Short Communication Confirmation of livia Identification on farnesiana and its First Report in Southern Tunisia

Rim Mkaouar, LGVRF-Gestion et Valorisation des Ressources Forestières, INRGREF, Université de Carthage, BP. 10, 2080 Ariana, Tunisia, Axel Hausmann, Bavarian State Collection of Zoology (ZSM), München, Germany, Staatliche Naturwissenschaftliche Sammlungen Bayerns (SNSB), München, Germany, and Mohamed Lahbib Ben Jamâa, LGVRF-Gestion et Valorisation des Ressources Forestières, INRGREF, Université de Carthage, BP. 10, 2080 Ariana, Tunisia ______ABSTRACT Mkaouar, R., Hausmann, A., and Ben Jamâa M.L. 2016. Confirmation of Deudorix livia identification on Acacia farnesiana and its first report in Southern Tunisia. Tunisian Journal of Plant Protection 11: 165-169.

In 2012, Deudorix livia was responsible for severe losses in production of trees in the regions of Gafsa and Gabes situated in Southern of Tunisia. D. livia was observed on some pods of Acacia farnesiana planted as windbreak around these orchards. Larvae of D. livia were observed drilling fruits to eat seeds. At present, the life cycle of D. livia is poorly studied. We confirmed its identification using molecular tools and reported for the first time its occurrence in Southern of Tunisia.

Keywords: Acacia farnesiana, Deudorix livia, pomegranate, Southern Tunisia ______

In 2006, a new pomegranate pest in pomegranate fruit production and to was reported as Virachola livia for the identify pests associated with this Acacia first time in the center and the southern of species. Investigations showed that Tunisia and it caused disastrous damages symptoms caused by V. livia on as high as 52% rotting in pomegranate pomegranate fruits (Fig. 1) were detected fruits (7). According to Gharbi (2), V. earlier on Acacia pods and were caused livia was noticed especially on by Deudorix livia (V. livia). pomegranate and on green pods of Acacia In 2013, DNA analysis of 17 farnesiana used as windbreak around individuals of Tunisian D. livia (V. livia) irrigated pomegranate orchards. In 2012, collected from Zerkine and Metwia a survey was conducted to study the role (Gabes), and Ksar (Gafsa) on A. of A. farnesiana in the huge losses farnesiana and pomegranate, were recorded performed to identify the species. PCR amplification and DNA sequencing were conducted at the CCDB (Canadian Center Corresponding author: Rim Mkaouar for DNA Barcoding, Guelph, Canada) Email: [email protected] following standard high-throughput protocols (5) that can be accessed on the Accepted for publication 30 April 2016 website (13). PCR amplification with a

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single pair of primers consistently reported for the first time in in covering the 658 bp region near the 5’ 2004. Moreover, Muller (10) reported that terminus of the mitochondrial D. livia is a migratory species which is Cytochrome c Oxidase I (COI) gene expanding its development to the North. including the standard 658 bp barcode Larvae of D. livia are polyphagous region for the kingdom (4). DNA and break through fruits of its host plants. extracts were stored at the CCDB, with This pest was also reported on aliquots already deposited in the DNA dactylifera, Acacia green pods, Prunus Bank facility of the ZSM (14). All salicina, Eriobotrya japonica, Punica sequences were deposited also in granatum (3), and occasionally on olive GenBank. Results of DNA analyses flowers (6). revealed that both A. farnesiana and This work is the first record of D. pomegranate fruits were attacked by D. livia in Tunisia and in North Africa on A. livia. farnesiana. In fact, observed D. livia In 1992, the larvae attack green pods, develop holes subgenus Virachola was considered as a then eat seeds (Fig. 2) until complete genus by some authors. Later, D. livia development, followed by prepupae (Fig. was firstly described and figured under 3) and pupa stages. Larvae of D. livia the name of Virachola (15). In Tunisia, in start to feed late in April. Adults (Fig. 4) 2006, this was firstly described emerge from mid-May till mid-July (8). as V. livia according to morphological D. livia is reported to have three descriptions of Hanna (3) and generations on A. farnesiana in Tunisia, morphological confirmation of Katbeh then, the adult migrates to its secondary Bader (6). However, in 2008, Virachola host pomegranate when Acacia pods was considered as a subgenus and called become dry, and a new generation Deudorix (15). In the present starts (8, 9). Nevertheless, in the Middle investigation, nineteen barcode sequences East, on both Acacia and Punica, this pest were obtained with the length of 658 bp. can develop between six and eight Nucleotide sequence was inscribed in the generations (1). Barcode of Life Data System in the public Currently, damages caused by D. dataset DS-LIVIA (DOI dx.doi.org/ livia seem to be more or less insignificant 10.5883/DS-LIVIA). due to the restricted number of its primary The pomegranate Playboy is host A. farnesiana and to its competition considered as a dangerous pest for the with Ectomyelois ceratoniae, the major pomegranate in the Arabian Peninsula threatening pomegranate trees in reaching Mediterranean coast of , Southern Tunisia. Israel, Lebanon and Syria (11, 12). It was ______RESUME Mkaouar R., Hausmann A. et Ben Jamâa M.L. 2016. Confirmation de l’identification de Deudorix livia sur Acacia farnesiana et son premier signalement au sud tunisien. Tunisian Journal of Plant Protection 11: 165-169.

En 2012, Deudorix livia a causé des dégâts importants aux vergers des grenadiers dans la région de Gafsa et Gabès située au sud tunisien. D. livia a été observé sur les gousses d’Acacia farnesiana plantées comme brise-vent autour de ces vergers. Les larves de D. livia ont été observées dans le fruit mangeant les graines d’Acacia. A présent, le cycle de vie de D. livia est mal étudié. Nous avons

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confirmé l’identification de ce ravageur en utilisant les outils moléculaires et nous avons signalé pour la première fois son occurrence au sud tunisien.

Mots clés: Acacia farnesiana, Deudorix livia, grenadier, sud tunisien ______

a b

Fig. 1. Damage of Deudorix livia on a pomegranate fruit (a) and Acacia green pod (b).

Fig. 2. Larvae of Deudorix livia eating a seed of Acacia farnesiana.

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Fig. 3. Prepuae of Deudorix livia.

Fig. 4. Adult of Deudorix livia.

______ملخص مكور، ريم وأكسل ھوسمان ومحمد الحبيب بن جامع .2016. تأكيد تشخيص حشرة Deudorix livia على أشجار السنط )Acacia farnesiana( وأول تقرير لوجودھا في الجنوب التونسي. Tunisian Journal of Plant Protection 11: 165-169.

في عام 2012 ، تسببت يرقات الحشرة Deudorix livia في خسائر فادحة على فاكهة الرمان في منطقت ْي قفصة وقابس الواقعت ْين في الجنوب التونسي. ولوحظت على قرون أشجار Acacia farnesiana التي غرست كمصدات الرياح حول هذه البساتين. وشوهدت يرقات D. livia داخل فاكهة Acacia تأكل البزور. ال توجد حتى اآلن دراسات معمقة حول الدورة الحياتية للحشرة D. livia. لقد أكدنا تشخيص هذه اآلفة باستخدام المواد الجزيئية وقدمنا أول تقرير لوجوده في الجنوب التونسي.

كلمات مفتاحيه: جنوب تونس، رمان، Acacia farnesiana ،Deudorix livia ______

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LITERATURE CITED 1. Avidov, Z. 1958. A study of a life history of du grenadier Virachola livia sur les brise-vents pomegranate butterfly (Virachola livia Klug.) d’Acacia au sud tunisien. Rev. Rég. Arid. 35: in Israel. Ktav. Rehov. 8: 153-167. 1749-1754. 2. Gharbi, N. 2010. Laboratory rearing of the 9. Mkaouar, R., Ben Jamâa, M.L., and Dhahri, S. pomegranate fruit butterfly Virachola livia on 2014. Some biological aspects of Virachola two host plant in Tunisia. Tunisian J. Plant livia on Acacia farnesiana in the South of Prot. 5: 195-199. Tunisia. Pages 48-50. In: Proceedings of the 3. Hanna, A.D. 1939. The pomegranate fruit 4th Medinsect meeting of the 7-03-14 IUFRO butterfly Virachola livia Klug: Morphology, group "Entomological Research in life history and control. Ministry of Mediterranean Forest Ecosystems" and the Agriculture, Egypt, 54 pp. next meeting of the 7-03-06 IUFRO group 4. Hebert, P.D.N., Ratnasingham, S., and De Waard, "Integrated Management of Forest Defoliating J.R. 2003. Barcoding animal life: cytochrome c ". April 9-14, 2014, Antalya, Turkey. oxidase subunit 1 divergences among closely 10. Muller, G.C., Kravchenko, V.D., Phillips, A., related species. Proc. R. Soc. Lond. B. 270 Shwarz-Tzachor, R., and Benyamini, D. 2005. Supplement 1: S96-S99. The first record of the Eremic Lycaenid 5. Ivanova, N.V., De Waard, J.R., and Hebert, Deudorix livia (Klug,1834) in Europe P.D.N. 2006. An inexpensive automation ( :). Atalanta 36: 109- friendly protocol for recovering high-quality 112. DNA. Mol. Ecol. Notes 6: 998-1002. 11. Tolman, T. 2001. of Europe. 6. Katbeh-Bader, A., Amr, Z.S., and Isma’el S. University Press Princeton, New Jersey, 536 2003. The butterflies of Jordan. J. Res. Lepid. pp. 37: 11-26. 12. Williams, J.G.A. 1971. Field Guide to the 7. Ksentini, I., Jardak, T., and Zeghal N. 2011. First Butterflies of Africa. Hought. Miff., Boston, report on Virachola livia Klug. (Lepidoptera : 38 pp. Lycaenidae) and its effects on different 13. http://www.dnabarcoding.ca/pa/ge/research/pro- pomegranate varieties in Tunisia. EOPP Bull. tocols. 41: 178-182. 14. http://www.zsm.mwn.de/dnabank/ 8. Mkaouar, R., Saadaoui E., and Ben Jamâa, M.L. 15. http://tolweb.org/Deudorix/112465/2008.04.14 2014. Premières observations sur le papillon

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