Molecular characterization of a Colombian Bacillus thuringiensis strain with activity against Tecia solanivora (Lepidoptera: Gelechiidae)
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The Guatemalan potato moth, Tecia solanivora (Lepidoptera: Gelechiidae), is one of the most important pests affecting potatoes in northern South America, causing crop losses ranging from 50 to 100%. In this work, we isolated a native strain of Bacillus thuringiensis (146-15801) with high activity against this moth, and we characterized its Cry genes. This strain was the most active out of eleven strains tested against T. solanivora in laboratory bioassays. Specific PCR and genome walk allowed the isolation of Cry genes showing 99 and 95% similarity to the sequences reported in Cry1Ac and Cry2 genes, respectively. Immunolocation experiments demonstrated that Cry1Ac and Cry2 bind to the midgut epithelial cells, suggest that these two proteins are involved in the insecticidal activity against T. solanivora. Results obtained in this work are relevant because native strains comprise a source of biological components for the development of new bioinsecticides, as well as new Cry genes that can be used for biological control.
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 fieldcollected strains of Bacillus thuringiensis. Applied and Environmental Microbiology 63 (12): 4883-4890.
BRADFORD, M. M. 1976. A rapid and sensitive method for the quantization of microgram quantities of protein utilizing the principle of protein-dye binding. Analytical Biochemistry 72: 248-254.
CRICKMORE, N.; ZEIGLER, D. R.; FEITELSON, J.; SCHNEPF, E.; VAN RIE, J.; LERECLUS, D.; BAUM, J.; DEAN, D. H. 1998. Revision of the nomenclature for the Bacillus thuringiensis pesticidal crystal proteins. Microbiology and Molecular Biology Reviews 62 (3): 807-813
DE MAAGD, R. A.; DIRK, B.; WILLEM, S. 1999. Bacillus thuringiensis toxinmediated insect resistance in plants. Trends in Plant Science 4 (1): 9-13.
DUNNETT, C. W. 1955. A multiple comparison procedure for comparing several treatments with a control. Journal of the American Statistical Association 50:1096-1121.
FINNEY, D. 1971. Probit analysis. Cambridge University Press (Londres) 400 p.
GILLILAND, A.; CHAMBERS, C.; BONE, E.; ELLAR, D. 2002. Role of Bacillus thuringiensis Cry1 δ-endotoxin binding in determining potency during Lepidopteran larval development. Applied and Environmental Microbiology 68: 1509-1515.
GROCHULSKI, P.; MASSON, L.; BORISOVA, S.; PUSZTAICAREY, M.; SCHWARTZ, J. L.; BROUSSEAU, R.; CYGLER, M. 1995. Bacillus thuringiensis CryIA(a) insecticidal toxin: crystal structure and channel formation. Journal of Molecular Biology 254 (3): 447-464.
HALL, T. A. 1999. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium Series 41: 95-98.
LONDOÑO, M.; MONTOYA, O. 1994. Cría de Tecia solanivora Povolny para ensayos biológicos. ICA-CORPOICA (Colombia). 35 p.
MacLEOD, A. 2005. Data sheets on quarantine pests: Tecia solanivora. European and Mediterranean Plant Protection Organization 35: 399-401.
ORDUZ, S.; ROJAS, W.; CORREA, M. M.; MONTOYA, A. E.; DE BARJAC, H. 1992. A new serotype of Bacillus thuringiensis from Colombia toxic to mosquito larvae. Journal of Invertebrate Pathology 59: 99-103.
PEFEROEN, M. 1997. Progress and prospects for field use of Bt genes in crops. Trends in Biotechnology 15 (5): 173-177.
POVOLNY, D. 1973. Scrobipalpopsis solanivora sp. n.–A new pest of potato (Solanum tuberosum) from Central America. Acta Universitatis Agriculturae, Facultas Agronomica 21: 143-146.
SAMBROOK, J.; RUSSELL, D. 2001. Molecular cloning. A laboratory manual. 3a ed. Cold Spring Harbor Laboratory Press. New York. 1252 p.
SCHNEPF, E.; CRICKMORE, N.; VAN RIE, J.; LERECLUS, D.; BAUM, J.; FEITELSON, J.; ZEIGLER, D.; DEAN, D. 1998. Bacillus thuringiensis and its pesticidal crystal proteins. Microbiology and Molecular Biology Reviews 62 (3): 775-806.
SIEBERT, P.; CHENCHIK, A.; KELLOGG, D.; LUKYANOV, K.; LUKYANOV, S. 1995. An improved PCR method for walking in uncloned genomic DNA. Nucleic Acids Research 23 (6): 1087-1088.
VALDERRAMA, A.; VELASQUEZ, N.; RODRIGUEZ, E.; ZAPATA, A.; ZAIDI, M.; ALTOSAAR, I.; ARANGO, R. 2007. Resistance to Tecia solanivora (Lepidoptera: Gelechiidae) in three transgenic andean varieties of Potato expressing Bacillus thuringiensis Cry1Ac protein. Journal of Economic Entomology 100 (1): 172-179.
ZEDDAM, J.; ORBE, K.; LÉRY, X.; DANGLES, O.; DUPAS, S.; SILVAIN, J. 2008. An isometric virus of the potato tuber moth Tecia solanivora (Povolny) (Lepidoptera: Gelechiidae) has a trisegmented RNA genome. Journal of Invertebrate Pathology 99 (2): 204-211.
- NORA RESTREPO, DIANA GUTIÉRREZ, MARÍA MERCEDES PATIÑO, ISABELLE THIÉRY, ARMELLE DELÉCLUSE, SERGIO ORDUZ, Cloning, expression and toxicity of a mosquitocidal toxin gene of Bacillus thuringiensis subsp. medellin , Revista Colombiana de Entomología: Vol. 23 No. 2 (1997)
- SERGIO ORDUZ, NORA RESTREPO, THAIS DÍAZ, JUAN S. ZULUAGA, WILLIAM ROJAS, FIVE NEW STRAINS OF LARVA PATHOGEN FUNGUS OF MOSQUITO, Lagenidium giganteum, OF COLOMBIA , Revista Colombiana de Entomología: Vol. 19 No. 3 (1993)
- SERGIO ORDUZ, MARGARITA CORREA, ASTRID ELENA MONTOYA, WILLIAM ROJAS, A NEW ISOLATION OF Bacillus thuringiensis FROM COLOMBIA, TOXIC FOR MOSQUITO LARVES , Revista Colombiana de Entomología: Vol. 16 No. 2 (1990)
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