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Türkiye’de İstalacı bir tür olan Nysius cymoides (Spinola, 1837) (Hemiptera: Lygaeidae)’in Moleküler Tanımlaması

Sahte çinç böceği, Nysius cymoides (Hemiptera: Lygaeidae), geniş bir konukçu çeşitliliğine sahip önemli bir zararlıdır. İlk olarak İtalya'da tespit edilen bu zararlı, şimdi Türkiye’de dahil olmak üzere tüm dünyada tarımsal zararlara yol açmaktadır. Nysius cymoides örnekleri, Temmuz 2020'de Edirne Keşan bölgesinde ayçiçeği tarlalarında yapılan sörvey araştırması sonucunda elde edilmiştir. Zararlının popülasyonundaki artış ve ayçiçeği ile mısır tarlarındaki zararı, bu türün doğru teşhis edilmesi zorunlu kılmıştır. Zararlının teşhisi ve moleküler karakterizasyonunun araştırılması için mitokondrial sitokrom oksidaz I (COI) barkod alanı kullanıldı. Ergin ve geç ergin öncesi dönem örnekleri stereozoom mikroskop altında incelenmiş ve morfolojik olarakta tanımlandı. COI genin bir parçası (659 bç) LCO1490/ HCO 2198 primerleri ile çoğaltıldı. Moleküler karakterizasyon için PCR ürünleri doğrudan dizilendi. Dizilim GenBank veri tabanına OL989232 erişim numarası ile kaydedildi. Moleküler analizler MEGA X yazılımı kullanılarak yapıldı. Sonuçta, nükleotid frekansları A = 33.81%, T/U = 35.79%, C = 16.03% ve G = 14.37% olarak belirlendi. ASAP ve ABGD kullanılarak uygulanan mesafeye dayalı tür sınırlama yöntemi, en düşük ASAP puanına (1.5) sahip bölmede 5 tür belirlendi. En iyi ASAP puanına sahip bu bölümün barkod boşluk mesafesi %1.7 olarak belirlendi. Eşik mesafesi %1.62 olarak belirlendi. Karşılaştırılan diziler arasında en yüksek nükleotid benzerliği, çalışmamız sonucu elde edilen dizi (OL989232) ile Nysius cymoides France (KJ541649.1) dizisi arasında %99.8 olarak tespit edildi. Bu çalışma ile Edirne’den toplanan Nysius cymoides örneklerinin ilk kez moleküler teşhisi gerçekleştirilmesidir.

Anahtar Kelimeler:

Nysius cymoides, , DNA barkod, , COI, , mtDNA, , Moleküler tanımlama

Molecular Identification of the Invasive Species Nysius cymoides (Spinola, 1837) (Hemiptera: Lygaeidae) in Turkey

The false chinch bug, Nysius cymoides (Spinola, 1837) (Hemiptera: Lygaeidae), is an important pest with a wide host range. It was first discovered in Italy but now causing agricultural damage worldwide, including Turkey. Specimens of Nysius cymoides were collected from surveys of the sunflower and corn fields in Malkara-Keşan province, in Edirne in July 2020. Because of the population increase of the pest and their damages to sunflowers and corn fields, the pest identification have become an important issue. We investigated molecular characterization and identification of the pest by using mitochondrial cytochrome oxidase I (COI) barcode region. Adult and late immature stage of the specimens were examined under a stereozoom microscope and morphologically identified as well. The partial COI gene (659 bp) was amplified by using LCO1490/ HCO 2198 and PCR products were sequenced directly for molecular characterization. The sequence was registered GenBank database with accession number OL989232. Molecular analyzes were performed using MEGA X software. As a result, the nucleotide frequencies were A = 33.81%, T/U = 35.79%, C = 16.03%, and G = 14.37%. The distance-based species limitation method applied by using ASAP and ABGD, identified 5 species in the partition with the lowest ASAP score (1.5). The barcode gap distance of this partition with the best ASAP score was determined as 1.7%. The threshold distance was determined as 1.62%. The highest nucleotide identity of the studied specimen (OL989232) was detected as 99.8% with the specimen of Nysius cymoides from France (KJ541649.1) in GenBank. The objective of this study was the first attempt to identify Nysius cymoides specimens collected in Edirne based on DNA barcode regions.

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Abacıgil, T.Ö., Varlı, V.S., Tezcan, S., 2010. Edremit (Balıkesir) Körfezi çevresindeki zeytin bahçelerinde kışlak tuzaklarla saptanan Heteroptera türleri. Türk. Entomol. Derg.34 (1): 105-115.
Ashlock, P.D., 1967. A generic classification of the Orsillinae of the world (Hemiptera-Heteroptera: Lygaeidae). Univ Calif Publ Entomol. 48:1–82.
Aukema, B., 2013. Nysius cymoides (Spinola, 1837). Fauna Europaea version 2017.06 [WWW document].
Badam, A. K., Mohagheg, M. A. J., Ostovan, H., 2006. Biofogy of the seed bug Nysius cymoides (Heteroptera: Lygaeidae) in Mazandaran canola fields. 17 th Iranian Plant Protection Congress, 2-5 Sept. Iran, 247 p.
Bandelt, H.J., Forster, P., Röhl, A., 1999. Median-joining networks for inferring intraspecific phylogenies. Molecular Biology and Evolution. 16(1): 37–48.
Becker, T., Knapp, M., 2004. A Powerful Strategy to Account for Multiple Testing in the Context of Haplotype Analysis. AJHG. 75(4): 561-570.
Bocchi, S., Cinquanta, D., Negri, M., Dioli, P., Limonta, L., 2016. Nysius cymoides (Spinola) on Chenopodium quinoa Willd. cultivated in Italy. Journal of Entomological and Acarological Research. 48: 332–334.
Bolu, H., 2012. Güneydoğu Anadolu Bölgesi antepfıstığı alanlarındaki böcek ve akar faunasının saptanması. Türk. Entomol Derg. 26 (3): 197-2008.
Demirel, N., 2009. Determination of heteroptera species on canola plants in Hatay province of Turkey. African Journal of Agricultural Research. 4: 1226–1233.
Folmer, O., Black, M., Hoeh, W., Lutz, R., Vrijenhoek, R., 1994. DNA primers for amplification of mitochondrial cytochrome c oxidase subunit I from diverse metazoan invertebrates. Molecular Marine Biology and Biotechnology. 3: 294-299.
Haouas, D., Gnidez, HG., Hafsi, C., Nhaili, H., Matocq, A., 2019. Nysius cymoides (Heteroptera, Lygaeidae), an emerging insect pest in Tunisia. Bulletin OEPP/EPPO Bulletin. 49 (2): 355–358.
Hebert, P.D.N, Cywinska, A., Ball, S.L, deWaard, J.R., 2003. Biological identifications through DNA barcodes. Proc R Soc Lond B Biol Sci. 270: 313-321.
Hori, K., 2000. Possible causes of disease symptoms resulting from the feeding of phytophagous Heteroptera, pp. 11-35.
Kumar, S., Stecher, G., Li, M., Knyaz, C., Tamura, K., 2018. MEGA X: Molecular Evolutionary Genetics Analysis across computing platforms. Molecular Biology and Evolution. 35: 1547-1549.
Kuyulu A., S. Yücel., Genç, H., 2019. Molecular identification of invasive drosophilid, Zaprionus tuberculatus Malloch (Diptera:Drosophilidae) in Çanakkale, Turkey. Turkish Journal of Zoology. 43 (5): 531-535.
Kuyulu, A., Genç, H., 2020. Genetic diversity of codling moth Cydia pomonella L. (Lepidoptera: Tortricidae) populations in Turkey. Turkish Journal of Zoology. 44(5): 462-471.
Matocq, A., Özgen, İ., 2010. A preliminary list of Heteroptera collected in Mardin and Siirt provinces from South-Eastern Anatolia of Turkey (Hemiptera). Munis Entomology & Zoology. 5: 1011-1019.
Mohaghegh Neyshabouri, J., 2009. A temperature-dependent developmental rate model for Nysius cymoides (Het.: Lygaeidae). https://agris.fao.org/agris-search/search.do?recordID=IR2011000266#
Mollashahi M, Sahragard A, Mohaghegh J, Hosseini R, Sabouri H., 2017. Two-sex life table of Nysius cymoides (Spinola) (Hem.: Lygaeidae) at different photoperiods under laboratory conditions. Plant Pest Res. 6(4):73–87.
Mutun, S., Atay, G., 2015. Phylogeography of Trigonaspis synaspis (Hymenoptera: Cynipidae) from Anatolia based on mitochondrial and nuclear DNA sequences. Eur. J. Entomol. 112(2): 259–269.F
Nei, M., Maruyama, T., Chakraborty, R., 1975. The bottleneck effect and genetic variability in populations. Evolution. 29:1–10.
Özgen, İ., 2012. The species of suborder Heteroptera (Hemiptera) on vineyards agroecosystems which found in Diyarbakır, Elazığ and Mardin provinces, Turkey. Mun. Ent. Zool. 7: 255-258.
Özgen, İ., Kaymak Kara, B., Miroğlu, S., Koç, İ., Dioli, P., 2020. A New Potential Pest of East and Southeastern Anatolia in Turkey: Nysius cymoides (Spinola, 1837) (Heteroptera, Lygaeidae). Munis Entomology & Zoology. 15 (1): 265-268.
Pentinsaari, M., Vos, R., Mutanen, M., 2017. Algorithmic single-locus species delimitation: Effects of sampling effort, variation and nonmonophyly in four methods and 1870 species of beetles. Molecular Ecology Resources. 17: 393– 404.
Pericart, J., 1998. Hemipteres Lygaeidae Euro-Mediterraneens, Generalites systematique:1. Pre-miere partie.- Faune de France 84 A, Paris, France.
Puillandre, N., Lambert, A., Brouillet, S. Achaz, G., 2012. ABGD, Automatic Barcode Gap Discovery for primary species delimitation. Mol. Ecol. 21: 1864–1877.
Puillandre, N., Brouillet, S., Achaz, G., 2021. ASAP: assemble species by automatic partitioning. Molecular Ecology Resources. 21(2): 609-620.
Rozas, J., Ferrer-Mata, A., Sánchez-DelBarrio, J.C., Guirao-Rico, S., Librado, P., Ramos-Onsins, S.E., Sánchez-Gracia, A., 2017. DNA sequence polymorphism analysis of large data sets. Molecular Biology and Evolution. 34. (12): 3299-3302.
Saitou, N., Nei, M., 1987. The neighbor-joining method: a new method for reconstructing phylogenetic trees. Molecular Biology and Evolution. 4: 406-425.
Sarafrazi A., Shiroudbakhshi R., Zahiri R., Zangeneh A., 2009. Nysius species fauna of Iran and their distribution on canola. In: 6th Asia-Pacific Congress of Entomology (APCE 2009), October 18–22, 2009, Beijing, China.
Scaccini, D., Furlan, L., 2019. Outbreak of Nysius cymoides on second crop soybean Glycine max and proposal for Integrated Pest Management. Bulletin of Insectology.72 (1): 29-34.
Scaccini, D., Furlan, L., 2021. Nysius cymoides (Hemiptera: Lygaeidae), a potential emerging pest: overview of the information available to implement integrated pest management. International Journal of Pest Management. 67(1): 73-88.
Sweet, M.H., 2000. Seed and chinch bugs (Lygaeoidea). In: Schaefer CW, Panizzi AR (eds) Heteroptera of economic importance. CRC Press, Boca Raton, 143–264.
Thompson, J.D., Higgins, D.G., Gibson, T.J., 1994. CLUSTAL W: Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research. 22 (22): 4673-4680.
Yasunaga T., 1990 A revision of the genus Adelphocoris Reuter (Heterptera, Miridae) from Japan. Japan Journal of Entomology. 58: 606–618.
Wipfli M.S., Wedberg J.L., Hogg D.B., 1990. Damage potentials of three plant bug (Hemiptera: Heteroptera: Miridae) species to birds foot trefoil grown for seed in Wisconsin. Journal of Economic Entomology. 83: 580–584.
Yazıcı, G., 2022. Türkiye'de Kültür Bitkilerinde Istilacı Tür Nysius cymoides (Spinola, 1837) (Hemiptera: Heteroptera: Lygaeidae)’ın Yeni Konukçuları ve Yayılma Alanları. KSÜ Tarım ve Doğa Dergisi. 25 (2): 267-273.
Yücel, S. ve H.Y. Genç., 2022. Genetic Diversity of Cotton Bollworm, Helicoverpa armigera (Lepidoptera: Noctuidae) Population in Çanakkale. COMU J. Agric. Fac. 10(1): 211-221.
Zhou, Z., Guo, H., Han, L., Chai, J., Che, X., Shi, F., 2019. Singleton molecular species delimitation based on COI-5P barcode sequences revealed high cryptic/undescribed diversity for Chinese katydids (Orthoptera: Tettigoniidae). BMC Evolutionary Biology. 19:79