EFFECT OF EGG PARASITOID, TRICHOGRAMMA CHILONIS,

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1 Sarhad J. Agric. Vol.28, No.2, 2012 EFFECT OF EGG PARASITOID, TRICHOGRAMMA CHILONIS, IN COMBINATION WITH CHRYSOPERLA CARNEA AND NEEM SEED EXTRACT AGAINST TOMATO FRUITWORM, HELICOVERPA ARMIGERA MUHAMMAD USMAN, MIAN INAYATULLAH, AMJAD USMAN KAMRAN SOHAIL and SYED FAHAD SHAH Department of Entomology, Agricultural University, Peshawar Pakistan. ABSTRACT To investigate the efficiency of Trichogramma chilonis, T. chilonis in combination with Chrysoperla carnea and against tomato fruitworm, Helicoverpa armigera, field experiments were carried out at the Research Farm of Agricultural University, Peshawar, Pakistan during summer The plot size was 8 x 5 meters. Five treatments were used i.e. trichocard having 300 parasitized (T 1 ), trichocard having 300 parasitized in combination with 45 instar larvae (T 2 ), trichocard having 300 parasitized and (T 3 ), trichocard having 300 parasitized in combination with 45 Chrysoperla larvae, (T 4 ) and control (T 0 ). The lowest number of H. armigera larvae (0.68 plant -1 ) recorded in T 4 was significantly lower than those of all other plots which were followed by T 3 and T 2 i.e and 0.97 plant -1 while the highest number of larvae plant -1 (1.40) was recorded in T 0. The yield of tomato was highest (11570 kg) recorded in T 4 and the lowest yield (7250 kg) was recorded in T 5. The percent weight loss was minimum (11.04%) in T 4 while maximum (20.45%) in T 0. Based on yield and percent fruit damage, treatment having trichocard having 300 parasitized in combination with Chrysoperla and is the most promising for effective management of H. armigera on tomato. Key Words: Egg parasitoid, Trichogramma chilonis, Chrysoperla carnea, neem, Helicoverpa armigera. Citation: Usman, M., M. Inayatullah, A. Usman, K. Sohail and S.F. Shah Effect of egg parasitoid, Trichogramma chilonis, in combination with Chrysoperla carnea and neem seed extract against tomato fruitworm, Helicoverpa armigera. Sarhad J. Agric. 28(2): INTRODUCTION Tomato, Lycopersicum escolentum Mill, is an important and widely used vegetable crop. It is very nutritive and delicious; very few vegetables can match its nutritional value. Tomato is one of the most important vegetable crops cultivated for its fleshy fruits and considered as important commercial and dietary vegetable crop. It is short duration crop and gives high yield, it is important from economic point of view and hence area under its cultivation is increasing day by day (Kevany et al., 2008). In Pakistan the fruit borer Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is a key pest of several crops. Tomato, cotton, pigeon pea, and chickpea are consistently suffered by this insect in different parts of the country. The fruit borer is destructive in larval stages and cause serious damage to tomato fruit. The moth (adult) generally appears during April-May and lay on leaves. The young larvae feed on leaves, flowers and bore in tomato fruit and complete most of its development inside the fruit. Larvae have 5-6 instars, all larval stages damage tomato fruit. During development the larvae may emerge from one fruit and enter another and cause infestation. The last larval instars come out from the tomato fruit, falls down to the soil and pupate in soil. A single caterpillar can damage up to 12 fruits and hence lowers the market value of the fruit and ultimately farmers bear great losses (Hussain and Bilal, 2007). Among the parasitoids the Trichogramma species are the most commonly used egg parasitoids in biological control program released on a large scale against lepidopterous pests. Adults of T. chilonis (Hymenoptera: Trichogrammatidae) are tiny wasps that kill the of hosts by laying one or more of their own inside the much larger of Lepidoptera. Recognizing the potential of the Trichogramma species as a biological control agents, entomologist in the early 1900 began to mass rear Trichogramma for insect control. Today Trichogramma species are the most widely used insect natural enemy in the world because their mass rearing is easy on one hand and they attack many important crop insect pests on other hand (Ayvaz et al., 2008). Chrysoperla or green lacewing belongs to the insect order Neuroptera and is found in most environments throughout the world. There are several species and the most common is C. carnea Stephen. The adults feed on pollen, nectar or honeydew of aphids, but the larvae are voracious predators (Easterbrook et al., 2006). Neem, a plant originated (botanical) pesticides which is environment friendly and is well known for its diverse pest control properties. It works as an insect feeding deterrent, but in various forms it also serves as a repellent, growth regulator, oviposition (egg deposition) suppressant, sterilant, or toxin. Neem caused higher reduction in the larvae of shoot and fruit borer Earias vittella and E. insulana and recorded the highest yields of

2 M. Usman et al. effect of egg parasitoid, trichogramma chilonis against tomato fruitworm and 4180 kg ha -1 in okra crop (Dhingra et al., 2008). In Pakistan tomato pests are usually controlled by using chemicals and no serious efforts have been made to use non chemical methods. Published information shows that efforts have been made in many countries of the world to control tomato fruit worm by using natural enemies including C. carnea and Trichogramma. In Khyber Pakhtunkhwa Inayatullah (2007) studied the effect of Trichogramma alone and in combination with C. carnea on the tomato fruit worm and reported significant reduction in fruit worm infestation and percent yield loss. The author, however did not record any significant increase in yield. In the present studies the effect of Trichogramma alone and the combined effect of Trichogramma with Chrysoperla and neem seed extract were studied. In the present research, we tried: i. To study the effect of T. chilonis alone and in combination with C. carnea, and against tomato fruit worm, H. armigera, at the Research Farm of the Khyber Pakhtunkhwa Agricultural University Peshawar, and ii. to study the effect of T. chilonis alone and in combination with C. carnea, and Neem extract on the yield of tomato. MATERIALS AND METHODS Experiments were conducted at the Research Farm of Agricultural University, Peshawar, Pakistan during summer The experiment was composed of 5 treatments each with 3 replications. The plot size was 8 x 5 m and between each treatment a 3 meters buffer zone was maintained. The distances between the replications were 2 meters. The experiment was designed as RCB Design. Rearing of Trichogramma and C. carnea The rearing of both Trichogramma and Chrysoperla was done at the research laboratory of the Department of Entomology, Agricultural University, Peshawar, Pakistan on standard diet by the procedure adopted by Inayatullah, (2007). Field Preparation and Release of Trichogramma and Chrysoperla The field was prepared for tomato with normal cultural practices. The experiment was conducted on wellprepared seedbed, ploughed twice for obtaining bumper crop. Transplantation was done in the month of March. The release of trichocards was started when the tomato crop reached the flowering stage. Total six releases were done at ten days interval throughout cropping season. The trichocards (from the laboratory culture) from which the emergence was expected in few hours were selected in laboratory; the required numbers of parasitized on card were counted. Trichocard having number of 300 parasitized, in combination with Chrysoperla and were applied to each plot except control. The trichocards were hooked up with the metal rod and were placed in the center of each plot. The lower portion of the rod was greased to prevent the access of ants and any other crawling insects. The required numbers of second of C. carnea were obtained from the Chrysoperla culture established in the entomology research laboratory and were taken to the field. Test tubes containing the second instar larvae were shaken on the leaves of plants at different sites in the same plot. Preparation of Neem Extract Mixture of half Kg crushed neem seed and 10g surf (detergent) were tightened in muslin cloth and placed in 5 liter water for 24h. This gave us 10% solution of using the following formula (Shah, 2004). V 1 C 1 =V 2 C 2 V 1 = volume of water which is required V 2 = volume of known quantity C 1 = given concentration C 2 = required concentration Data Collection Ten plants plot -1 were selected randomly and the number of larvae on all the selected plants were counted and recorded at one week interval. This process was repeated for all the six releases. The mature tomato fruit were collected separately from each plot. Weight and number of damaged tomato fruit were recorded for each plot separately. The total yield of tomato was determined by adding the yields of tomato per picking of each plot. Yield = Total weight of tomato collected from each plot Percent weight loss = wt. of damage fruit x 100 Total wt of tomato

3 Sarhad J. Agric. Vol.28, No.2, Treatments T 1 Trichocard having 300 parasitized T 2 Trichocard having 300 parasitized in combination with 45 instar larvae T 3 Trichocard having 300 parasitized in combination with Neem extract T 4 Trichocard having 300 parasitized in combination with 45 instar larvae and neem extract (no trichocards) T 0 RESULTS AND DISCUSSION Data collected on 1 st week after the treatment indicated that the lowest number of H. armigera larvae plant -1 recorded in treatments T 3 and T 4 i.e. recorded 0.50 and The treatments T 1 and T 2 on par with T 0 control which recorded 0.90 larvae plant -1. The treatments T 3 and T 4 were significantly different from the T 0 Table I. Table I Field layout of the experiment T 1 T 2 T 3 T 4 T 0 Rep 1 + Rep 2 + Rep 3 + Data in Table II recorded on 2 nd week revealed the lowest number (0.63) of H. armigera larvae plant -1 was recorded in T 4 followed by T 3 i.e larvae plant -1. The number of H. armigera larvae plant -1 recorded on T 0 (0.93 larvae plant -1 ) was at par with T 1 and T 2. Significant difference was recorded between the T 3, T 4 and T 0. The number of H. armigera larvae plant -1 recorded on 3 rd week was highest in T 1 i.e. recorded 1.03 larvae plant -1 followed by 0.97 and 0.87 at T 2 and T 3 whereas the lowest number of 0.73 larvae plant -1 was recorded in T 4. The number of H. armigera larvae plant -1 recorded in T 0 was All the treatments were significantly different with the T 0. Table II Effect of Bio agents and on the infestation of tomato fruit worm, Helicoverpa armigera, plant -1 recorded on different dates during 2009 Treatments 15 th April 22 nd April 29 th April 5th May 12 th May 19 th May 26 th May 3 rd June 10 th June Mean T a 1.03a 1.20b 1.33b 1.50b 1.37b 1.13b 0.97b 0.67b 1.12b T a 0.93a 0.97c 1.23b 1.40b 1.20b 0.90bc 0.70c 0.67b 0.97c T b 0.83ab 0.87cd 0.93c 1.10c 0.97c 0.90bc 0.77c 0.67b 0.85d T b 0.63b 0.73d 0.67d 0.90d 0.77d 0.80c 0.67c 0.50b 0.68e T a 0.93a 1.40a 1.63a 1.83a 1.67a 1.67a 1.57a 1.00a 1.40a Means followed by the same letters within each column are not significantly different at 5 % level of probability (LSD Test). Results in Table II on 4 th week indicated that the lowest number of H. armigera larvae was recorded in T 4 i.e larvae plant -1 which was followed by 0.93 larvae plant -1 at T 3. The number of larvae recorded on T 1 and T 2 were 1.33 and 1.23 larvae plant -1 as compared to the T 0 where the number of larvae recorded 1.63 larvae plant -1. Statistical analysis of the data showed significant difference between the treatments and T 0. Data recorded on 5 th week after the application showed the highest (1.50) number of H. armigera larvae plant -1 in T 1 followed by 1.40 and 1.10 larvae plant -1 in T 2 and T 3 as compared to T 4 where the number of H. armigera larvae was recorded lowest (0.90 larvae plant -1 ). In T 0 the number of H. armigera larvae was 1.83 larvae plant -1. Data collected on 6 th week revealed that the lowest number of H. armigera larvae plant -1 was recorded in T 4 i.e larvae plant -1 followed by 0.97 larvae plant -1 at T 3. The number of larvae recorded in T 1 and T 2 and T 0 were 1.37, 1.20 and 1.67, respectively. The number of H. armigera larvae on 7 th week was highest (1.13 larvae plant -1 ) in T 1 as compared to T 0 where the number of larvae recorded was 1.67 larvae plant -1. The number of larvae plant recorded in T 2 on par with T 3 and T 4. On 9 th week the highest number of larvae plant -1 was recorded in control i.e larvae plant -1. The number of larvae 0.70 plant -1 recorded in T 2 on par with T 3 and T 4. On 10 th week the number of larvae 0.67 larvae plant -1 recorded in T 1 was also on par with T 2, T 3 and T 4. In T 0 the

4 M. Usman et al. effect of egg parasitoid, trichogramma chilonis against tomato fruitworm 256 number of larvae recorded was 1.00 larvae plant -1. In both weeks all the treatments were significantly different from the control. Results concluded from overall mean of the data indicated that the lowest number of H. armigera larvae was recorded in T 1 i.e. recorded 1.12 larvae plant -1 followed by 0.97 and 0.85 larvae plant -1 in T 2 and T 3 as compared to T 4 where the number of larvae was recorded lowest (0.68 larvae plant -1 ). In control the number of larvae recorded was 1.40 larvae plant -1. The treatments T 1, T 2, T 3 and T 4 were significantly different from the control. Yield and Percent Weight Loss The mean yield of tomato fruit recorded in T 1, T 2, T 3 and T 4 were 8428, 9039, and kg respectively, as compared to control plot where the yield of tomato fruit was 7250 kg. Table II shows that highest yield (11570 kg ha -1 ) was recorded in T 4 which was non-significant from the yield recorded in T 3 (10010 kg/ha) while significantly higher than T 1, T 2 and T 0. Percent weight loss of tomato fruit recorded in T 1, T 2, T 3 and T 4 were 15.18, 13.65, and 11.04%, respectively as compared to T 0 plot where the percent weight loss of tomato fruit was 20.45%. Statistical analysis of the data showed that the lowest percent damage (11.04%) recorded in T 4 was non-significant from T 3 (12.59%) and significant with all other treatments. Table III Effect of Bio agents and on tomato yield and percent weight loss by H. armigera during 2009 Treatments Yield (Kg) % weight loss T Trichogramma parasitized / hectare 8428 BC B T Trichogramma parasitized with 66000/ha Chrysoperla 9039 BC BC T Trichogramma parasitized with AB CD T Trichogramma parasitized with 66000/ha Chrysoperla, A D T C A Means followed by the same letters are not significantly different at 5 % level of probability (LSD Test). The above results show that Trichogramma parasitism, Trichogramma in combination with Chrysoperla and neem (T 4 ) does have an impact on population of tomato fruit worm. In the present experiment Trichogramma in combination with Chrysoperla and (T 4 ) decreases the population of tomato fruit worm. These results are in conformity with those of Reddy and Manjunatha (2000) who used Trichogramma chilonis alone and in combination with plant extracts and synthetic pesticides and reported positive results in controlling lepidopterous pests of field crops. Shah (2008) after conducting field experiments also reported that Chrysoperla carnea and Trichogramma chilonis are the best bio-control agents for controlling tomato fruit worm in tomato crop. For significant reduction in pest population however, application of 50 2 nd instar C. carnea larvae and 400 Trichogramma wasps were significant in reducing the pest population. Despite application of bio-control agent the pest survived, though in significantly less number, to cause some damage. Workers around the world could not achieve 100% control with Trichogramma, Chrysoperla and. This includes Shah (2008) who tested Trichograma, Chryosperla and respectively, against H. armigera. In the present studies the mean number of tomato fruit worm recorded show that the application of Trichogramma in combination with Chrysoperla and neem (T 4 ) significantly help in reducing the infestation of tomato fruit worm. Although the treatment with the Trichogramma in combination with Chrysoperla and neem (T 4 ) significantly reduced the number of tomato fruit worm, it also put a positive impact on yield. Shah (2008) achieved significant increase in yield. In the present experiment, the reason for this low-significant reduction may be that due to less number of pest larvae, they faced little competition and distributed well in the field plant -1 and caused damage. Figures of percent weight loss show that all the treatments are better than control and application of Trichogramma in combination with Chrysoperla and neem can be recommended. Release of Trichogramma in combination with Chrysoperla and neem put a positive impact on reducing fruit worm infestation, yield and percent weight losses. CONCLUSION AND RECOMMENDATIONS Trichogramma and C. carnea in combination with neem seed extract work well against tomato fruit worm and can be incorporated into and IPM strategy directed towards managing tomato fruit worm and can also be used in the biological control programme. Despite the application of these bio-control agents losses due to fruit worm were still high. More work should be done to enhance the efficacy of these bio-control agents. Cannibalism exists in C. carnea, it is impossible to rear two or more C. carnea in a test tube which is extremely laborious, costly and time consuming. Work should be done to minimize or modify this cannibalism behavior of C. carnea.

5 Sarhad J. Agric. Vol.28, No.2, REFERENCES Ayvaz, A., E. Karasu, S. Karaborklu and S. Yilmaz Dispersal ability and parsitizion performance of egg parasitoid Trichogramma evanescene Westwood (Hymenoptera: Trichogrammatidae) in field and storage conditions. Turk. J. Biol. 32: Dhingra, S., S. Walia, J. Kumar, S. Singh, G. Singh and B.S. Parmar Field efficacy of azadirachtin-a, tetrahydroazadirachtin-a, NeemAzal and endosulfan against key pests of okra (Abelmoschus esculentus) J. Pest Mgt. Sci. 64(11): Easterbrook, M. A., J.D. Fitzgerald and M.G. Solomon Suppression of aphids on strawberry by augmentative releases of larvae of the lacewing Chrysoperla carnea (Stephens). Biol. Sci. Tech. 16(9): Hussin, B. and S. Bilal Efficacy of different insecticides on tomato fruit borer Helicoverpa armigera. J. Entomol. 4(1): Inayatullah, M of tomatoes fruit worm Helicoverpa armigera (Noctuidae: Lepidoptera) using Trichogramma and Chrysoperla. 2 nd Tech. Rep. HEC Funded Project. Deptt. Entomol, Agric. Univ. Peshawar, Pakistan. pp Kevany, B.M., M.G. Taylor and H.J. Klee Fruit-specific suppression of the ethylene receptor LeETR4 results in early-ripening tomato fruit. J. Pl. Biotech. 6(3): Nabil, E. Wakeil and S. Vidal Using Chrysoperla carnea in combination with Trichogramma species for controlling Helicoverpa armigera. J. Appld. Entomol. 124(5-6): Reddy, G. V. P and M. Manjunatha Laboratory and field studies on the integrated pest management of Helicoverpa armigera in cotton, based on performance trap catch threshold level. J. Appld. Entomol. 125(4-5): Shah, A. R Impact of neem oil and neem seed extract on tomato fruit worm (H. armigera) and its natural enemies. M.Sc (Hons) Thesis, Agric. Univ. Peshawar, Pakistan. pp Shah, R Impact of the use of predators and parasitoids on population of tomato fruit worm (Helicoverpa armigera) Lepidoptera: Noctuidae at Dargai. M.Sc (Hons) Thesis. Agric. Univ. Peshawar, Pakistan. pp

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