Allelopathic Effects of Some Medicinal Plants Extracts on Seed

Germination and Growth of Common Weeds in Mashhad Area

H. Arouiee*, S. Quasemi , M. Azizi and H. Nematy

Horticultural Science Department, College of Agriculture,

Ferdowsi University of Mashhad, IRAN

Abstract
Allelopathy can be regarded as a component of biological control in which plants are used to
reduce the vigour and development of other plants. Allelopathy refers to the direct or indirect
chemical effects of one plant on the germination, growth, or development of neighbouring plants.
The allelopathic effects of leaf extracts of Thymus vulgaris, Lavadula sp, Rosmarinus officinalis
and Eucalyptus citriodora at five levels (0, 10, 20, 30 and 100 %) on seed germination and some
growth characteristics of Solanum nigrum and Amaranthus retroflexus were investigated. Seed
germination factors of the weeds showed different reactions. Highest seed germination was in
control treatment. Increasing in the level of treatments cause reduction in seed germination percent
and MDG increased. The internode length and plant height, node number and leaves total
chlorophyll content decreased with increasing rate of extracts. There were significant different
between the medicinal plants that used. Lavand extract exhibited higher inhibitory effects than
others and eucalyptus had lowest influence on seed germination and growth parameters. There
were significant different between all treatments and control.

Keywords: Alleopathy, Medicinal Plants, Seed Germination, Allelochemicals

1. Introduction
Using of chemical compounds are decreasing due to their remaining harmful effects in agriculture
products. So, the researchers are searching new ways for natural compounds instead of chemical
compounds. In this way, the use of effective materials of herbal has considered as natural
compounds and analytic in nature. Sometimes a single chemical produced by one organism or
plant is harmful to another but beneficial to a third organism or plant [1-4].
Allelopathy is defined as direct or indirect interaction, whereby chemicals and their
breakdown metabolites released by one plant or organism influence the physiological processes of
another neighboring plants and/or organisms [5-7], and is one of the most controversial of
ecological interactions [8]. In general, the chemical interactions that occur among living organisms
including plants, insects and microorganisms are called allelopathy, and organic compounds
involved in allelopathy are called allelochemicals. The release of allelochemicals from plants
occurs by volatilization, leaching from leaves, exudation from roots and degradation of dead plant
parts. All parts have been shown to contain allelochemicals but leaves and roots are the most
important sources [1-4]. Nowadays, using of allelopathy natural compounds have consider as
replacement for synthetic compounds for fight against weeds. On the contrary of some artificial

139The 8th International Symposium on Biocontrol and Biotechnology

pesticides, generally natural compounds are solution in water and non-halogen and because of
their little stability in environment, make little toxicity surface. The inhibitory effect of black
walnut on neighboring plants or associated plants species is one of the oldest examples in research
and study of allelopathy [9]. Many plants are implicated in allelopathic release of different active
substances, such as (pHBA) (Camelina allysum) that influences flax growth [10], jouglon (Juglans
nigra and J. regia) [9, 11]. Other allelochemicals are phenolic acids and associated compounds,
these are the most common growth inhibitors produced by living plants or released from decaying
plant parts by microbial action or leaching. Challa and Rarvindra [7] reported that Cyperus
rotundus Linn., Cynodon dactylon (L) Pers., Echinochloa colonum (L) Link., Portulaca oleracea
L. and Lagasca mollis Cav. were the major infesting weeds in all cropping seasons and
considerably reduced the yield of vegetables at the India Institute of Horticultural Research,
Bangalore,s trial fields. Results from studies that used “test plants” such as lettuce and radish are
difficult to generalize to other species. Therefore, study of allelopathic effects of some medicinal
plants on weeds and other plants species would be useful not only as a guide for organic culture
but also for rotation programming in medicinal plants production. Fuji and companions [12]
showed some of medicinal plants are part of strong allelopathic plants. Azizi and Fuji [13] studied
on effect of essential oil germination inhibition and the extract some of medicinal plants on
Amaranthus retroflexus and Portulaca oleracea. Alan and Odkins studied the role of medicinal
plants as natural herbicide. They concluded some of medicinal plants have prevention ability of
growth of weeds. Juglun,s herbicide property proved of black walnut and Persian walnut [9]. There
were many researches that indicate existing artimisin in Artimisia sp. as natural herbicide. In
addition to obtained results by researchers, using in organic cultivation can be useful guide for
rotation program choice. The present work was aimed at finding out the allelopathic effects of
garden thyme, rosemary, eucalyptus and lavender on seed germination and growth of Amaranthus
retroflexus and Solanum nigrum, on the viability of seeds and growth of the weeds.

2. Materials and Method

In this work, we selected different parts (herbs and leaves) of some medicinal plants for
preparation of water extract, such as Thymus vulgaris, Lavadula sp, Rosmarinus officinalis and
Eucalyptus citriodora flowering stages. The experimental parts of plant being tested were
collected freshly and dried at room temperature (22-25 ºC) in dark condition. After appropriate
drying, the parts were kept in an air tight box until extraction. Then, 5 g of dried sample was
homogenised in 100 ml of distilled water by blender and filtered. The treatments were applied in
five levels (0, 10, 20, 30, and 100%). Dry residue of the extract determined as described in the
Hungarian pharmacopea. The filtrate was used in experiments diluting with distilled water or
without diluting. There were 5 treatments for application on seeds of different weeds (Table 1).
The seeds of Amaranthus retroflexus and Solanum nigrum were obtained from seed bank of
Ferdowsi University of Mashhad. The seeds were surface sterilized with 1% sodium hypochlorite
then washed with distilled water three times. Then 50 seeds were placed in a Petri dish that
furnished with sheets of filter paper moistened with distilled water and water extract of plant parts
as control and treatments, respectively. The seed dormancy was broken by dense sulfuric acid
(H2SO4, 98%) (on common amaranth seed) and potassium nitrate (KNO3, 0.2%) (on night shade
seed). Then we keep them with relevant extract, so that filter paper steep, but seeds don’t plunge in
solution. Daily count of germination seeds were done until 10 days. The studying factors included:
Germination Percentage (GP), Mean Daily Germination (MDG), MPUG, Germination Rate (GR),
and some plants growth parameters.

140The 8th International Symposium on Biocontrol and Biotechnology

Statistical methods:
Two factorial independent experiments on basis of completely randomized design with two factors
containing medicinal plants and different concentrations of extract, in three replications, were
used. Duncan َ◌ s multiple range test was used for comparing the means of seed germination
percent and rate. Each Petri dishes set as experimental plot. In this research, the used software for
data analysis and drawing of diagrams is MSTATC and EXCEL, respectively.

3. Results and Discussion

The results indicated that the treatments influenced on the qualitative and quantitative
characteristics of tested weeds, so that there were significant different between treatments (Tables
1 and 2). Mean comparison of the data indicated that there were significant different between all
treatments and control. Lavender was more effective and eucalyptus had lowest effects. Between
concentration of the extract at 100 % had more effective than others. Increasing in the levels of
extract accompanied with declined in the germination and growth of both weeds. Germination
percent, germination rate, MDG and MPUG influenced by plant extracts in comparison of control,
so that the all germination parameters decreased (Tables 3 and 4). Kocacaliskan and Terzi [14]
indicated that jouglone and leaf extracts of walnut inhibited seed germination and seedling growth
of tomato, cucumber, garden cress and alfalfa. Rietveld [15] shown that seed germination and
seedling growth of several species were sensitive to juglone. Similar results were indicated that
juglone inhibited significantly from the germination of cucumber [16], tomato and bean [17],
alfalfa [18].

Table 1. Analysis variance of allelopathic effects of of some medicinal plants extracts on seed
germination and growth of Solanum nigrum in Mashhad conditions

MS
Characteristics FA FB AB Error

df 3 4 12 40
GP 193.86** 13897.10** 56.92** 5.07
MPUG 0.98** 70.90** 0.29** 0.03
MDG 12.26** 19.18** 5.60** 1.36
Plant Length 120.26** 68.26** 24.002** 3.69
Chlorophyll Content 108.59** 89.91** 12.70* 5.17
ns: Non significant; * Significant at P ≤ 5; ** Significant at P ≤ 1

The results indicated that plant length had different responses to the treatments. Plants
length decreased in Lavender and thyme extract treatment. Increasing in the levels of extract
accompanied with plant length declined but there were not significant different between treatments
(Figure 1- b,d). The highest plant length in rosemary and eucalyptus extract belong to 20 and 10
% of treatments, respectively. Leaf chlorophyll content in the control was highest in compare to
other treatments. Increasing in concentration of extract, chlorophyll content increased, too (Figure
2). There were many researches that the allelopathy is a kind of stress. It may be increase the
amount of leaf chlorophyll content for increasing efficiency of photosynthesis of the plants [19].
Challa and Ravindra [7] reported that Portulaca leaf leachates reduced the root growth of knol-

141The 8th International Symposium on Biocontrol and Biotechnology

 khol, and onion. Leaf leachates of Portulaca oleracea and Cyperus caused 63% and 50 %
reduction of root growth in radish.

Table 2. Analysis variance of allelopathic effects of of some medicinal plants extracts on seed
germination and growth of Amaranthus retroflexus in Mashhad conditions

MS
Characteristics FA FB AB Error
Df 3 4 12 40
GP 390.64** 15255.83** 109.47** 4.73
MPUG 1.991** 77.825** 0.558** 0.02
MDG 21.933** 111.099** 24.838** 0.3
Plant Length 9.032ns 24.070* 13.252ns 7.92
Chlorophyll Content 3.993ns 79.668** 9.145ns 6.69
ns: Non significant; * Significant at P ≤ 5; ** Significant at P ≤ 1

Table 3. Effects of different levels of some medicinal plants extract on seed germination
parameters of Solanum nigrum

Treatments (Extract %)

Medicinal Plants 10 20 30 100 0

Germination Percent (%)

Thyme 21 c 4.33 e 2.33 e 1e 83 a

83 a
Rosemary 12.33 d 11.67 d 3.67 e 3e
83 a
Eucalyptus 25 b 21 c 9d 1.67 e
83 a
Lavand 10.33 d 1e 1e 1e

Mean Daily Germination (MDG)

Thyme 1.48 c 0.29 e 0.14 e 0.05 e 5.91 a

5.91 a
Rosemary 0.86 d 0.81 d 0.24 e 0.19 e
5.91 a
Eucalyptus 1.76 b 1.48 c 0.62 d 0.10 e
5.91 a
Lavand 0.72 d 0.5 e 0.5 e 0.5 e

142The 8th International Symposium on Biocontrol and Biotechnology

 Table 3 (cont.) Effects of different levels of some medicinal plants extract on seed germination
parameters of Solanum nigrum

Mean Period of Ultimate Germination (MPUG)

Thyme 2.47 b 1.43 c 3.43 b 0.1 f 3.91 b

3.91 b
Rosemary 2.88 b 2.51 b 1.1 c 1.43 c
3.91 b
Eucalyptus 3.13 b 3.66 b 6.77 a 0.43 e
3.91 b
Lavand 2.68 b 0.1 f 0.1 f 0.1 f

Germination Rate (GR)

Thyme 5.84 b 1.94 de 0.87 ef 0.5 f 17.23 a

Rosemary 3.35 d 3.38 d 1.83 e 1.33 e 17.23 a

Eucalyptus 5.98 b 4.38 c 1.33 e 0.83 ef 17.23 a

Lavand 3.07 d 0.5 f 0.5 f 0.5 f 17.23 a

Table 4. Effects of different levels of some medicinal plants extract on seed germination
parameters of Amarantus retroflexus

Treatments (Extract %)

Medicinal Plants 10 20 30 100 0

Germination Percent (%)

Thyme 19.67 d 15 e 1g 1g 87 a
87 a
Rosemary 35‫ذ‬ 19.67 d 16.33 de 1g
87 a
Eucalyptus 25.67 c 1g 1g 1g
87 a
Lavand 8.33 f 1g 1g 1g

Mean Daily Germination (MDG)

Thyme 1.43 d 1.1 e 0.1 g 0.1 g 6.24 a

6.24 a
Rosemary 2.53 b 1.43 d 1.19 de 0.1 g
6.24 a
Eucalyptus 1.86 c 0.1 g 0.1 g 0.1 g
6.24 a
Lavand 0.62 f 0.1 g 0.1 g 0.1 g

143The 8th International Symposium on Biocontrol and Biotechnology

 Table 4 (cont.) Effects of different levels of some medicinal plants extract on seed germination
parameters of Amarantus retroflexus

MPUG
0.1 f 0.1 f
Thyme 9.8 a 8.92 b 4.22 e
4.22 e
Rosemary 4.75 e 8.81 b 5.9 d 0.1 f
0.1 f 0.1 f 4.22 e
Eucalyptus 7.26 bc 0.1 f
0.1 f 0.1 f 4.22 e
Lavand 10.15 a 0.1 f

Germination Rate (GR)

Thyme 1.41 e 1.38 e 0.5 g 0.5 g 13.87 a

13.87 a
Rosemary 4.85 b 1.79 d 2.06 d 0.5 g
13.87 a
Eucalyptus 2.52 c 0.5 g 0.5 g 0.5 g
13.87 a
Lavand 0.87 f 0.5 g 0.5 g 0.5 g

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Plant Length (cm)

23 24
b b
22
22 20
b
18
21 b 16
d d d d
14
b
20 12
10
bc
19 8
6
18 4
2
17 0
10% 20% 30% 100% ‫ﺷﺎھد‬ 10% 20% 30% 100% ‫ﺷﺎھد‬

Concentration Concentration

a) Rosemary b) Thyme

28 24
26 a b
22
24
20
22 b
20 b 18
18 16
cd d d
16 14 d d
d
14 12
12 10
10 8
8
6
6
4
4
2 2
0 0
10% 20% 30% 100% ‫ﺷﺎھد‬ 10% 20% 30% 100% ‫ﺷﺎھد‬
‫ﻏﻠظت ﻋﺻﺎره اﺳطوﺧودوس‬
Concentration Concentration
c) Eucalyptus d) Lavand

Figure 1. Effects of different levels of some medicinal plants extract on

plants length of Solanum nigrum

145The 8th International Symposium on Biocontrol and Biotechnology

Leaf Chlorophyll Content

38
36 ab 40
bc ab ab 38 a
34 ab
32 36 ab
30 34
28 d 32 cd
26 30
28 d
24
‫ﻣﯾزان ﮐﻠروﻓﯾل‬

22 26

‫ﻣﯾزان ﮐﻠروﻓﯾل‬
20 24
18 22
16 20
14 18
12 16
10 14
8 12
6 10
4 8
2 6
0 4
2
10% 20% 30% 100% ‫ﺷﺎھد‬ 0
10% 20% 30% 100% ‫ﺷﺎھد‬

Concentration Concentration
a) Rosemary b) Thyme

36
34 ab ab ab abc 28
32 d
30 d d
28 d
26
24 d
d
22
20
18 26
16
14
12
10
8
6
4 24
2
0 10% 20% 30% 100% ‫ﺷﺎھد‬

10% 20% 30% 100% ‫ﺷﺎھد‬

‫ﻏﻠظت ﻋﺻﺎره اﮐﺎﻟﯾﭘﺗوس‬

Concentration Concentration
c) Eucalyptus d) Lavand

Figure 2. Effects of different levels of some medicinal plants extract on leaf chlorophyll content
of Solanum nigrum

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146The 8th International Symposium on Biocontrol and Biotechnology

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147The 8th International Symposium on Biocontrol and Biotechnology