African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12267

Full Length Research Paper

Genotyping isolates of the entomopathogenic fungus Beauveria bassiana sensu lato by multi-locus polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) analysis

Shun-chang Pu, Ming-jun Chen, Zhong-you Ma, Ling Xie, Zeng-zhi Li and Bo Huang*
Anhui Provincial Key Laboratory of Microbial Pest Control, Anhui Agricultural University, Hefei 230036, China.
Email: [email protected]

  •  Accepted: 27 April 2010
  •  Published: 31 July 2010

Abstract

Multi-locus denaturing gradient gel electrophoresis (DGGE) analysis was developed to investigate the genotypes of Beauveria bassiana sensu lato. Sensitive tests indicated all isolates with one or more nucleotide differences at EF-1α and Bloc could be distinguished by DGGE except for one pair of strains that differed at four nucleotide positions. Ten, twelve and five genotypes were identified at the EF-1α, Bloc and ITS locus, respectively, among seventeen isolates, which together differentiated 13 genotypes. These results demonstrated that multi-locus DGGE is a potentially useful molecular marker for genotyping, identifying and tracking the fates of experimentally released strains of B. bassiana sensu lato. Moreover, by multi-locus DGGE for scanning B. bassiana  sensu lato isolates with different multi-locus sequences, genetic diversity of B. bassiana  sensu lato was effectively investigated with substantially reduced time and cost in subsequent DNA sequencing.

 

Key words: Denaturing gradient gel electrophoresis, multi-locus sequences, genotyping, sensitive tests.

Abbreviation

DGGE, Denaturing gradient gel electrophoresis; RAPD, random amplified polymorphic DNA; ISSR, inter simple sequence repeat; PCR-SSCP,polymerase chain reaction single-strand conformation polymorphism; SSR, simple sequence repeat.