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The molecular diversity of different isolates of Beauveria bassiana (Bals.) Vuill. as assessed using intermicrosatellites (ISSRs)

Cellular & Molecular Biology Letters volume 12, pages 240–252 (2007)Cite this article

Abstract

Inter-microsatellite PCR (ISSR-PCR) markers were used to identify and to examine the genetic diversity of eleven Beauveria bassiana isolates with different geographic origins. The variability and the phylogenetic relationships between the eleven strains were analyzed using 172 ISSR-PCR markers. A high level of polymorphism (near 80%) was found using these molecular markers. Seven different isolates showed exclusive bands, and ISSR primer 873 was able to distinguish between all the strains. The dendrogram obtained with these markers is robust and in agreement with the geographical origins of the strains. All the isolates from the Caribbean region were grouped together in a cluster, while the other isolates grouped in the other cluster. The similarity exhibited between the two clusters was less than 50%. This value of homology shows the high genetic variability detected between the isolates from the Caribbean region and the other isolates. ISSR-PCR markers provide a quick, reliable and highly informative system for DNA fingerprinting, and allowed the identification of the different B. bassiana isolates studied.

Abbreviations

AFLP:

amplified fragment length polymorphism

EST:

expressed sequence tag

ISSR:

inter simple sequence repeat

ITS:

internal transcribed spacer

PCR:

polymerase chain reaction

RAPD:

random amplified polymorphic DNA

RFLP:

restriction fragment length polymorphism

Rp:

resolving power

SSCP:

single-strand conformation polymorphism

SSR:

simple sequence repeat

References

  1. Padjama, V. and Kaur, G. Use of the fungus Beauveria bassiana (Bals.) Vuill (Moniliales: Deuteromycetes) for controlling termites. Curr. Sci. 81 (6) (2001) 645–647.

    Google Scholar 

  2. Ludwig, S.W. and Oetting, R.D. Efficacy of Beauveria bassiana plus insect attractants for enhanced control of Frankliniella occidentalis (Thysanoptera: Thripidae). Florida Entomologist 85 (2002) 270–272.

    Article  Google Scholar 

  3. Estrada, M.E. [Aplicación de Beuaveria bassiana (Bals.) Vuill. En la lucha biológica contra Diatraea saccharalis (Fabricius)]. Revista Caña de Azúcar del Fondo Nacional de Investigaciones Agropecuarias de Venezuela. 23 (1997) 12–19.

    Google Scholar 

  4. Rehner, S.A. Phylogenetics of the insect pathogenic genus Beauveria. In: Insect — fungal associations ecology and evolution. (Vega, E.F. and Blackwell, M. Eds.). Oxford University Press, New York, N.Y., 2005, 3–27.

    Google Scholar 

  5. Rehner, S.A. and Buckley, E. A Beauveria phylogeny inferred from nuclear ITS and EF1-α sequences: evidence for cryptic diversification and links to Cordyceps teleomorphs. Mycologia 97 (2005) 84–98.

    Article  CAS  PubMed  Google Scholar 

  6. Roddam, L.F. and Rath, A.C. Isolation and characterization of Metarhizium anisopliae and Beauveria basssiana from subantartic macquarie island. J. Invert. Pathol. 69 (1997) 285–288.

    Article  Google Scholar 

  7. Liu, H., Skinner, M., Bridge, M. and Parker, B.L. Characterization of Beauveria bassiana and Metarhizium anisopliae isolates for management of tarnished plant bug, Lygus lineolaris (Hemiptera: Miridae). J. Invert. Pathol. 82 (2003) 139–147.

    Article  Google Scholar 

  8. Wright, S.P., Carruthers, R.I., Bradley, C.A., Jaronski, S.T., Lacey, L.A., Wood, P. and Galaini-Wraight, S. Pathogenicity of the entomopathogenic fungi Paecilomyces spp. and Beauveria bassiana against the silver leaf whitefly, Bermisia argentifolii. J. Invert. Pathol. 71 (1998) 217–226.

    Article  Google Scholar 

  9. Castrillo, L.A. and Brooks, W.M. Differentiation of Beauveria bassiana isolates from the darkling beetle, Alphitobius diaperinus, using isozyme and RAPD analyses. J. Invert. Pathol. 72 (1998) 190–196.

    Article  Google Scholar 

  10. Hegedus, D.D., Pfeifer, T.A., Mulik, D.S. and Kachatourians, G.G. Characterization and structure of the mitochondrial small rRNA gene of the entomopathogenic fungus Beauveria basssiana. Genome 41 (1998) 471–476.

    Article  CAS  PubMed  Google Scholar 

  11. Castrillo, L.A., Wiehmann, B.M. and Brooks, W.M. Genetic variation in Beauveria bassiana populations associated with the darkling beetle, Alphitobius diaperinus. J. Invert. Pathol. 73 (1999) 269–275.

    Article  CAS  Google Scholar 

  12. Wang, C.S., Li, Z.Z. and Butt, T.M. Molecular studies of co-formulated strains of the entomopathogenic fungus, Beauveria bassiana. J. Invert. Pathol. 80 (2002) 29–34.

    Article  CAS  Google Scholar 

  13. Couteaudier, Y. and Viaud, M. News insights into populations structure of Beauveria bassiana with regard to vegetative compatibility groups and telomeric restriction fragment length polymorphisms. FEMS Microbial Ecology 27 (1997) 175–182.

    Article  Google Scholar 

  14. Berretta, M.F., Lecuona, R.E., Zandommeni, R.O. and Grau, O. Genotyping isolates of the enthomopathogenic fungus Beauveria bassiana by RAPD with fluorescent label. J. Invert. Pathol. 71 (1998) 145–150.

    Article  CAS  Google Scholar 

  15. Coates, B.S., Hellmich, R.L. and Lewis, L.C. Beauveria bassiana haplotype determination based on nuclear rDNA internal transcribed spacer PCRRFLP. Mycol. Res. 106 (2003) 40–50.

    Article  CAS  Google Scholar 

  16. Van, B.A. The role of short sequence repeats in epidemiologic typing. Current Opin. Microbiol. 2 (1999) 306–311.

    Article  Google Scholar 

  17. Enkerli, J., Widmer, F., Gessier, C. and Keller, S. Strain-specific microsatellite markers in the entomopathogenic fungus Beauveria brongniartii. Mycol. Res. 105 (2001) 1079–1087.

    CAS  Google Scholar 

  18. Viaud, M., Couteaudier, Y., Levis, C. and Riba, G. Genome organization in Beauveria bassiana: Electrophoretic karyotype, gene mapping, and telomeric fingerprint. Fungal Genet. Biol. 20 (1996) 175–183.

    Article  CAS  Google Scholar 

  19. Neuveglise, C. and Brigoo, Y. Identification of group-I introns in the 28s rDNA of the entomopathogenic fungus Beauveria brongniartii. Curr. Genet. 27 (1994) 38–45.

    Article  CAS  PubMed  Google Scholar 

  20. Poprawski, T.J., Riba G., Jones W.A. and Ajoun, A. Variation in isoesterase profiles of geographical opulations of Beauveria bassiana (Deuteromycotina: Hyphomycetes) isolated from Silona weevils (Coleoptera: Curculionidae). Environ. Entomol. 17 (1988) 275–279.

    Google Scholar 

  21. Mugnai, L., Bridge P.D. and Evans, H.C. A chemotaxonomic evaluation of the genus Beauveria. Mycol. Res. 92 (1989) 199–209.

    Article  Google Scholar 

  22. Williams, J.G.K., Kubelik, A.R., Livak, K.J., Rafalski, J.A. and Tingey, S.V. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Res. 18 (1990) 6531–6535.

    Article  CAS  PubMed  Google Scholar 

  23. Welsh, J. and McClelland, M. Fingerprinting genomes using PCR with arbitrary primers. Nucleic Acids Res. 18 (1990) 7213–7218.

    Article  CAS  PubMed  Google Scholar 

  24. Cravanzola, F., Piatti, P., Bridge, P.D. and Ozino, O.I. Detection of genetic polymorphism by RAPD-PCR in strains of the entomopathogenic fungus Beauveria brongniartii isolated from European cockchafer (Melolontha spp.). Lett. Appl. Microbiol. 25 (1997) 289–294.

    Article  CAS  Google Scholar 

  25. Piatti, P. Cravanzola, F., Bridge, P.D. and Ozino, O.I. Molecular characterization of Beauveria brongniartii isolates obtained from Melolontha melolontha in Valle d’Aosta (Italy) by RAPD-PCR. Lett. Appl. Microbiol. 26 (1998) 317.

    Article  CAS  Google Scholar 

  26. Burt, A., Carter, D.A., Koenig, G.L., White, T.J. and Taylor, J.W. Molecular markers reveal cryptic sex in the human pathogen Coccidioides immitis. Proc. Natl. Acad. Sci. U.S.A. 93 (1996) 770–773.

    Article  CAS  PubMed  Google Scholar 

  27. Geiser, D.M., Pitt, J.I. and Taylor, J.W. Cryptic speciation and recombination in the aflatoxin-producing fungus Aspergillus flavus. Proc. Natl. Acad. Sci. U.S.A. 95 (1998) 388–393.

    Article  CAS  PubMed  Google Scholar 

  28. Taylor, J.W., Jacobson, D.J. and Fisher, M.C. The evolution of asexual fungi: reproduction, speciation and classification. Annu. Rev. Phytopathol. 37 (1999) 187–246.

    Article  Google Scholar 

  29. Bidochka, M.J., Kamp, A.M., Lavender, T.M., Dekoning, J. and De Croos, J.N.A. Habitat association in two genetic groups of the insect-pathogenic fungus Metarhizium anisopliae: uncovering cryptic species? Appl. Environ. Microbiol. 67 (2001) 1335–1342.

    Article  CAS  PubMed  Google Scholar 

  30. Bidochka, M.J., Menzies, F.V. and Kamp, A.M. Genetic groups of the insect-pathogenic fungus Beauveria bassiana are associated with habitat and thermal growth preferences. Arch. Microbiol. 178 (2002) 531–537.

    Article  CAS  PubMed  Google Scholar 

  31. Steenkamp, E.M. and Wingfield, B.D. 2003. Cryptic speciation in Fusarium subglutinans. Mycologia 94 (2003) 1032–1043.

    Article  Google Scholar 

  32. Uma Devi, K., Reineke, A., Nageswara Rao Reddy, N., Uma Maheswara Rao, C. and Padmavathi, J. Genetic diversity, reproductive biology, and speciation in the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin. Genome 49 (2006) 495–504.

    Article  PubMed  CAS  Google Scholar 

  33. Zietkiewicz, E., Rafalski, A. and Labuda, D. Genome fingerprinting by simple sequence repeat (SSR)-anchored polymerase chain reaction amplification. Genomics 20 (1994) 176–183.

    Article  CAS  PubMed  Google Scholar 

  34. Kantety, R.V., Zeng, X.P., Bennetzen, J.L. and Zehr, B.E. Assessment of genetic diversity in Dent and Popcorn (Zea mays L.) inbred lines using intersimple sequence repeat (ISSR) amplification. Mol. Breed. 1 (1995) 365–373.

    Article  CAS  Google Scholar 

  35. Pejic, I., Ajmone-Marsan, P., Morgante, M., Kozumplick, V., Castiglioni, P., Taramino, G. and Motto, M. Comparative analysis of genetic similarity among maize inbred lines detected by RFLPs. Theor. Appl. Genet. 98 (1998) 1248–1255.

    Article  Google Scholar 

  36. Fang, D.Q., Roose, M.L., Krueger, R.R. and Federicé, C.T. Fingerprinting trifoliate orange germplasm accessions with isozymes, RFLPs and intersimple sequence repeat markers. Theor. Appl. Genet. 95 (1997) 211–219.

    Article  CAS  Google Scholar 

  37. Nagaoka, T. and Ogihara, Y. Applicability of inter-simple sequence repeat polymorphisms in wheat for use as DNA markers in comparison to RFLP and RAPD markers. Theor. Appl. Genet. 94 (1997) 597–602.

    Article  CAS  Google Scholar 

  38. Prevost, A. and Wilkinson, M.J. A new system of comparing PCR primers applied to ISSR fingerprinting of potato cultivars. Theor. Appl. Gen. 98 (1999) 107–112.

    Article  CAS  Google Scholar 

  39. Martín, J.P. and Sánchez-Yélamo, M.D. Genetic relationships among species of the genus Diplotaxis (Brassicaceae) using inter-simple sequence repeat markers. Theor. Appl. Genet. 101 (2000) 1234–1241.

    Article  Google Scholar 

  40. Métais, I., Aubry, C., Hamon, B. and Jalouzot, R. Description and analysis of genetic diversity between commercial bean lines (Phaseolus vulgaris L.). Theor. Appl. Genet. 101 (2000) 1207–1214.

    Article  Google Scholar 

  41. Matos, M., Pinto-Carnide, O. and Benito, C. Phylogenetic relationships among Portuguese rye based on isozyme, RAPD and ISSR markers. Hereditas 134 (3) (2001) 229–236.

    Article  CAS  PubMed  Google Scholar 

  42. Fernández, M.E., Figueiras, A.M. and Benito, C. The use of ISSR and RAPD markers for detecting DNA polymorphism, genotype identification and genetic diversity among barley cultivars with known origin. Theor. Appl. Genet. 104 (2002). 845–851.

    Article  PubMed  CAS  Google Scholar 

  43. Jana, T., Sharma, T.R. and Singh N.K. SSR-based detection of genetic variability in the charcoal root rot pathogen Macrophomina phaseolina. Mycol. Res. 109 (2005) 81–86.

    Article  CAS  PubMed  Google Scholar 

  44. Adamek, L. Submerse cultivation of the fungus Metarhizium anisopliae (Metsch.) Sorok. Folia Microbiológica 10 (1965) 255–257.

    CAS  PubMed  Google Scholar 

  45. Sneath, P.H.A. and Sokal, R.R. Numerical Taxonomy. The principles and practice of numerical classification. W.H. Freeman and Co, San Francisco, U.S.A. (1973).

    Google Scholar 

  46. Rohlf, F.J. NTSYS-pc, numerical taxonomy and multivariate analysis system. Applied biostatistical Inc, New York, U.S.A. (1993).

    Google Scholar 

  47. Felsenstein, J. Confidence limits on phylogenies: an approach using the bootstrap. Evolution 39 (1985) 783–791.

    Article  Google Scholar 

  48. Yap, I.V. and Nelson, B.J. WinBoot: a program for performing bootstrap analysis of binary data determine the confidence limits of UPGMA-based dendrograms. Int. Rice Res. Inst. (IRRI) (1996) 1-22.

  49. Goldwin, I.D., Aitken, A.B. and Smith, L.W. Application of Inter simple sequence repeats (ISSR) markers to plant genetics. Electrophoresis 18 (1997) 1524–1528.

    Article  Google Scholar 

  50. Glare, T.R. and Inwood, A.I. Morphological and genetic characterization of Beuvaria spp. from New Zealand. Mycol. Res. 102 (1998) 250–256.

    Article  Google Scholar 

  51. Wang, C., Li, Z., Typas, M.A. and Butt, T.M. Nuclear large subunit rDNA group I intron distribution in a population of Beavaria bassiana strains: phylogenetic implications. Mycol. Res. 107 (2003) 1189–1200.

    Article  CAS  PubMed  Google Scholar 

  52. De Muro, M.A., Mehta, S. and Moore, D. The use of amplified fragment length polymorphism for molecular analysis of Beauveria bassiana isolates from Kenya and other countries and their correlation with host and geographic origin. FEMS Microbiol. Newslett. 229 (2003) 249–257.

    Article  CAS  Google Scholar 

  53. Mohan, M.C. Phenotypic and molecular characterization of Beauveria bassiana (Bals.) Vuill. isolates suitable for integrated pest management of cotton boll worm (Helicoverpa armigera Hub.) in semi-arid tropics. Ph.D. dissertation, Andhra University, Visakhapatnam, India. (2002).

    Google Scholar 

  54. Padmavathi, J., Uma Devi, K., Uma Maheswara Rao, C. and Nageswara Rao Reddy, N. Telomere fingerprinting for assessing chromosome number, isolate typing and recombination in the entomopathogen, Beauveria bassiana (Balsamo) Vuillemin. Mycol. Res. 107 (2003) 572–580.

    Article  CAS  PubMed  Google Scholar 

  55. Budak, H., Shearman, R.C., Parmaksiz, I. and Dweikat, I. Comparative analysis of seeded and vegetative biotype buffalograsses based on phylogenetic relationship using ISSRs, SSRs, RAPDs, and SRAPs. Theor. Appl. Genet. 109 (2004) 280–288.

    Article  CAS  PubMed  Google Scholar 

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Authors and Affiliations

  1. Departamento de Genética, Facultad de Biología, Universidad Complutense, 28040, Madrid, Spain

    M. Elena Estrada, Manuel V. Camacho & César Benito

  2. Programa de Fitomejoramiento, Instituto Nacional de Investigaciones de la Caña de Azúcar (INICA), La Habana, Cuba

    M. Elena Estrada

Authors
  1. M. Elena Estrada
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  2. Manuel V. Camacho
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  3. César Benito
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Correspondence to César Benito.

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Estrada, M.E., Camacho, M.V. & Benito, C. The molecular diversity of different isolates of Beauveria bassiana (Bals.) Vuill. as assessed using intermicrosatellites (ISSRs). Cell Mol Biol Lett 12, 240–252 (2007). https://doi.org/10.2478/s11658-006-0069-4

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  • DOI: https://doi.org/10.2478/s11658-006-0069-4

Key words

  • Beauveria bassiana
  • Strain identification
  • Inter-microsatellite
  • Phylogenetic relationships

Cellular & Molecular Biology Letters

ISSN: 1689-1392

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