A PCR-based tool for cultivation-independent detection and quantification of Metarhizium clade 1

摘要: The entomopathogenic fungus Metarhizium anisopliae and sister species are some of the most widely used biological control agents for insects. Availability specific monitoring quantification tools essential investigation environmental factors influencing their distribution. Naturally occurring as well released strains in environment traditionally monitored with cultivation-dependent techniques. However, detection may be limited due to lack a defined reliable range such methods. Cultivation-independent PCR-based offer high throughput analyses target taxa various environments. In this study cultivation-independent method was developed, which allows clade 1, is formed by majus, guizhouense, pingshaense, anisopliae, robertsii Metarhiziumbrunneum, formerly included M. cryptic complex. This based on use clade-specific primers, i.e. Ma 1763 2097, that positioned within internal transcribed spacer regions 1 2 nuclear ribosomal RNA gene cluster, respectively. BLAST similarity searches empirical specificity tests performed non-target species, bulk soil DNA samples, demonstrated diagnostic tool targeted 1. Testing primer pair qPCR assays validated complex samples significantly correlated quantification. new will allow highly rapid environment. Habitat densities can then analyzed habitat preferences greater detail using techniques genetic typing isolates.