Abstract

  There is an ever growing need for control of insects of agricultural and health importance. The practice of using chemicals for this purpose has been flawed due to many untoward fall outs including insect resistance, non-target toxicity and non-biodegradability. A promising and viable practice is the use of biological agents such as Bacillus thuringiensis (Bt) that are efficacious as well as safe. Bacillus thuringiensis (Bt) was first isolated in the early 1900s, and interest in it and its used have soared as a result of its insecticidal capability. Bt is a Gram-positive, endospore-forming organism that produces crystal proteins that serve as toxins to select insects. Thus, its use as a bioinsecticide has exploded prompting the search for newer isolates and the establishment of culture collections of Bt strains in many laboratories. The purpose of the work presented here was to assess the biochemical profile of 72 strains collected in Middle Tennessee as well as those of two standard strains, B. t. israelensis (BtI) and B. t. kurstaki (BtK) as a method for their characterization. The approach was to determine the biochemical profile of each strain using the Biolog Identification System, which allowed for the assessment of the metabolic activity of each strain in the presence of 95 different substrates. It was found that the strains clustered into two major groups having common substrates, and these groups gave two and three sub-clusters. Thus, it was concluded that the strains of this study had varying metabolic ability for the substrates provided in the Biolog Gen III Microplates; however, there were certain substrates that were commonly used by the strains used in this study.

Keywords: Bacillus thuringiensis; Biolog Identification System; Gen III Microplates; Biochemical Profile

References

1. Alper Mehlika., et al. “Distribution, occurrence of cry genes, and lepidopteran toxicity of native Bacillus thuringiensis isolated from fig tree environments in Aydın Province”. Turkish Journal of Agriculture and Forestry 38.6 (2014): 898-907.
2. Armada E., et al. “Native plant growth promoting bacteria Bacillus thuringiensis and mixed or individual mycorrhizal species improved drought tolerance and oxidative metabolism in Lavandula dentata plants”. Journal of Plant Physiology 192 (2016): 1-12.
3. Badran, Ahmed H., et al. “Continuous evolution of Bacillus thuringiensis toxins overcomes insect resistance”. Nature 533.7601 (2016): 58.
4. Bassi, Daniela., et al. “Transcriptome analysis of Bacillus thuringiensis spore life, germination and cell outgrowth in a vegetable-based food model”. Food Microbiology 55 (2016): 73-85.
5. Ben-Dov Eitan. "Bacillus thuringiensis subsp. israelensis and its dipteran-specific toxins”. Toxins 6.4 (2014): 1222-1243.
6. Blackburn, Michael B., et al. “Phylogenetic distribution of phenotypic traits in Bacillus thuringiensis determined by multilocus sequence analysis”. PloS one 8.6 (2013): e66061.
7. Buentello-Wong S., et al. “Characterization of Cry Proteins in Native Strains of Bacillus thuringiensis and Activity Against Anastrepha ludens1”. Southwestern Entomologist 40.1 (2015): 15-25.
8. Chouikha Iman and B Joseph Hinnebusch. "Silencing urease: a key evolutionary step that facilitated the adaptation of Yersinia pestis to the flea-borne transmission route”. Proceedings of the National Academy of Sciences 111.52 (2014): 18709-18714.
9. Deng, Chao., et al. “Division of labour and terminal differentiation in a novel Bacillus thuringiensis strain”. The ISME Journal 9.2 (2015): 286.
10. Ejiofor AO and T Johnson. "Physiological and molecular detection of crystalliferous Bacillus thuringiensis strains from habitats in the SouthCentral United States”. Journal of Industrial Microbiology and Biotechnology 28.5 (2002): 284-290.
11. El-Ghany., et al. “Molluscicidal activity of Bacillus thuringiensis strains against Biomphalaria alexandrina snails”. Beni-Suef University Journal of Basic and Applied Sciences 6.4 (2017): 391-393.
12. Fagerlund Annette., et al. “Sin R controls enterotoxin expression in Bacillus thuringiensis biofilms”. PLoS One 9.1 (2014): e87532.
13. Gassmann Aaron J., et al. “Field-evolved resistance by western corn rootworm to multiple Bacillus thuringiensis toxins in transgenic maize”. Proceedings of the National Academy of Sciences 111.14 (2014): 5141-5146.
14. Guinebretière Marie-Hélène., et al. “Bacillus cytotoxicus sp. nov. is a novel thermotolerant species of the Bacillus cereus group occasionally associated with food poisoning”. International Journal of Systematic and Evolutionary Microbiology 63.1 (2013): 31-40.
15. Hollensteiner Jacqueline., et al. “Bacillus thuringiensis and Bacillus weihenstephanensis inhibit the growth of phytopathogenic Verticillium species”. Frontiers in Microbiology 7 (2017): 2171.
16. Jia Yanhua., et al. “A genetically modified broad-spectrum strain of Bacillus thuringiensis toxic against Holotrichia parallela, Anomala corpulenta and Holotrichia oblita”. World Journal of Microbiology and Biotechnology 30.2 (2014): 595-603.
17. Jiménez Guillermo., et al. “Description of Bacillus toyonensis sp. nov., a novel species of the Bacillus cereus group, and pairwise genome comparisons of the species of the group by means of ANI calculations”. Systematic and Applied Microbiology 36.6 (2013): 383-391.
18. Kamar Rita., et al. “Pathogenic potential of Bacillus cereus strains as revealed by phenotypic analysis”. Journal of Clinical Microbiology 51.1 (2013): 320-323.
19. Kim Min-Ju., et al. “Various enterotoxin and other virulence factor genes widespread among Bacillus cereus and Bacillus thuringiensis strains”. Journal of Microbiology and Biotechnology 25.6 (2015): 872-879.
20. Marchlewicz Ariel., et al. “Bacillus thuringiensis B1 (2015b) is a gram-positive bacteria able to degrade naproxen and ibuprofen”. Water, Air, and Soil Pollution 227.6 (2016): 197.
21. Masri Leila., et al. “Host–pathogen coevolution: the selective advantage of Bacillus thuringiensis virulence and its cry toxin genes”. PLoS Biology 13.6 (2015): e1002169.
22. Mukherjee Krishnendu., et al. “Experimental evolution of resistance against Bacillus thuringiensis in the insect model host Galleria mellonella results in epigenetic modifications”. Virulence 8.8 (2017): 1618-1630.
23. Oves Mohammad., et al. “Biosorption of heavy metals by Bacillus thuringiensis strain OSM29 originating from industrial effluent contaminated north Indian soil”. Saudi Journal of Biological Sciences 20.2 (2013): 121-129.
24. Palma Leopoldo., et al. “Bacillus thuringiensis toxins: an overview of their biocidal activity”. Toxins 6.12 (2014): 3296-3325.
25. Palma Leopoldo., et al. “Draft genome sequences of two Bacillus thuringiensis strains and characterization of a putative 41.9-kDa insecticidal toxin”. Toxins 6.5 (2014): 1490-1504.
26. Pardo-Lopez., et al. “Bacillus thuringiensis insecticidal three-domain Cry toxins: mode of action, insect resistance and consequences for crop protection”. FEMS Microbiology Reviews 37.1 (2013): 3-22.
27. Peng Dong-hai., et al. “The expression and crystallization of Cry65Aa require two C-termini, revealing a novel evolutionary strategy of Bacillus thuringiensis Cry proteins”. Scientific Reports 5 (2015): 8291.
28. Shu Changlong., et al. “Use of a pooled clone method to isolate a novel Bacillus thuringiensis Cry2A toxin with activity against Ostrinia furnacalis”. Journal of Invertebrate Pathology 114.1 (2013): 31-33.
29. Surhio Muhammad Ali., et al. “Complete degradation of dimethyl phthalate by biochemical cooperation of the Bacillus thuringiensis strain isolated from cotton field soil”. RSC Advances 4.99 (2014): 55960-55966.
30. Zhu Lei., et al. “Genomic and transcriptomic insights into the efficient entomopathogenicity of Bacillus thuringiensis”. Scientific Reports 5 (2015): 14129.

Citation

Citation: Fahdah Alshammari., et al. “Determination of the Biochemical Profile of Bacillus thuringiensis Strains Isolated from Middle Tennessee". Acta Scientific Microbiology 3.3 (2020): 01-05.

Copyright

Copyright: © 2020 Fahdah Alshammari., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.