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Growth, Moulting and Metamorphosis Inhibitory Activity of Calotropis Procera Extract Against Poikilocerus Pictus Fabr

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Growth, Moulting and Metamorphosis Inhibitory Activity of Calotropis Procera Extract Against Poikilocerus Pictus Fabr MAY 2014 – JULY 2014, Vol. 4, No. 3; 2165-2174. E- ISSN: 2249 –1929 Journal of Chemical, Biological and Physical Sciences An International Peer Review E-3 Journal of Sciences Available online atwww.jcbsc.org Section B: Biological Sciences CODEN (USA): JCBPAT Research Article Growth, Moulting and Metamorphosis Inhibitory Activity of Calotropis Procera Extract against Poikilocerus Pictus Fabr *Surjeet Singh Jat1, R.C. Saxena1 and Jiya Lal Solanki2 *1Pest Control & Ayurveda Drug Research Laboratory, Department of Zoology, S.S.L. Jain College, Vidisha (M.P.), India 2Department of Chemistry, Government College Badwah, Khargone, (M.P.), India Received: 27 April 2014; Revised: 31 May 2014; Accepted: 06 June 2014 Abstract: The extract isolated from plant Calotropis procera, when applied topically on Poikilocerus pictus at three different doses out of 10 insects, certain could not immersed into the adult and remains half ecdysed which were found to be quite dose dependent as the dose increases from 10-30 µg, the number of half ecdysed adult also got increased. Moreover, ecdysed adult was inversely proportionate to the doses. The nymphal maturation period which was 15±1 days in controlled got decreased to 12±2 days at 30 µg extract treatment. When 5 different dose of extract were applied on the insect. First two concentrations caused 100% mortality of egg i.e. fertility was found to be almost scant as compared to the control group where it showed 94% egg hatching. Similarly mortality was found to be inversely proportional to the dosage the concentration caused delayed metamorphosis with several deformities. The effect of extract on fecundity and fertility was also witnessed to be decreased as against the control group. In the present study, IR spectrum exhibited characteristics absorption bands at 2075 cm-1 and 1636.13 cm-1 for un-saturation and mass spectrum. Finally, on the basis of interpretation of graphs, the structure of compound pentacyclic triterpinic acetate was elucidated. Keywords: Calotropis procera, Poikilocerus pictus, Phytoecdysone. 2165 J. Chem. Bio. Phy. Sci. Sec. B, May 2014 – July 2014; Vol.4, No.3; 2165-2174. Growth … Jat et al. INTRODUCTION Phytoecdysone are mimics of hormones used by the arthropod (insects) and crustacean (crabs/lobsters) families in the moulting process known as ecdysis. Hoffmeister1 isolated a compound named “ecdysone” from the plant extract, that’s why they are called phytoecdysone and are quite abundant in plants. When insects eat the plants with these chemicals they may prematurely moult, lose weight or suffer other metabolic damage and die. Chemically, phytoecdysone are classified as triterpenoids, the group of compounds that includes triterpene saponins, phytosterols and phytoecdysteroids. Phytoecdysteroids are steroidal compounds produced by plants that also interfere with insect ecdysis.2 About 5–6% of plants species, including primitive groups such as ferns, contain measurable amounts of phytoecdysteroids.3 Some plants or fungi that produce phytoecdysteroids include Achyranthes bidentata4, Tinospora cordifolia5, Pfaffia paniculata6, Leuzea carthamoides7, Serratula coronata8, Cordyceps and Asparagus.9 Phytoecdysteroid are structural analogs of the insect moulting steroid hormone ecdysone occurring in plants. Plants comprise rich sources of ecdysteroids in high concentration and with broad structural diversity. Plants contain not only the steroidal compounds but also have another type of compounds known as Juvenile hormone analogue (JHA) which simply called “Juvabione”, the term coined by Stall10. More than 2000 plants species belonging to different family have been reported in India containing JH analogue, ecdysone and Anti-juvenile hormones. The grasshopper Poekilocerus pictus (Fabr.) of order Orthoptera and family Acrididae is a well-known pest of Calotropis plants. Exposures of sub-lethal dose of the insecticides greatly affect the growth and development of gonads of insects11-15. JHAs have already been developed as effective insect control agents16 and are currently receiving a significant amount of attention at every stage in the life cycle of insects regulating moulting, metamorphosis, development, reproduction and many of the physiological and biochemical processes associated with these compounds disrupting ecdysteroid biosynthesis, further metabolism or mode of action could make of effective insect control agents. Moreover, Morsy et al.17 have isolated some alkaloids, phytosteroids and resinous substances from Calotropis procera which when applied topically to the 3rd instar larvae which killed them before maturation. That’s why it was thought important to seek out phytoecdysone as antagonists from Calotropis procera plant for applying it on Poikilocerus pictus insects which were reared for the study of various aspects of their life cycle viz. growth, moulting and metamorphosis. MATERIALS AND METHODS The whole plant Calotropis procera R. Br. of family Asclepiadaceae was used as plant materials for the isolation of active principles to see the effect on Poikilocerus pictus. Grasshopper Poekilocerus pictus (Order Orthoptera and family Acrididae) is a well-known pest of Calotropis plants. The plant after proper identification and authentification were shade dried in the lab and powdered material was prepared of 40-60 mesh size. Soxhlet extraction method as per Harborne18 was followed for the extraction and isolation of plant materials and powdered material of 40-60 mesh size of the whole plant of Calotropis procera were loaded in the Soxhlet and petroleum ether, benzene, methanol and chloroform were used as solvent. Then, 6-8 cycles were run with different-2 solvents to get a good percentage yield (Table 1). The obtained semi solid crude extract were evaporated in the water bath to get solid extract. Then TLC and column chromatography was done for the further purification (Table 2). Purified fractions were sent to SAIF, CDRI Lucknow for the spectral analysis. Obtained purified extract was applied topically on the nymph of P. pictus to see the effect on growth, moulting and metamorphosis of Poikilocerus pictus. 2166 J. Chem. Bio. Phy. Sci. Sec. B, May 2014 – July 2014; Vol.4, No.3; 2165-2174. Growth … Jat et al. Table-1: Percentage yields of crude extract isolated by Soxhlet apparatus. Plant Solvent Wt. of dried Volume of Weight of % yield name powder solvent extract Calotropis P. ether 50 gm 500 ml 4.12 8.24 procera Benzene 50 gm 500 ml 5.05 10.1 Chloroform 50 gm 500 ml 6.10 12.22 Table-2: TLC of extract of plant C. procera with Rf value & color characteristics. S. No. Solvent Spots Rf Color characterization System Value Visual light UV light Iodine chamber Plant Extract CHCL3: Spot 1 0.28 Yellow Green yellow Brown MeOH Spot 2 0.85 Light green Green Dark brown (97:3) Spot 3 0.92 Green Black-green Black Authentic CHCL3: Similar to 0.81 light Green Dark green Brown marker value MeOH (97:3) Spot 2 RESULTS AND DISCUSSION As the result mentioned in (Table 3) described the effect of extract of plant Calotropis procera on Poikilocerus pictus when applied topically at three different doses out of 10 insects taken in each group treated as well as controlled due to effect of plant extract many of them could not immersed into the adult and remains half ecdysed. In the treatment group the number of half ecdysed were found to be quite dose dependent as the dose increases from 10-30 µg, the number of half ecdysed adult also got increased. Regarding ecdysed adult, it was inversely proportionate to the doses. The nymphal maturation period which was 15±1 days in controlled got decreased to 12±2 days at 30 µg extract treatment. This clearly indicates that the extract is quite effective on insects for reducing the ecdysed adult as well as the nymphal maturation period. When standard extract of Juvavione (5 µg) was applied topically the maturation period of nymph got decreased one day more i.e. 11±2 (Graph 1). Table-3: Effect of Calotropis procera extracts on Poikilocerus pictus when applied topically. Treatment P. Pictus Number of half Number of Nymph maturation (n=10 each) ecdysed adult ecdysed adult period in days Control group 10 1 9 15±1 Not treated Calotropis procera 10 2 8 14±1 extract 10 µg Calotropis procera 10 4 6 13±1 extract 20 µg Calotropis procera 10 5 5 12±2 extract 30 µg Juvavione 5 µg 10 3 7 11±2 2167 J. Chem. Bio. Phy. Sci. Sec. B, May 2014 – July 2014; Vol.4, No.3; 2165-2174. Growth … Jat et al. Nymph Maturation Period in days 15 10 5 0 Nymph maturation period Control Treated in days Treated with C. Treated with C. procera with C. Treated procera 10µg procera with 20µg 30µg Juvavione 5 µg Graph-1: Effect of C. procera extracts on Poikilocerus pictus during growth, moulting and metamorphosis. 100 90 80 70 60 50 Percentage of egg hatching 40 Percentage of mortality 30 20 10 0 Control Treated Treated Treated Treated Treated with C. with C. with C. with C. with C. procera procera procera procera procera 50 µg 40 µg 30 µg 20 µg 10 µg Graph-2: Effect of different doses of Calotropis procera extract upon development stages of Poikilocerus pictus. 2168 J. Chem. Bio. Phy. Sci. Sec. B, May 2014 – July 2014; Vol.4, No.3; 2165-2174. Growth … Jat et al. Table-4: Effect of Calotropis procera on development of Poikilocerus pictus. Food Total No. of nymph No. of nymph found died No. of adult emerged Control 9 1 8 Treated* 10µg 8 1* 7 Treated* 20µg 8 2 6 Treated* 30µg 9 4 5 *Leaves of cotton smeared with extract of Calotropis procera Results mentioned in (Table 4) showed the effect of pure extract of Calotropis procera upon the developmental stages of Poikilocerus pictus when 5 different doses in µg were applied on the insect. First two concentrations caused 100% mortality of egg i.e. fertility was found to be almost scant as compared to the control group where it showed 94% egg hatching.
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