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Title: Multiplexed microsatellite markers for seven Metarhizium species

item MAYERHOFER, JOHANNA - Agroscope
item LUTZ, ANDY - Agroscope
item WIDMER, FRANCO - Agroscope
item Rehner, Stephen
item LEUCHTMAN, ADRIAN - Eth Zurich
item ENKERLI, JÜRG - Agroscope

Submitted to: Journal of Invertebrate Pathology
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/21/2015
Publication Date: 11/1/2015
Citation: Mayerhofer, J., Lutz, A., Widmer, F., Rehner, S.A., Leuchtman, A., Enkerli, J. 2015. Multiplexed microsatellite markers for seven Metarhizium species. Journal of Invertebrate Pathology. 132:132-134.

Interpretive Summary: Crop-feeding insects annually cause billions of dollars in agricultural losses. Although some beneficial fungi can be used to control some pest insects under certain conditions, little is known about the reasons for this variable effectiveness and their frequent inability to persist in agricultural environments. Because one method of determining the differences among closely related fungi is to examine their molecular (i.e., DNA) differences, in this study we screened 34 DNA markers known as polymorphic microsatellite or SSR markers in fungi with potential use as biocontrol agents. We discovered an optimal set of 15 highly variable markers that showed the greatest reliability and sensitivity in differentiating among individual genotypes among fungal species that are common in agricultural soils, including species developed as biological control agents. The results are significant because they provide improve molecular methods for determining subtle differences among these fungi. This information will be used by researchers discovering and implementing integrated pest management strategies of field crop pests.

Technical Abstract: Cross-species transferability of 41 previously published simple sequence repeat (SSR) markers was assessed for 11 species of the entomopathogenic fungus Metarhizium. A collection of 65 Metarhizium isolates including all 54 used in a recent phylogenetic revision of the genus were characterized. Between 15 and 34 polymorphic SSR markers produced scorable PCR amplicons in seven species, including M. anisopliae, M. brunneum, M. guizhouense, M. lepidiotae, M. majus, M. pingshaense, and M. robertsii. To provide genotyping tools for concurrent analysis of these seven species fifteen markers grouped in five multiplex pools were selected based on high allelic diversity and easy scorability of SSR chromatograms.