BIHAREAN BIOLOGIST 11 (1): 15-19 ©Biharean Biologist, Oradea, Romania, 2017 Article No.: e161201 http://biozoojournals.ro/bihbiol/index.html

An outline on biology and behavior of Bracon variator Nees, 1812 (Hym.: ), an ectoparasitoid of Cydia johanssoni Aarvik & Karsholt, 1993 (Lep.: ) from Iran

Jabbar VALIPOUR, Hassan Ali VAHEDI* and Abbas Ali ZAMANI

Department of Plant Protection, Agricultural College, Razi University, Kermanshah, Iran. *Corresponding author, H.A. Vahedi, E-mail: [email protected]

Received: 28. May 2015 / Accepted: 21. December 2015 / Available online: 10. January 2016 / Printed: June 2017

Abstract. Bracon variator Nees, is a larval ectoparasitoid of Cydia johanssoni Aarvik & Karsholt, a major seed feeder of Stinking bean trefoil, Anagyris foetida L. (Leguminosae) at forest habitats in Gilan-e Gharbe, Kermanshah, Western parts of Iran. This investigation was conducted during spring 2014, in order to determine the most important of pest larvae, the rate of parasitism and to study some biological aspects. The ectoparsitoid prefers 3rd to 5th instar larvae. Host development was arrested immediately upon parasitism. Each adult female laid three eggs close to host larvae, inside seed case. It had two generations per year. Both aestivation and hibernation of the parasitoid occur in the form of prepupa in cocoon inside infested seeds. Adult activity of first and second generation was observed in the begining and ending of May, respectively. The average of parasitism rate of second generation was calculated 18.77±3.80. Cydia johanssoni larvae, as a host of Bracon variator, has been recorded for the first time.

Key words: stinking bean trefoil, Anagyris foetida, Cydia johanssoni, Bracon variator, ectoparasitoid, Kermanshah, biological control.

Introduction strobilella L. (attacking spruce seed, Picea spp.), C. latiferreana Walsingham (attacking oak fruits) and C. caryana Fitch The Braconidae constitutes one of the most species-rich hy- (hickory shuckworm, an important pest of pecan) (Oboyski menopterous families, with 48 subfamilies, more than 1050 2011). On the other hand, C. succedana Denis & Schiffermul- genera and about 17,600 species described worldwide. The ler is beneficial because it provides some biological control family has two major lineages: (a) the cyclostome braconids, of gorse, Ulex europaeus (Leguminosae) which is a serious most of which are idiobiont ectoparasitoids of concealed weed in some areas of the temperate Atlantic coast of and Coleoptera larvae although many are Europe and the British Isles including Ireland. It has become koinobiont endoparasitoids of Diptera and and naturalized elsewhere in Europe, North Africa and the Mid- (b) the noncyclostome braconids which are all endoparasi- dle East (Sixtus 2004, Sixtus et al. 2007, Hill et al. 2008). toids and are generally koinobionts, typically attacking the There are various ecological factors involved in the early instars of their hosts (Shaw & Huddleston 1991, La- search and discovery of concealed hosts by braconid species. chaud & Gabriela 2012). Braconids therefore vary greatly in Some species tend to be specialized to a particular host; for their biology (Yu et al. 2012, Quicke & Van Achterberg 1990, example, Hyssopus pallidus Askew (: Braconi- Sharkey 1993). dae), a larval parasitoid of the codling moth C. pomonella L., Bracon variator is a highly oligophagous gregarious ecto- is able to enter infested apples through the calyx or the tun- parasitoid that attacks the larvae of a wide range of , nel made by the host larvae and parasitize them (Mattiacci et such as Lepidoptera. For example, B. variator paralyses the al. 1999). larvae of Hadena bicruris Hufnagel (Lepidoptera: Noctuidae) The city of Gilan-e Gharb is located in west of Kerman- before depositing on average 3 eggs on fourth- or fifth-instar shah province at an altitude of 800-2500 meters and has the larvae (rarely third-instar) in the Netherlands (Elzinga 2005, typical continentally-influenced Mediterranean climate of Elzinga et al. 2005). Like Cydia johanssoni, Hadena bicruris is a western Iran, with a mean annual precipitation of about 300- seed specialist, feeding on Silene latifolia Poir. (Caryophylla- 320 mm in the last 5 years (Based on meteorological statis- ceae) seeds (Jelmer et al. 2007). tics, Kermanshah, Iran). The dry season starts in May and Several orders, for example Lepidoptera, attacks continues until mid-October, lasting about 5.5 months. Stink- the seeds before they are shed by the parent plant (Ramirez ing bean trefoil, Anagyris foetida L. (Leguminosae) is a shrub & Traveset 2010, Zhang et al. 1997, Andersen 1988). Cydia found in the alkaline soil habitat, hot dry summer Mediter- Hübner 1825 (Lep.: Tortricidae) is a genus with 231 species ranean climate and well-drained; with light woodland or no and subspecies and is found worldwide except Antarctica; shade. Leaf, stem and root extract of this plant is cathartic larvae of many Cydia species feed within the fruits, seeds and vermifuge in medicinal volumes (Zargari 1997). Its and flowers, or under the bark or within the fleshy stems of wood is used in producing a variety of baskets (Akan 2013). at least 65 host-plant species including angiosperms and The studied region covers an area of about 930 hectares. conifers (Oboyski 2011). Many Cydia species are economi- The height of each stinking bean trefoil shrub varies between cally important to agriculture and forestry ecosystems. For 1-3 m. Unusually, growth begins in early November, so that example, C. pomonella L. is a major economic pest of apples it is green during the winter, senescing in mid-June (per- orchards, often causing 25-50% losses (Beşleagă et al. 2013). sonal observations). The most common tree in the area is the The larvae of C. fagiglandana feed inside the nuts of beech, oak, Quercus persica L., with some wild pistachio, Pistacia Fagus sylvatica (Leguminosae) causing high seed mortality in khinjuk Stocks, Prunus serotina Ehrh., Crataegus azarolus L. South Sweden beech forests (Nielsen 1977, Nilsson & Wastl- and Amygdalus scoparia L.. jung 1987); other econonomically important species are C. The first-instar larvae of Cydia johanssoni Aarvik and nigricana Fabricius (pea moth, attacking legume crops), C. Karsholt (Lep.: Tortricidae) penetrate into the seed pod in 16 J. Valipour et al. late April, usually killing the seed (Valipour et al. 2015). Results Morphologically, C, johanssoni is superficially similar to C. blackmoreana Walsingham, which is known from Morocco, This is first record of B. variator from Kermanshah, Iran and Syria and Iraq (Razowski 2003); but C. johannsoni differs is the first report of this species as a parasitoid of C. johans- from C. blackmoreana in having broader forewings, darker soni larvae. hindwings and a different pattern of markings on the fore- Description of female: Body 4 mm long; head orange wing (Aarvik & Karsholt 1993); there are also some differ- with black spots; ratio of head width to its height 6:5; ovi- ences in the female genitalia (Trematerra & Aarvik 1998). positor 2 mm long, black and longer than metasoma; an- Here, we present some biological and behavioral obser- tenna 2 mm long, with 23-25 flagellomeres; forewings 3.5 vations on the parasitoid of C. johanssoni attacking Anagyris mm long (Fig. 2); mandibles yellow, with apical part brown; foetida in Gilan-e Gharbe, Iran, in order to consider possible mesosoma orange, metanotum black and ratio of mesosoma integrated pest management of this moth. length to its width 19:9; propodeum black, without carina; wings brown with dark brown veins; length of 3-SR vein of forewing 1.6 times length of r vein. Materials and methods

This research was performed in Gilan-e Gharbe, Kermanshah, Iran (Fig. 1). During the study, infested pods of A. foetida were opened and the parasitized instars of C. johanssoni were collected. In order to study the biology and behavior of the parasitoid on parasitized lar- vae, infested seed capsules were kept in transparent cylindrical plas- tic containers, each 10 cm in diameter and 7 cm height, each with mesh openings, under natural laboratory conditions. Adult parasi- toid were collected in natural environment by an aspirator and kept in 75% ethyl-alcohol sent to Dr. Corenlis van Achterberg Figure 2. Adult female of (Department of Terrestrial Zoology, Naturalis Biodiversity Center, B. variator, with long ovi- The Netherlands) for identification. Voucher specimens were depos- positor. ited in laboratory of entomology, agricultural college of Razi Univer- sity, Kermanshah province.

Biology: First generation adult appeared in the first ten days of May, with a second generation appear- ing about a fortnight later but adult wasps were most abun- dant in early June. Adult activity was greatest during the hottest part of the day (12.00-14.00 hrs). The parasitoid popu- lation was greatest on infested pods with C. johanssoni larvae (Fig. 3).

Figure 1. Map showing the location of A. foetida forest in Gilan-e Gharbe, Kermanshah, Western parts of Iran.

In order to determine parasitism rate of second generation of this parasitoid, (because the time of first generation had been ex- pired), during first third of June, at least 100 infested seed pods by of C. johanssoni were collected daily and the number of parasi- tized larvae was counted and parasitism rate was calculated. The parasitoid was visible on the outside of the caterpillars. In order to observe parasitism behavior, several live female were collected along with pods containing unparasitized lar- vae and transported to laboratory. One adult female wasp was re- leased into a petri dish with a infested pod and oviposition observed. Figure 3. B. variator: Adult female activity (oviposition and feeding) A digital camera, model Canon, PowerShot S3 IS and a digital sony during early June, with high density on infested pod of stinking EXwave HAD camera connected to a stereomicroscope, model Blue bean trefoil. Light, were used to take video films and photograps of the behav- iour. Immediately after oviposition, the seed capsules were dissected, Oviposition began after a lengthy search period. The fe- under a stereomicroscope to observe the number of parasitoid eggs male wasp first inspected the drilling location into the pod, laid. using her antennae by tapping the pod. She then moved so that her mesothorasic legs straddled the drilling position,

An outline on biology and behavior of Bracon variator, an ectoparasitoid of Cydia johanssoni from Iran 17

The oviposition (Fig. 4 botton). is not an easy task. The tip of the ovipositor almost always gets stuck to tiny irregu- larities of the pod surface. When she looses her balance, re- starts oviposition from the beginning. since the exact point of drilling is crucial and must be recalculated for accuracy. The eggs were white and bacilli-form, with a diameter approxi- mately equal to the tip of a lab needle and eggs were ovi- posited directly onto the host larvae (Fig. 4 above). The in- cubation period is 18 hours at 33 °C and relative humidity of 14 percent. Upon hatching, the parasitoid larva perforated the caterpillar’s cuticle by its mandibles and fed on exter- nally the body hemolymph for 4-5 days. Upon seed matura- tion, the parasitoid maturated and larva secreted a webbed cocoon within the seed capsule and then pupated. pupal pe- riod of the first generation lasted about 10 days. Adults of second generation left the seed pod in late May via a hole made by adult. Adults lived as free-living adults for up to 3- 4 days until they mated and oviposited. When the larvae of C. johanssoni were paralysed, their movement was reduced. As a result of feeding by the parasitoid larvae, the moth

Figure 4. The bacilli-form eggs of B. variator oviposited onto the larva was weakened and eventually was reduced to the head host larvae (above) and adult oviposation into pod (bottom) capsule and body cuticle (Fig. 5B, see arrow). The free living adult wasps fed on semiliquid resin of pods produced by the 1st larval instar of C. johanssoni when it penetrates in the pod (Fig. 5A). Each pod of A. foetida con- tains 1-12 seeds (usually 6-7), and sometimes whole seeds of a pod are infested by caterpillars; Each seed often is fed by one caterpillar. The parasitoid aestivation and hibernation was as a prepupa in a cocoon inside infested seeds. There are 2 overlapping generations during a year in natural condi- tions (first generation is complete and the second is incom- plete), because of the diapause of the parasitoid larvae go into at the end of the second generation. Additionaly, para- sitism activity of B. variator was also observed on an un- known pyralid seed feeder (Lepidoptera: Pyralidae), this was the only other host record for B. variator. The unknown

Figure 5. .Infested pod with black spot (symptom points) and se- pyralid larva was green in color with large body, larger than miliquid resin: Adult parasitoide feeding on semiliquid resin of C. johanssoni larva and had a very low population in the pod (A) and developed larvae of B. variator inside the seed ( B) study area (Fig. 6). The average percentage of parasitism by the second gen- eration of B. variator on C. johanssoni larvae was 18.77± 3.80 (Table 1).

Table 1. The average percentage of parasitism by the second genera- tion of B. variator on C. johanssoni larvae, 2014.

Date Total number Number of parasitized Parasitism of C. johanssoni larvae of C. johanssoni rate (%) Larvae by B. variator 1 June 2014 120 9 7.5 2June 2014 105 7.5 7.14 3 June 2014 120 9 7.5

4 June 2014 120 7 5.83 Figure 6. B. variator: parasitoid larvae feeding upon Pyralid seed 5 June 2014 135 22 16.29 feeder larva. 6 June 2014 110 20 18.18

7 June 2014 130 29 22.3 lifted her abdomen and inserted her ovipositor almost verti- 8 June 2014 110 38 34.54 cally into the pod. Almost always each caterpillar had three 9 June 2014 100 33 33 10 June 2014 110 39 35.45 parasitoid eggs laid on itself (Figs 5A and 6); rarely, it was Mean 116 21.35 18.77±3.80 noted that more than 3 eggs were laid on host up to a maxi- mum of six.

18 J. Valipour et al.

Since almost the whole A. foetida shrubs are matured, parasitoid. We gratefully acknowledge the help of Dr Chris Hodgson their leaves and pods are shed after the 10th June, our data (Department of Biodiversity and Biological Systematics, The has stopped. General distribution of this wasp includes: National Museum of Wales, Cathay's Park, Cardiff) for helping and his comments on this manuscript. We are grateful also for our China, Central Asia, Mongolia, Siberia, Russia, Crimea, Iran, colleagues' support and help in Department of Natural Resources of Romania, Yugoslavia, Hungry, Finland, Sweden, Italy, Bel- Gilang-e Gharbe during the implementation of this research. gium, Poland, Austria, Germany, France, England, Switzer- land and turkey (Yu et al. 2012).

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