J. Expt. Biosci. 5(1):29-34, January 2014
ISSN 2223-9626 (Online), ISSN 2077-3358 (Print)
USE OF PLANT EXTRACTS TO PROTECT MUNGBEAN SEEDS FROM THE ATTACK OF Callosobruchus chinensis Hafeza Khatun1*, Afroza Sultana2 and Shefali Begum1 *Corresponding author: E-mail:
[email protected] Abstract The aim of this investigation was to determine the efficacy of four different kinds of plants extracts, garlic, onion, castor seeds and neem seeds to protect the stored mungbean seeds (Vigna radiata) from the attack of Callosobruchus chinensis. The efficacy of plant extracts were assessed at different doses (10µml, 15µml and 20µml) on the oviposition rate, adults emergence and longevity of Callosobruchus chinensis. It was found that the oviposition rate, adults emergence and longevity of Callosobruchus chinensis on mungbean seeds were reduced due to treated with garlic, onion, castor and neem seeds extracts. In all plant extracts of garlic, onion, castor and neem seeds their effectiveness increased with the increase of doses. Among the four different kinds of plant extracts neem seeds extract showed maximum efficacy in terms of oviposition rate, number of adult emergence and adult longevity of Callosobruchus chinensis.. Key words: Plants extract, Callosobruchus chinensis, Vigna radiata, and Oviposition
Introduction The stored grain pulses are very important item of human food to the teeming population of Bangladesh. They contain high percentage of protein, starch and several types of vitamins. The common people in our country depend on this supplemental protein for their growth and development as they are not able to buy higher protein. As they are good substitutes for fish, meat, eggs etc and at the same time cheap, they are called “poor man’s meat”. After harvesting the dried seeds are stored in the ware house or bamboo made duli for daily consumption and for sowing in the next season. Generally these are heavily infested in the stored condition by Bruchids. Bruchids breed rapidly in storage and by the time they are detected. Among these Bruchids, been weevil C. chinensis, commonly known as “Dhora”, belonging to the family Bruchidae of the order Coleoptera is the major pest in storage as well as in the field (Shinoda and Yoshida, 1989). Gujar and Yadav (1978) recorded 55% to 60% weight loss of stored beans and 46% to 60% loss in protein content by infestation by the beetles. In dried stored beans, large holes through which adult have escaped are easily visible. The adult emerge out from the seed rendering these pulses unfit for human consumption and sowing. (Parker, Talekar and Skinner, 1995). Pesticides are mainly used for successful storage, although these chemicals are hazardous to human health, wild life and the environment. This dilemma has made it mandatory to search for alternative methods of controlling the insect pests that are not only effective but also health wise acceptable. One solution to these problems might be to replace synthetic chemicals by the compounds which occur naturally in plants. Such chemical substances may be toxic to insects in varying degrees and are therefore of potential selective advantages in deterring those enemies (Fraenkel 1969). It was reported that some plant extract inhibited multiplication of insect population (Sangappa, 1977; Ali et al., 1983; Ahmed et al., 1993). Repellant and antifeedant effects of Azadiracta indica were reported by Jilani and Malik (1973) and Leuschner (1975). According to Ivabijaro (1983) and Jilani (1983) the Azadiractin, a component of the neem tree, (A. indica and Melia azedarach), is a promising alternative to synthetic insecticides because of their repellent and 1
Department of Zoology, University of Dhaka 2Department of Zoology (Entomology), Jagannath University, Dhaka-1100, Bangladesh.
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antifeedant effects. The nontoxicity of neem oil has been reported by Larson (1989). Golob and Webley (1980) have screened many vegetable oils for use in preventing post harvest losses due to insect infestation. Plant materials are easily available and cheap, it will be of much helpful to control grain pests and thus the use of chemical insecticides could be avoided. A large number of studies have been carried out on the effectiveness of different kinds of plant extracts against Callosobruchus spp. (Al Lawati et al. , 2002. Chiranjeevi and Sudhakar, 1996, Das, 1989, Khaire et al., 1993, Kachare et al.,1994, Ketker 1986, Pandey et al., 1986, Rajapakse and Senanayake, 1997, Rouf et al., 1996,Tanzubil, 1987). But very little work has been done in Bangladesh (Naumann and Isman , 1995, and Rahman and Rahman, 2004). With this view in mind the present work was undertaken to study the effects of different parts of plants extract on oviposition, adult emergence and longevity of Callosobruchus chinensis on green mung pulse Phaseolus aureus. Materials and Methods The experiment was conducted at the Entomology laboratories of the Department of Zoology. All the experiments were conducted in the incubator at 25°C and humidity 65 ±5%. Plant extracts For this experiment four types of plant parts were used. Garlic (Allium sativum), Onion (Allium cepa), Castor seed (Ricinus communis) and Neem seed (Azadirachta indica). Preparation of extract Garlic,onion,castor seed and neem seeds were crushed separately using mortar and pestle making paste like substance. No water was needed for making garlic and onion pastes.When 200 gm castor seed and neem seeds were separately pounding to paste with the pestle and mortar, 5 to 10 ml cold water was added at a time. The paste was then shifted to a piece of fine muslin cloth and was pressed to collect 20 ml of the crude extract from each of the plant parts utilized. Preparation of seed For the experiment the green mung pulse was collected from local market. Seeds were cleaned and disinfected by keeping the seeds at 70oC for about 18 hours in incubator. Testing of plant parts against test insects To start the experiment some adult beetles were brought to the laboratory from the godown of Tongi bazar. The rearing medium selected for stock culture was mung pulse. So they were reared for three or more generations to obtain sufficient numbers. The stock cultures were set up by placing about 100 adults of both male and female beetle in petridishes. Each contains 200 gm of mung seeds. The petri dishes were closed to prevent the escape of the adult beetle. Then the pulse beetles were allowed to ovipositon on the mung seeds for about 24 hours to obtain sufficient number of egg. Then the adults were removed to another petridish containing 200 gm of seeds. The petridishes were marked with date and kept in incubator with 250C temperature and 65 ±5% relative humidity. For one measurement 5 petridishes were needed, each petridish known as choice chamber and 4 plastic cups were placed in each petridish. Among these, 2 control cup and 2 treatment cup. Control cup marked with C and treatment cup with T each cup with 100 seeds. Control cup with normal seed where treatment cup with extract mixed seeds. Plant extract was added to the the treatment cup by micropipette with different doses such as10µ ml, 15µ ml and 20µ ml and mixed properly with the seed surfaces. Control and treatment cups were arranged serially in alternate order in opposite direction in a choice chamber. Thus there were 5 choice chamber in the oviposition preference test experiments with each type of plant extract used for everyone measurement. In each choice chamber, 5 pair
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J. Expt. Biosci. 5(1):29-34, January 2014
ISSN 2223-9626 (Online), ISSN 2077-3358 (Print)
newly emerged adults were released from rearing petridishes. The choice chamber was kept in incubator at 25o C temperature. After 20-30 days adults emerged. The number of adult C. chinensis emerged from treated seeds were counted, recorded and compared with the number emerged from their respective controls. The design was CRD and the data were analysed statistically and mean separation was done by LSD test. Results and Discussion Effect on oviposition The number of eggs laid on pulses treated with plant extract was significantly lower than their respective controls. Among the four plant extracts the neem seed extract (10, 15& 20µ ml) treated seeds showed the lowest number of eggs. When the pulses were treated with castor seed extract (10, 15& 20µ ml) treated seeds showed the highest number of eggs. (Table-1). In case of garlic and onion extract treated seeds the lowest number of adult emerged with the dose20µ ml. Effect on adult emergence The highest percentage of adult emergence was found in the control in comparison with their respective treatments. The lowest number of adults emerged in the neem treated pulses. In case castor seed extracts adult emergence significantly differed at p< 0.05 whereas the oviposition preference and longevity were insignificant at p ≥0.05 for all doses (10, 15, 20µ ml). In the same way neem seed extract showed insignificant at p ≥0.05 for the rate of oviposition, adult emergence and longevity. Effect on longevity The mean longevity of emerged adults of pulses treated with garlic, onion, castor and neem extract (10, 15& 20µ ml) was significantly lower than the respective controls. On control pulses the longevity was 14 to 18 days where as on the treated seed it was 7 to 10 days. In case of neem seed extract treated seed were significantly lower than the other plants extract and highest longevity of emerged adult found in garlic extract treated pulses for 10 µml extracts. Table 1. Mean number of eggs laid on the control seeds and treated seeds with crude extracts of garlic, onion, castor seed & neem seed (10, 15 and 20 µml) Plant extracts 10 µml Garlic Onion Castor Neem
Control 50.5±5.3563 33.5±3.9334 28.7±3.1625 40.3±3.4674
Treated 8.2±1.5405 11.4±1.2754 16.2±3.9715 2.6±0.7333*
No. of eggs laid Doses 15 µml Control Treated 24.3±3.786 3.3±0.6506 30.3±4.885 11±2.4875 21.6±6.231 20.4±4.876 35.7±4.531 3.1±0.679*
20 µml Control 6.7±2.5431 16.6±5.5561 42.9±6.1218 33.2±3.8724
Treated 1.6±0.6863* 7.2±2.2597* 14.4±2.757* 1.3±0.1448*
* Significant at α=0.05
Several workers have reported growth and development inhibition properties of plant extracts on pulse beetle, C. chinensis. Much of this literature refers to the effects of neem oil. Ketker (1986) reviewed the effect of neem oils , along with three other non edible oils against C. chinensis. It was found that the growth index with neem was the lowest; neem also had the greatest ovicidal effect. Reddy et al. (1994) reported that mung bean seeds treated neem oil halted the embryonic development of C. chinensis, protecting stored seed for a period of 12 months. Rajapakse and Senanayake (1997) showed that seeds treated with neem oil significantly reduced oviposition of
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C. chinensis. The antifeedent activity of neem seed extracts has been reported for numerous insects pest (Warthen,1989). Neem extracts have been shown to repel or deter several homopteran pests, including leaf hoppers and plant hoppers on rice (Heyde et al., 1984; Saxena and khan, 1987). Schmutterer (1987) observed that the seed kernel of neem has fecundity inhibiting quality. Jilani and Malik (1973) reported that the neem seeds exhibited maximum repellency against insects in comparison with leaves and flowers. Tikku et al. (1981) reported 100% control of the emergence of Bruchus chinensis from the grains treated with castor extract. Dohary et al., (1988) reported a high dose (1 ml/100g) soybean, neem and sesame oils caused 100% adult mortality of C. chinensis and C. maculatus after one day of exposure. Rouf et al.(1996) reported that mixing of neem leaf powder with lentil seeds resulted in reduced oviposition and adult emergence in the same beetle. Chiranjeevi and Sudhakar (1996) revealed that neem seed powder, mixed with mungbean seed, completely prevented the development of C. chinensis. It indicates a significant potential for plant extracts as a possible source of natural products that could be used as an alternative to synthetic insecticides. Table 2. Mean number of adults emerged from control and treated seeds with crude extract of garlic, onion, castor seed and neem seed (10, 15 and 20 µml). Plant extracts 10 µml Garlic Onion Castor Neem
Control 48.9±5.2777 31.1±3.7072 25.1±2.8380 36±3.6147
Treated 5.3±1.3988* 5±1.0719 7.6±2.4367 0.7±0.2603*
No of adult emerged Doses 15 µml Control Treated 23.4±3.6642 1.9±0.4333* 28.8±4.9324 5.1±1.0159 18.8±5.4970 3.9±1.1685 33.6±4.796 0.8±0.2905*
20 µml Control 5.7±2.7553 16±5.2345 39.4±6.0391 30±3.3399
Treated 1±0.4714* 3.7±0.8825 1.8±0.512* 0.2±0.133*
* Significant at α=0.05
Table 3. Mean longevity of emerged adults on control & treated seed with crude extracts of garlic, onion, castor seed and neem seed (10, 15 and 20 µml). Plant extracts
Garlic Onion Castor Neem
10 µml Control Treated 17.6±0.6531 8.6±0.6* 15.8±0.5537 8.2±0.6960 15.8±0.8137 8.25±1.416 16.4±0.653 8±1.9777*
Longevity of emerged adult Doses 15 µml Control Treated 16±0.7888 9.33±1.0795* 14.4±1.0241 8.2±0.6289* 16±2.521 8.25±1.3354 17.6±0.498 8±1.9321*
20 µml Control Treated 15.25±2.3676 7±1.8378* 16.25±2.6925 8.5±1.422* 16.4±0.9333 8±1.545* 17±0.5374 7±2.1134*
* Significant at α=0.05
It may be concluded that the efficacy of different kinds of plant extracts in terms of oviposition, adult emergence and longevity, neem seed extract is most effective in all(10µml, 15µml, 20µml) doses. In all plant extracts ( garlic, onion, castor seed, neem seed) effectiveness increased with the increase of doses. In present study 20µml dose was found the most effective one. So neem seed > garlic> castor seed> onion > are most promising as seed protectents. It indicates that plant extracts could be used as an alternative to synthetic insecticides.
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J. Expt. Biosci. 5(1):29-34, January 2014
ISSN 2223-9626 (Online), ISSN 2077-3358 (Print)
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