Strain-specific detection of introduced Beauveria bassiana in agricultural fields by use of sequence-characterized amplified region markers

• Published in: Journal of invertebrate pathology Vol. 82; no. 2; pp. 75 - 83
• Main Authors: Castrillo, L.A., Vandenberg, J.D., Wraight, S.P.
• Format: Journal Article
• Language: English
• Published: Amsterdam Elsevier Inc 01.02.2003; Elsevier
• Subjects: Agriculture; Animals; Beauveria bassiana; Biological and medical sciences; Biological control; Control; Entomopathogenic fungus; Fundamental and applied biological sciences. Psychology; Microbial control; Mitosporic Fungi - genetics; Mitosporic Fungi - isolation & purification; Molecular markers; Pest Control, Biological; Phytopathology. Animal pests. Plant and forest protection; Polymerase Chain Reaction; Polymorphism, Single-Stranded Conformational; Protozoa. Invertebrates; Sequence-characterized amplified region markers; Species Specificity; Strain detection; Beauveria bassiana; Molecular markers; Entomopathogenic fungus; Microbial control; Strain detection; Sequence-characterized amplified region markers; Entomopathogen; Biological control; Molecular marker; Strain specificity; Random amplified polymorphic DNA marker; Fungi; Polymerase chain reaction; Gene amplification; Pest; Microbiological control; Fungi Imperfecti; Biopesticide; Detection; Microbial insecticide; Thallophyta
• ISSN: 0022-2011; 1096-0805
• DOI: 10.1016/S0022-2011(02)00190-8

Field studies on the efficacy and persistence of an introduced strain of Beauveria bassiana for insect control require detection assays to differentiate the non-native strain from indigenous populations. In this study we developed strain-specific molecular markers based on polymerase chain reaction amplification of sequence-characterized amplified regions (SCAR) in combination with dilution plating on semi-selective medium to detect and estimate density of propagules of a commercial strain of B. bassiana (strain GHA) in field samples. Using random amplified polymorphic DNA (RAPD) analysis, unique fragments that distinguished GHA from other strains of B. bassiana were obtained. Three amplicons, OPA-14 0.44, OPA-15 0.44, and OPB-9 0.67, generated with RAPD primers were cloned and sequenced and used as bases for designing SCAR primers OPA14 F/R 445, OPA15 F/R 441, and OPB9 F/R 677, respectively. All three SCAR primers were highly sensitive, capable of detecting 100 pg B. bassiana GHA genomic DNA, and thus could be used to detect varying levels of the fungus in the field.