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

Transcription

1 International Journal of Botany and Research (IJBR) ISSN Vol. 3, Issue 4, Oct 2013, 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,

2 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, , 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. Per centage mortality Number of dead larvae Number of larvae introduced x 100 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 (LC 30 ), median lethal concentration (LC 50 ) and upper fiducial limit (LC 90 ) were calculated using probit analysis (Finney, 1971).

3 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 LC 50 and LC 90 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 LC 50 value of % 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 LC 50 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 = ; df = 1; p = 0.000; p < ) which is followed by aqueous extracts (F = ; df =1; p = 0.000; p < ), ethanol extracts (F = ; df = 1; p = ; p < 0.001) and chloroform extracts (F = ; df = 1; p = ; 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 ± %) (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 ± %) 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 = ; p = 0.000; p < 0.05) followed by A. conyzoides (F= ; df = 4; p = 0.000; p < 0.05).

4 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 ( %) (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.

5 Mortality(in % Growth Inhibitory Activities of Ageratum conyzoides Linn and Artemesia vulgaris Linn of Asteraceae 17 against Spodoptera litura Fab (Lepidoptera: Noctuidae) Ageratum conyzoides Lethal concentration ( 24, 48, 72 hrs) Chloroform Aqueous Ethanol Water A Figure 1: Probit Analysis Parameters of Extracts of Selected Plants at Different Hours of Treatment on Third Instar Larvae of S. Litura B I 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

6 18 F. Brisca Renuga II III IV V 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 REFERENCES 1. Abbot, W. S. (1925). A method of computing the effectiveness of insecticides. Journal Economic Entomology, 18, Abudulai B.., Shepard, M. & Mitchell, P. L. (2001). Parasitism and predation on eggs of Leptoglossus phyllopus (L.) (Hemiptera: Coreidae) in cowpea: impact of endosulfan sprays. J. Agr. Urban Entomol., 18, Aydin, M. H. & Gurkan. M. O. (2006). The Efficacy of spinosad on different strains of Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae). Turk. J. Biol., 30, Anjoo Kambooj & Ajay Kumar Saluja. (2008). Ageratum conyzoides L.: A review on its phytochemical and pharmacological profile. Bioresource Technology, 89 (2),

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