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International Journal of Botany and Research (IJBR)

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International Journal of Botany
and Research (IJBR)
ISSN 2277-4815
Vol. 3, Issue 4, Oct 2013, 13-20
© TJPRC Pvt. Ltd.

GROWTH INHIBITORY ACTIVITIES OF AGERATUM CONYZOIDES LINN AND

ARTEMESIA VULGARIS LINN OF ASTERACEAE AGAINST SPODOPTERA LITURA FAB
(LEPIDOPTERA: NOCTUIDAE)

F. BRISCA RENUGA
Associate Professor, Holy Cross College (Autonomous), Nagercoil, Tamil Nadu, India

ABSTRACT

Plant extracts have offered many beneficial uses in agriculture. The worldwide efforts in search for natural
products and their analogues in the crop protection market have been remarkably successful, foremost in the field of insect
control. In the present study, two plants from Asteraceae families are examined for their insecticidal activities against an
important pest namely Spodoptera litura (Fab.).

The chloroform, ethanol, water and aqueous extracts of these plants alter the developmental time and
morphogenesis of S. litura larvae. Ageratum conyzoides and Artemesia vulgaris cause maximum of biological and
morphogenesis effects. Maximum of prepupation and larval deformities were observed in the larvae treated with
Asteraceae plants. This confirms the interference of Juvenile hormone (JH) mimics of Asteraceae plants with the endocrine
system.

KEYWORDS: Ageratum conyzoides, Artemesia vulgaris, Spodoptera litura, Morphogenesis

INTRODUCTION

In recent years, the use of synthetic organic insecticides in crop insect pest control programme around the world
has resulted in damage to the environment, pest resurgence, pest resistance to insecticides and lethal effects on non-target
organisms. These negative impacts of chemical insecticides have forced scientists to search of alternate techniques for the
management of insect pests (Abudulai et al., 2001). Botanical insecticides such as are often effective alternatives to
organophosphates or other neurotoxins for pest control due to multiple modes of action. These include toxicity, antifeedant
and anti-oviposition effects Sutherland et al., 2003).

While plant chemicals may produce toxic effects when ingested by insects, antifeeding activity may determine the
extent of insect herbivory. Several papers have been published on the entomotoxic properties of crude extracts from
different plant species ( Tapondiou et al., 2005; Ulrichs et al., 2008; Baskar et al., 2009).

Recently, the discovery of insecticide activity of phototoxins present in Asteraceae species has stimulated the
interest in this plant family as part of the search for new plant derived insecticides. Ageratum conyzoides is a common
annual herbaceous weed with long history of traditional medicinal use in many countries especially in the tropical and sub
tropical regions

A wide range of chemical compounds including alkaloids, cumarins, flavonoids, chromenes, benzofurans, sterols
and terpenoids have been isolated from this species. Extracts and metabolites from this plant have been found to possess
pharmacological and insecticidal activities (Anjoo Kamboj and Ajay Kumar Saluja, 2008).

The Ageratum and Artemesia are rapidly spreading plants and is a major problem for environmentalist, ecologist,
14 F. Brisca Renuga

farmers and animal scientists. On the other hand, the larvicidal activity of weed plants that is found in vast areas on plains
as well as on hilly regions is not attempted so far. Weed plants that grow in large numbers in a vast area makes such areas
uncultivable as well as unsuitable as a fodder for the cattle. A number of such weed plants growing in vast areas is
observed on the hilly region of the Nilgiris district of Tamil Nadu.

MATERIAL AND METHODS

Rearing of Insects
Spodoptera litura was collected from castor, Recinus communis L. (Euphorbiaceae) in and around Coimbatore
District, Tamil Nadu, India and reared under laboratory condition (29 ±1°C temperature; 65% – 70% RH and 11L and 13D
photoperiods) on castor leaves in plastic containers of 21.0 X 28.0 X 9.0 cm size. Laboratory emerged third and fifth instar
larvae were used for the experiments.

Extraction and Preparation of Plant Extracts

Aerial parts of Ageratum conyzoides and Artemesia vulgaris were collected from Coonoor, Nilgris, Tamil Nadu at
the height of about 1200 MSL. Selected plant parts were washed thrice with tap water and once with distilled water, dried
at room temperature and crushed to fine powder.

Five hundred grams of powder was sequentially partitioned in chloroform, ethanol and water. For the aqueous
extract 300g of plant powder was mixed with one litre of distilled water and lyophilized for 8hrs and then kept for 24 hrs.
The extract was prepared by filtering.

One gram of crude plant extracts were redissolved in their respective solvents to prepare 10 per cent stock
solution with 5ml of an adjuvant of 0.05 % Teepol. This stock solution was used to prepare different concentrations of
0.025, 0.05, 0.075, 1 0.25, 0.5, 0.75, 1, 2.5, 4, 5 , 6 and 7 percent of chloroform, ethanol, water and aqueous extracts were
prepared.

Bioassay

Leaf dip method was followed for assessing the pesticidal activity of plants. Ten gram of castor leaves was soaked
in different concentration of the plant extracts separately. For the control, leaves were soaked in respective solvents and
adjuvant. Treated leaves were air dried for five minutes and were supplied to the fifth instar larvae (6 hrs starved) for three
days continuously.

A minimum of 10 larvae / concentration were used for each experimental categories and was replicated six times.
If any mortality was observed in the control then the mortality data was subjected to Abbot’s formula (Abbot, 1925) in
order to find out corrected percent mortality using the following formula.

Number of dead larvae

Per centage mortality  x 100
Number of larvae introduced

Number in T after treatment

Corrected per centage mortality  (1  ) x 100
Number in C after treatment

Where, T- experimental category and C – control category.

The lower fiducial limit (LC30), median lethal concentration (LC50) and upper fiducial limit (LC90) were calculated
using probit analysis (Finney, 1971).
Growth Inhibitory Activities of Ageratum conyzoides Linn and Artemesia vulgaris Linn of Asteraceae 15
against Spodoptera litura Fab (Lepidoptera: Noctuidae)

Developmental Studies

Newly moulted larvae were reared on fresh castor leaves until fourth moulting. Six hours starved fifth instar
larvae were weighed and fed with leaves treated with 1: 1 dilution of LC 50 concentration of chloroform, ethanol, water and
aqueous extracts of the plants with 0.05% Teepol as an adjuvant continuously for 72 hrs. Control leaves were treated with
respective solvent alone with 0.05% adjuvant.

The uneaten leaves were replaced with fresh treated leaves (10 larvae / concentration; five replicates). After 72
hrs, the larvae were fed with untreated castor leaves still prepupation. The larval period and pupal duration were recorded.

Statistical Analysis

The data was analyzed by completely randomized, one-way Analysis of Variance (ANOVA) and the means were
separated using Tukey Multiple Range Test (TMRT) significances are expressed at 5% level. All the statistical analyses
were performed using the statistical system of SPSS version 11.5. LC50 and LC90 values were calculated using probit
analysis (Finney, 1971).

RESULTS

Chloroform, ethanol, water solvent and aqueous extract of the selected plant were toxic to the larvae of
Spodoptera litura, although there were some differences between extracts (Figure 1A & B). Ethanol solvent extracts of
both plants were highly toxic followed by aqueous, chloroform and water. Of the two plants A. vulgaris is highly toxic
(Figure 1 A) than A. conyzoides with lowest LC50 value of 0.140 % at 72 hrs (Figure 1 A).

Decreasing trend of susceptibility with increasing hours of exposure is also noticed in the third instar larvae of S.
litura as the LC50 value is decreasing with increasing exposure time. High mortality was observed in the ethanol solvent
extract treated categories.

In A. vulgaris treated categories larval period is extended. Water solvent extract of A. conyzoides exhibits
maximum extension of larval period. Similarly the extension of pupal period was observed in all treated categories except
the ethanol extract of A. vulgaris and chloroform extract of A. conyzoides (Figure 2 II A & B). The highest percent of
extension was found in the aqueous extract of A. vulgaris.

Of the four solvents of four plants, the effects of water extracts is highly significant on larval (F = 1200.054; df =
1; p = 0.000; p < 0.0001) which is followed by aqueous extracts (F = 329.83; df =1; p = 0.000; p < 0.0001), ethanol
extracts (F = 122.65; df = 1; p = 0.0004; p < 0.001) and chloroform extracts (F = 46.632; df = 1; p = 0.0024; p < 0.01).
The same level of significance was recorded for the pupal duration of S. litura.

Minimum prepupation was recorded in A. conyzoides ethanol extract (72.0 ± 0.20 %) (Figure 3, I E) followed by
water (73.3 ± 0.071 %) (Figure 3, I W) and chloroform extracts (74.6 ± 0.04 %) (Figure 3, I C). Pupation was also highly
reduced to 68.0 ± 0.22 % in A. conyzoides ethanol and water (68.0 ± 0.245%) (Figure 3, II E & W). Emergence of adult
Spodoptera litura was highly reduced by water extracts of A. vulgaris (61.0 ± 0.11%) and is followed by aqueous extracts
(64.0 ± 0.245%). Formation of deformed adult was higher in A. conyzoides ethanol (20 ± 0.374 %) and chloroform (9.3 ±
0.032%) (Figure 3, V E & C). In the present study, no larval or pupal intermediate were observed, however, considerable
changes occur in the morphological character like prepupation, deformed pupae and deformed adults S. litura were
produced. A. vulgaris caused highly significant morphological changes (df = 4; F = 2258.68; p = 0.000; p < 0.05) followed
by A. conyzoides (F= 583.63; df = 4; p = 0.000; p < 0.05).
16 F. Brisca Renuga

DISCUSSIONS

Noctuid moths from genera Spodoptera are polyphagous pests causing economic damage in several agricultural
crops throughout the world (EI-Aswad et al., 2003). Broad spectrum insecticides have been used for its control has resulted
in development of resistance to many of the registered pesticides for its control (Kranthi et al., 2002; Aydin and Gurkan,
2006). In this scenario newer types of insecticides originating from natural products, targeting Spodoptera litura could be
useful alternative for integrated pest management.

In the present work high mortality showed by ethanol extracts than the aqueous extracts might be due to the
presence of more polar compounds in ethanol than the aqueous extracts. Audrey and Isman (2004) also suggested the use
of complex mixtures as pest control agents could be advantageous as natural mixtures may act synergistically. These
complex mixtures occur in plants may show greater bioactivity compared to the individual constituents when they are
separated in the laboratory (Chen et al., 1995) and insect resistance is much less likely to develop with mixtures of
compounds (Feng and Isman, 1995; Ateyyat et al., 2009). The mortality observed in the four solvent extracts of A.
conyzoides (32.03%) and A. vulgaris (32.25%) of Asteraceae plants does not agree with earlier reports which showed high
mortality at very low concentration (0.013 – 0.10%) (Bouda, 2001; Moreira et al., 2007) which may be due to variation
in the active principle of the selected plants. Pascual and Robleodo (1999) demonstrated that the geographical variations
also varying the phytochemical composition of plants, for example high altitude plants accumulate more flavanoids than
others.

Enhancement and reduction in the larval period could be related to the disruption of endocrine systems controlling
moulting and which may be due to disruption in the synthesis and release of ecdysteroids or related of hormones essential
for growth. Earlier reports also confirm the interference of selected plants with the hormonal system (Lange et al., 1983;
Padmaja and Rao, 1999). Maximum pupation and larval deformities were also observed in the larvae treated with
Asteraceae plants. This confirms the interference of Juvenile hormone (JH) mimics of Asteraceae plants with the endocrine
system (Kamal and Mehra, 1991; Saxena, et al., 1994). Juvenile hormone mimics play a crucial role in the regulation of
the molting and metamorphosis. Both larval and pupal periods have been prolonged by A. conyzoides.

The observation of discolouration, oozing out of fluid, softening of body and unclear segmentation in the treated
categories may be due to the fact that the Asteraceae plants have strong insecticidal effects by interfering reproductive
cycle by interfering with the hormones (Onyilagha et al., 2004). Larsen et al. (1993) reported that a great variation was
expected in the activity of Asteraceae plants as they exhibit great structural diversity of compounds. From the present study
it is concluded that all the four plants can be used as biopesticide and further studies are essential to identify the active
compounds present in them for a promising plant - based biopesticide which is safer to human beings and beneficial
organisms and has minimum threat to environment.

CONCLUSIONS

Artemesia vulgaris and Ageratum conyzoides showed insecticidal and growth regulatory effects that caused
changes in the developmental duration of S. litura. The chloroform, ethanol, water and aqueous extracts of these plants
altered the biology and morphogenesis of S. litura larvae. From the present study it is concluded that all the four plants can
be used as biopesticide and further studies are essential to identify the active compounds present in them for a promising
plant - based biopesticide which is safer to human beings and beneficial organisms and has minimum threat to
environment.
Growth Inhibitory Activities of Ageratum conyzoides Linn and Artemesia vulgaris Linn of Asteraceae 17
against Spodoptera litura Fab (Lepidoptera: Noctuidae)

Ageratum conyzoides

Mortality(in %
4

0
30 50 90 30 50 90 30 50 90
Lethal concentration ( 24, 48, 72 hrs)

Chloroform Ethanol
Aqueous Water

A B
Figure 1: Probit Analysis Parameters of Extracts of Selected Plants at Different Hours of Treatment on Third
Instar Larvae of S. Litura

II

Figure 2: Biological Effects of Ethanol (A), Chloroform (B), Aqueous (C), and Water (D) (in Days) Extract of
Selected Plants on S. litura Larval (I) and Pupal Duration; CON – Control; AGE – A. conyzoides; ART - A. vulgaris

I
18 F. Brisca Renuga

II

III

IV

Figure 3: Morphogenetic Effects of Water ( W) Chlorofom ( C) , Aqueous (A) and Ethanol (E) Extract of Selected
Plant on S. Litura Prepupation (I), Pupation (II), Deformed Pupae (III), Adult Emergence (IV) and Deformed Adult
(V) ( in %); CON - Control; AGE – A. Conyzoides; ART- A. Vulgaris
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