Abstract
Insecticidal crystal proteins (ICPs) produced by Bacillus thuringiensis (Bt) exhibit strong toxicity. Soil bacteriophages destroy the ICPs in nature. Also, environmental pH, temperature, and ultraviolet (UV) radiation shorten the ICPs’ validity and infectivity. To enhance the validity of ICPs of Bt, the soil Bt phages and the environmental parameters such as soil pH, temperature, and UV should be subjected to continuous evaluation.
In this study, five Bt bacteriophages were isolated, characterized, and named BtØ3, BtØ5, BtØ7, BtØ9, and BtØ11. Electron microscopy investigation showed that the five phages have an icosahedral head and a long contractile tail. In addition, the restriction endonuclease BamHI enzyme cleaves the phage genomic DNA suggesting that all five phages have double-stranded DNA (dsDNA) belonging to the order Caudovirales. The various inter simple sequence repeat restriction patterns suggested that the five phages genetically are not similar, and similarity metrics analysis placed the five phages into two clusters.
The reported lytic activity of phages against Bt was as follows: BtØ7 (100%), BtØ9 (100%), BtØ3(83%), BtØ5(83%), and BtØ11(75%). Moreover, the phages were 17% more effective in lysing Bt than the commercial antibiotics.
Bt phages isolated from this study highlighted the importance of regular assessment of soil conditions and the lytic potentials of naturally occurring Bt phages to protect Bt sp from being attacked or destroyed, and to calculate the exact Bt dose concentration of successful application in pest control, this will enhance the environmental health, food security, and crop safety.
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Data Availability
All data sets are presented in the main manuscript.
Abbreviations
- Bt:
-
Bacillus thuringiensis
- ICP:
-
Insecticidal crystal protein
- CFU:
-
Colony-forming unit
- SM:
-
Sodium chloride, Magnesium sulfate, and gelatin used for routine manipulation of phage suspensions
- %:
-
Percent
- LB:
-
Luria broth
- NA:
-
Nutrient Agar
- Hrs:
-
Hours
- ISSRs:
-
Inter simple sequence repeats
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Acknowledgements
We thank Dr. Rashid I. H. Ibrahim, Department of Biological Sciences, College of Science, King Faisal University, Saudi Arabia for proofreading the manuscript and language editing.
Funding
This work was supported by the Deanship of Scientific Research, Vice Presidency for Graduate Studies and Scientific Research, King Faisal University, Saudi Arabia [Project No. GRANT2435].
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A.A.H. planned, designed, and executed the experimental work of phage isolation & characterization, and wrote the preliminary results. I.M. constructed the experiments of gene typing using ISSRs-PCR analysis. E.A. set the work idea, revised the results, wrote, peer viewed, edited the manuscript in its ready-to-publish version, and landed the grant to support this work. All authors have read and agreed to the published version of the manuscript.
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Hassan, A.A., Mohamed, I. & Afkar, E. Characterization of Bacillus thuringiensis bacteriophages: morphogenesis, lytic potentials and inter simple sequence repeat analysis. Biologia 78, 3625–3635 (2023). https://doi.org/10.1007/s11756-023-01501-8
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DOI: https://doi.org/10.1007/s11756-023-01501-8