International Journal of Zoological Investigations Vol. 3, No. 1, 15-20 (2017) ______

International Journal of Zoological Investigations

Contents available at Journals Home Page: www.ijzi.net

ISSN: 2454-3055

Parasitoid Wasps Attacking the American Serpentine Leafminer Liriomyza trifolii in Kafr EL-Shiekh, Egypt

Rowfida A. Bassiony1,2,*, Abou-attia F.A.3, Samy M. A.2, Asmhan E. Youssef3 and Ueno T.1

1 Institute of Biological Control, Faculty of Agriculture, Kyushu University, Fukuoka, Japan 2 Plant Protection Research Institute, ARC, Sakha, Kafr El-Sheikh, Egypt 3 Economic Entomology Department, Faculty of Agriculture, Kafrelsheikh University, Egypt

*Corresponding author

Received: 30th March 2017 Accepted: 25th April 2017 ______Abstract: Dipteran leafminers of the genus Liriomyza are important pests of many vegetable crops worldwide. In Egypt, broad bean yield had been severely harmed in recent years due to dipteran leafminer infestations. Biological control with is known as an effective measure to reduce leafminer damage. In the present study, we first examined the composition of species and parasitism rates of parasitoid wasps attacking L. trifolii to highlight the importance of parasitoids in regulating leafminer populations on bean fields in Kafr EL-Shiekh, Egypt. Then, the effects of plant species, weather factors and planting dates on the parasitism rate of parasitoid wasps were investigated, in order to examine how the above -mentioned factors influence the impact of the parasitoids on leafminer populations. Two parasitoid wasps Opius dissitus and Diglyphus isaea were commonly found on broad bean (faba bean) and French bean fields. These parasitoids were detected throughout the growing season of bean plants, though the parasitism rates were relatively low. Parasitism rates were significantly higher in French bean than in broad bean, suggesting host plant preference in the parasitoid species. Temperature and precipitation appeared not to affect the parasitism rates.

Key words: Biological control, host plant preference, pest management, ______

Introduction Dipteran leafminers belonging to Legumes are of special importance as staple the genus Liriomyza (Diptera: Agromyzidae) food all over the world and are cultivated in are a group of herbivores whose larvae many countries (Mounir et al., 2015). In settle in plant leaflets, feeding on mesophyll Egypt, broad bean or faba bean Vicia faba L. tissue (Johnson et al., 1983; Parrella et al., and snap bean Phaseolus vulgaris L. are 1985; Salvo and Valladares, 2007). At major bean crops; the former bean is one of present, more than 20 Liriomyza species are the oldest crops and occupies the third most of economic importance as pest important grain legumes whereas the latter (Anonymous, 2009). The American is notably a pioneer Egyptian export crop serpentine leafminer L. trifolii (Burgess) is (Nassib et al., 1991; Anil et al., 2013). one of the most major pests of many vegetables, ornamental and horticultural However, it is not known whether the plants worldwide (Spencer, 1972). Control predominant parasitoid is D. isaea or the of leafminers is thus essential to stable other parasitoid in other regions of the production of marketable vegetables (Salvo country. Parasitoid fauna of the leafminers and Valladares, 2007; Liu et al., 2009). in many region of Egypt still remains In Egypt, leafminers including L. unclear. trifolii cause economically serious damage Our previous study has shown that in and loss of many vegetables including bean Kafr EL-Shiekh Governorate, L. trifolii is a crops (Abul-Nasr and Assem, 1961; Shahein serious pest of bean plants and exploring and El-Maghraby, 1988). Such leafminers effective parasitoid agents are on strong are often difficult to control solely by demand (Rowfida et al., in prep.). Thus, the chemical (Parrella et al., 1984). aim of the present study is to determine the Instead, biological control with parasitoids species composition and parasitism rate of or integrated pest management with parasitoid wasps that attack L. trifolii on biocontrol measure can be a promising bean plants in Kafr EL-Shiekh, Egypt. The measure to manage the leafminers (Salvo present study also examines how bean and Valladares, 2007; Liu et al., 2009). More species and abiotic factors could affect the than 140 species of parasitoids have been parasitism rate of the parasitoids detected. reported to attack Liriomyza leaf-miners, Materials and Methods and such natural enemies are often the most reliable agents of mortality of Liriomyza The field study was carried out in order to leafminers (Liu et al., 2009). Several species obtain parasitoids attacking L. trifolii. For this purpose, four different fields of broad of leafminer parasitoids have been commercially mass-reared and been used bean with different planting dates were set up and field sampling was made from the against Liriomyza in greenhouses or in the field (Liu et al., 2009). earliest growing stage of bean to just before Although parasitoids are useful in harvest. Sampling was initiated when bean plants were one month old, and continued leafminer management, the species composition is greatly variable depending once per week until the beans were harvested. Thus, sampling was done on the season and region, and, hence, it is important to determine parasitoid species from15/12/2015 to 15/03/2016, from 29/12/2015 to 16/03/2016, and from that are effective in the region and/or 12/01/2016 to 22/03/2016 for each of the season concerned (Tran et al., 2007; Mujica and Kroschel, 2011; Ueno and Tran, 2015). three fields of broad bean var. Giza 3 and Also, effective species of parasitoids may one field of Snap bean var. Giza 6 from 15/12/2015 to 15/03/2016 in Kafr El- depend on crop species or variety (Martin et Sheikh Governorate. al., 2006; Musundire et al., 2012). In Alexandria, Egypt, D. isaea has recently been In each study field, four points were shown as the most predominant species and randomly selected, and 25 leaflets were positively correlated with the leafminer collected from each collection point. When population (Naema and Hegazi, 2014). collecting sample leaflets, the cross-

16 diameter method was used from each plant; Results and Discussion one leaf was picked from all different plant In the present study, leafminers in our height levels. The collection was made at a region were identified as Liriomyza trifolii. weekly basis. Collected leaflets were Two parasitoid species were obtained; they returned to the laboratory, and the numbers were identified as Opius dissitus and of leafminer larvae were examined. The Diglyphus isaea, and were collected from sampled leaflets from each replicate were all broad bean and French bean fields. The placed separately into Petri dishes (10 cm diam.), and were kept in a growth chamber at 25 ± 2 ºC, 70 ± 5% RH, with a photoperiod of 14:10 h (L: D) for the emergence of adult leafminer and parasitoids. The samples in each Petri dish were checked every day for leafminer pupae and larval parasitoids. When leafminer pupae were found, they were carefully separated from the leaflets, and transferred into a new Petri dish (10 cm diam.) to record their Figure 1: The effect of planting date on Opius dissitus origin. The number of pupae from each parasitism rate on both bean crops replicate was recorded. The pupae were mean parasitism rates for each field were; placed in the same growth chamber for 6.4% vs 2.0% for broad bean field 1, 4.7% vs further development into adults. The 2.1% for field 2, 8.7% vs 1.6% for field 3, while 18.3% vs 3.6% for French bean numbers of emerged adults of L. trifolii and field (shown as Opius vs Diglyphus). The of its parasitoids, from the pupae were recorded and were identified. Likewise, larval parasitoids emerging were collected and were identified. The Liriomyza spp. were identified using external morphology and male aedegous (Spencer, 1972). Also, the parasitoids were identified according to the keys given by Chao-Dong and Da-Wei (2001), Gates et al. (2002) and Yefremova et al. (2011). Parasitism rates were calculated as Figure 2: The effect of planting date on Diglyphus isaea parasitism rate on both bean crops the number of parasitoids/total number of parasitism rates were then statistically leafminer larvae + parasitoids (for analyzed after arcsine square root parasitoids emerging from leafminer larvae) transformation with a paired t-test with the and the number of parasitoids/number of four fields as x-grouping. The analysis leafminer pupae (for parasitoids emerging from host pupae). Data were analyzed using revealed that the mean parasitism rates of Opius parasitoids were significantly higher JMP software (SAS Institute). than those of Diglyphus (n = 187, F = 4.80,

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P = 0.003) while the parasitism rates O. dissitus is a solitary koinobiont differed among the fields (F = 7.68, P< parasitoid wasp attacking Liriomyza spp., 0.0001). Therefore, Opius parasitoids were and is reported from Africa, and Asia more abundant than Diglyphus parasitoids. (Neuenschwander, 1987; Bordat, 1995). Both of the two parasitoids were Like other members of koinobiont detected in all fields throughout the growing parasitoids of leafminers, the female season (Figs. 1 and 2). Parasitism rates of O. parasitoid attacks the larval stage of host while the offspring parasitoid emerges from dissitus were much higher in French bean field than in broad bean fields (Fig. 1). Opius the host pupae (Bordat, 1995). This species parasitoids also showed much greater may be an effective biocontrol agent in fluctuation in parasitism rates. However, rather dried environments; in Senegal of those of D. isaea were uniformly low in all Africa, only O. dissitus is known to be relatively abundant among 9 species of study fields (Fig. 2). Unfortunately, we had only one French bean field in the present parasitoids released to control leafminers study, and we do not know whether French (Neuenschwander, 1987). bean is a preferred plant for Opius The other parasitoid species detected parasitoids. Plant preference in leafminer in our study was D. isaea. This parasitoid is paprasitoids is not well understood, and this an idiobiont larval parasitoid attacking the will be a subject in our future study. larval stage of leafminer hosts, and is known To examine whether precipitation as an effective agent in controlling leafminer and temperature could affect the parasitism populations. D. isaea is one of several rates, statistical models (least square model) parasitoids that are commercially available were constructed with three independent for leafminer control (Liu et al., 2009). Although this parasitoid is often found variables (precipitation, temperature and abundant and this also holds true in sampling date) and a random effect (field). For Diglyphus, any of factors were not Alexandria, Egypt (Neama and Hegazi, significant (P> 0.05) while sampling date 2014), parasitism by D. isaea was uniformly only was found significant for low throughout the season in our study. Opiusparasitism (P = 0.008). In fact, Opius O. dissitus is the most abundant parasitism was generally higher in earlier parasitoid detected throughout the growing growing seasons (Fig. 1). season. Therefore, this parasitoid can be the most promising biocontrol agent in our In our study, we found two major parasitoids attacking the pest leafminer, region in Egypt. Future studies will focus on i.e., O. dissitus and D. isaea. In the previous how we can effectively incorporate O. two independent studies made in Egypt, dissitus into pest management strategy in both Opius and Diglyphus species have bean fields. Also, in the present study, parasitism been found major among the parasitoids of Liriomyza pests (Shahein and El- rates were generally low for both the Maghraby, 1988; Naema and Hegazi, species. Weather or climate conditions may also be an explanation; however, 2014). temperature and precipitation were not

18 significant factors influencing parasitism in Hemiptarsenus varicornis (Hymenoptera: the present study. Because chemical ). J. Fac. Agric., Kyushu Univ. 52: 57-61. pesticide applications were frequently made Ho THG and Ueno T. (2011) The effects of honey as a dietary supplement on the survivorship and the surrounding vegetable fields, nutrition-storing capacity of Hemiptarsenus contamination of pesticides may harm the varicornis (Hymenoptera: Eulophidae), a parasitoid of Liriomyza (Diptera: Agromyzidae) leafminer parasitoids (Tran and Ueno, leafminers. Int. J. Trop. Sci. 78: 78-84. 2012), weakening their activity in our fields. Johnson MW, Welter SC, Toscano NC, Ting IP and Alternatively, lack or shortage of Trumble JT. (1983) Reduction of leaflet appropriate food sources like nectar may be photosynthesis rates by mining activity of the reason; it is known that leafminer (Diptera: Agromyzidae). J. Econ. Entomol. 76: 1061-1063. parasitoids can enhance their performance Liu TX, Kang L, Heinz KM and Trumble J. (2009) and reproductive success when Biological control of Liriomyza leafminers: carbohydrate sources are available (Ho and progress and perspective. CAB Reviews: Ueno, 2007, 2011). In any case, future Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources 4: 1-16. studies should also address how we can Martin V, Valladares G and Salvo A. (2006) A improve the activity of leafminer parasitoids tritrophic analysis of host preference and in the field. performance in a polyphagous leafminer. Entomol. Exp. Appl. 121: 93-190. References Mounir AM, Abo El-Yazid A, Orabi IOA, Zahran AA and Abul-Nasr S and Assem AH. (1961) A , El-Oksh II. (2015) Effect of sowing date, gamma Liriomyza bryoniae (Kalt.), attacking irradiation and intracultivar differences on cucurbitaceous plants in Egypt. Bull. Entomol. Soc. growth, pod characteristics and some endogenous Egypt. 45: 401-403. plant growth regulators in snap Beans. World J. Anil KS, Bharati RC, Naresh CM and Anitha P. (2013) Agric. Sci. 11: 380-390. An assessment of Broad bean (Vicia faba L.) Mujica N and Kroschel J. (2011) Leafminer current status and future prospect. African J. (Diptera: Agromyzidae) occurrence, distribution, Agric. Res. 8: 6634-6641. and parasitoid associations in field and vegetable Anonymous. (2009) List of pests recommended for crops along the Peruvian coast. Environ. Entomol. regulation as quarantine pests. EPPO. 40: 217-230. http://www.eppo.org/ QUARANTINE/listA2.htm. Musundire R, Chabi-Olaye A, Salifu D and Krüger K. Bordat D. (1995) Morphometric, biological and (2012) Host plant-related parasitism and host behavioral differences between Hemiptarsenus feeding activities of Diglyphus isaea varicornis (Hym., Eulophidae) and Opius dissitus (Hymenoptera: Eulophidae) on Liriomyza (Hym., ) parasitoids of Liriomyza huidobrensis, Liriomyza sativae, and Liriomyza trifolii (Dipt., Agromyzidae). J. Appl. Entomol. 119: trifolii (Diptera: Agromyzidae). J. Econ. Entomol. 423-427. 105: 161-168. Chao-Dong Z and Da-Wei H. (2001) A taxonomic Neama AA and Hegazi EM. (2014). Parasitoids of study of Eulophidae from Zhejiang, China leafminers Liriomyza spp. (Diptera: Agromyzidae) (Hymenoptera: Chalcidoidoidea). Acta attacking Faba bean in Alexandria, Egypt. Zootaxonomica Sinica 26: 533-547. Egyptian J. Biol. Pest Cont. 24: 301-305. Gates MW, Hearaty JM, Schauff ME, Wagner DL, Nassib AM, Khalil SA, Hussein AHA. (1991) Faba bean Whitfield JB and Wahl DB. (2002) Survey of the production and consumption in Egypt. In: parasitic Hymenoptera on leafminers in “Present Status and Future Prospects of Faba . J. Hym. Res. 11: 213-270. Bean Production and Improvement in the Mediterranean Countries”, CIHEAM, Zaragoza, Ho THG and Ueno T. (2007) Improving parasitoid Spain, pp 127-131. performance by improving adult food quality: a case study for the leafminer parasitoid Neuenschwander P. (1987) Introduction of exotic parasitic wasps for the control of Liriomyza trifolii

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(Diptera: Agromyzidae) in Senegal. Tropical Pest Tran DH, Tran TTA, Mai LP, Ueno T and Takagi M. Manag. 33: 290-297. (2007) Seasonal abundance of Liriomyza sativae (Diptera: Agromyzidae) and its parasitoids on Parrella, M, Keil C and Morse J. (1984) vegetables in southern Vietnam. J. Fac. Agric., resistance in Liriomyza trifolii. Calif. Agric. 38: 22- Kyushu Univ. 52: 49-55. 23. Tran DH and Ueno T. (2012) Toxicity of insecticides Parrella MP, Jones VP, Youngman RR and Lebeck LM. to Neochrysocharisokazakii, a parasitoid of (1985) Effect of leaf mining and leaf’s tippling of Liriomyza leafminers on vegetables. J. Fac. Agric., Liriomyza spp. on photosynthetic rates of Kyushu Univ. 57: 127-131. . Ann. Entomol. Soc. Am. 78: 90- 93. Ueno T and Tran DH. (2015) Neochrysocharis okazakii (Hymenoptera: Eulophidae) as a major Salvo A and Valladares GR. (2007) Leafminer of stone leek leafminer Liriomyza parasitoids and pest management. Cien. Inv. Agric. chinensis (Diptera: Agromyzidae) in Central 34: 167-185. Vietnam. Psyche article ID 179560. Shahein A and El-Maghraby MMA. (1988) Studies on Yefremova ZH, Civelek S, Boyadzhiyev PDO and Eskin the hymenopterous parasitoids of Liriomyza A. (2011) A review of Turkish Diglyphus Walker trifolii (Burgess) (Dipt., Agromyzidae) on broad (Hymenoptera: Eulophidae), with description of a beans in Egypt. J. App. Entomol. 106: 377-380. new species. Ann. Soc. Entomol. Fr. (n.s.) 47: 273- Spencer KA. (1972) Diptera. Agromyzidae, 269. Handbooks for the Identification of British ,

Roy. Entomol. Soc., London, London, X(5g): 1-136.

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