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

Abstract

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.

Keywords

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

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

Abstract

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.

Thanks

We thank to Dr. Pınar TURHAN for providing the specimens. We also thank to Paul Skelley for the identification of the specimen in Division of Plant Industry Florida Department of Agriculture and Consumer Services.

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