Donus GENCER, Aydin YESİLYURT, Mustafa GÜLLÜ, İsmail DEMİR, Remziye NALCACIOGLU

Yaban tip ve rekombinant omurgasız iridesan virüslerinin beş yaygın zararlı üzerindeki böcek öldürücü aktiviteleri

Omurgasız iridesan virüs 6 (IIV6), çeşitli zararlı böcek türlerini düşük oranda enfekte edebilir. Rekombinant DNA teknolojisi ile oluşturulan yeni özelliklere sahip virüsler biyolojik kontrol ajanları olarak etkin bir şekilde kullanılabilir. Daha önce IIV6 157L ORF’si yerine birisi yeşil floresan protein geni (rCIV-Δ157L-gfp) ve diğeri de gfp ile birleştirilmiş bir akrep Androctonus australis (Linnaeus, 1758) böcek toksin geni (rCIV-Δ157L/gfp-AaIT) ihtiva eden iki rekombinant IIV6 inşa edilmişti. Bu çalışmada, yaban tip IIV6 ve iki rekombinant virüs, Helicoverpa armigera (Hübner, 1805) (Lepidoptera: Noctuidae), Spodoptera littoralis (Boisduval, 1883) (Lepidoptera: Noctuidae), Lymantria dispar (Linnaeus, 1758) (Lepidoptera: Erebidae), Euproctis chrysorrhoea (Linnaeus, 1758) (Lepidoptera: Erebidae) ve Tenebrio molitor (Linnaeus, 1758) (Coleoptera: ‎Tenebrionidae) larvalarında enfeksiyon oluşturma yeteneği açısından değerlendirildi. Çalışma 2018 ve 2019 yılları arasında Karadeniz Teknik Üniversitesi Biyoloji Bölümü'nde gerçekleştirildi. Her bir böcek larvasını enjekte etmek için virüslerin beş farklı konsantrasyonu (103, 104, 105, 106 ve107 TCID50/ml) kullanıldı. rCIV-Δ157L/gfp-AaIT ile enfekte olmuş S. littoralis dışında tüm larvalar felç oldu. rCIV-Δ157L/gfp-AaIT ile enfekte olmuş böcek larvalarının LC50'si sırasıyla, S. littoralis, T. molitor, L. dispar, H. armigera ve E. chrysorrhoea üzerinde 0.3 x 107, 0.7 x 105, 0.2 x 105, 0.15 x 105, 0.7 x 104 TCID50/ml olarak belirlenmiştir. En yüksek virüs konsantrasyonlarına göre hesaplanan LT50 değerleri, rCIV-Δ157L/gfp-AaIT için S. littoralis, T. molitor, L. dispar, H. armigera ve E. chrysorrhoea'da sırasıyla 10.5, 6.2, 4.7, 7.5 ve 5 gün olarak bulundu. Bu çalışma, rekombinant IIV6'nın Lepidoptera ve Coleoptera takımlarına ait bazı böceklerde patojeniteyi artırdığını göstermiştir.

Anahtar Kelimeler:

Omurgasız iridesan virüs, insektisidal aktivite, rekombinant virüs

Insecticidal activities of wild type and recombinant invertebrate iridescent viruses on five common pests

Invertebrate iridescent virus 6 (IIV6) can infect a broad range of pest insect species. Viruses with new features created by recombinant DNA technology can be used effectively as biological control agents. Previously, recombinants have been constructed: IIVs harboring green fluorescent protein gene (gfp) in place of IIV6 157L ORF (rCIV-Δ157L-gfp) and a scorpion Androctonus australis (Linnaeus, 1758) insect toxin gene (AaIT) fused with gfp (rCIV-Δ157L/gfp-AaIT). In this study, wild type IIV6 and the two recombinants, were evaluated for their ability to cause infections on Helicoverpa armigera (Hübner, 1805) (Lepidoptera: Noctuidae), Spodoptera littoralis (Boisduval, 1883) (Lepidoptera: Noctuidae), Lymantria dispar (Linnaeus, 1758) (Lepidoptera: Erebidae), Euproctis chrysorrhoea (Linnaeus, 1758) (Lepidoptera: Erebidae) and Tenebrio molitor (Linnaeus, 1758) (Coleoptera: ‎Tenebrionidae) larvae. This study was performed at Karadeniz Technical University, Department of Biology during 2018 and 2019. Five different concentrations (103, 104, 105, 106 and 107 TCID50/ml) of viruses were used to inject each insect larvae. All larvae, infected with rCIV-Δ157L/gfp-AaIT, became paralyzed, except S. littoralis. The LC50 of insect larvae infected by rCIV-Δ157L/gfp-AaIT were determined as 0.3 x 107, 0.7 x 105, 0.2 x 105, 0.15 x 105, 0.7 x 104 TCID50/ml on S. littoralis, T. molitor, L. dispar, H. armigera and E. chrysorrhoea, respectively. LT50 values, calculated according to the highest virus concentrations, were found as 10.5, 6.2, 4.7, 7.5 and 5 d on S. littoralis, T. molitor, L. dispar, H. armigera and E. chrysorrhoea, respectively, for rCIV-Δ157L/gfp-AaIT. This study showed that recombinant IIV6 has increased pathogenicity on some insects from Lepidoptera and Coleoptera.

Keywords:

Invertebrate iridescent virus, insecticidal activity, recombinant virus,

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