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1. Development of novel betabaculovirus (HycuGV-Hc1) as a biopesticide (HycuGV-TR61) and its efficacy on the fall webworm, Hyphantria cunea Drury (Lepidoptera: Erebidae) larvae

Author

Zeynep Bayramoglu, Donus Gencer, and Ismail Demir

Subjects
Hyphantria cunea, Betabaculovirus, Biological insecticide, Efficacy, Agriculture
Abstract

Abstract Background The aim of this study was to develop an oil formulation from a local betabaculovirus. Hyphantria cunea Drury (Lepidoptera: Erebidae) granulovirus (HycuGV-Hc1) was isolated from the infected larvae to test its efficacy on the pest. The oil formulation was prepared by mixing the viral suspension with sunflower oil and some adjuvants and named HycuGV-TR61. Crude virus and the formulations were carried out on third instar H. cunea larvae using 1 × 104–8 OBs /ml concentrations and were exposed to temperatures (28, 35, and 42 °C) and UV-B light at different periods (0, 1, 3, 5 h). Results The mortality rate, which was 50% at the lowest concentration (1 × 104 OB/ml), reached 99.86% at the highest concentration (1 × 108 OB/ml). LC50 values of fresh and old forms were calculated as 0.64 × 104 and 0.87 × 104 OBs/ml, respectively. Application of shelf life showed that there was non-significant change in the pathogenic activity of the formulation with time. In the experiments, it was observed that the activity decreased as the temperature and time of exposure increased. Significantly difference in larval mortality was observed when fresh and old formulations were exposed to 0, 1, 3 and 5 h to UV-B, (old: 96.7, 86, 80 and 60%; fresh: 97.1, 90, 85 and 62%, respectively). Conclusions The results revealed superior aspects of HycuGV-TR61, which was developed as a local viral biopesticide, its resistance to abiotic factors and its potential to be used in pest control.

Published
2023
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2. Comparison of the potential activities of viral and bacterial chitinases

Author

Suleiman Abulikemu, Aydin Yesilyurt, Donus Gencer, Mehtap Usta, and Remziye Nalcacioglu

Subjects
Bacteria, Virus, Chitinase, Potential activity, Agriculture
Abstract

Abstract Background Chitin, a long-chain polymer of N-acetylglucosamine, is a major structural component of the insect exoskeleton and the peritrophic membrane (PM). Chitinases are able to effectively break down glycosidic bonds of chitin polymer thus can be used in agriculture to control plant pathogen insects. These enzymes can be synthesized by higher plants, animals, protista, bacteria, and viruses. Results In this study, viral and bacterial chitinases were compared for their potential activity on a laboratory test insect. The genes encoding chitinases of Autographa californica nucleopolyhedrovirus (AcNPV) and Cydia pomonella granulovirus (CpGV) were amplified from genomic DNAs by PCR and cloned into the pET-28a (+) expression vector. The chitinase proteins of these 2 viruses (AcNPV-Chi, CpGV-Chi) and Serratia marcescens chitinase C (ChiC) protein which was previously cloned were overexpressed in Escherichia coli. Expressed proteins were purified and confirmed by western blot analysis as 50, 63, and 68 kDa for AcNPV, CpGV, and S. marcescens chitinases, respectively. Enzyme activities of the chitinases were confirmed. Chitinases were also compared to each other in silico. The insecticidal effects of these proteins were evaluated on Galleria mellonella L. larvae. Bioassays were performed on the 3rd instar larvae for each chitinase protein in triplicate. The results showed that although there were differences in enzymatic activities and domain organizations, all 3 microbial chitinases produced almost the same level of insecticidal activity on the test insect. LC50 and LT50 values were compatible with the mortality results. These results were a preanalysis for comparing the effects of microbial chitinases. Conclusion Potential activity experiments should be carried out on more insects to provide detailed information on the insecticidal effects of bacterial and viral chitinases.

Published
2021
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3. Determination of fungal pathogens of Hypera postica (Gyllenhall) (Coleoptera: Curculionidae): isolation, characterization, and susceptibility

Author

Birgul Yucel, Celalettin Gozuacik, Donus Gencer, Ismail Demir, and Zihni Demirbag

Subjects
Hypera postica, Microbial control, Beauveria bassiana, Isolation, Susceptibility, Agriculture
Abstract

Abstract Background Fungal pathogens of Hypera postica (Gyllenhall) (Coleoptera: Curculionidae) were collected from the vicinities of Adana and Igdir in Turkey. The pathogenicity of the fungal isolates against the pest were investigated. According to morphologic (colony morphology, spore shape) and molecular (sequences of ITS1-5.8S ITS2 region and EF1-α, Bloc, and RPB1 genes) characterizations, the isolates were identified as Beauveria bassiana (HpI-2, HpI-6, HpI-7, HpI-10, HpA-3, HpA-4, HpA-5) and Beauveria pseudobassiana (HpI-4). All these strains were isolated from H. postica for the first time. In order to determine pathogenesis of all isolates on the target pest, bioassays were conducted against larvae and adults, as screening of (1 × 107 conidia/ml) and dose-response (1 × 105, 1 × 106, 1 × 107, 1 × 108, conidia/ml), under laboratory conditions. The fungal isolates, closely related to each other, yielded significantly varied mortalities on larvae and adults. H. postica larvae were found more susceptible than adults to the fungal isolates in all tests. The highest mortality rates (100 and 98%) for larvae and adults, respectively, were obtained by B. bassiana strain HpA-5 within 14 days at 1 × 108 conidia/ml concentration. The median lethal concentration (LD50) of HpA-5 required to kill the larvae and adults of H. postica at concentrations of 2.37 × 104 and 1.4 × 105 conidia/ml, respectively. These results are promising; therefore, the B. bassiana strain HpA-5 can potentially be used against H. postica.

Published
2018
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4. Comparison of the potential activities of viral and bacterial chitinases

Author

Aydin Yesilyurt, Remziye Nalcacioglu, Suleiman Abulikemu, Donus Gencer, and Mehtap Usta

Subjects
0301 basic medicine, Plant Science, medicine.disease_cause, Microbiology, 03 medical and health sciences, chemistry.chemical_compound, Chitin, medicine, Escherichia coli, Potential activity, Expression vector, 030102 biochemistry & molecular biology, Ecology, biology, Bacteria, fungi, Chitinase, Agriculture, biology.organism_classification, Virus, Galleria mellonella, Autographa californica, 030104 developmental biology, chemistry, Insect Science, Serratia marcescens, biology.protein, Agronomy and Crop Science
Abstract

Background Chitin, a long-chain polymer of N-acetylglucosamine, is a major structural component of the insect exoskeleton and the peritrophic membrane (PM). Chitinases are able to effectively break down glycosidic bonds of chitin polymer thus can be used in agriculture to control plant pathogen insects. These enzymes can be synthesized by higher plants, animals, protista, bacteria, and viruses. Results In this study, viral and bacterial chitinases were compared for their potential activity on a laboratory test insect. The genes encoding chitinases of Autographa californica nucleopolyhedrovirus (AcNPV) and Cydia pomonella granulovirus (CpGV) were amplified from genomic DNAs by PCR and cloned into the pET-28a (+) expression vector. The chitinase proteins of these 2 viruses (AcNPV-Chi, CpGV-Chi) and Serratia marcescens chitinase C (ChiC) protein which was previously cloned were overexpressed in Escherichia coli. Expressed proteins were purified and confirmed by western blot analysis as 50, 63, and 68 kDa for AcNPV, CpGV, and S. marcescens chitinases, respectively. Enzyme activities of the chitinases were confirmed. Chitinases were also compared to each other in silico. The insecticidal effects of these proteins were evaluated on Galleria mellonella L. larvae. Bioassays were performed on the 3rd instar larvae for each chitinase protein in triplicate. The results showed that although there were differences in enzymatic activities and domain organizations, all 3 microbial chitinases produced almost the same level of insecticidal activity on the test insect. LC50 and LT50 values were compatible with the mortality results. These results were a preanalysis for comparing the effects of microbial chitinases. Conclusion Potential activity experiments should be carried out on more insects to provide detailed information on the insecticidal effects of bacterial and viral chitinases.

Published
2021

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

Author

Aydin Yesilyurt, Mustafa Güllü, Remziye Nalcacioglu, Ismail Demir, and Donus Gencer

Subjects
Fen, law, Insect Science, Science, fungi, Recombinant DNA, Wild type, Omurgasız iridesan virüs,insektisidal aktivite,rekombinant virüs, Zoology, Biology, Invertebrate iridescent virus,insecticidal activity,recombinant virus, Invertebrate, law.invention
Abstract

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., 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.

Published
2020

7. Characterization of a novel baculovirus isolate from Malacosoma neustria (Linnaeus, 1758) (Lepidoptera: Lasiocampidae) in Samsun and its pathogenicity in different hosts

Author

Remziye Nalçacioğlu, Aydin Yesilyurt, Ismail Demir, Oğuzhan Yanar, Donus Gencer, and OMÜ

Subjects
education.field_of_study, Veterinary medicine, food.ingredient, biology, Population, Helicoverpa armigera, biology.organism_classification, mortality, Lepidoptera genitalia, Alphabaculovirus, food, Lasiocampidae, Insect Science, Hyphantria, Noctuidae, identification, Baculovirus, Malacosoma neustria, education, Lymantria
Abstract

Malacosoma neustria (Linnaeus, 1758) (Lepidoptera: Lasiocampidae) causes economic losses in apple, pear, plum, willow, oak and other economically important trees. In this study, an alphabaculovirus was isolated from the larval population of M. neustria from Samsun in the central Black Sea Region of Turkey between 2015 and 2016. Electron microscope analysis of occlusion bodies (OBs) obtained from dead larvae showed that the nucleocapsids of a new isolate (ManeNPV-T4) are multiply enveloped. The Kimura two-parameter analysis and the phylogenetic tree were performed based on concatenated nucleotide and amino acid sequences of the partial lef-8, lef-9 and polh genes from ManeNPV-T4 isolate compared to those of other 51 baculoviruses. Insecticidal activity tests against third instar M. neustria larvae produced 48 to 100% mortalities. The LC 50 of ManeNPV-T4 was 0.78 x 10 3 OBs/ml in M. neustria . Additionally, the isolate caused mortalities lower than 50% in Spodoptera exigua (Hubner, 1808) (Lepidoptera: Noctuidae) , Lymantria dispar (Linnaeus, 1758) (Lepidoptera: Erebidae) , Hyphantria cunea (Drury, 1773) (Lepidoptera: Arctiidae) and Helicoverpa armigera (Hubner, 1805) (Lepidoptera: Noctuidae) larvae. Consequently, the new nucleopolyhedrovirus is infectious on M. neustria larvae and other lepidopterans.

Published
2019

8. Characterization and pathogenicity of Beauveria bassiana strains isolated from Galleria mellonella L. (Lepidoptera: Pyralidae) in Turkey

Author

Dönüş Gençer and Zeynep Bayramoğlu

Subjects
Galleria mellonella, Beauveria bassiana, Biological control, Potential, Agriculture
Abstract

Abstract Background The greater wax moth, Galleria mellonella L. (Lepidoptera: Pyralidae), is among the most important wax pests economically. In the larval stage, the pest feeds on honeycomb wax and seriously damages the combs that were left unattended by the bees. Recently, the interest in ecologically safer alternatives to chemical insecticides has increased due to the significant success achieved with entomopathogenic microorganisms in the control of several lepidopteran pest larvae with minimum or no harm to the bees, natural rivals, animals, and humans. The current study was conducted to investigate the pathogenicity of two entomopathogenic fungus isolates (Beauveria bassiana G-A and G-B) isolated from dead G. mellonella larvae and their efficacy in pest control under laboratory conditions. Results Morphological and molecular identification revealed that the two isolates corresponded to B. bassiana species G-A and G-B strains. The response to the inoculation with the two fungal strains was conducted on G. mellonella larvae at 1 × 105–9 conidia/ml concentrations. The concentrations led to 96.54 and 89.66% mortality in G-A and G-B B. bassiana isolates at the highest concentration on day 10, respectively. LC50 was calculated between 0.2 × 106 (0.03 – 1.6) and 0.6 × 106 (0.07 – 6.1) conidia/ml. Conclusion The present study findings demonstrated that these isolates had the potential for G. mellonella control and B. bassiana isolates were a safe alternative to chemical control and could be recommended for use to protect stored wax products.

Published
2022
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