Comparative Analysis of Genetic Diversity Among Bacillus Thuringiensis and Bacillus Sphaericus and Their Fusants Using Molecular Markers
Biochemistry and Molecular Biology
Volume 3, Issue 5, September 2018, Pages: 63-70
Received: Nov. 10, 2018; Accepted: Dec. 7, 2018; Published: Jan. 10, 2019
Views 686      Downloads 101
El-Kawokgy Tahany Mohamed Abou El-Fath, Microbial Genetics Department, National Research Centre, Giza, Egypt
Ibrahim Shafik Darwish, Molecular Genetics and Genome Mapping Department, Agricultural Genetic Engineering Research Institute (AGERI), Agricultural Research Center (ARC), Giza, Egypt
Ashraf Gamil Attallah, Microbial Genetics Department, National Research Centre, Giza, Egypt
Article Tools
Follow on us
Ten genetically stable fusants strains were obtained as a result of protoplast fusion technique between Bacillus sphaericus (Bs) and Bacillus thuringiensis (Bt), six of them produced from, Bs GHAI Bt 66 Ia experiment (fusants no. 1, 2, 16, 26, 31 and 34) and four fusants produced from Bs GHAI Bt I977experiment (fusants no.1, 2, 17 and 25). Two molecular marker systems, four ISSR primers (Inter-Simple Sequence Repeat) and seven SCoT primers (Start Codon Targeted Polymorphism) were employed for detection of genetic variation between the parents and their fusants. Four ISSR primers and seven SCoT detected 55and 97 amplicons, among which 92.696% and 81.948% bands were polymorphic respectively. The genetic similarity values among the species were calculated based on ISSR and SCoT profiles which ranged from 0.50 to 0.83 and 0.59 to 0.90 respectively. Based on the marker analysis, the three parents and the ten fusants were clustered into three major groups for ISSR primer and SCoT primers. Cluster analysis were performed to construct a dendrogram of the Bacillus Strains and their fusants and also reveals high genetic variation among the parents and their fusants. It could be concluded that each type of the two molecular marker approaches of DNA analysis could identify the different between the parents, and all of their fusants under investigation have probably originated from closely related ancestors and possess high degree of genetic similarity.
Bacillus Thuringiensis, Bacillus Sphaericus, Genetic Diversity, ISSR and SCoT Marker
To cite this article
El-Kawokgy Tahany Mohamed Abou El-Fath, Ibrahim Shafik Darwish, Ashraf Gamil Attallah, Comparative Analysis of Genetic Diversity Among Bacillus Thuringiensis and Bacillus Sphaericus and Their Fusants Using Molecular Markers, Biochemistry and Molecular Biology. Vol. 3, No. 5, 2018, pp. 63-70. doi: 10.11648/j.bmb.20180305.11
Copyright © 2018 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License ( which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Fraser, M. P., Yue, Z. W. and Buzcu, B. (2003). Source apportionment of fine particulate matter in Houston, TX, using organic molecular markers. Atmos. Environ. 37, 2117–2123.
Hibbett, D. S., Binder, M., Bischoff, J. F., Blackwell, M., Cannon, P. F., Eriksson, O. E., Huhndorf, S., James, T., Kirk, P. M., Lücking, R., et al. (2007). A higher-level phylogenetic classification of the Fungi. Mycol. Res. 111, 509–547.
Zhang, Y., Yan, H., Jiang, X., Wang, X., Huang, L., Xu, B., Zhang, X. and Zhang, L. (2016). Genetic variation, population structure and linkage disequilibrium in Switchgrass with ISSR, SCoT and EST-SSR markers. Hereditas 153.
Petrovičová, L., Balážová, Ž., Vivodík, M. and Gálová, Z. (2017). Detection genetic variability of secale cereale l. By scot markers. Potravinarstvo Slovak J. Food Sci. 11, 197–202.
Gorji, A. M., Poczai, P., Polgar, Z. and Taller, J. (2011). Efficiency of Arbitrarily Amplified Dominant Markers (SCOT, ISSR and RAPD) for Diagnostic Fingerprinting in Tetraploid Potato. Am. J. Potato Res., Springer-Verlag 88, 226–237.
Ng, W. L. and Tan, S. G. (2015). Inter-Simple Sequence Repeat (ISSR) markers: Are we doing it right? ASM Sci. J. 9, 30–39.
Abdel-Mawgood, A. L. (2012). DNA Based Techniques for Studying Genetic Diversity. Genet. Divers. Microorg. 30.
Anne, C. (2006). Choosing the right molecular genetic markers for studying biodiversity: from molecular evolution to practical aspects. Genetica, Kluwer Academic Publishers 127, 101–120.
Shafiei-Astani, B., Ong, A. H. K., Valdiani, A., Tan, S. G., Yien, C. Y. S., Ahmady, F., Alitheen, N. B., Ng, W. L. and Kuar, T. (2015). Molecular genetic variation and structure of Southeast Asian crocodile (Tomistoma schlegelii): Comparative potentials of SSRs versus ISSRs. Gene 571, 107–116.
Shen, J., Ding, X., Liu, D., Ding, G., He, J., Li, X., Tang, F. and Chu, B. (2006). Intersimple Sequence Repeats (ISSR) Molecular Fingerprinting Markers for Authenticating Populations of Dendrobium officinale. Biol. Pharm. Bull. 29, 420–422.
Iruela, M., Rubio, J., Cubero, J. I., Gil, J. and Millán, T. (2002). Phylogenetic analysis in the genus Cicer and cultivated chickpea using RAPD and ISSR markers. Theor. Appl. Genet. 104, 643–651.
El-Kawokgy, T. M. A., Zowail, M. E. M. and Hegazy, W. K. (2004). Genetic improvement of Bacillus thuringiensis as a biocontrol agent against Biomphalaria alexandrina snail. Mansoura Univ. J. Agric. Sci. Egypt.
El-Kawokgy, T. M. A., Hussein, H. A., Aly, N. A. H. and Mohamed, S. A. H. (2014). Highly toxic and broad-spectrum insecticidal local Bacillus strains engineered using protoplast fusion. Can. J. Microbiol. 61, 38–47.
Hussein H. A, Kawokgy, T. A. M, Aly, N. A. H and Mohamed, S. A. H. (2014) Genetic modification of Bacillus Strains with a wide spectrum of insecticidal activities. J Genet Eng Biotechnol 8, 33–42.
Attallah, A. G., Abo-Serreh, N. and Abd-El-Aal, S. K. (2014). Molecular Characterization of Beauvaria sp. with Inter Simple Sequence Repeat (ISSR) and RAPD Markers. Int. J. ChemTech Res. 6, 1407–1415.
Ruas, P. M., Ruas, C. F., Rampim, L., Carvalho, V. P., Ruas, E. A. and Sera, T. (2003). Genetic relationship in Coffea species and parentage determination of interspecific hybrids using ISSR (Inter- Simple Sequence Repeat) markers. Genet. Mol. Biol. 26, 319–327.
Hatti, A. D., Taware, S. D., Taware, A. S., Pangrikar, P. P., Chavan, A. M. and Mukadam, D. S. (2010). Genetic-diversity-of-toxigenic-and-non-toxigenic-Aspergillus-flavus-strains-using-ISSR-markers. African Journal of Biotechnology Vol. 2 (7), pp. 194–197.
Ali, B. A. (2003). Genetics similarity among four breeds of sheep in Egypt detected by random amplified polymorphic DNA markers. Afr. J. Biotechnol. 2, 194-197.
Williams, J. G. K., Kubelik, A. R., Livak, K. J., Rafalski, J. A. and Tingey, S. V. (1990). DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18, 6531–6535.
Omear, H. A. (2009). Using The RAPD Markers To Analyze Variation Among Some Species Of The Genus Alternaria, PhD Thesis, MSc. thesis-Ccollage of Science–University of Tikrit. Iraq.
Kernodle, SP, RE Cannon and JG Scandalios, Kernodle, SP, and RE Cannon and JG Scandalios. (1993). Concentration of primer and template qualitatively affects product in RAPD-PCR. Biotechniques 1, 362–364.
Abdulateef, S. M., Aljubori, M. H. and Abdulbaqi, N. J. (2014). Genetic Diversity Among Some Aspergillus flavus Isolates by Using Inter simple sequence repeats (ISSR). Iraqi J. Sci. 55, 8.
Raghunathachari, P., Khanna, V. K., Singh, U. S. and Singh, N. K. (2000). RAPD analysis of genetic variability in Indian scented rice germplasm (Oryza sativa L.). Curr. Sci. 79, 994–998.
Saker, M. M., Youssef, S. S., Abdallah, N. A., Bashandy, H. S. and Sharkawy, M. El. (2005). Genetic analysis of some Egyptian rice genotypes using RAPD, SSR and AFLP 9. African Journal of Biotechnology Vol. 4 (9), 882-890.
Abdulateef, S. M., Aljubori, M. H. and Abdulbaqi, N. J. (2014). Genetic Diversity Among Some Aspergillus flavus Isolates by Using Inter simple sequence repeats (ISSR). Iraqi J. Sci. Vol 55, No.3A,:986-993.
Sawant, S. V., Singh, P. K., Gupta, S. K., Madnala, R. and Tuli, R. (1999). Conserved nucleotide sequences in highly expressed genes in plants. J. Genet. 78, 123–131.
Collard, B. C. Y. and Mackill, D. J. (2009). Start Codon Targeted (SCoT) Polymorphism: A Simple, Novel DNA Marker Technique for Generating Gene-Targeted Markers in Plants. Plant Mol. Biol. Report. 27, 86–93.
Gupta, P. K., Varshney, R. K., Sharma, P. C. and Ramesh, B. (1999). Molecular markers and their applications in wheat breeding. Plant Breed. 118, 369–390.
Joshi, A. K. and Schabes, Y. (1997). Tree-Adjoining Grammars. In Handbook of Formal Languages: Volume 3 Beyond Words (Rozenberg, G., and Salomaa, A., eds.), pp 69–123, Springer Berlin Heidelberg, Berlin, Heidelberg.
Sommer, R. and Tautz, D. (1989). Minimal homology requirements for PCR primers. Nucleic Acids Res. 17, 6749.
Kwok, S., Kellogg, D. E., McKinney, N., Spasic, D., Goda, L., Levenson, C. and Sninsky, J. J. (1990). Effects of primer-template mismatches on the polymerase chain reaction: Human immunodeficiency virus type 1 model studies. Nucleic Acids Res. 18, 999–1005.
Ophel, K. and Kerr, A. (1990). Agrobacterium vitis sp. nov. for Strains of Agrobacterium biovar 3 from Grapevines. Int. J. Syst. Evol. Microbiol. 40, 236–241.
Mackill, D. J. (1995). Classifying Japonica Rice Cultivars with RAPD Markers. Crop Sci. 35, 889–894.
Kaushik, A., Saini, N., Jain, S., Rana, P., Singh, R. K. and Jain, R. K. (2003) Genetic analysis of a CSR10 (indica) × Taraori Basmati F3 population segregating for salt tolerance using ISSR markers. Euphytica 134, 231.
EL-Shaer, H. F; Attallah, A. G; and AL-Namouly, Sh.D. (2014) Genetic Diversity Based on SCoT and ISSR Markers in Agrobacterium isolated from Egyptian 5, 1605–1616.
Science Publishing Group
1 Rockefeller Plaza,
10th and 11th Floors,
New York, NY 10020
Tel: (001)347-983-5186