African Journal of Agricultural Science and Technology (AJAST) Vol. 3, Issue 7, pp. 310-315. June, 2015 http://www.oceanicjournals.org/ajast ISSN 2311-5882 ©2015 Oceanic Journals Full Length Research Paper Synergetic effects of various plant extracts as bio-pesticide against Wheat Aphid (Diurophous noxia L.) (Hemiptera: Aphididae) Habib Ali1*, Muhammad Qasim2, Hafiz Sohaib Ahmed Saqib1, Muhammad Arif1 and Saif-ul Islam1 1 Department of Plant Protection, Fujian Agriculture and Forestry University, Fuzhou, China, 350002. 2 Department of Agricultural Entomology, University of Agriculture, Faisalabad-Pakistan, 35040. *Corresponding author. E-mail: habib_ali1417@yahoo.com Accepted 6th July, 2015 Human health and environment are greatly affected by extensive used of synthetic insecticide, which is why the alternative way of pesticides, such as botanical pesticide consumption increase with the passage of time to control of insect pests. The insecticidal impact of Moringa (Moringa oleifera), Neem (Azadirachta indica) and Euclyptus (Eucalyptus globules) leaves extracts were investigated on Diurophous noxia (Hemiptera: Aphididae). This study was aimed to evaluate the efficiency of bio-pesticide instead of synthetic chemicals against D. noxia. Experimental results showed that the percentage of mortality fluctuate with the fluctuation of concentration with time intervals and have great impact on the population of D. noxia. Single, doubled and tripled combinations of leaves extract were tested at two different concentrations (25 and 50%). The results suggested and concluded that from all the application of single botanicals A. indica give better result as compared to other single application while application of double combination M. olerifera with A. indica give significance result instead of other double mixture, meanwhile the triple combination (M. olerifera, A. indica and E. globules) performed excellent result as the others triple mixtures. The study revealed that combine used of M. olerifera, A. indica and E. globules leaves extract were very effective against D. noxia and surge as an alternative way of pesticides instead of synthetic chemicals. Key words: Wheat aphid, plant extract, mortality, M. olerifera, A. indica, E. globule. INTRODUCTION Wheat is the staple food of Pakistan, which is being used 129 kg per head annually and this crop covers more than eighty percent of country. In 2008-2009, wheat were cultivated over an area of 8.459 million hectares giving 23.03 m. tones production (Zeb et al., 2011). Wheat is great source of income in the rural areas of Pakistan and as well as in the world. Yield per acre of wheat in Pakistan is much lower than the other wheat growing countries. Many factors contribute to the low yield of wheat in Pakistan including, method of cultivation, improper irrigation facilities, low level of soil fertility and particularly insect pest attacks. ` All over the world, wheat is faced with many problems related to insect pest attacks such as weeds, pathogens and insects, which are main causal agents in yield reduction (25–50%) (Pimentel et al., 1991; Oerke, 2006). Thus, wheat is drastically being affected by sucking insect pests and dominated by wheat aphid, Diuraphis noxia (Mordvilko) (Homoptera: Aphididae). There are six species of wheat aphid that include Russian wheat aphid (D. noxia), the English grain ear aphid (Sitobion avenae), common green wheat aphid (Schizaphis graminum), bird cherry-oat aphid (Rhopalosiphum padi), and rose grain aphid Afr. J. Agric. Sci. Technol. 311 (Metopolophium dirhodum) which were reported to damage wheat but D. noxia was reported most damaging and dangerous specie (Toit, 1986; Prinsloo et al., 2007). It attacks specially at early stage of wheat and causes significant loss almost twenty percent at the end of harvest. The population of D. noxia is greater from last few years and now it becomes a regular pest of Pakistan (Aheer et al., 2008). Management of D. noxia was being done by commercial insecticides consistently and blindly, but the applications of chemicals caused some nonrecoverable problems, like development of new resistant insect’s strains, accumulation of pesticide residues in food chain and on the other hand increasing the cost of these pesticides. Thus, some plant extracts integrated in the control measures to limit the populations of wheat aphid across the globe, by repellence, mortality, and affecting the fecundity (Jilani and Su, 1983; Kim et al., 2003). Moringa is much beneficial in daily life, due to certain values, such as; antispasmodic (JU et al., 2014), diuretic, antihypertensive, cholesterol lowering (Mehta et al., 2003), antioxidant, antidiabetic, hepatoprotective, antibacterial and antifungal activities (Farjana et al., 2003). Pulverized leaves of M. oleifera were applied to Tribolium castaneum (Herbst) (Anita et al., 2012; Saleem et al., 2014) and Aedes aegypti (Ferreira et al., 2009), which caused 100% mortality to larvae as well as adults within nine days. The tree of neem, A. indica, is a multipurpose plant that is cultivated in almost all parts of the world. Three hundred elements are reported in the extract of A. indica, which are very effective against insects, termites, and nematodes and known as biological regulator against insects’ pests as well as environmental friendly with respective to the ecosystem (El Shafie and Basedow, 2003). The active compound interferes with the feeding activity of pests, interrupt molting process, and growth postponed (Ahmed and Grainge, 1985; Mordue et al., 2010). Genus eucalyptus is an aromatic plants belonging to a family Myrtaceae, which originated from Australia with seven hundred species identified all over the world (Brooker and Kleinig, 2006), and is regarded as one of the most-extensively planted pulpwood species. E. globules is also a multipurpose plant and has variety of composition (Brooker and Kleinig, 2006). The genus Eucalyptus do not only provide firewood for biomass and minimize the concentration of environmental carbon dioxide level but is also used direct or indirect for the control of insect pest, and known as allopathic property to the tree (Liu et al., 2008). In general, the plant secondary metabolites including phenolics, tannins and even monoterpenes are considered to have co-evolved with herbivory (Vourch et al., 2002; Bailey et al., 2004; Foley and Moore, 2005). There is no combine study reporting the use of ` different natural plants extracts (M. oleifera, A. indica and E. globules) as a bio-pesticide against D. noxia. Therefore, the present study was carried out to investigate the synergetics effect of M. oleifera and other plants extract as bio-pesticide. MATERIALS AND METHODS Experiments were carried out during 2013-14 at Entomological Research Center, University of Agriculture, Faisalabad, Pakistan. Experimental area was divided into two parts, untreated plots or control plots with the size of 5 × 3 m and treated plots with the size of 5 × 3 m sprayed with Neem, Euclyptus and Moringa with two different concentration (at 25% con. 1/4 plant extracts with 3/4 distilled water and at 50% conc. 2/4 plants extracts with 2/4 distilled water) for the control of aphid, one spray at flowering or milky stage (growth stage (GS) 69, (Tottman, 1987) and the other spray at maturing stage of wheat. The plots were arranged in a completely randomized block design with split plot arrangement. All agronomic practices were uniformly carried out according to farmers’ practices in the study area. The planned study was conducted with the objective to find effectiveness of some botanical bio-pesticides against wheat aphid. It were comprised of eight treatments viz. T1= Moringa leafs extracts, T2= Neem leafs extract, T3= Eucalyptus leafs extracts, T4= Moringa leafs extract+ Neem leafs extract, T5= Moringa leafs+ Eucalyptus leafs extracts, T6= Neem + Eucalyptus leafs extracts, T7= Moringa + Neem + Eucalyptus leafs extracts, T8= control were applied (Table 1). Two sprays, one at milky stage and other at maturing stage at different concentrations (25 and 50%) were applied. All the recommended agronomic practices were carried out during experimentation. Fresh and healthy leaves of three allelopathic tree species viz., M. oleifera, A. indica and E. globules were collected from University of Agriculture and surrounded areas of Faisalabad, Pakistan. They were chosen due to their availability in the wheat production regions, their medicinal properties (Van Wyk and Gericke, 2000) and their insect repelling properties (Bruce et al., 2002; Zehnder et al., 2007; Halbert et al., 2008; Görür et al., 2009). Fresh leaves from each of 3 test tree species were soaked in sterilized distilled water for 24 h. After soaking, leaves were grind through rotary shaker for extraction, and extract was filtered through a double layered muslin cloth followed by Whatman No. 1 filter paper and used in the further experiments immediately. For recording the aphid’s population, wheat plants/plot were randomly to count aphids from leaves and spikes in the field. The average number of aphids/plant was calculated before and after application of plants extracts. At the economic threshold level of Habib et al. 312 Table 1. Experimental design of conducted study. Experimental design of conducted study T1 Moringa leaves Extract (MLE) @ 25 and 50% conc. T2 Neem leaves Extract (NLE) @ 25 and 50% conc. T3 Eucalyptus leaves Extract (ELE) @ 25 and 50% conc. T4 Moringa + Neem (MLE+ NLE) @ 25 and 50% conc. T5 Moringa + Eucalyptus (MLE+ ELE) @ 25 and 50% conc. T6 Neem+ Eucalyptus leaves Extract (NLE+ ELE) @ 25 and 50% conc. T7 Moringa+ Neem+ Eucalyptus leaves Extract (MLE+ NLE+ ELE) @ 25 and 50% conc. T8 Control wheat aphid, these bio-pesticides were applied twice once at milky stage and other in mature stage as foliar application with manual sprayer. The sprays were applied at two different concentrations; 25 and 50%. Data was collected 24 h before and 24, 48, 72 h and one week after application of bio-pesticide. For recording the aphid’s population 15 randomly selected plants and density of aphid’s plant-1 with mean values of spike/tiller and leaf/tiller were collected. For aphid’s control, the above bio-pesticides were sprayed on the wheat plants surface with the given concentrations. The calculation of percentage mortality was achieved with the help of the formula; % Mortality = Pre treatment − Post treatment × 100 Pre treatment Data was analyzed with the help of MSTAT-C computer program (Freed and Scott, 1986) and means were compared at 5% significance level. RESULTS AND DISCUSSION Significant results were obtained with the application of plants extracts with regard to mortality and dormant effect on population buildup of wheat aphid. Pretreatment data were recorded before 24 h for the application of bio-pesticides (Table 2). The obtained results revealed that different concentration of plants extracts as bio-pesticides varied greatly with regard to insect mortality. All plant extracts were observed to be effective against wheat aphid, but in case of single application of plant extracts, Neem Leaf Extract (NLE) performed better and give significant results than all other single extracts (Moringa Leaf Extract, MLE and Eucalyptus Leaf Extract, ELE). Table 2. ANOVA for the population of wheat aphid per spike before 24 h of extracts application. Source Blocks Treatments Error Total DF 2 7 14 23 Stander error, 0.556. ` SS 0.3333 7.625 13 20.958 MS 0.1667 1.0893 0.9286 F P 1.2 0.377 According to single applications of bio-pesticide on leafs, for example, MLE average mortality% of D. noxia were recorded 58.37, 45.87, 33.37, 20.87 and 47.95, 55.37, 41.97, 22.25%: ELE, 50, 35.43, 22.93, 14.65 and 43.97, 51.78, 38.38, 21.73% at 25 and 50% concentrations respectively but NLE performed better results, that is, 62.5, 50, 37.5, 25 and 73.52, 52.93, 44.10, 23.50% as compared to other single application at both concentrations after 24, 48, 72 h and one week of time intervals of applications, respectively. With the application of double plant extracts mortality% were recorded in MLE + ELE , 50, 37.5, 25, 14.62 and 55.98, 59.81, 46.42, 29.55%: NLE+ ELE, 56.25, 45.81, 33.37, 16.68 and 59.98, 54.46, 41.06, 24.34% but with the combination of MLE+ NLE, 58.37, 47.93, 35.43, 18.75 and 64.92, 47.78, 38.10, 20.36% % give significance results as compare to other double combinations at 25 and 50% concentrations on spikes after 24, 48, 72 h and one week of time intervals of application respectively. Similarly, application of plant extracts in combination of three, MLE+ NLE+ ELE performed better and excellent results than application of plant extracts in alone or combination at 25 and 50% concentration at both sprays. Due to combined application of three plant extracts as bio-pesticide average mean mortality of wheat aphid were 64.62, 52.06, 39.62, 29.18 and 82.34, 61.73, 49.27, 32.41% on leaves after 24, 48, 72 h and one week of application respectively (Figure 1). Significant results were obtained on spikes with the application of plants extracts with regard to mortality with different concentrations. Pretreatment data were recorded on spikes before 24 h for the application of bio-pesticides (Table 3). Table 3. ANOVA for the population of wheat aphid per leafs per tiller before 24 h of extracts application. Source Blocks Treatments Error Total DF 2 7 14 23 SS 7.5833 1.1667 7.0833 15.833 MS 3.7917 0.1667 0.506 F P 0.3 0.928 Stander error, 0.4107. According to single applications of bio-pesticide on spikes, MLE average mortality % of D. noxia were Afr. J. Agric. Sci. Technol. 313 Figure 1. Mortality of wheat aphids on leafs at the concentration of 25 and 50% solution, each colure of 1st line indicate the 25% and each colure of 2 nd row indicate the 50% concentration of solution, while different colors show different time intervals. Figure 2. Mortality % of wheat aphids on spikes at the concentration of 25 and 50% solution each colure of 1st line indicate the 25% and each colure of 2 nd row indicate the 50% concentration of solution, while different colors show different time intervals). recorded 44.23, 25.65, 15.02, 9.46 and 51.61, 25.80, 12.90, 9.67: ELE 34.51, 13.27, 7.96, 2.58 and 48.38, 21.51, 4.32, 2.15: at 25 and 50% concentrations but NLE performed better results, that is, 47.79, 27.42, 16.80, 11.20 and 67.85, 57.13, 43.74, 26.95 as compare to other single application at both ` concentrations after 24, 48, 72 h and one week of time intervals of applications respectively. With the application of double plant extracts mortality % were recorded in MLE + ELE , 55.57, 43.45, 18.18, 15.15 and 55.93, 41.93, 17.22, 13.97% : NLE+ ELE, 41.58, 19.46, 9.71, 6.02 and 65.17, 54.46, 41.06, Habib et al. 24.34% but with the combination of MLE+ NLE, 61.07, 38.92, 29.20, 23.27 and 73.21, 64.29, 51.78, 33.91% give significance results as compare to other double combinations at 25 and 50% concentrations on spikes after 24, 48, 72 h and one week of time intervals of application respectively. Similarly, application of plant extracts in combination of three, MLE+ NLE+ ELE performed better than application of plant extracts in alone or combination at 25 and 50% concentration at both sprays. Due to combined application of three plant extracts as biopesticide average mean mortality of wheat aphid as 65.48, 46.89, 37.17, 31.03 and 75.89, 65.17, 51.78, 33.91% on spikes recorded after 24, 48, 72 h and one week of application respectively (Figure 2). Our results are in comparison with Kim et al. (2003) in which 90% mortality of Sitophilus oryzae were found with application of different plants extracts viz Acrorus gramineus rhizhome, A. indica, Acrorus calamus and IIIicium verum after 3 or 4 days of treatments. Elhag (2000) studied the effect of 9 different plant extracts against Callobrouchus maculates and the highest mortality were observed with application of A. indica (76.8%), Heliotropium bacciferum (59.2%) and citrous peel (58.6) of plants extracts. Similarly, Chitra et al. (1991) and Chandel et al. (1987) studied the toxicity of different plants extracts viz, 0.5% aqueous crude leaf extract of A. squamosa, 1% aqueous neem leaf extract and 1% of petroleum extract of Eucalyptus globulus and leaf extract of Vinca rosea petroleum, extract of Parthenium hysterosphorius and whole plant extract of yellow berries night shade, Solanum xanthocarpum were used against brinjal spotted leaf beetle, Henosepilachna vigintioctopuncata and found 100% protection against II and III instar grubs of this beetle. Conclusion The result showed that use of plant extracts alone or combination as bio-pesticides at 50% concentration were highly effective against wheat pests (wheat aphid). More important was the primary antifeedant effects of bio-pesticides were persistent for a period of at least 7 days with little or no deficiency. The result revealed that maximum mortality % of aphids were recorded in T7= Moringa + Neem+ Eucalyptus leafs extracts at 50% concentration on leafs (82.34, 61.73, 49.27 and 32.41%) and on spikes (75.89, 65.17, 51.78 and 33.91%) after 24, 48, 72 h and one week of application respectively, While minimum mortality % were recorded in T8= Untreated plots (0%) followed by T3= Eucalyptus leafs extracts, T6= Neem+ Eucalyptus leafs extracts at both sprays and at both concentrations respectively. From the results of the experiments, it can be concluded and suggested that the combined use of bio- ` 314 pesticides (M. oeifara + A. indica + E. globolus) at 50% concentration are very effective against wheat aphid. Moreover, these plants extracts can be used as an alternative tool to conventional synthetic insecticides and are friendly to human health as well as to the environment. The use of these bio-pesticides could play an important role in integrated pest management programs in future. 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