Biopesticidal effect of Tinospora crispa extracts against flea beetles, Phyliotera sinuata ateph Nor Aziyah B.1,2, Norain I.2, Nor Aimi A.W. 2, Lim B.T. 2, Wan Zarina W.K. 2 and Siti Nur Amirah Diana F. 2 1Faculty
of Applied Sciences, Universiti Teknologi MARA, 40450 Shah Alam, Selangor, MALAYSIA 2Department of Applied Sciences, Universiti Teknologi MARA, 13500 Permatang Pauh, Penang, MALAYSIA Email :
[email protected]
ICCE-2013. ANTWERP, BELGIUM. 8-10 JULY 2013
INTRODUCTION The usage of synthetic insecticides to control insect pests has led to both insecticide resistance and environmental persistence. Alternatively, phytochemicals or plant derived pesticides which are ecofriendly, non-toxic to non target organisms, non persistent in nature have been used. Therefore, researchers world over are engaged in a mission to hunt for novel phytochemicals that could potentially be used in the management of insect-pests. Tinospora crispa (family Menispermaceae), known as ‘akar patawali’ or ‘seruntum’ is an indigenous plants which grows wild in Malaysia. The plant has been widely used in Asia and Africa as herbal remedy for a long time. In traditional medicine, a decoction from the stems of T. crispa has been used for anti-inflammation, reducing thirst, increasing appetite, antipyretics, and maintaining good health. The stem of T. crispa have been reported as one of the most promising biopesticides which is effective against green leafhopper, brown planthoppers, small mottled willow moth and diamondback moth.
OBJECTIVES to evaluate the insecticidal activity of T. crispa stems extracts against Phyliotera sinuata ateph in mustard plant (Brassica juncea rugosa). to identify the active components in the biopesticidal active extract of T. crispa .
METHODOLOGY Preparation of Extracts Sample Preparation (Fresh stems were washed, dried and grinded into powder)
Leaves and stems of T. crispa
Preparation of Samples
Extraction of sample (Sample powder were extracted with pet. ether as the solvent by Soxhlet apparatus. The process was repeated by using chlorofom, ethanol and ethyl acetate as the solvent)
0.5 g of crude extract was weighed and 10 drops of tween 20 was added to the extract and mixed using mortar and pestle. 1.0 L distilled water was added to the mixture and shake to get a homogenous solution. The commercial pesticide, cyperin (0.5 mL) dissolved in 500 mL distilled water is used as positive control (CCP) and distilled water is used as the negative control. The solution of negative and positive control samples was also added with 10 drops of tween 20 and shaked well.
Biopesticidal Studies Side by side, Brassica juncea rugosa plants were grown in micro plot sized 3x4 meter in the experimental field. The experiments were laid out in randomized complete block design (RCBD) with six treatments and four replicates. A 500 mL-hand sprayer (WARDS, USA) was used. The tank of the sprayer was filled with 250 mL of test solution. One-weekold plant seedlings in plastic pots of 25 cm (diameter) x 20 cm (height) (one seedling/pot) were used. Quick sprays, with approximately 5 mL of the test solution, were applied for each pot.
Phytochemical Screening Solvent removal (The solvent used in the extraction was removed using rotary evaporator)
Crude extract
Thin layer chromatography was carried out on Merck 5554 Kieselgel 60F254 pre-coated UV sensitive aluminum foil plates. Ethyl acetate: chloroform (5 mL: 95 mL) solvent system was used and the spots were detected by spraying with Dragendorf reagent, anisaldehyde in concentrated sulfuric acid and 10% sulfuric acid in ethanol, followed by heating on a hot plate. Ultra violet (UV) sensitive TLC plates were also visualized under UV (254–365nm) light using UVGL-58 Handheld UV Lamp. The FT-IR spectrums of the active extract was recorded by Nicolet 6700 FT-IR Spectrometer. GCMS analysis was carried out using Perkin Elmer, Clarus 600 Gas Chromatograph.
RESULTS
% reduction for each spray of treatments FTIR spectrum of the ethanol extract of T. crispa
Table 1: Student Newman Keuls (SNK) comparison analysis between different treatments (a : significantly different, p < 0.05).
DISCUSSION & CONCLUSION The overall results of this study showed that EETC obtained from Tinospora crispa were effective and significant (p < 0.05) in reducing the population of Phyliotera sinuata ateph in mustard plant. The percentage reduction of EETC is as good as CCP, a commercial pesticide, therefore these bio-pesticides can be used as potential source of sustainable eco-friendly botanical pesticide to protect from Phyliotera sinuata ateph. Phytochemical screening revealed the presence of alkaloid, terpenoid and glycosides in the extracts.
REFERENCES
TLC plate of the ethanol extract of T. crispa
Abate T. and Ayalew G. Progresses on Vegetables Pest Management Research In: Proceeding of the 2nd International Horticulture Workshop, (Hearth, E. and Lemma Dessalegn eds). FAO, Addis Ababa, Ethiopia, 187-193, (1992). Norain I., Syazwani A. S., Nor Aziyah B, Siti Nur Amirah Diana F., Lim B.T., Wan Zarina W. K. The effect of Tinospora crispa extract against Spodoptera exigua on Spinacia oleracea. Malaysian Journal of Fundamental and Applied Sciences, 9 (2), 110-114, (2013). Prijono D. Botanical insecticides for organic farming. National symposium on organic farming: integration of traditional and innovative agricultural techniques, Bogor, 457-476, (2004).
GCMS analysis suggested that Ethyl pentadecanoate; 2-Propenoic acid, dodecyl ester; 1,2Benzenedicarboxylic acid; Hexadecanoic acid, bis(2ethylhexyl) ester; Butyl 2,4-dimethyl-2-nitro-4pentanoate; Oxalic acid, decyl 2-ethylhexyl ester; 1Eicosanol; 1-Tetradecanol and 1-Octacosanol were among the volatile components of EETC.
