56 B 1.17 A 1.74 A 0.41 A
Xanthium strumarium 28.56 B 3.30 B 4.30 A 59.33 A 1.40 B 2.57 C 0.51 B
Our results are in agreement with the findings of
(Shahrokhi et al., 2011), who reported the negative
effect of Redroot Amaranth essential oil on the
growth of wheat seedlings.
Redroot Amaranth extract at concentration level of
100 reduced CA of Fennel, Anjowan, and Cumin as
compared to control group (table 2) and Redroot
Amaranth had the most negative impact on CA (table
4).
Also, Lamb's Quarters extract at concentration level
of 100, reduced the CA to 100% in Fennel and Cumin
as compared to control group. In the Lamb's Quarters
extract, the main cause of allelopathic effects is
Chlorogenic acid in the extract in which phytotoxin
has been identified as the main compound (Mallik et
al., 1994).
Redroot Amaranth and Rough Cocklebur extracts also
contain high levels of phenolic compounds. Phenolic
compounds are the main inhibitor of cell division
which ultimately reduces radicle and seedling growth
(Mao et al., 2006). A higher CA ratio was obtained in
Ajwain as compared to Fennel and Cumin.
Total water capacity
Seedling water content was significantly affected by
different concentrations of weed extracts so that
Redroot Amaranth and Rough Cocklebur extracts at
concentration level of 100 reduced total water
capacity to 100%, as compared to control group (table
2). Redroot Amaranth had the most negative impact
on CA (table 4). Our results were confirmed by
(Smitht et al., 2001) who investigated the allelopathic
effects of Redroot Amaranth extract on seedling
growth of Carya illinoinensis.
The extracts at concentration level of 50 had no
significant effect on seedling water content (table 2).
Conclusion
The results of this experiment showed that under
laboratory conditions, Lamb's Quarters, Redroot
Amaranth and Rough Cocklebur extracts affected the
seed germination of Ajwain, Fennel and Cumin but
Redroot Amaranth extract had the most negative
J. Bio. & Env. Sci. 2014
89 | Shams et al
impact on germination characteristics of the studied
medicinal species.
According to the obtained results, the need for the
weed control of the aforementioned weeds before
cultivation and during seedling growth and
establishment becomes more and more clear. In
addition to the farm experiments in order to find the
allelopathic potential of the mentioned weeds, the
investigation of allelopathic effects of other weeds on
germination characteristics (properties) of the
studied medicinal species under both laboratory and
farm conditions seems necessary.
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Allelopathic effects of weeds extract on seed germination of Trachyspermum copticum L., Foeniculum vulgare and Cuminum cyminum

  • 1. J. Bio. & Env. Sci. 2014 82 | Shams et al RESEARCH PAPER OPEN ACCESS Allelopathic effects of weeds extract on seed germination of Trachyspermum copticum L., Foeniculum vulgare and Cuminum cyminum Mostafakemal Shams1* , Ehsan Zandi Esfahan2 , Mahdi Ramezani3 , Mahnaz Ghandkanlou1 1 Department of agriculture, Payame Noor University, P.O. Box 19395- 3697, Tehran, Iran 2 Rangeland Research Division, Research Institute of Forests and Rangelands, P.O. Box 13185-116, Tehran, Iran 3 Science and Research Branch, Islamic Azad University, P.O. Box 14778. Tehran, Iran Article published on July 08, 2014 Key words: Allelopathic effects, weed species, Trachyspermum copticum L., Foeniculum vulgare, Cuminum cyminum Abstract Seed germination stage is one of the most basic growth stages in medicinal plants that may be adversely affected by the chemicals released from other plants in the soil environment. The present study was performed to investigate the allelopathic effects of weeds, including (Xanthium strumarium, Amaranthus retroflexus and Chenopodium album) as common weeds in medicinal plant's farms, on seed germination parameters, total dry weight and allometry coefficient of Trachyspermum copticum L., Foeniculum vulgare and Cuminum cyminum L. in laboratory conditions. Treatments included extracts of all organs of three weed species, applied on the seeds of the medicinal plants at three concentrations of zero (distilled water), 50 and 100% by volume. The results showed that the extracts of mentioned weed species had a significant inhibitory effect on germination characteristics of the studied medicinal plants. The greatest negative impact on seed germination was recorded for the extract of Amaranthus retroflexus, but Trachyspermum copticum L. showed a better resistance to the extract of Amaranthus retroflexus as compared to Foeniculum vulgare and Cuminum cyminum. *Corresponding Author: Mostafakemal Shams  shams100@inbox.com Journal of Biodiversity and Environmental Sciences (JBES) ISSN: 2220-6663 (Print) 2222-3045 (Online) Vol. 5, No. 1, p. 82-90, 2014 http://www.innspub.net
  • 2. J. Bio. & Env. Sci. 2014 83 | Shams et al Introduction The world's need for medicinal plants as medicines, cosmetics and as a food supplement for improving human welfare is rising day by day. Allelopathic is the interaction of produced plant chemicals on each other. Allelopathy is one of the major problems in most agricultural lands so that many weed species affect the germination and growth of other plants by producing secondary metabolites (Anowar Razvy et al., 2011; Mizutani, 1999) and the Production of secondary metabolites in plants depends on environmental conditions (Shams et al., 2013). The reduction of plant yield is not only due to weed interference in field but allelopathic agents can play an effective role in this regard (Iqbal and Wright, 1999). In the farms, Xanthium strumarium (Rough Cocklebur), Amaranthus retroflexus (Redroot Amaranth) and Chenopodium album (Lamb's Quarters) as weed species cause a sharp decline in the yield of cultivated plants (Azizi and Fuji, 2006). Allelopathic activity of Redroot Amaranth has been fully proven. Saponins, phenolics, chlorogenicacid, scopoline and benzoic acid are allelopathic compounds of Redroot Amaranth (Inderjit and Duke, 2003). According to the new researches, seven sesquiterpens have been reported for Redroot Amaranth. The inhibitory effects of these compounds on the germination of Lettuce at minimum concentration have been demonstrated by biochemical studies (D'Abrosca et al., 2006). Aldehydes, alkaloids, apo-carotenoids, flavonoids, chlorogenic acid and saponins have been reported by the studies conducted on Lamb's Quarters (Della Greca et al., 2004). The aqueous extract of the roots and shoots of Lamb's Quarters reduced the growth of coleoptiles in Maize (El-Khatib et al., 2004). According to the results reported by (Mallik et al., 1994), new compounds were identified in Lamb's Quarters, including chlorogenic acid and seven phenolic compounds, having a toxic effect on the germination of Radish. Rough Cocklebur is also one of the weeds, threatening medicinal plants so that it severely competes in the fields through changing the physiological characteristics of medicinal plants (Wassom et al., 2002). (Cutler and Cole, 1983) reported that potassium carboxyactractyloside and hypoglyaemic, extracted from plant residues of Rough Cocklebur, strongly inhibited the growth of hypocotyls of wheat. The extracts of Rough Cocklebur also inhibited the germination of pepper, pumpkin and white clover (Kadioglu, 2004). In general, Xanthium strumarium (Rough Cocklebur), Amaranthus retroflexus (Redroot Amaranth) and Chenopodium album (Lamb's Quarters) are common weeds having high effect on the growth and germination of crops (Azizi and Fuji, 2006). There is little information on the mechanism between their allelopatic substance and medicinal plants growth in farms. The fight against weeds requires spending high costs but it can be done easily through recognizing the allelopathic agents and its mechanism. Therefore, the current study was aimed to investigate the harmful effects of secreted allelopathic substances and the extracts of (Chenopodium album, amaranthus retroflexus and Xanthium strumarium) on seed germination and growth of Trachyspermum copticum L., Foeniculum vulgare and Cuminum cyminum.
  • 3. J. Bio. & Env. Sci. 2014 84 | Shams et al Materials and methods The present research was conducted at the laboratory of medicinal plants, Faculty of Agriculture of Arak University in 2008. The experiment was performed in a completely randomized design with four replications. Treatments were included extracts of Xanthium, Amaranthus retroflexusand Chenopodium album at three concentration levels of 0, 50, and 100%. The medicinal species studied in this research were as follows: Trachyspermum copticum L., Foeniculum vulgare and Cuminum cyminum. The aqueous extracts of weeds were prepared from all plant parts (seeds, stems, leaves, and roots). The plants were collected from the field and then were dried in an electric oven at 60 °C and were powdered by grinding. 100 ml distilled water was added to 100 grams of powder of each plant and then it was placed on a shaker for 72 hours to prepare the main solution (100%). The treatments included the solutions of 100% (100% main solution), 50% (50% main solution+50% distilled water), and distilled water was considered as control. To prevent fungal infection, solution containers, the petri dishes, and seeds were disinfected by soaking in sodium hypochlorite 10% for 10 minutes and to remove the effect of sodium hypochlorite they were washed five times with distilled water. Then, 100 seeds of medicinal plants were placed on Whatman filter paper No. 2, inside the Petri dishes and placed into germinator with a temperature of 23+2 °C. In this experiment, the traits presented in Table 2 were measured and germinated seeds were counted daily. A 2-mm radicle was considered as seed germination and evaluation ended when the number of germinated seeds did not differ in two consecutive counts. The length of radicle and plumule was measured on the eleventh day. Data analysis was performed by SPSS15 software and mean comparisons were done by Duncan's Multiple Range Test at a significance level of 5%. Results and discussion Germination percentage According to the results of ANOVA, the effect of the type of extract and different levels of extract was significant on germination percentage. Table 1. Definition and formula related to the variables related to seed germination used in this study. N (the number of germinated seeds), D (the number of days after germination), SFW (seedling fresh weight), SDW (seedling dry weight), RL (radicle length) and PL (plumule length). Variable Formula References UG Ultimate Germination The maximum number of seeding's that germinated during the experiment. GR Germination rate (Saxena et al., 1996) MPUG Mean Period of Ultimate Germination (Saxena et al., 1996) IP Inhibitory percent (Saxena et al., 1996) SVI Seed vigour index SVI= UG × Mean (RL+PL) (Biradar et al., 2007) TWC Total water capacity (Samad et al., 2008) CA Coefficient of allometry Radicle length/Plumule length (Saxena et al., 1996)
  • 4. J. Bio. & Env. Sci. 2014 85 | Shams et al Mean comparison of the effects of Lamb's Quarters, Redroot Amaranth and Rough Cocklebur extracts showed that Lamb's Quarters at concentration level of 50 had the lowest effect on the reduction of seed germination percentage compared to Redroot Amaranth while at the same concentration Redroot Amaranth and Rough Cockleburhad the highest effect. Seed germination was reduced to 100% by the extract of Redroot Amaranth and Rough Cockleburat concentration level of 100 but a reduction of 95% was recorded for Lamb's Quarters extract, indicating a lower inhibitory effect. Therefore, according to the obtained results, the increased extract concentration caused decreased germination percentage of the studied medicinal plants. Our results are in agreement with the findings of (D'Abrosca et al., 2006; Mallik et al., 1994). Previous researches indicate that weed extracts with a negative impact on the induction of germination hormones such as gibberellin and the activity of specific enzymes including amylase and proteinase which are essential for the germination process cause reduced germination percentage (Kruse et al., 2000). Reduced germination percentage caused by Rough Cocklebur extract is due to the chlorogenic acid and phenolic in the extract (Mallik et al., 1994). In addition, the inhibitory effects of Rough Cocklebur extract on germination percentage may be due to the toxic effects of folic acid in the extract (Einhelling, 2008). Therefore, the cultivation of medicinal plants is not recommended on the farms with the mentioned weeds. Table 2. Effect of Chenopodium album, Amaranthus retroflexus and Xanthium strumarium extracts at concentration levels of 0, 50 and 100 on Carum copticum, Foeniculum vulgare andPortulaca oleraceae seed germination. Data followed by a different letter were significantly different (P≤ 0.05) according to the Duncan Multiple Range Test. Plant Weed Substance level UG GR MPUG IP SVI CA TWC T.copticum Ch. album 0 71 D 11.9 F 7.5 BCD 30 B 2.6 EF 4.6 B 0.70 B 50 20 BC 1.7 C 13.2 EF 72 C 1.4 BC 0.8A 0.90 C 100 16 B 1.1 BC 9.8 CDE 77 C 1.39 B 0.82A 0.93 C A. retroflexus 0 70 D 11.9 F 7.5 BCD 0.0 A 3.0 F 4.8 B 0.92 C 50 20 BC 1.3 C 16.5 F 72 C 1.5 BC 1.3 A 0.95 C 100 0.0 A 0.0 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A X. strumarium 0 70 D 11.9 F 7.5 BCD 0.0 A 3 F 4.8 B 0.92 C 50 26 C 1.5 C 5.1 ABC 63 A 1.7 BCD 1.3 A 0.93 C 100 0 A 0.0 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A F.vulgare Ch. album 0 70 D 8.5 E 10.1 CDE 0.0 A 3.0 F 4.8 B 0.92 C 50 18 B 1.5 B 13.4 EF 75 C 2.2 DE 4.6 B 0.89 C 100 0 A 0.0 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A A. retroflexus 0 70 D 8.5 E 10.1 CDE 0.0 A 3.0 F 4.8 B 0.92 C 50 0 A 0.0 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A 100 0 A 0.0 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A X. strumarium 0 70 D 8.5 E 10.1 CDE 0.0 A 3.0 F 4.8 B 0.92 C 50 21 BC 2.2 B 3.4 AB 71 C 1.9 CD 7.4 C 0.88 A 100 0 A 0.0A 0.0 A 100 D 0.0 A 0.0 A 0.00 A C.cyminum Ch. album 0 70 D 5.5 D 12.6 DEF 0.0 A 3.0 F 4.8 B 0.92 C 50 0 A 0.0 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A 100 0 A 0.00 A 0.0A 100 D 0.0 A 0.0 A 0.00 A A. retroflexus 0 70 D 5.5 D 12.6 DEF 0.0 A 3.0 F 4.8 B 0.92 C 50 0 A 0.00 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A 100 0 A 0.00 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A X. strumarium 0 70 D 5.56 D 12.6 DEF 0.0 A 3.0 F 4.8 B 0.92 C 50 0 A 0.00 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A 100 0 A 0.00 A 0.0 A 100 D 0.0 A 0.0 A 0.00 A
  • 5. J. Bio. & Env. Sci. 2014 86 | Shams et al In this research, the highest and the lowest germination percentage caused by different concentrations of weed extracts were related to Ajwain and Cumin, respectively (table 3). This indicates that the seeds of Ajwain have a better resistance against the germination inhibiting factors in the weed extracts. Germination rate The results of mean comparisons with Duncan’s Multiple Range Test showed that the extracts of Redroot Amaranth and Rough Cockleburat concentration level of 100 dramatically reduced the germination rate (GR). Lamb's Quarters extract at low concentration had less impact on reducing the GR compared to the control group, indicating the lower allelopathic effects of Lamb's Quarters extract compared to Redroot Amaranth and Rough Cocklebur (table 2). According to the results, the highest (maximum) GR in control group (distilled water) was related to Ajwain while the lowest GR was recorded for Cumin. The same was found for different concentrations of weed extract. Ajwain showed a better resistance against high concentrations of weed extracts in comparison with Fennel and Cumin. (ZH and SG, 2012) showed that small seeds were more exposed to allelochemicals due to having a large contact surface and are more vulnerable to the allelopathic effects of weeds. Our results showed that seed size had no effective role in this regard since the seeds of Ajwain were smaller than that of Fennel and Cumin (Table 3). However, the highest resistance against allelochemicals was related to Ajwain and it is probably due to its specific physiological properties. As seen in Table 2, allelopathic effects not only reduce the germination but also cause a delay in germination, affecting the species competition (T. Nilsen and M. Orcutt, 1996). Allelochemichals in the extract lead to the slowing of plant vital processes due to the reduction of seed respiration which ultimately reduce the GR. Mean Period of Ultimate Germination According to the results of Table 2, significant differences were found among the germination onset of the seeds affected by different concentrations of weed extracts. Among the treatments, Rough Cocklebur and Redroot Amaranth extracts at concentration level of 100% increased the mean period of ultimate germination (MPUG) (table 2), Our results are in consistence with the findings of (Rezaei et al., 2008), who studied the effect of Redroot Amaranth extract on the germination of Rapeseed. In other words, Rough Cocklebur had the most impact on MPUG compared to Lamb's Quarters (table 4). Therefore, MPUG increased by increase of weed extract concentration. This may be due to the reduced respiration caused by allelochemicals in the extract, ultimately leading to reduced energy for vital plant processes such as germination. Among the studied species, the lowest MPUG was recorded for Ajwain (Table 3) because Ajwain showed higher resistance against allelochemicals and for this reason, its high germination rate reduced the MPUG. Inhibitory percent Triterpens are one of the allelopathic compounds in Redroot Amaranth. Evidence has shown that terpens inhibit cell elongation and cell division and also are considered as germination inhibiting factors. Sesquiterpenes, with nerolidol skeleton, are one of the germination inhibiting factors in lettuce (D'Abrosca et al., 2006). The Inhibitory percent (IP) of Redroot Amaranth extract increased by increasing the concentration of
  • 6. J. Bio. & Env. Sci. 2014 87 | Shams et al the extract as compared to control group, germination was inhibited to 100% in the seeds of Ajwain, Fennel, and Cumin (Table 2). This is due to the presence of sesquiterpens in Redroot Amaranth extract and this compound is considered as the most important reason for allelopathic effects of Redroot Amarant (D'Abrosca et al., 2006). Seed germination of the studied medicinal species decreased by increasing the concentration of Rough Cocklebur extract as Rough Cocklebur extract at concentration level of 100 increased the IP up to 100% in Ajwain, Fennel, and Cumin compared to control group (Table 2). Table 3. Mean values of seed germination characteristics of T. copticum, Foeniculum vulgare and Cuminum cyminum caused by weed extracts. Data followed by a different letter were significantly different (P≤ 0.05) according to the Duncan Multiple Range Test. Plant UG GR MPUG IP SVI CA TWC T. copticum 32.56 A 4.59 A 7.46 A 57.11 A 1.62 A 2.05 A 0.69 A Foeniculum vulgare 27.67 B 3.24 B 5.23 B 60.67 A 1.46 B 2.93 A 0.50 B Cuminum cyminum 23.33 C 1.84 C 4.20 C 66.67 A 1.00 C 1.60 B 0.31 C Also, Lamb's Quarters extract at concentration level of 100 reduced the seed germination of Fennel and Cumin up to 100% as compared to control group but Ajwain was less affected (70%). The allelopathic effect of Lamb's Quarters extract may be due to flavonoids in its extract, reducing seed germination by inhibition of energy transmission. The studied medicinal species showed different resistance against IP of different weed extracts and the highest resistance was obtained for Ajwain compared to Fennel and Cumin. Seed vigor index Seedling length and germination percentage of the studied medicinal species decreased by increasing the concentration of weed extracts. As a result, seed vigor index (SVI) decreased, indicating that seedling resistance depends on the toxicity of weed extract. According to the obtained results, different concentrations of weed extracts significantly affected SVI so that weed extracts at high concentration level had the most impact on the reduction of SVI compared to control group. The extract of Redroot Amaranth had the most impact on reducing the seed viability of the study medicinal species. This result is in agreement with the findings of (D'Abrosca et al., 2006) who studied the effect of Redroot Amaranth extract on the seed viability of lettuce. Among the weed extracts at low concentration level, the lowest impact on SVI of medicinal species was recorded for Lamb's Quarters extract (Table 4). The highest SVI was obtained for the seeds of Ajwain, resulting from its genetic and physiological characteristics (Table 3). Seedling fresh weight According to the results, different concentrations of weed extract significantly affected seedling fresh weight. The lowest seedling fresh weight was obtained for Rough Cocklebur and Redroot Amaranth extracts, caused by the concentration level of 100 (Table 2). Reduced seedling fresh weight of Corn and Soya, caused by Lamb's Quarters extract, also has been reported (El-Khatib et al., 2004). Reduced absorption of storage nutrients required for growth is of the factors reducing seedling fresh weight. Allelopathic compounds are effective on cell division and the synthesis of proteins and hormones, resulting in the decrease of cell growth and ultimately the
  • 7. J. Bio. & Env. Sci. 2014 88 | Shams et al reduction of seedling fresh weight (Shahrokhi et al., 2011). Phenolic compounds in the weed extracts cause reduced ATP production by decreasing mitochondrial respiration. Also, phenolics have the ability to change the mitochondrial membrane and prevent the transmission of required energy for vital growth processes. This irregularity followed by a series of physiological effects leads to reduced growth and ultimately seedling fresh weight (Yang et al., 2002). Coefficient of allometry (Radicle length/Plumule length) Previous results indicate the negative effect of Redroot Amaranth extract on Coefficient of allometry (CA). In the current experiment, CA decreased by increasing the concentration of Lamb's Quarters, Rough Cocklebur, and Redroot Amaranth extracts (table 2). Table 4. Mean values effect of Chenopodium album, Amaranthus retroflexus and Xanthium strumarium extracts on seed germination. Data followed by a different letter were significantly different (P≤ 0.05) according to the Duncan Multiple Range Test. Weed UG GR MPUG IP SVI CA TWC Chenopodium album 29.44 B 3.36 B 7.40 C 61.56 C 1.51 B 2.27 B 0.58 C Amaranthus retroflexus 25.56 A 3.02 A 5.19 B 63.56 B 1.17 A 1.74 A 0.41 A Xanthium strumarium 28.56 B 3.30 B 4.30 A 59.33 A 1.40 B 2.57 C 0.51 B Our results are in agreement with the findings of (Shahrokhi et al., 2011), who reported the negative effect of Redroot Amaranth essential oil on the growth of wheat seedlings. Redroot Amaranth extract at concentration level of 100 reduced CA of Fennel, Anjowan, and Cumin as compared to control group (table 2) and Redroot Amaranth had the most negative impact on CA (table 4). Also, Lamb's Quarters extract at concentration level of 100, reduced the CA to 100% in Fennel and Cumin as compared to control group. In the Lamb's Quarters extract, the main cause of allelopathic effects is Chlorogenic acid in the extract in which phytotoxin has been identified as the main compound (Mallik et al., 1994). Redroot Amaranth and Rough Cocklebur extracts also contain high levels of phenolic compounds. Phenolic compounds are the main inhibitor of cell division which ultimately reduces radicle and seedling growth (Mao et al., 2006). A higher CA ratio was obtained in Ajwain as compared to Fennel and Cumin. Total water capacity Seedling water content was significantly affected by different concentrations of weed extracts so that Redroot Amaranth and Rough Cocklebur extracts at concentration level of 100 reduced total water capacity to 100%, as compared to control group (table 2). Redroot Amaranth had the most negative impact on CA (table 4). Our results were confirmed by (Smitht et al., 2001) who investigated the allelopathic effects of Redroot Amaranth extract on seedling growth of Carya illinoinensis. The extracts at concentration level of 50 had no significant effect on seedling water content (table 2). Conclusion The results of this experiment showed that under laboratory conditions, Lamb's Quarters, Redroot Amaranth and Rough Cocklebur extracts affected the seed germination of Ajwain, Fennel and Cumin but Redroot Amaranth extract had the most negative
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