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Amplification and restriction endonuclease digestion of the Pr1 gene for the detection and characterization of Metarhizium strains

Published online by Cambridge University Press:  01 March 1997

SORAYA C. M. LEAL ,
DAVID J. BERTIOLI ,
TARIQ M. BUTT ,
JOHN H. CARDER ,
PAUL R. BURROWS  and
JOHN F. PEBERDY
SORAYA C. M. LEAL
Affiliation:
Entomology and Nematology Department, IACR-Rothamsted, Harpenden, Hertfordshire AL5 2JQ, U.K.
DAVID J. BERTIOLI
Affiliation:
Present address: CENARGEN/EMBRAPA, SAIN Parque Rural, CP02372, Brasilia, DF, Brazil. Entomology and Nematology Department, IACR-Rothamsted, Harpenden, Hertfordshire AL5 2JQ, U.K.
TARIQ M. BUTT
Affiliation:
Entomology and Nematology Department, IACR-Rothamsted, Harpenden, Hertfordshire AL5 2JQ, U.K.
JOHN H. CARDER
Affiliation:
Plant Pathology and Microbiology Department, Horticulture Research International, Wellesbourne, Warwick CV35 9EF, U.K.
PAUL R. BURROWS
Affiliation:
Entomology and Nematology Department, IACR-Rothamsted, Harpenden, Hertfordshire AL5 2JQ, U.K.
JOHN F. PEBERDY
Affiliation:
Department of Life Science, Nottingham University, Nottingham NG7 2RD, U.K.
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Abstract

A method is described for identifying strains of Metarhizium suitable for use in field experiments. It involves the restriction endonuclease digestion of a PCR product derived from the Pr1 protease gene and the analysis of the fragments by electrophoresis. Using this technique, 40 Metarhizium strains produced 15 different profile types and were clustered into four groups. Correlation between the profile of restriction fragments and geographic origin was observed for certain groups of strains. This PCR strategy allowed the identification of fungal strains, using as samples spores scraped from the surface of single insects killed by the fungus or single whole dead insects with external mycelium. The sequence of the Pr1 PCR product from two strains revealed that Pr1 gene has at least three introns.

Type
Research Article
Information
Mycological Research , Volume 101 , Issue 3 , March 1997 , pp. 257 - 265
Copyright
© The British Mycological Society 1997

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