Isolation, characterization and serotype classification of Bacillus thuringiensis from different soil samples of Fars province
Subject Areas :
Molecular Microbiology
Elham Moazamian
1
,
Nima Bahador
2
,
Manochehr Rasouli
3
,
Negar Azarpira
4
1 - Islamic Azad University, Science and Research Branch, Fars, Iran
2 - Islamic Azad University, Science and Research Branch, Fars, Iran
3 - Department of Immonology, Professor Alborzi Clinical Microbiology Research Center, Shiraz
University of Medical Sciences, Shiraz, Iran
4 - Organ Transplant Research Center, Shiraz University of Medical Sciences, Shiraz, Iran
Received: 2010-03-16
Accepted : 2010-04-14
Published : 2010-04-01
Keywords:
SDS-PAGE,
Bacillus thuringiensis,
Cry protein,
Serotyping,
Abstract :
Background and Objective: Bacillus thuringiensis (Bt) is a bacterium best known for its production of crystal-like bodies comprised of one or more crystal proteins in sporulation phase, which can be toxic to insects, nematodes and cancer cells. The purpose of this study is isolation, characterization and comparison of Cry proteins in the isolated strains obtained in different parts of Fars province. Material and Methods: 25 soil samples were collected from six different habitats in Fars province using seven different methods. Characterization was performed based on biochemical tests, protein crystal morphology by phase-contrast microscope and variation of cry protein toxin using SDS-PAGE. Results: The tests showed a highest count of Bt strains in soil samples obtained from Shiraz city. The Bt colonies were classified according to biochemical tests such as: sucrose and salicin fermentation, lecithinase production and esculin hydrolyzing. Conclusion: Results showed that although isolated strains have a variety of protein bands with different sizes, the proteins with 34KD was observed following proteinase K digestion in all the samples.
References:
Vidal-Quist JC, Castanera P, Gonzalez-Cabrera J. Diversity of Bacillus thuringiensis strains isolated from citrus orchards in Spain and evaluation of their insecticidal activity against Ceratitis capitata. Microbiol Biotechhnol. 2009; 19(8):749-759.
Yu J, Tan L, Liu Y. Phylogenetic analysis of Bacillus thuringiensis based of PCR amplified fragment polymorphism of flagellin genes. Curr Microbiol. 2002; 45: 139-143.
Mizuki E, Park YS, Saitoh H, Yamashita S, Akaot H, Higuchi K, Ohba M. Parasporin, a human leukemic cell-recognizing parasporal protein of Bacillus thuringiensis. Clin Diag lab Imm. 2000; 7(4): 625-634.
Kondo S, Mizuki E, Akao T, Ohba M. Antitrichomonal strains of Bacillus thuringiensis. Parasitol Res. 2002; 88: 1090-1092.
Uemori A, Maeda M, Yasutake K, Ohgushi A, Kagoshima K, Mizuki E, Ohba M. Ubiquity of parasporin-1 producers in Bacillus thuringiensis natural populations of Japan. Naturwissenschaften. 2007; 94:34-38.
Yamashita S, Katayama H, Saitoh H, Akao T, Park YS, Mizuki E, Ohba M, Ito A. typical three-domain Cry proteins of Bacillus thuringiensis strain A1462 exhibit cytocidal activity on limited human cancer cells. J Biochem. 2005; 138 (6): 663-672.
Uemori A, Ohgush A, Yasutake K, Maeda M, Mizuki E, Ohba M. Parasporin-1Ab, a novel Bacillus thuringiensis cytotoxin preferentially active on human cancer cells In Vitro. Anticancer Research. 2008; 28: 91-96.
Apaydin O, Yenidunya AF, Harsa S, Gunes H. Isolation and characterization of bacillus thuringiensis strains from different grain habitats in Turkey. World J Microbi Biotechnol. 2005; 21: 285-292.
Santana MA, Moccia CC, Gillis AE. Bacillus thuringiensis improved isolation methodology from soil samples. J Microbiol Met.2008; 75: 357-358.
Salehi Jouzani, G, Pourjan Abad A, Seifinejad A, Marzban R, Kariman K, Maleki B. Distribution and diversity of Dipteran-specific cry and cyt genes in native Bacillus thuringiensis strains obtained from different ecosystems of Iran. J Ind Microbiol Biotechnol. 2008; 35: 83-94.
Rampersad J, Ammons D. Bacillus thuringiensis isolation method utilizing a novel stain, low selection and high throughput produced atypical results. BMC Microbiol. 2005; 24:5:52.
Chatterjee SN, Bhattacharya T, Dangar TK, Chandra G. Ecology and diversity of Bacillus thuringiensis in soil environment. Afr J Biotech. 2007; 6 (13): pp. 1587-1591.
Travers RS, Martin PAW, Reichelderfer CF. Selective process for efficient isolation of Bacillus spp. Appl. Environ. Microbiol. 1987; 53: 1263-1266.
Lee DW, Akao T, Yamashita S, Katayama H, Maeda M, Saitoh H, Mizuki E, Ohba M. Non insecticidal parasporal proteins of a Bacillus thuringiensis isolate exhibit a preferential cytotoxicity against human leukemia T cells. Biochem Biophys comm. 2000; 272: 218-223.
Aramideh S, SaferalizadehMH, PourmirzaAA, Rezazadeh BariM, KeshavarziM, Mohseniazar M. Characterization and pathogenic evaluation of Bacillus thuringiensis isolates from West Azerbaijan province-Iran. AJMR. 2010; 4 (12):1224-1229.