Fluorescent Random Amplified Microsatellites (F-RAMS) analysis of mushrooms as a forensic investigative tool

Area of Science:

• Mycology
• Molecular Biology
• Genetics

Background:

• Accurate identification of mushroom species, particularly hallucinogenic varieties, is crucial for research and safety.
• Microsatellite markers offer potential for high-resolution genetic profiling.
• Fluorescent Random Amplified Microsatellites (F-RAMS) is a technique for DNA fingerprinting.

Purpose of the Study:

• To evaluate the effectiveness of F-RAMS for species and sub-species level profiling of hallucinogenic mushrooms.
• To compare the performance of two different fluorescently-labeled primers in mushroom identification.

Main Methods:

• Profiling of 15 Amanita rubescens samples and 22 other mushroom samples (Amanita, Psilocybe) using F-RAMS.
• Utilized two 5'degenerate primers: 5'-6FAM-SpC3-DD (CCA)5 and 5'-6FAM-SpC3-DHB (CGA)5, targeting distinct microsatellite regions.
• Analysis of amplicon similarities to construct a dendrogram for species clustering.

Main Results:

• The primer 5'-6FAM-SpC3-DHB (CGA)5 demonstrated superior reliability in grouping same-species samples and clustering related species.
• High intra-specific variation was observed within the Amanita rubescens samples using both primers.
• Generated amplicons for A. rubescens were categorized into discriminant, private, and marker classes based on their individualizing potential.

Conclusions:

• F-RAMS, particularly with the 5'-6FAM-SpC3-DHB (CGA)5 primer, is a viable method for high-resolution profiling of hallucinogenic mushrooms.
• The technique can distinguish between closely related species and reveal significant genetic diversity within a single species.
• Further application of F-RAMS can aid in understanding mushroom population genetics and taxonomy.