Abstract
Despite the successful application of toxins from Bacillus thuringiensis as biological control agents against pests, pests are showing resistance against an increasing number of Bacillus thuringiensis toxins due to evolution; thus, new toxins with higher toxicity and broad-spectrum activity against insects are being increasingly identified. To find new toxins, whole genome sequencing of the novel B. thuringiensis strain Bt S3076-1 was performed, and ten predicted toxic genes were identified in this study, including six cry genes, two tpp genes, one cyt gene and one vip gene, among which six were novel toxins. Subsequently, SDS‒PAGE analysis showed that the major proteins at the spore maturation stage were approximately 120 kDa, 70 kDa, 67 kDa, 60 kDa and 40 kDa, while active proteins after trypsin digestion (approximately 70 kDa and 40 kDa) exhibited LC50 values of 149.64 μg/g and 441.47 μg/g against Spodoptera frugiperda and Helicoverpa armigera larvae, respectively. Furthermore, pathological observation results showed that the peritrophic membrane of Spodoptera frugiperda and Helicoverpa armigera larvae was degraded. These findings will provide an experimental reference for further research on the insecticidal activity, toxicity spectrum and synergism of these toxins in Bt S3076-1.
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Abbreviations
- Bt:
-
Bacillus thuringiensis
- °C:
-
Degree centigrade
- RH:
-
Relative humidity
- pH:
-
Potential of hydrogen
- rpm:
-
Revolution per minute
- h:
-
Hour/hours
- min:
-
Minutes
- g:
-
Gram/grams
- µg:
-
Microgram
- ml:
-
Millilitres
- µl:
-
Microlitres
- M:
-
Molar
- mM:
-
Millimolar
- V:
-
Ultraviolet
- Kb:
-
Kilobase
- bp:
-
Base pair
- kDa:
-
Kilo Dalton
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Acknowledgements
This work was supported by grants Guangxi Minzu University Introduction of Talent Research Start-up Fund (2020KJQD23), Guangxi Natural Science Foundation (Grant No.2022GXNSFBA035446), The Basic Ability Enhancement Program for Young and Middle-aged Teachers of Guangxi (2022KY0159).
Funding
This study was funded by grants Guangxi Minzu University Introduction of Talent Research Start-up Fund (2020KJQD23), Guangxi Natural Science Foundation (Grant No.2022GXNSFBA035446), The Basic Ability Enhancement Program for Young and Middle-aged Teachers of Guangxi (2022KY0159).
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TY completed the strain culture, and proteome identification and their analysis conducted the prokaryotic expression of the predicted proteins, performed the larvae bioassays experiments, and wrote and revised the whole manuscript. ZW and LL complete genome sequenced and made genome-wide analysis of strain Bt S3076-1. MJ and XF were involved in project design and manuscript modification. WH participated in the bioassay analysis. YZ is responsible for writing, revising, and finalizing the manuscript. All authors have read and approved the final manuscript.
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Yang, T., Wu, Z., Li, L. et al. Identification and analysis of toxins in novel Bacillus thuringiensis strain Bt S3076-1 against Spodoptera frugiperda and Helicoverpa armigera (Lep.: Noctuidae). Arch Microbiol 205, 168 (2023). https://doi.org/10.1007/s00203-023-03490-3
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DOI: https://doi.org/10.1007/s00203-023-03490-3