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2016 Host Utilization of tbe Endoparasitoid, Cotesia glomerata L. (Hymenoptera: Braconidae) in Different Instars of Pieris brassicae L. (: Pieridae) M. I. Ullah University of Sargodha, [email protected]

Muhammad Arshad University of Sargodha

S. Ali University of Sargodha

Yasir Iftikhar University of Sargodha

Jaime Molina-Ochoa Universidad de Colima, [email protected]

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Ullah, M. I.; Arshad, Muhammad; Ali, S.; Iftikhar, Yasir; Molina-Ochoa, Jaime; and Foster, John E., "Host Utilization of tbe Endoparasitoid, Cotesia glomerata L. (Hymenoptera: Braconidae) in Different Instars of Pieris brassicae L. (Lepidoptera: Pieridae)" (2016). Faculty Publications: Department of Entomology. 549. http://digitalcommons.unl.edu/entomologyfacpub/549

This Article is brought to you for free and open access by the Entomology, Department of at DigitalCommons@University of Nebraska - Lincoln. It has been accepted for inclusion in Faculty Publications: Department of Entomology by an authorized administrator of DigitalCommons@University of Nebraska - Lincoln. Authors M. I. Ullah, Muhammad Arshad, S. Ali, Yasir Iftikhar, Jaime Molina-Ochoa, and John E. Foster

This article is available at DigitalCommons@University of Nebraska - Lincoln: http://digitalcommons.unl.edu/entomologyfacpub/ 549 Egyptian Journal of Biological Pest Control, 26(3), 2016, 625-629 Host Utilization of tbe Endoparasitoid, Cotesia glomerataL. (Hymenoptera: Braconidae) in Different Instars of Pieris brassicae L. (Lepidoptera: Pieridae)

UHabl, M. I.; M. Arsbad1; S. AlP; Y. Iftikbar2; J. Molina-Ocboa3,4 and J. E. Foster4 'Dept. of Entomology, University of Sargodha, Sargodha 40100 Pakistan, [email protected] 2Dept. of Plant Pathology, University ofSargodha, Sargodha 40100 Pakistan 3Universidad de Colima, Coordinaci6n General de Investigaci6n Cientifica, Centro Universitario de Investigaci6ny Desarrollo Agropecuario, Facultad de Medicina Veterinaria y Zootecnia, Crucero de Tecoman, autopista Colima­ Manzanillo, Km. 40, Tecoman, Colima 28930, Mexico. 4Department of Entomology, University of Nebraska-Lincoln, Lincoln, NE 68583 US (Received: June 20, 2016 and Accepted: July 27, 2016)

ABSTRACT

Pieris brassicae L. (Lepidoptera: Pieridae) is a destructive pest of brassicaceous plants around the world. Biological control of this pest has attained much attention being environment friendly. Parasitism rate and growth of the endoparasitoid , Cotesia giomerata L. (Hymenoptera: Braconidae) was studied on first (L I) and second (L2) larval instars of P. brassicae. Adult fresh mass and developmental time ofthe parasitoid was higher when it parasitized Ll than L2 of host larvae. Thus, host instars contributed significant variations to parasitoids adult mass accumulation during their development. Parasitized host caterpillars in L2 grew significantly larger than in L I due to less parasitism rate. Similarly, the development of C. glomerata was faster in Lias compared to L2. Larval mortality of the host and parasitoid was also significantly higher in case of Ll. It is indicated from the study that C. glomerata has a promising role managing P. brassicae larvae at early stages.

Key words: Cotesia glomerata, Pieris brassicae, Parasitism rate, Developmental time, Fresh mass.

INTRODUCTION Similarly, C. glomerata can impose (100%) losses of its host larvae (Spieth and Schwarzer, 200). Many brassicaceous plants and their hybrids are the most important commercial crops worldwide The objective of this study was to observe the host (Hopkins et al., 2009). Vegetables belonging utilization and development of C. glomerata at two to family Brassicaceae are important source for different instars of P. brassicae along with adult human diet, consumed by a large number of people parasitoid mass, developmental time and pupal in the world, especially in Asia aDd Europe (Cartea survival. The growth and development of parasitized et al., 2011). herbivores like Pieris brassicae and un-parasitized caterpillars were also compared to L. (Lepidoptera: Pieridae) are the major pests that evaluate the effects of parasitism. cause significant losses of economic importance (Tanaka et al., 2007). The caterpillars are herbivores MATERIALS AND METHODS feed only on the order Capparales plants that produce a toxin named 'glucosinolates' (OS) (Harvey et al., Insect culture 2010). P. brassicae caterpillars and C. glomerata cocoons were collected from the cabbage field near For safer environment and to avoid the issues of University College of Agriculture, University of resistance against synthetic chemicals, biological Sargodha, Pakistan. Host and parasitoids were reared control of insect pests is considered a suitable method at 25±I°C and 65±5 RH in laboratory at the to keep the insect damages below threshold levels Department of Entomology. Caterpillars of P. (Metspalu et al., 2003). Traditionally, biological brassicae were maintained on Brassica oleracea var. control of P. brassicae depends on entomopathogenic capitata and C. glomerata was reared on 1st instar of viruses, bacteria, fungi, parasitoids and predators P. brassicae caterpillars to maintain its culture. Adult (van Driesche et al., 2003). Braconid parasitoid wasps were fed on 10% sugar solution given on species of CQJesia parasitize the pierids cotton wool and allowed for egg laying. Afterwards, effectively. Among these, Cotesia glomerata L. parasitoid cocoons were collected and transferred into (Hymenoptera: Braconidae) is the most important new cages for further experiments. parasitoid of P. brassicae. It is a cosmopolitan endoparasitoid of many lepidopteran larvae, Growth line of parasitized and un-parasitized especially P. brassicae. Typically, it produces a larvae of P. brassicae single brood of cocoons in cluster form with an Thirty P. brassicae 1st (Ll) and 2nd instar (L2) average of 25-30 wasps (Ou et al., 2003). This larvae were collected from the insect culture and koinobiont parasitoid attacks its hosts and kills them placed on central leaves of cabbage plants in at 4th or 5th instars (Harvey et al., 1994). ventilated glass cages (45 x 43 x 43cm) at 626

temperature of 25± loe. One plant of cabbage was attained 396.94mg and 401.2lmg of 20 day old kept in each glass for each instar. After 2 hours of larvae, respectively. Overall larval body-mass of L2 releasing the larvae, 10 males and 10 females adult was observed higher when compared to Ll. The parasitoids were released in each cage. In one results showed that larvae of C. glomerata exerted experiment, wasps were removed from cages after 24 significant effect on the growth of host larvae as hrs and in another experiment wasps were removed compared to un-parasitized larvae. The parasitized L2 after 48 hrs to check parasitism rate at different larvae attained the maximum body-mass of 42.31 mg, durations. After cocoon formation, parasitoid when larvae were 18 days old. Overall, larval body­ cocoons were collected from the cages and placed in mass ofLl larvae did not increase 20mg (Fig. 1). It is Petri plates until adult emergence. Male and female obvious that the nutritional requirements of adult wasps were separated and weighed to measure koinobiont parasitoids are highly intensive to cause their fresh masses. Number of days was recorded heavy weight losses in their host larvae because of from parasitism initiation to adult emergence to food shortage for them (Harvey et al., 2004 and calculate the developmental time from the host Mironids and Savoopoulou-Soultani, 2009). caterpillars. Host larval mortality, total number of Obviously, C. glomerata utilized its host larvae for its parasitoid cocoons and number of adult emergence development and caused significant host mortality. from cocoons were also counted. The experiments Growth and development of host larvae during were replicated three times. parasitism is a very interesting phenomenon that enables the stabilization of stored food in body of Measurement of parasitoids fate parasitoid because increased brood size require more Twenty Ll and L2 caterpillars of P. brassicae food (Harvey et a/., 20 10). were placed individually in glass vials. Ten pairs of wasp adults (male and female) were released in each Development of C. glomerata glass vial (Harvey et al., 2010). Ovipositor insertion Adult wasp development was faster when into host larval body was visualized to confirm the parasitized LI host caterpillars as compared to L2. parasitism and parasitized larvae were transferred The growth and development of parasitoid was more into clean glass cages. Fresh leaves of cabbage were when developed in L I larvae as compared to L2 provided to parasitized larvae. Data of host larval larvae of the host. It is due to fact that amount of death, parasitoid pupation, parasitoid cocoons that resources was higher in Ll instars for utilization by failed to adult formation and total number of parasitized larvae as compared to L2 instar. parasitoid ad\llt emergence were recorded. The experiment was replicated three ti'!les. Adult fresh mass of C. glomerata was significantly affected (F = 45.5. P

_ Ll (unparasitlzed) l2 (un parasl t ized) _____ L~ (Pa rasitized) -4t-l2 (parasit ized) 450 50 w 400 45 ~ 350 40 35 .§: 300 30 :11 250 25 E 200 20 I 1 5 0 15 11 100 10 :z: 50 5 o . .... - ,:::! , .. • , ~ . , . :~-" o 2 4 6 8 10 12 14 16 18 20 22 24 26 Hoe. ege (days)

Fig. (I): qt:()~~~li~c::__ ()fpllI1lsi.tized al1~!l.I1~Pll~ll s i. tizc::~ P.~rassicae cate1l>.illars.

_ l1 (unparasJ t lzed) ,.,.. L2 (unparasft i zed) -"-Ll. (pa r .sitlze d) ...... - L2 (parasltized)

~SO so ... 400 I 4 5 ~ 350 40 3S l300 3 0 I:: 2S 2 0 I '1S0 15 1,·00 .10 50 5 0 2 4 6 H 10 .1_2 1. 4 1. 6 .1.8 2 () 22 2.4 26 Hos. age (day.)

Fig. (2): Fresh mass of adult male and female C. glomerata developed from P. Brassicae L 1 and L2 caterpillars. Numbers in brackets shows the total number of individuals.

_ I ~ l . (uopau ..... siti2f'!'.d) t.2: (\Jnpara $ftl~4'!!d ) -+-l.1. (Par.uitiz,t!!'d) ____ t.2 (p2llr ... sltl7.~ d) 4 50 5 0 w 400 45 :! 350 40 35 .§:300 1 30 250 I 'I :IS e 200 20 ~ 150 .!! 1 .1.5 • I 100 10 50 I 5 0 ~ .. a:. o 2 4 6 8 10 18 20 22 2 4 26

Fig. (3): Development time of male and female C. glomerata in P. brassicae Ll and L2 caterpillars after different time intervals.

_ L1 (unparasltlzed) _. L2 (unpanlSltized) _Ll (Parasit ized) __ililt__ L2 (parasitize d) 450 SO ... 400 45 ." +I 350 40 35 I 300 3 0 := 250 ... 25 E 200 20 I 150 15 100 10 1 SO 5 o o 2 4 6 8 10 12 14 16 18 2 0 22 24 2 6 Host age (day.)

Fig. (4): Fate of parasitized Ll and L2 caterpillars of P. brassicae when parasitized by C. glomerata females for different time intervals. 628

_ Ll (un parasitized) L2 (unparasltlzed) _L1 (Parasitized) -W-- L2 (parasitized) 50 :I 45 40 3S 30 iiliIII 250 1.. .. I 2S E 200 1 20 150 ..!!~ i, 15 ~ 100 1

:z: T A.•. A···· ····· ",M..... ,.... ~'''.·-··r··· """""'····1 ~· S: t -= , -~ ., .: " I:., I~ . 1 2 4 6 8 10 12 14 16 18 20 22 24 26 Host age (days)

Fig. (5): Percentage of male and female C. glomerata adults emerged from cocoons developed from P. brassicae parasitized caterpillars (Ll and L2) at different time intervals. determine the size and growth of parasitoids (Hu and ACKNOWLEDGEMENT Vinson, 2000). The authors are thankful to Prof. Dr. Muhammad Fate of parasitized larvae, emergence % and sex Afzal, Dean Faculty of Agriculture, University of ratio Sargodha, Pakistan for initial review of this Host larval mortality was the highest in Ll manuscript. caterpillars as compared to L2. Furthermore, larval mortality was higher when host larvae were REFERENCES parasitized for 48 hrs than those parasitized for 24 hrs but with insignificant difference. Percentage of host Cartea, M.E., M. Francisco, P. Soengas and P. pupation was observed less (4.44%) in Ll larvae as Velasco 2011. Phenolic Compounds in Brassica compared to L2 ones (10%) when parasitized for Vegetables. Molecules. 16:251-280. 24 hrs (Fig. 4). Percentage of parasitoid adult Gu, H., Q. Wang and S. Dom 2003. Super-parasitism emergence from cocoons was also greater in L 1 host in Cotesia glomerata: response of host and larvae as compared to L2. Interestingly, percentage consequences for parasitoids. Ecol. Entomol., of male adult parasitoids was higher than the females 28:422-431. in both host larval instars used for parasitism assay Harvey, J.A., E. H. Poelman and R. Gols 2010. (Fig. 5). Contribution of host larval instars was Development and host utilization in Hyposoter also significant towards variations in parasitoid eben in us (Hymenoptera: Ichneumonidae), a cocoon formation after successful parasitization of solitary endo-parasitoid of Pieris rapae and P. P. brassicae. Percentage of parasitoid cocoon brassicae caterpillars (Lepidoptera: Pieridae). formation was much higher in Ll caterpillars as BioI. Control, 53:312-318. compared to L2. This fact is supported by Harvey, J.A., T. M. Bezemer, J. A. Elzinga and M. R. the variations in host mortality, host development Strand . 2004. Development of the solitary and parasitoid development in both host larval endoparasitoid Microplitis demolitor: host quality instars. Furthermore, percentage of adult emergence does not increase with host age and size. Ecol. from cocoons was greatly depended upon the host Entomol., 29:35-43. larval instars. It might be possible that parasitoid Harvey, J.A ., 1. F. Harvey and D. J. Thompson 1994. larvae show better physiological integration when Flexible larval growth allows use of a range of parasitize initial instar of the host larvae as compared host sizes by a parasitoid wasp. Ecol. 75 :1420- to later ones (Wang and Liu. 2002). One possible 1428. explanation is also that later instars larvae of host had Harvey, J.A.,M. A. Jervis,G. J. Z. Gols,N. Jiang and strong immune response against parasitoids as L. E. M. Vet 1999. Development of the parasitoid, compared to early instars larvae (Harvey et al., 1999 Cotesiarubecula (Hymenoptera: Braconidae) in and 2010). Pierisrapae and P. brassicae (Lepidoptera: PyraLidae): evidence for host regulation. J. Insect In conclusion, C. glomerata is a successful PIJysio., 45 : 173-182. biological control agent that can be used as a principal Hopkins, R. J., N. M. van Dam and J. J. A. van Loon parasitoid against P. brassicae but its efficacy 2009. Role of glucosinolates in insect- plant depends greatly upon larval instar. relationships and multitrophic interactions. Annu. 629

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