Fluorescent amplified fragment length polymorphism analysis of Bacillus anthracis, Bacillus cereus, and Bacillus thuringiensis isolates

• Published in: Applied and Environmental Microbiology Vol. 70; no. 2; pp. 1068 - 1080
• Main Authors: Hill, K.K, Ticknor, L.O, Okinaka, R.T, Asay, M, Blair, H, Bliss, K.A, Laker, M, Pardington, P.E, Richardson, A.P, Tonks, M
• Format: Journal Article
• Language: English
• Published: Washington, DC American Society for Microbiology 01-02-2004
• Subjects: amplified fragment length polymorphism; animal pathogenic bacteria; Animals; Bacillus anthracis; Bacillus anthracis - classification; Bacillus anthracis - genetics; Bacillus cereus; Bacillus cereus - classification; Bacillus cereus - genetics; Bacillus thuringiensis; Bacillus thuringiensis - classification; Bacillus thuringiensis - genetics; bacterial insecticides; Biological and medical sciences; Deoxyribonuclease EcoRI - metabolism; Deoxyribonucleases, Type II Site-Specific - metabolism; Deoxyribonucleic acid; DNA; DNA Fingerprinting - methods; DNA, Bacterial - analysis; DNA, Bacterial - genetics; entomopathogenic bacteria; Fluorescence; food pathogens; Fundamental and applied biological sciences. Psychology; Genetics; Genetics and Molecular Biology; Humans; Microbiology; Phylogeny; Polymorphism, Restriction Fragment Length; Serotyping; soil bacteria; Soils; strains; Bacillus anthracis; Bacillus cereus; Bacillales; Bacillus thuringiensis; Fluorescence; Bacteria; Isolate; Bacillaceae; Fragmentation; Polymorphism
• ISSN: 0099-2240; 1098-5336
• DOI: 10.1128/AEM.70.2.1068-1080.2004

DNA from over 300 Bacillus thuringiensis, Bacillus cereus, and Bacillus anthracis isolates was analyzed by fluorescent amplified fragment length polymorphism (AFLP). B. thuringiensis and B. cereus isolates were from diverse sources and locations, including soil, clinical isolates and food products causing diarrheal and emetic outbreaks, and type strains from the American Type Culture Collection, and over 200 B. thuringiensis isolates representing 36 serovars or subspecies were from the U.S. Department of Agriculture collection. Twenty-four diverse B. anthracis isolates were also included. Phylogenetic analysis of AFLP data revealed extensive diversity within B. thuringiensis and B. cereus compared to the monomorphic nature of B. anthracis. All of the B. anthracis strains were more closely related to each other than to any other Bacillus isolate, while B. cereus and B. thuringiensis strains populated the entire tree. Ten distinct branches were defined, with many branches containing both B. cereus and B. thuringiensis isolates. A single branch contained all the B. anthracis isolates plus an unusual B. thuringiensis isolate that is pathogenic in mice. In contrast, B. thuringiensis subsp. kurstaki (ATCC 33679) and other isolates used to prepare insecticides mapped distal to the B. anthracis isolates. The interspersion of B. cereus and B. thuringiensis isolates within the phylogenetic tree suggests that phenotypic traits used to distinguish between these two species do not reflect the genomic content of the different isolates and that horizontal gene transfer plays an important role in establishing the phenotype of each of these microbes. B. thuringiensis isolates of a particular subspecies tended to cluster together.