Molecular Characterisation of Soil-Dwelling Bacillus thuringiensis using Transcriptional Regulator, XRE Gene and the Crystal Protein, cry2 gene
DOI:
https://doi.org/10.47430/ujmr.2161.019Keywords:
Bacillus thuringiensis, transcriptional regulator, XRE, crystal protein, cry2Abstract
Bacillus thuringiensis (Bt) is the organism that is used most frequently in biological pest management, which is distinguished by the capacity to possess crystalline inclusions throughout the sporulation phase. There is an increasing need to use biological control in controlling plant pathogens due to the inherent advantages. However, the detection of Bt has become more time consuming and cumbersome due to the numerous available crystal genes. The goal of the study was to isolate strains of Bacillus thuringiensis from the soil, characterise the isolates using the transcriptional regulator, XRE gene and the crystal proteins cry2gene and compare the efficiency of these two biomarkers in identifying Bt species. Five different Bacillus thuringiensis strains were isolated from soil samples in Zaria, Nigeria. Polymerase chain reaction was used to detect the existence of the cry2 and XRE genes. Four (80%) of the five isolates harboured the XRE genes, while none (0%) harboured the cry2 genes. This observation is a likely indication that the XRE gene is a reliable biomarker in the identification of Bt isolates from environmental samples. In order to ensure speed and reproducibility in the detection of Bt from environmental samples, molecular techniques targeting the XREgene are recommended.
References
Adeyemo, I. A., Abdul-Wahab, S. O., & Obadofin, A. A. (2018). Biocontrol potential of Bacillus thuringiensis isolated from soil samples against mosquito larvae. Ife Journal of Science, 20(2), [Page range?]. ?] DOI: https://doi.org/10.4314/ijs.v20i2.8
Bartoszewicz, M., & Marjanska, P. S. (2017). Milk-originated Bacillus cereus sensu lato strains harbouring Bacillus anthracis-like plasmids are genetically and phenotypically diverse. Food Microbiology, 64, 23–30. DOI: https://doi.org/10.1016/j.fm.2017.05.009
Bello, A., & Hussaini, I. M. (2017). Detection of multiple cry genes in Bacillus thuringiensis isolated from different soil types in Zaria. Microbiology Research Journal International, 20(2), 1–6. ?] DOI: https://doi.org/10.9734/MRJI/2017/30397
Ben-Dov, E., Zaritsky, A., Dahan, E., Barak, Z., Sinai, R., Manasherob, R., Khamraev, A., Troitskaya, E., Dubitsky, A., Berezina, N., & Margalith, Y. (1997). Extended screening by PCR for seven cry-group genes from field-collected strains of Bacillus thuringiensis. Applied and Environmental Microbiology, 63(12), 4883–4890. ?] DOI: https://doi.org/10.1128/aem.63.12.4883-4890.1997
Bergey, D. H. (2004). Bergey’s manual of determinative bacteriology (9th ed.). Williams & Wilkins.
Carozzi, N. B., Kramer, V. C., Warren, G. W., Evola, S., & Koziel, M. G. (1991). Prediction of insecticidal activity of Bacillus thuringiensis strains by polymerase chain reaction product profiles. Applied and Environmental Microbiology, 57(11), 3057–3061. ?] DOI: https://doi.org/10.1128/aem.57.11.3057-3061.1991
Crickmore, N. (2020). Bacillus thuringiensis toxin nomenclature.
Crickmore, N., Zeigler, D. R., Schnepf, E., Van Rie, J., Lereclus, D., Baum, J., Bravo, A., & Dean, D. H. (2011). Bacillus thuringiensis toxin nomenclature.
Dharmender, K., Kamla, C., & Boora, K. S. (2008). Characterization of native Bacillus thuringiensis strains by PCR-RAPD based fingerprinting. Indian Journal of Microbiology, 48(1), 124–127. ?]
Eswarapriya, B., Gopalsamy, B., Kameswari, B., Meera, R., & Devi, P. (2010). Insecticidal activity of Bacillus thuringiensis IBT-15 strain against Plutella xylostella. International Journal of PharmTech Research, 2(3), 2048–2053.
Fiuza, L. M. (2015). Thuringiensin: A toxin from Bacillus thuringiensis. Bt Research, 6(4), 1–12.
Frankenhuyzen, K. V. (2009). Insecticidal activity of Bacillus thuringiensis crystal proteins. Journal of Invertebrate Pathology, 101(1), 1–16. DOI: https://doi.org/10.1016/j.jip.2009.02.009
Helgason, E., Økstad, O. A., Caugant, D. A., Johansen, H. A., Fouet, A., Mock, M., Hegna, I., & Kolstø, A. B. (2000). Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis—One species on the basis of genetic evidence. Applied and Environmental Microbiology, 66(6), 2627–2630. DOI: https://doi.org/10.1128/AEM.66.6.2627-2630.2000
Jain, D., Sunda, S. D., Sanadhya, S., Nath, D. J., & Khandelwal, S. K. (2017). Molecular characterization and PCR-based screening of cry genes from Bacillus thuringiensis strains. 3 Biotech, 7(1), 4. DOI: https://doi.org/10.1007/s13205-016-0583-7
Kaur, S. (2002). Potential for developing novel Bacillus thuringiensis strains and transgenic crops and their implications for Indian agriculture. Agricultural Biotech Net, 4(120), 1–10.
Liang, H., Liu, Y., Zhu, J., Guan, P., Li, S., Wang, S., & Zheng, A. (2011). Characterization of cry2-type genes of Bacillus thuringiensis strains from soil-isolated of Sichuan basin, China. Brazilian Journal of Microbiology, 42(1), 140–146. DOI: https://doi.org/10.1590/S1517-83822011000100018
Liu, Y., Lai, Q., Göker, M., Meier-Kolthoff, J. P., Wang, M., Sun, Y., Wang, L., & Shao, Z. (2015). Genomic insights into the taxonomic status of the Bacillus cereus group. Scientific Reports, 5, 14082. DOI: https://doi.org/10.1038/srep14082
Lone, S. A., Yadav, R., Malik, A., & Padaria, J. C. (2016). Molecular and insecticidal characterization of Vip3A protein producing Bacillus thuringiensis strains toxic against Helicoverpa armigera (Lepidoptera: Noctuidae). Canadian Journal of Microbiology, 62(2), 179–190. DOI: https://doi.org/10.1139/cjm-2015-0328
Lone, S. A., Malik, A., & Padaria, J. C. (2017). Characterization of lepidopteran-specific cry1 and cry2 gene harbouring native Bacillus thuringiensis isolates toxic against Helicoverpa armigera. Biotechnology Reports, 15, 27–32. DOI: https://doi.org/10.1016/j.btre.2017.05.001
Rasko, D. A., Altherr, M. R., Han, C. S., & Ravel, J. (2005). Genomics of the Bacillus cereus group of organisms. FEMS Microbiology Reviews, 29(2), 303–329. DOI: https://doi.org/10.1016/j.fmrre.2004.12.005
Roh, J. Y., Choi, J. Y., Li, M. S., Jin, B. R., & Je, Y. H. (2017). Bacillus thuringiensis as a specific, safe, and effective tool for insect pest control. Journal of Microbiology and Biotechnology, 17(4), 547–559.
Rolle, R. L., Ejiofor, A. O., & Johnson, T. L. (2005). Determination of the plasmid size and location of d-endotoxin genes of Bacillus thuringiensis by pulse field gel electrophoresis. African Journal of Biotechnology, 4(6), 580–585. DOI: https://doi.org/10.5897/AJB2005.000-3106
Schünemann, R., Knaak, N., & Fiuza, L. M. (2014). Mode of action and specificity of Bacillus thuringiensis toxins in the control of caterpillars and stink bugs in soybean culture. ISRN Microbiology, 2014, 135675. DOI: https://doi.org/10.1155/2014/135675
Stefani, F. O. P., Bell, T. H., Marchand, C., de la Providencia, I. E., Yassimi, A. E., St-Arnaud, M., & Hijri, M. (2015). Culture-dependent and -independent methods capture different microbial community fractions in hydrocarbon-contaminated soils. PLOS ONE, 10(6), e0128272. DOI: https://doi.org/10.1371/journal.pone.0128272
Travers, R. S., Martin, P. A. W., & Reichelderfer, C. F. (1987). Selective process for efficient isolation of soil Bacillus species. Applied and Environmental Microbiology, 53(6), 1263–1266. ?] DOI: https://doi.org/10.1128/aem.53.6.1263-1266.1987
Vilas-Bôas, G. T., & Lemos, M. V. F. (2004). Diversity of cry genes and genetic characterization of Bacillus thuringiensis isolated from Brazil. Canadian Journal of Microbiology, 50(9), 605–613. DOI: https://doi.org/10.1139/w04-052
Wei, S., Chelliah, R., Park, B. J., Kim, S. H., Forghani, F., Cho, M. S., Park, D. S., Jin, Y. G., & Oh, D. H. (2019). Differentiation of Bacillus thuringiensis from Bacillus cereus group using a unique marker based on real-time PCR. Frontiers in Microbiology, 10, 883. DOI: https://doi.org/10.3389/fmicb.2019.00883
Willey, J. M., Sherwood, L. M., & Woolverton, C. J. (2008). Prescott, Harley, and Klein’s microbiology (7th ed.). McGraw-Hill Higher Education.
Downloads
Published
Issue
Section
License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.









