Insecticidal property of Calotropis Gigantea Against Papaya Mealybug (Paracoccus Marginatus) on Aila by IJIRST

IJIRST â&#x20AC;&#x201C;International Journal for Innovative Research in Science & Technology| Volume 4 | Issue 1 | June 2017 ISSN (online): 2349-6010

Insecticidal Property of Calotropis Gigantea against Papaya Mealybug (Paracoccus Marginatus) on Ailanthus Excelsa R. Sumathi Research Officer I Division of Bioprospecting Institute of Forest Genetics and Tree Breeding, Coimbatore, Tamil Nadu

Dr. D. Rajasugunasekar Scientist E Genetics and Tree Breeding Division Institute of Forest Genetics and Tree Breeding Coimbatore, Tamil Nadu

D. Suresh Babu Research Scholar Division of Bioprospecting Institute of Forest Genetics and Tree Breeding Coimbatore, Tamil Nadu

Dr. N. Senthilkumar Scientist E Division of Bioprospecting Institute of Forest Genetics and Tree Breeding Coimbatore, Tamil Nadu

Dr. S. Murugesan Scientist G Division of Bioprospecting Institute of Forest Genetics and Tree Breeding Coimbatore, Tamil Nadu

Abstract Management of invasive alien insect, the papaya mealy bug, Paracoccus marginatus is of great importance because it even threatened tree species of economic importance. Secondary metabolites of the plants, including herbs and weeds are used as suitable alternative pesticide/insecticide and Calotropis gigantea is one of an ideal plant species for the extraction of pest resistant molecules. The present study focused on the exploration for an alternate ecofriendly plant based insecticide. The bioefficacy of the extracts of the flowers of Calotropis gigantea tested against papaya mealy bug infestation in Ailanthus excelsa found to have insecticidal activity (90-95%) at 2000 ppm within 24 hours of treatment. Individual phytochemical profiling of active botanicals revealed significant variation among pink and white colour flower for tannin, saponins, free fatty acids and alkaloids quantity. HPLC analysis also revealed high amounts of the bioactive compounds such as syringic acid, syringaldehyde and hydroxy benzoic. The insecticidal potential of the flower extracts of C. gigantea may be due to the presence of the individual phenolic compounds which are identified and quantified in significant amount. Keywords: Paracoccus marginatus, Acacia nilotica, Calotropis gigantea, insecticide, phytochemical _______________________________________________________________________________________________________ I.

INTRODUCTION

Ailanthus excelsa Roxb. (Simaroubaceae), Indian Tree of Heaven is a fast growing multipurpose indigenous lofty deciduous tree species, extensively cultivated in India due to its commercial and economic importance. It is one of the top priority species for safety match industry in Tamilnadu and there is a tremendous scope to develop the species as fodder tree in near future. Though A.excelsa is cultivated in large scale there is a shortage of the raw material supply to industries due to heavy infestation of papaya mealy bug Paracoccus marginatus which causes heavy damage in A.excelsa both in nursery and plantations as well [1]. Invasive alien insect, the papaya mealy bug Paracoccus marginatus is a native to Mexico and/or Central America [2]. It is reported to be introduced accidently in India during 2008 and noticed in papaya, Carica papaya L. [3]. It is an invasive polyphagous [4] exotic sucking pest and due to its sudden outbreak and extensive damage, attained the status of a serious pest on a wide range of host plants. It caused havoc in agricultural and horticultural crops and infesting more than 60 host plants including economically important tree crops. Chemical measures to control the outbreak of papaya mealybug was not significantly success mainly due to the development of pesticide residue on the produce resulted in ecocidal effects. Hence ecofriendly non chemical method of pest management is of greater importance which is an alternative pest controls measure without the negative effects of synthetic insecticides. Plant derived insecticides encompasses an array of chemical compounds which protect the plants from the attack of insect-pests and moreover botanical insecticides exhibit a limited effect on beneficial insects, rarely toxic to mammals and man, and development of resistance in insects is limited [5]. The plant extracts with allelochemicals could improve the biodegradability of

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Insecticidal Property of Calotropis Gigantea against Papaya Mealybug (Paracoccus Marginatus) on Ailanthus Excelsa (IJIRST/ Volume 4 / Issue 1/ 039)

insecticide treatments and thereby protect the environment. In recent years, attempts are being made to replace some of the harmful synthetic pesticides utilized in large scale, by the pest resistant secondary metabolites of the plants, including herbs and weeds as suitable alternatives [6]. [7] reported that Calotropis gigantea is an ideal plant species for the extraction of pest resistant molecules. Hence C.gigantea is tested against the papaya mealybug causes severe damage to the A.excelsa cutting kept in the nursery mother bed for further propagation to understand its insecticidal potential to develop alternate biopesticide formulation. II. MATERIALS AND METHODS Preparation of Plant Extract Flower samples of C. gigantea (both white and pink colour) flowers were collected from Poosaripalayam (E 11º00’41’’ N 76º56’01’’), Coimbatore. The flowers were washed thoroughly in tap water and shade dried at room temperature and the dried samples were ground into fine powder. Dry powder of flower samples (10g) were subjected to hot extraction for 6hrs in soxhlet apparatus with methanol. For cold extraction the dry powder of flower samples (10g) were soaked overnight in methanol at room temperature. The extracts were concentrated in rotary evaporator and the concentrated extracts were subjected to freeze drying in a lyophilizer till dry powder was obtained. Finally the extracted powder was resuspended in methanol at the concentration of 100mg/ml (w/v). The cold and hot methanol extracts were stored at 4°C for further analysis. For HPLC analysis the methanol extract was filtered over anhydrous sodium sulphate followed by purification through filtration via Varian Bond Elut C18 solid phase extraction column. The purified samples were also stored in sterile vials at 4°C for further chromatographic analysis. 20 µl of this solution was employed for HPLC analysis. Phytochemical Quantification Hot and cold methanol extracts of both white and pink colour flower samples of C. gigantea were subjected to phytochemical analysis. Soluble protein was measured according to [8], carbohydrate quantification using [9], free fatty acids by [10], tannins are quantified using Vanillin hydrochloride method [11], Flavonoids were estimated by the method of [12], saponins content was determined by the method described by [13] and alkaloids by [14]. High Pressure Liquid Chromatography Profiling High Pressure Liquid Chromatography (HPLC) is chemistry based tool used to analyze and quantify the mixtures of chemical compounds and to find the amount of chemical compound within a mixture of other chemicals. HPLC instrument with L-4000 UV detector, L-6200 Intelligent pump and RP-C18 column from Hitachi with Data Ace workstation was used in the present study. The solvent system containing methanol and 0.4% acetic acid was used as the mobile phase in the ratio 80:20 and the flow rate was maintained at 0.5 ml/min at a wavelength of 290 nm, 20μl of the sample was injected into the instrument for analysis. Biopesticidal Study Ailanthus excelsa saplings collected from Moolanur (MC-9, MC-10, MC-12, MC-13, MC-14, MC-18) and Idikarai (I-8, I-9, I-12, I-18) were found infested with mealy bug. The said materials were used for the present study and sprayed with methanol extracts of flowers of C. gigantea. III. RESULT Quantitative Determination of the Chemical Constituents Plants are endowed with potential to produce a range of secondary metabolites like alkaloids, terpenoids, flavonoids, phenols, glycosides, sitosterols and tannins which are known to protect the plants from the attack of insect-pests [15]. The phytochemical analysis of this species exhibited the presence of alkaloids, total phenolics, total flavonoids, tannins, saponins, carbohydrates and free fatty acids. No significant variation was observed between hot and cold extracts. However in hot extracts significant variation was observed between pink and white colour flower for tannin, saponins, free fatty acids and alkaloids content quantity (Table 1 & Fig 1). Pink colour flower found to have comparatively low tannin, saponins, free fatty acids and alkaloid than white colour flower. Carbohydrate content was found to be low in white flower than pink colour flower. Table - 1 Phytochemical quantification of hot and cold methanol extracts of Calotropis gigantea (white and colour flowers) Calotropis gigantea S.No Phytochemicals Hot extract (mg/g) Cold extract (mg/g) Colour flower White flower Colour flower White flower 1 Flavonoids 8.34 8.12 7.98 7.56 2 Carbohydrate 39.09 27.37 32.15 37.94 3 Phenols 9.77 9.78 9.74 9.78 4 Tannin 11.19 23.52 25.36 24.81 5 Saponins 3.28 10.04 10.04 10.04 6 Free fatty acids 32.54 46.00 20.20 19.64

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Insecticidal Property of Calotropis Gigantea against Papaya Mealybug (Paracoccus Marginatus) on Ailanthus Excelsa (IJIRST/ Volume 4 / Issue 1/ 039)

Alkaloids

6.90

8.28

8.32

8.18

Fig. 1: Phytochemical profile of hot and cold methanol extracts of Calotropis gigantea (white and colour flowers)

HPLC Quantification and identification of plant botanicals is of great importance in the field of insect pest management. The presence of wide range of phytochemical constituent indicates the plant could be used in a multitude ways which may be beneficiary to the population [16]. Over all high amounts of the phyto constituents viz., syringic acid, syringaldehyde and hydroxy benzoic were comparatively recorded in white flower than colour flower (Table 2 & Fig 2). Vast variation in syringic acid content was observed between both hot and cold extracts of the flowers. Hot extract showed high content of syringic acid (0.336 mg/ml) than cold extract (0.007 mg/ml). Syringic acid was recorded in both the flowers and in all the extracts, whereas syringaldehyde was identified in hot methanol extract of both the flowers (Fig 2a, 2c) and hydroxy benzoic acid in cold methanol extract of both the flowers (2b, 2d). Table - 2 HPLC quantification of phytochemicals presents in C. gigantea flower extracts Syringic acid Syringaldehyde Hydroxy benzoic acid Extracts (mg/ml) (mg/ml) (mg/ml) White flower hot methanol 0.366 0.03 White flower cold methanol

0.007

0.0676

Colour flower hot methanol

0.337

0.012

Colour flower cold methanol

0.004

0.0517

Fig. 2a: HPLC chromatogram of hot methanol extract of white flower Fig. 2b: HPLC chromatogram of cold methanol extract of white flower

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Insecticidal Property of Calotropis Gigantea against Papaya Mealybug (Paracoccus Marginatus) on Ailanthus Excelsa (IJIRST/ Volume 4 / Issue 1/ 039)

Fig. 2c: HPLC chromatogram of hot methanol extract of Fig. 2d: HPLC chromatogram of cold methanol extract of colour flower colour flower Fig. 2: HPLC chromatogram of methanol extracts of white and colours C. gigantean

Bioefficacy of Calotropis gigantea white flower against papaya mealy bug infestation in Ailanthus excelsa Bioefficacy of hot methanol extract of Calotropis gigantea white flower against mealy bug revealed that 10,000 ppm was not effective in controlling the mealy bug infestation whereas 20,000 ppm was found effective (90-95%) within 24 hrs of treatment. Further infestation was not noticed till 20 days and after 20 days clones MC-10 and MC-12 were found susceptible to mealy bug attack, whereas Idikarai clones are resistance to further insect attack. Over all high amount of the phyto constituentsâ&#x20AC;&#x2122; viz., syringic acid, syringaldehyde and hydroxy benzoic acid were recorded comparatively more in white flower than pink flower and hot extract of the flower showed higher biopesticidal activity against mealybug infestation than the cold extract.

Fig. 3: Bioefficacy of hot and cold methanol extracts of Calotropis gigantea flowers (white and colour) against mealy bug infestation in Ailanthus excelsa cutting in nursery bed

IV. DISCUSSION Realizing the adverse effects of chemical insecticides, attention has now been diverted in favour of non-chemical methods for pest management [17]. Biopesticides are plant based substance or mixture of substances intended to prevent, repel and destroy any insect and pests. Calotropis species is highly potential special classes of plant resource that can avoid or repel the grazing animals [18]. The previous workers have reported the presence of effective phytochemical constituents which are responsible for its biological activities especially its insecticidal activity, in the various parts of Calotropis gigantea especially in the leaves [19]. Insecticidal activity of many plants and their compounds against different groups of insects has also been reported by many workers

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Insecticidal Property of Calotropis Gigantea against Papaya Mealybug (Paracoccus Marginatus) on Ailanthus Excelsa (IJIRST/ Volume 4 / Issue 1/ 039)

[20-24]. In the present study it is observed that C. gigantea flowers are enriched with phytochemicals, hence considered as an efficient biopesticide. Calotropis procera had been reported as a source of pest resistant molecules. The presence of bioactive compounds such as triterpenoids, flovonoids, alkaloids and quinines in the ethyl acetate extracts of A. aspera were responsible for its higher percentage of antifeedant activity against fourth instar larvae of Henosepilachna vigintioctopunctata [25]. These results are in accordance with the research findings that quinine remirol and cyperquinone isolated from the plants of the family Cyperaceae had strong antifeedant activity against S. litura which indicated that botanicals offer antifeedant activity against targeted pests [26]. This indicated that the active botanicals present in plants extracts inhibit larval feeding behaviour or make the food unpalatable resulting in feeding deterrence. [27] also found that the extracts of C. gigantea have shown the growth inhibiting effect on X. campestris concluding that Calotropis sp. may be used for generating biopesticides. V. CONCLUSION The use of plant extracts as insecticide could improve the biodegradability of insecticide treatments and therefore decrease the quantity of toxic insecticide residues. In the present study C. gigantea flowers found effective in controlling the mealy bug infestation within 24 hrs of treatment and further infestation was not noticed, hence considered as an efficient biopesticide. The presence of high amount of the effective phytochemical constituents such as syringic acid, syringaldehyde and hydroxy benzoic may be responsible for its biological activities especially biopesticidal activity against mealybug. REFERENCES [1] [2] [3] [4] [5] [6] [7] [8] [9] [10] [11] [12] [13] [14] [15] [16] [17] [18] [19] [20] [21] [22] [23] [24] [25] [26] [27]

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