Isolation of Bacillus zhangzhouensis OBB Liu et al. (Caryophanales: Bacillaceae) from native Polyphylla fullo (L., 1758) (Coleoptera: Scarabaeidae) larvae, PCR-based detection of cry1 gene and evaluation of its biological control potential

Abstract

Bacillus zhangzhouensis OBB Liu et al. (Caryophanales: Bacillaceae) isolate was obtained from Polyphylla fullo (L., 1758) (Coleoptera: Scarabaeidae) larvae in the Microbiology Laboratory of the Biology Department of the Faculty of Science of Atatürk University in 2024. Additionally, the presence of the cry1 gene was identified. After endospore staining of the isolate, the presence of crystal protein was detected by phase contrast microscopy and SEM analysis. As a result of PCR, only the presence of the cry1 gene was detected and confirmed. The total protein contents were compared with those of Bacillus thuringiensis (Berliner, 1915) (Bacteria: Bacillaceae) by performing SDS-PAGE analysis using a crystal protein and spore mixture. B. zhangzhouensis OBB showed bands ̴ 250 kDa and ̴ 80 kDa, while B. thuringiensis showed bands corresponding to ̴ 70 kDa and ̴ 45 kDa. Probit analysis was used to determine the LC50 value of the isolates, and the Abbott method was used to determine the mortality percentages of the larvae. Spore-crystal mixtures of B. thuringiensis and B. zhangzhouensis OBB isolates were tested against P. fullo larvae at doses of 1000, 2000, and 4000 ppm. The highest mortality rate was determined in the spore-crystal mixture of B. zhangzhouensis OBB isolate at 4000 ppm dose.

Keywords

• Bacillus zhangzhouensis OBB
• biopesticides
• cry genes
• Polyphylla fullo
• SDS-PAGE

Yerel Polyphylla fullo (L., 1758) (Coleoptera: Scarabaeidae) larvalarından Bacillus zhangzhouensis OBB Liu et al. (Caryophanales: Bacillaceae)'nin izolasyonu, cry1 geninin PCR tabanlı tespiti ve biyolojik kontrol potansiyelinin değerlendirilmesi

Öz

Bacillus zhangzhouensis OBB Liu et al. (Caryophanales: Bacillaceae) izolatı, 2024 yılında Atatürk Üniversitesi Fen Fakültesi Biyoloji Bölümü Mikrobiyoloji Laboratuvarında Polyphylla fullo (L., 1758) (Coleoptera: Scarabaeidae) larvalarından elde edilmiştir ve cry1 geninin varlığı tanımlanmıştır. İzolatın endospor boyaması yapıldıktan sonra faz kontrast mikroskobu ve SEM analizi ile kristal protein varlığı tespit edilmiştir. PCR sonucunda sadece cry1 geninin varlığı tespit edilmiştir ve doğrulanmıştır. Kristal protein ve spor karışımı kullanılarak SDS-PAGE analizi ile Bacillus thuringiensis (Berliner, 1915) (Bacteria: Bacillaceae) ile arasındaki total protein içerikleri karşılaştırılmıştır. Bacillus zhangzhouensis OBB ̴ 250 kDa ile ̴ 80 kDa arasında bantlar gösterirken, B. thuringiensis, 70 kDa ve ̴ 45 kDa'ya karşılık gelen bantlar göstermiştir.İzolatların LC50 değerini belirlemek için probit analizi, larvaların ölüm yüzdelerini belirlemek için ise Abbott yöntemi kullanılmıştır. B.thuringiensis ve B. zhangzhouensis OBB izolatlarının spor-kristal karışımları (1000, 2000 ve 4000 ppm) dozlarında P. fullo larvalarına karşı test edilmiştir. En yüksek ölüm oranı B. zhangzhouensis OBB izolatının 4000 ppm dozunda spor-kristal karışımında belirlenmiştir.

Anahtar Kelimeler

• Bacillus zhangzhouensis OBB
• biyopestisitler
• cry genleri
• Polyphylla fullo
• SDS-PAGE

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