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http://dx.doi.org/10.4014/jmb.0810.595

Diversity of Bacillus thuringiensis Strains Isolated from Citrus Orchards in Spain and Evaluation of Their Insecticidal Activity Against Ceratitis capitata

J.C., Vidal-Quist (Centro de Proteccion Vegetal y Biotecnologia, IVIA, Ctra. Moncada-Naquera km)
Castanera, P. (Departamento Biologia de Plantas, CIB, CSIC, C/ Ramiro de Maeztu)
Gonzalez-Cabrera, J. (Centro de Proteccion Vegetal y Biotecnologia, IVIA, Ctra. Moncada-Naquera km)

Publication Information

Journal of Microbiology and Biotechnology / v.19, no.8, 2009 , pp. 749-759 More about this Journal

Abstract

A survey of Bacillus thuringiensis (Berliner) strains isolated from Spanish citrus orchards has been performed, and the strains were tested for insecticidal activity against the Mediterranean fruit fly Ceratitis capitata (Wiedemann), a key citrus pest in Spain. From a total of 150 environmental samples, 376 isolates were selected, recording a total B. thuringiensis index of 0.52. The collection was characterized by means of phase-contrast microscopy, SDS-PAGE, and PCR analysis with primer pairs detecting toxin genes cry1, cry2, cry3, cry4, cry5, cry7, cry8, cry9, cry10, cry11, cry12, cry14, cry17, cry19, cry21, cry27, cry39, cry44, cyt1, and cyt2. Diverse crystal inclusion morphologies were identified: bipyramidal (45%), round (40%), adhered to the spore (7%), small (5%), and irregular (3%). SDS-PAGE of spore-crystal preparations revealed 39 different electrophoresis patterns. All primer pairs used in PCR tests gave positive amplifications in strains of our collection, except for primers for detection of cry3, cry19, cry39, or cry44 genes. Strains containing cry1, cry2, cry4, and cry27 genes were the most abundant (48.7%, 46%, 11.2%, and 8.2% of the strains, respectively). Ten different genetic profiles were found, although a total of 109 strains did not amplify with the set of primers used. Screening for toxicity against C. capitata adults was performed using both spore-crystal and soluble fractions. Mortality levels were less than 30%. We have developed a large and diverse B. thuringiensis strain collection with huge potential to control several agricultural pests; however, further research is needed to find out Bt strains active against C. capitata.

Keywords

Bacillus thuringiensis; Mediterranean fruit fly; cry; cyt; biological control; PCR; SDS-PAGE; phase-contrast microscopy; bioassays;

Citations & Related Records

Times Cited By Web Of Science : 6 (Related Records In Web of Science)

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