Bioactivities of cry gene positive Bacillus thuringiensis (Berliner) (Bacillales: Bacillaceae) strains on Ephestia kuehniella Zeller, 1879 and Plodia interpunctella (Hübner, 1813) (Lepidoptera: Pyralidae)

Yıl 2016, Cilt: 40 Sayı: 4, 365 - 375, 10.10.2016

Hatice Güneş , Mehlika Alper , Bekir Çöl , Hilal Tunca

Öz

Bacillus thuringiensis is the bacterium most commonly used for biopesticide production due to parasporal crystal formation during its growth cycle. As a consequence of repeated use, B. thuringiensis biopesticides may cause the development of resistance in the pests. Therefore, it is necessary to explore new B. thuringiensis strains with a certain degree of bioactivity. In this study (2012-2013), the bioactivity of native B. thuringiensis strains from the Aegean Region of Turkey were tested against second instar larvae of Ephestia kuehniella and Plodia interpunctella. The bioactivity of 21 B. thuringiensis strains with cry1, cry2 or cry9 gene was determined as percent mortality according to Abbott’s formula. The highest mortality rates were 42 and 63% in E. kuehniella and P. interpunctella, respectively. These mortality rates were equal to or 1.8 times greater than that of B. thuringiensis subsp. kurstaki. In addition, plasmid profiles of B. thuringiensis strains changed between 5-18 kb. Moreover, SDS-PAGE analysis of the most toxic strains indicated the presence of Cry1 and Cry2 proteins. Two different cry2 gene profiles containing either cry2Aa1 or combination of cry2Aa1 and cry2Ab2 genes were detected by PCR analysis. In addition, partial DNA sequence analysis of cry2A genes indicated phylogenetic differences among the toxic strains and B. thuringiensis subsp. kurstaki. As a result, these B. thuringiensis strains may be used to control both E. kuehniella and P. interpunctella as alternative biopesticides in cases of insect resistance to  currently used B. thuringiensis preparations.

Anahtar Kelimeler

Bacillus thuringiensis, bioactivity, Ephestia kuehniella, Plodia interpunctella

cry gene pozitif Bacillus thuringiensis (Berliner) (Bacillales: Bacillaceae) izolatlarının Ephestia kuehniella Zeller, 1879 ve Plodia interpunctella (Hübner, 1813) (Lepidoptera: Pyralidae) üzerindeki biyoaktiviteleri

Öz

Bacillus thuringiensis üreme döngüsü sırasında kristal oluşturması nedeniyle biyopestisit üretimi için en çok kullanılan bakteridir. Bacillus thuringiensis biyopestisitlerinin tekrarlayan kullanımları zararlılarda direnç gelişimine neden olabileceğinden, belirli düzeyde biyoakiviteye sahip yeni B. thuringiensis izolatlarının araştırılmasına ihtiyaç vardır. Bu çalışmada, Ege Bölgesin’den elde edilen doğal B. thuringiensis izolatlarının biyoaktivitesi Ephestia kuehniella ve Plodia interpunctella’nın ikinci dönem larvalarına karşı 2012-2013 yıllarında araştırılmıştır. cry1, cry2 ya da cry9 geni taşıyan 21 B. thuringiensis izolatının biyoaktivitesi Abbott formülüne göre yüzde ölüm olarak belirlenmiştir. En yüksek ölüm oranları, E. kuehniella ve P. interpunctella’ ya karşı sırasıyla %42 ve %63 bulunmuştur. Bu ölüm oranları, B. thuringiensis subsp. kurstaki’ninkine eşit veya B. thuringiensis subsp. kurstaki’ninkinden 1.8 kat daha yüksektir. Buna ek olarak, B. thuringiensis izolatlarının plazmit profilleri 5-18 kb arasında değişmiştir. Ayrıca, en toksik izolatların SDS-PAGE analizi Cry1 ve Cry2 proteinlerinin varlığını göstermiştir. PCR analizi ile ya cry2Aa1 veya cry2Aa1 ve cry2Ab2 genlerinin kombinasyonunu içeren iki farklı cry2 gen profili belirlenmiştir. Ayrıca, cry2A genlerinin kısmi DNA sekans analizi, toksik izolatlar ve B. thuringiensis subsp. kurstaki arasındaki filogenetik farklılıkları göstermiştir. Sonuç olarak, bu B. thuringiensis izolatları bilinen B. thuringiensis preperasyonlarına karşı böcek direnci olması durumunda alternatif biyopestisitler olarak hem E. kuehniella hem de P. interpunctella’yı kontrol etmek için kullanılabilir.

Anahtar Kelimeler

Bacillus thuringiensis, biyoaktivite, Ephestia kuehniella, Plodia interpunctella

Kaynakça

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