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Diptera: Tephritidae -..::Egyptian Journal of Plant Protection Research Institute Egypt. J. Plant Prot. Res. Inst. (2020), 3 (1): 493 - 501 Egyptian Journal of Plant Protection Research Institute www.ejppri.eg.net Evaluating the insecticidal efficiency of some legumes extracts against Bactrocera zonata (Diptera: Tephritidae) Mahenaz, A.A. Gab Alla and Basant, S. M. El-Banna Plant Protection Research Institute, Agricultural Research Center, Sabahia, Alexandria, Egypt. ARTICLE INFO Abstract: Article History Insecticidal and biological activity of three legumes; Lupinus Received: 17 / 2 / 2020 luteus L., Glycine max L. and Cicer arietinum; seeds ethanolic extracts Accepted: 31/3/2020 against adult and egg stages of peach fruit fly Bactrocera zonata Keywords (Saunders) (Diptera: Tephritidae) was investigated under laboratory Control, Bactrocera conditions. The phytochemical screening of ethanolic extract of the zonata, Lupinus luteus tested three legumes showed the presence of alkaloids, steroids, , Glycine max and flavonoids, terpenoids, tannins and saponins in G. max extract and Cicer arietinum. absence of saponins in C. arietinum . Also, terpenoids, flavonoids and tannins are absent in L. luteus ethanolic extract. Ethanolic extracts of the tested legume seeds achieved variable toxicity against adult and egg stages of B. zonata. Ethanolic extracts of G. max and C. arietinum -1 seeds (LC50 = 366.2 and 437.3 mg L , respectively) were more toxic against insect adults than L. luteus seeds ethanolic extract (LC50 = 627.5 mg L-1) after 8 days of exposure. Ethanolic extract of C. arietinum achieved the higher ovicidal activity (Egg hatchability reached to 0% at 1000 mg / L) compared to the other extracts. G. max and C. arietinum seeds ethanolic extracts were the most potent on the adult fecundity with 2.0 and 0.0 egg laid / female at 1000 mg / L, respectively compared to 49.0 egg / female in control. The tested ethanolic extracts of the three tested plant seeds reduced the adult longevity of B. zonata and the effect was concentration dependent. Results of the present study suggest that, plant extracts can be an effective tool in integrated pest management programs for the control of fruit flies. Introduction The peach fruit fly Bactrocera South and South- East Asia (Kapoor, zonata (Saunders) (Diptera: Tephritidae) 1993 and Duyck et al., 2004). The is a serious pest of fruits in Egypt and presence of this insect reduces the quality many world regions. It attacks a wide of fruits and subsequently negatively range of fruit and vegetable hosts (White affects their exportation (Shehata et al., and Elson-Harris, 1994). Saafan et al. 2008). Damage is caused mainly by the (1993) recorded B. zonata in many larvae, which feed during the growth and orchards in several Egyptian development of the fruit (Stonehouse et Governorates. Peach fruit fly originates in al., 2005). Therefore, the control of 493 Gab Alla and El-Banna, 2020 larvae is difficult using a single control tested against Bactrocera zonata adult. measure (Dhillon et al., 2005). Effects on the fecundity and fertility of Control methods of this insect are ranged treated adults were also evaluated. from foliage and soil spraying by the Materials and methods specific insecticide, bait-application, 1.Tested insect: B. zonata pupae were male annihilation techniques, releases of obtained from the Department of Pests sterilized flies and parasitoids, and Horticultural Crops, Plant Protection cultural controls (Khan et al., 2017). The Institute Research, Agricultural Research unwise use of synthetic insecticides Center. Emerged adults were introduced resulted in many environmental to cages of 30 cm- 30 cm - 30 cm size. problems, health problems, development Cages were enclosed with mesh screens of insecticide resistance and natural and have cloth sleeves at front and back enemies toxicity (Victor, 2009). sides for the introduction of the food Therefore, there is a need to search about (hydrolyzed yeast and sugar of ratio 1:3) more effective and safe alternative (Khan et al., 2016) and oviposition trays methods to these synthetic insecticides. (El-Minshawy et al., 1999). The insect Between these alternatives are the plant larvae were reared under the laboratory natural products. conditions (25 °C ± 2; 65% ± 5 RH.) on a Plants synthesize many aromatic semi-artificial diet (Wheat bran 500 gm, secondary metabolites, like phenols, molasses 250 gm, dried yeast 150 gm, phenolic acids, quinones, flavones, citric acid 4 gm and sodium benzoates 6 flavonoids, flavonols, tannins and gm in 1liter water) according to Awad coumarins which can negatively affect (1993). insects (Cowan, 1999). These compounds 2.Tested plants: can be used as repellents, feeding Seeds of three legumes; L. luteus, G. deterrents, toxins, defense chemicals and max and C. arietinum, ethanolic extracts growth regulators. Many are also were used to study its potential biocidal effective against biting Diptera, activity against the tested insect B.zonata. especially those volatile components Dried legume seeds were purchased from (Gurjar et al., 2012). It is documented local markets, Alexandria Governorate. that, the leaf extracts of eucalyptus The plants were identified (Eucalyptus globulus) shows efficacy by=Department of Horticultural against Bactrocera cucurbitae (Ali et al., Crops,Agricultural Research Center, 2011). In addition, Mahmoud and Shoeib Alexandria. (2008) showed that low concentrations of 3.Plant extraction: neem can be applied effectively as The legumes seeds=were sterilant and oviposition deterrent for the pulverized into fine=powder using a peach fruit fly populations. Neem blocks grinding mill. The extraction of the the ovarian development and can be used investigated plants was carried out as safe alternative of insecticides for the according to the method of (Mbatchou et control of a species (Mahfuza et al., al., 2011). Powder of each of L. luteus , 2007). In the present study, the toxic G.. max and C . arietinum seeds (200g) effects of ethanolic extract of three were soaked in (700 ml) of ethanol 98% legume plant seeds; Lupinus luteus L. (L. for two weeks with intermittent shaking. luteus), Glycine max L (‎G. max) and The extracts were separately evaporated Cicer arietinum L (C. arietinum), were to dryness at room temperature to obtain 494 Egypt. J. Plant Prot. Res. Inst. (2020), 3 (1): ): 493 - 501 the crude extracts (ethanol extracts). This added to 2 ml of distilled water and procedure was repeated 10 times. The boiled for 3:5 minutes. The resultant resulting crude extracts were stored in mixture was filtered, allowed to cool with glass vials pack closed at (2-4°C) until the filtrate shaken=vigorously. Honey used for phytochemical and bioassay comb froth higher than the aqueous layer assessment experiments. was taken as strongly positive for 4. Qualitative phytochemical screenings: saponins. The extracts obtained from 5.Insecticidal activity of legumes ethanol extract, were subjected to extract: preliminary phytochemical analysis tests The=bioassay=experiments=were= to identify the main chemical groups such carried=out=using=the=extract=of the as alkaloids, steroids, flavonoids, investigated plants to test its potential saponins and tannins according to biocidal activity against B. zonata. Series Mbatchou et al. (2011). of concentrations of each botanical 4.1.Alkaloids test (Wagner’s test): extract from the three legumes were One ml of each extract was treated prepared in ethanol and tested against B. with drops of Wagner’s reagent (Dissolve zonata adults. Tween-twenty was used as 2 g of iodine and 6 g of potassium iodide an emulsifier. Twenty adults of B. zonata in 100 ml distilled water) and observed (Males and Females) 10 days old gravid for the formation of reddish brown flies (Rehman et al., 2009a) put in plastic precipitate. container containing food (hydrolyzed 4.2. Flavonoids test (Willistatter test): yeast and sugar of ratio 1:3) and water. To an ethanol solution of each (Sultana et al., 2013). The tested extract extract, a piece of magnesium ribbon was was put as a layer on the banana peel in added, followed by dropwise addition of the plastic container. Emulsifier and concentrated HCl. Colors ranging from ethanol solution without extract was orange to red indicated flavonoids. added to a piece of banana peel and kept 4.3. Terpenoids and steroids test as untreated control. Each treatment in (Liebermann Buchart test): addition to control was replicated three A small quantity of each extract was time. The plastic container covered with dissolved in trichloromethane, and a muslin cloth and tight with rubber bands minimum volume of concentrated and kept under laboratory conditions. The sulphuric acid added to its content. A adult mortality and number of laid eggs blue or green color or a mixture of these /female was recorded after 2, 4, 6 and 8 two shades was taken as positive test for days of treatment. The eggs removed and terpenoids compounds and the formation counted from banana peel by pin and of dark pink or red color taken as positive placed in a petri dish having wet black test for Steroids compounds. cloth to calculate egg hatchability%. 4.4. Tannins test: Mortality percentages were calculated Each plant extract (0.2 ml) was re- and subjected to probit analysis according extracted with ethanol. The solution to Finney (1971). The concentration obtained was later treated with 5% ferric which cause 50% mortality (LC50) and chloride. A blue-black or blue-green the time required for 50% mortality appearance was taken as positive test for (LT50), confidence limits and slope ± SE tannins. were calculated. 4.5. Saponins test: 6. Statistical analysis: A small portion of each extract was 495 Gab Alla and El-Banna, 2020 Data were subjected to one-way present in the G. max L ethanolic extract. ANOVA test (Duncan’s Multiple Alkaloids, steroids and saponins were Comparison Range Tests at 0.05% level). identified in L. luteus extracts. C. The means were separated through arietinum ethanolic extract contains Tukey’s HSD (Honest Significant alkaloids, steroids, flavonoids, terpenoids Difference) test at a significance level of and tannins.
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