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Characterization of a Novel Bacillus thuringiensis Phenotype Possessing Multiple Appendages Attached to a Parasporal Body

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Abstract

Bacillus thuringiensis is a bacterium best known for its production of crystal-like bodies comprised of one or more Cry-proteins, which can be toxic to insects, nematodes or cancer cells. Although strains of B. thuringiensis have occasionally been observed with filamentous appendages attached to their spores, appendages in association with their parasporal bodies are extremely rare. Herein we report the characterization of Bt1-88, a bacterial strain isolated from the Caribbean that produces a spore–crystal complex containing six long appendages, each comprised of numerous thinner filaments approximately 10 nm in diameter and 2.5 μm in length. Each of the multi-filament appendages was attached to a single, small parasporal body located at one end of the bacterial spore. Biochemical tests, 16S rDNA gene sequencing, and the identification of two Cry proteins by partial protein sequencing (putatively Cry1A and Cry2A), unambiguously identified Bt1-88 as a strain of B. thuringiensis. Bt1-88 represents the second reported strain of B. thuringiensis possessing a parasporal body/appendage phenotype characterized by one or more long appendages, comprised of numerous filaments in association with a parasporal body. This finding suggests that Bt1-88 is a member of a new phenotypic class of B. thuringiensis, in which the parasporal body may perform a novel structural role through its association with multi-filament appendages.

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References

  1. Chaufax J, Marchal M, Gilois N, Jehanno I, Buisson C (1997) Investigation of natural strains of Bacillus thuringiensis in different biotypes throughout the world. Can J Microbiol 43:337–343

    Article  Google Scholar 

  2. Crickmore N, Zeigler DR, Feitelson J, Schnepf E, Van Rie J, Lereclus D, Baum J, Dean DH (1998) Revision of the nomenclature for the Bacillus thuringiensis pesticidal crystal proteins. Microbiol Mol Biol Rev 62:807–813

    CAS  PubMed  Google Scholar 

  3. Akiba T, Abe Y, Kitada S, Kusaka Y, Ito A, Ichimatsu T, Katayama H, Akao T, Higuchi K, Mizuki E, Ohba M, Kanai R, Harata K (2009) Crystal structure of the parasporin-2 Bacillus thuringiensis toxin that recognizes cancer cells. J Mol Biol 386:121–133

    Article  CAS  PubMed  Google Scholar 

  4. Rampersad J, Khan A, Ammons D (2003) A Bacillus thuringiensis isolate possessing a spore-associated filament. Curr Microbiol 47:355–357

    Article  CAS  PubMed  Google Scholar 

  5. Smirnova TA, Kulinich LI, Galperin MY, Azizbekyan RR (1991) Subspecies-specific haemagglutination patterns of fimbriated Bacillus thuringiensis spores. FEMS Microbiol Lett 90:1–4

    Article  CAS  Google Scholar 

  6. DesRosier JP, Lara JC (1981) Isolation and properties of pilli from spores of Bacillus cereus. J Bacteriol 146:613–619

    Google Scholar 

  7. Mizuki E, Ohba M, Ichimatsu T, Hwang SH, Higuchi K, Saitoh H, Akao T (1998) Unique appendages associated with spores of Bacillus cereus. J Basic Microbiol 38:33–39

    Article  CAS  PubMed  Google Scholar 

  8. Sthalteim T, Granum PE (2001) Characterization of spore appendages from Bacillus cereus strains. J Appl Microbiol 91:839–845

    Article  Google Scholar 

  9. Rode LJ, Crawford MA, Williams MG (1967) Clostridium spores with ribbon-like appendages. J Bacteriol 93:1160–1173

    CAS  PubMed  Google Scholar 

  10. Pope L, Yolton DP, Rode LJ (1967) Appendages of Clostridium bifermentans spores. J Bacteriol 94:1206–1215

    CAS  PubMed  Google Scholar 

  11. Pope L, Rode LJ (1969) Spore fine structure in Clostridium cochlearium. J Bacteriol 100:994–1001

    CAS  PubMed  Google Scholar 

  12. Samsonoff WA, Hashimoto T, Conti SF (1970) Ultrastructural changes associated with germination and outgrowth of an appendage-bearing clostridial spore. J Bacteriol 101:1038–1045

    CAS  PubMed  Google Scholar 

  13. Rampersad J, Ammons D (2005) A Bacillus thuringiensis isolation method utilizing a novel stain, low selection and high throughput produced atypical results. BMC Microbiol 5:52–62

    Article  PubMed  Google Scholar 

  14. Reyes A, Ibarra JE (2004) Fingerprinting of Bacillus thuringiensis type strains and isolates by using Bacillus cereus group-group repetitive extragenic palindromic sequence-based PCR analysis. Appl Environ Microbiol 71:1346–1355

    Article  Google Scholar 

  15. Heddi A, Grennier AM, Khatchadourian C, Charles H, Nardon P (1999) Four intracellular genomes direct weevil biology: nuclear, mitochondrial, principal endosymbiont, and wolbachia. Evolution 96:6814–6819

    CAS  Google Scholar 

  16. Hall TA (1999) BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symp Ser 41:95–98

    CAS  Google Scholar 

  17. Tamura K, Dudley J, Nei MK (2007) MEGA4: molecular evolutionary genetics analysis (MEGA) software version 4.0. Mol Biol Evol 24:1596–1599

    Article  CAS  PubMed  Google Scholar 

  18. Thompson JD, Higgins DG, Gibson TJ, Clustal W (1994) Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680

    Article  CAS  PubMed  Google Scholar 

  19. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    CAS  PubMed  Google Scholar 

  20. Felsenstein J (1985) Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39:783–789

    Article  Google Scholar 

  21. Altschul SF, Gish W, Miller W, Myers EW, Lipman DJ (1990) Basic local alignment search tool. J Mol Biol 215:403–410

    CAS  PubMed  Google Scholar 

  22. Laemmli JK, Favre M (1973) Maturation of the head of bacteriophage T4 I. DNA packing agents. J Mol Biol 80:575–599

    Article  CAS  PubMed  Google Scholar 

  23. Reynolds ES (1963) The use of lead citrate at a high pH as electron-opaque stain in electron microscopy. J Cell Biol 17:208–212

    Article  CAS  PubMed  Google Scholar 

  24. Rampersad J (2006) A study on the isolation of Bacillus thuringiensis from the environment: a critical look at library construction. PhD thesis, The University of the West Indies, Trinidad and Tobago

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Acknowledgments

LIRA and RCC thanks financial support for this research, from Secretaría de Investigación y Posgrado, Instituto Politécnico Nacional, through grants SIP 20070722, SIP 20082599 and SIP 20090912. AVS was supported by scholarships from CONACYT and PIFI. LIRA and RCC are recipients of CONACYT, COFAA and EDI fellowships. JNR thanks the Faculty Research Council at the University of Texas- Pan American for financial support of work described in this Report. We would like to acknowledge the technical assistance of Dr. Klaus Linse, University of Texas, Austin, in performing N-terminal protein sequencing.

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Authors and Affiliations

  1. Departamento de Microbiología, Escuela Nacional de Ciencias Biológicas del I.P.N, Carpio y Plan de Ayala S/N, Casco de Santo Tomas, Mexico DF, 11340, Mexico

    Antonio Ventura-Suárez, Ramón Cruz-Camarillo & Luz I. Rojas-Avelizapa

  2. Central de Microscopía, Escuela Nacional de Ciencias Biológicas del I.P.N, Carpio y Plan de Ayala S/N, Casco de Santo Tomas, Mexico DF, 11340, Mexico

    Edgar O. López-Villegas

  3. Department of Chemistry, University of Texas–Pan American, 1201 West University Drive, Edinburg, TX, 78539, USA

    Joanne Rampersad & David R. Ammons

  4. Laboratorio de Bioinsecticidas, CINVESTAV-Irapuato. Km 9.6 Lib. Norte Carretera Irapuato-León, Irapuato, Guanajuato, 36500, Mexico

    Jorge E. Ibarra

Authors
  1. Antonio Ventura-Suárez
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  2. Ramón Cruz-Camarillo
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  3. Joanne Rampersad
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  4. David R. Ammons
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  5. Edgar O. López-Villegas
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  6. Jorge E. Ibarra
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  7. Luz I. Rojas-Avelizapa
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Corresponding author

Correspondence to Luz I. Rojas-Avelizapa.

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Ventura-Suárez, A., Cruz-Camarillo, R., Rampersad, J. et al. Characterization of a Novel Bacillus thuringiensis Phenotype Possessing Multiple Appendages Attached to a Parasporal Body. Curr Microbiol 62, 307–312 (2011). https://doi.org/10.1007/s00284-010-9678-2

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  • DOI: https://doi.org/10.1007/s00284-010-9678-2

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