ORIGINAL ARTICLE
Isolation, identification and pathogenicity assessment of a new isolate of entomopathogenic fungus, Beauveria bassiana in Iran
 
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Department of Plant Protection, Faculty of Agriculture, Urmia University, Urmia, Iran
CORRESPONDING AUTHOR
Safavi Seyed Ali
Department of Plant Protection, Faculty of Agriculture, Urmia University, Urmia, Iran
 
Journal of Plant Protection Research 2010;50(2):158–163
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ABSTRACT
A new isolate (BEH) of entomopathogenic fungus, Beauveria bassiana was isolated from soil using DOC2 selective medium. This isolate was characterized by conidiophores consisting of whorls and dense clusters of short conidiophorous cells with one-celled spherical conidia. Conidial length and width were 2.27±0.22 μm and 1.85±0.32 μm, respectively with length/width ratio of 1.23. Colonies on SDYA medium were normally white to pale yellow and sometimes red pigmented in reverse. Because of importance of this pathogen in biocontrol programs around the world and difficulties with morphological identification, a molecular technique was developed to assist complementary identification of the fungus. Pr1, a pathogenicity-related alkaline cuticle-degrading serine protease, with defined sequence in B. bassiana was amplified using PCR technique. The presence of this gene in isolated fungus (BEH) with 744 bp sequence length, as visualized on agarose gel affirmed the data from morphological studies that the new isolate (BEH) pertained to entomopathogenic fungus, B. bassiana. Pathogenicity of new isolate against Tenebrio molitor and its recovering was the other confirmation that the isolated fungus belonged to B. bassiana, using further light microscope studies.
CONFLICT OF INTEREST
The authors have declared that no conflict of interests exist.
 
REFERENCES (40)
1.
Almeida J.E.M., Alves S.B., Pereira R.M. 1997. Selection of Beauveria spp. isolates for control of the termite Heterotermes tenuis (Hagen, 1858). J. Appl. Entomol. 121: 539–543.
 
2.
Bidochka M.J., Khachatourians G.G. 1987. Purification, properties of an extracellular protease produced by the entomopathogenic fungus Beauueria bassiana. Appl. Environ. Microbiol. 53 (7): 1679–1684.
 
3.
Bidochka M.J., Khachatourians G.G. 1990. Identification of Beauveria bassiana extracellular protease as a virulence factor in pathogenicity toward the migratory grasshopper, Melanoplus sanguinipes. J. Invertebr. Pathol. 56: 362–370.
 
4.
Bidochka M.J., McDonald M.A., St. Leger R.J., Roberts D.W. 1994. Differentiation of species and strains of entomopathogenic fungi by random amplification DNA (RAPD). Curr. Genet. 25: 107–113.
 
5.
Boucias D.G., Boucias D., Pendland J.C. 1998. Principles of Insect Pathology. Springer, 537 pp.
 
6.
Brady B.L.K. 1979a. Beauveria bassiana. CMI descriptions of pathogenic fungi and bacteria, No. 602. p. 1964–1995. Commonwealth Mycological Institute, Kew, Surrey, England.
 
7.
Brady B.L.K. 1979b. Beauveria brogniartii. CMI descriptions of pathogenic fungi and bacteria, No. 602, Commonwealth Mycological Institute, Kew, Surrey, England.
 
8.
Charnley A.K. 1997. Entomopathogenic fungi and their role in pest control. p. 185–201. In: “The Mycota IV Environmental and Microbial Relationships” (D.T. Wicklow, B.E. Soderstorm, eds.). Springer, Berlin, Germany.
 
9.
Charnley A.K. 2003. Fungal pathogens of insects: cuticle degrading enzymes and toxins. Adv. Botanical Res. 40: 241–321.
 
10.
Charnley A.K., St. Leger R.J. 1991. The role of cuticle-degrading enzymes in fungal pathogenesis in insects. p. 267–287. In: “Fungal Spore Disease Initiation in Plants and Animals” (E.T. Cole, H.C. Hoch, eds.). Plenum Press, New York, 555 pp.
 
11.
Chrzanowska J., Banas J., Kolaczkowska M. 2001. Purification and characterization of Beauveria bassiana proteinases. Acta Biotechnol. 21: 73–81.
 
12.
Clarkson J.M., Charnley A.K. 1996. New insights into the mechanisms of fungal pathogenesis in insects. Trends Microbiol. 4 (5): 197–203.
 
13.
Feng M.G., Poprawski T.J., Khachatourians G.G. 1994. Production of formulation and application of the entomopathogenic fungus Beauveria bassiana for the insect control: current status. Biocontrol Sci. Technol. 4: 3–34.
 
14.
Glare T.R., Inwood A.J. 1998. Morphological and genetic characterisation of Beauveria spp. from New Zealand. Mycol. Res. 102 (2): 250–256.
 
15.
Gupta S.C., Leathers T.D., El-Sayed G.N., Ignoffo C.M. 1992. Insect cuticle-degrading enzymes from the entomopathogenic fungus Beauveria bassiana. Exp. Mycol. 16: 132–137.
 
16.
Hajek A.E., St. Leger R.J. 1994. Interactions between fungal pathogenesis and insect hosts. Annu. Rev. Entomol. 39: 293–322.
 
17.
Hassan A.E.M., Charnley A.K. 1989. Ultrastructural study of the penetration by Metarhizium anisopliae through Dimilin-affected cuticle of Manduca sexta. J. Invertebr. Pathol. 54: 117–124.
 
18.
Hegedes D.D., Khachatourians G.G. 1993. Construction of cloned DNA probes for the specific detection of the entomopathogenic fungus Beauveria bassiana in grasshoppers. J. Invertebr. Pathol. 62: 233–240.
 
19.
Hegedes D.D., Khachatourians G.G. 1996. Identification and differentiation of the entomopathogenic fungus Beauveria bassiana using polymerase chain reaction and single-strand conformation polymorphism analysis. J. Invertebr. Pathol. 67: 289–299.
 
20.
Joshi L., St. Leger R.J., Bidochka M.J. 1995. Cloning of a cuticledegrading protease from the entomopathogenic fungus, Beauveria bassiana. FEMS Microbiol. Lett. 125: 211–218.
 
21.
Kim H.K., Hoe H.S., Suh D.S., Kang S.C., Hwang C., Kwon S.T. 1999. Gene structure and expression of the gene from Beauveria bassiana encoding bassiasin I, an insect cuticle-degrading serine protease. Biotechnol. Lett. 21: 777–783.
 
22.
Kosir J.M., MacPherson J.M., Khachatourians G.G. 1991. Genomic analysis of a virulent and a less virulent strain of the entomopathogenic fungus Beauveria bassiana, using restriction fragment length polymorphisms. Can. J. Microbiol. 37: 534–541.
 
23.
Maurer P., Couteaudier Y., Girard P.A., Bridge P.D., Riba G. 1997. Genetic diversity of Beauveria bassiana and relatedness to host insect range. Mycol. Res. 101: 159–164.
 
24.
Moino J.R., Alves S.B., Pereira R.M. 1998. Efficacy of Beauveria bassiana (Balsamo) Vuillemin isolates for control of storedgrain pests. J. Appl. Entomol. 122: 301–305.
 
25.
Möller E.M., Bahnveg G., Sandermann H., Geiger H.H. 1992. A simple and efficient protocol for isolation of high molecular weight DNA from filamentous fungi, fruit bodies, and infected plant tissues. Nucleic Acid Res. 20 (22): 6115–6116.
 
26.
Mugnai L., Bridge P.D., Evans H.C. 1989. A chemotaxonomic evaluation of the genus Beauveria. Mycol. Res. 92: 199–209.
 
27.
Paterson I.C., Charnley A.K., Cooper R.M., Clarkson J.M. 1994. Specific induction of a cuticle-degrading protease of the insect pathogenic fungus Metarhizium anisopliae. Microbiology 140: 185–189.
 
28.
Samson R.A., Evans H.C., Latge J.P. 1988. Atlas of Entomopathogenic Fungi. Springer-Verlag, Berlin, 187 pp.
 
29.
Shimazu M., Sato H. 1996. Media for selective isolation of an entomogenous fungus, Beauveria bassiana (Deutromycotina: Hyphomycetes). Appl. Entomol. Zool. 31 (2): 291–298.
 
30.
Shimizu S., Tsuchitani Y., Matsumoto T. 1993. Production of an extracellular protease by Beauveria bassiana in the haemolymph of the silkworm, Bombyx mori. Lett. Appl. Microbiol. 16: 291–294.
 
31.
St. Leger R.J., Allee L.L., May R., Staples R.C., Roberts D.W. 1992. World-wide distribution of genetic variation among isolates of Beauveria spp. Mycol. Res. 96: 1007–1015.
 
32.
St. Leger R.J., Charnley A.K., Cooper R.M. 1986a. Cuticle-degrading enzymes of entomopathogenic fungi: Mechanisms of interaction between pathogen enzymes and insect cuticle. J. Invertebr. Pathol. 47: 295–302.
 
33.
St. Leger R.J., Charnley A.K., Cooper R.M. 1986b. Cuticle-degrading enzymes of entomopathogenic fungi: Synthesis in culture on cuticle. J. Invertebr. Pathol. 48: 85–95.
 
34.
St. Leger R.J., Cooper R.M., Charnley A.K. 1987. Distribution of chymoelastases and trypsin-like enzymes in five species of entomopathogenic deuteromycetes. Arch. Biochem. Bioph. 258: 123–131.
 
35.
St. Leger R.J., Joshi L., Bidochka M.J., Roberts D.W. 1996. Construction of an improved mycoinsecticide overexpressing a toxic protease. Proc. of the National Academy of Sciences of the United States of America 93: 6349–6354.
 
36.
Townsend R.J., Glare T.R., Willoughby B.E. 1995. The fungi Beauveria spp. causes grass grub population collapse in some Waikato pastures. Proc. of the 48th New Zealand Plant Protection Conference 48: 237–241.
 
37.
Urtz B.E., Rice W.C. 2000. Purification and characterization of a novel extracellular protease from Beauveria bassiana. Mycol. Res. 104: 180–186.
 
38.
Viaud M., Couteaudier Y., Levis C., Riba G. 1996. Genome organization in Beauveria bassiana: electrophoretic karyotype, gene mapping and telomeric fingerprint. Fungal Genet. Biol. 20: 175–183.
 
39.
Wang C., Shah F.A., Patel N., Li Z., Butt T.M. 2003. Molecular investigation on strain genetic relatedness and population structure of Beauveria bassiana. Environ. Microbiol. 5 (10): 908–915.
 
40.
Zacharuk R.Y. 1970. Fine structure of the fungus Metarhizium anisopliae infecting three species of larval Elateridae. III. Penetration of the host integument. J. Invertebr. Pathol. 15: 372–396.
 
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