Efficient PCR - based Approach for Rapid Identification of Earth Star Mushrooms Employing Species-Specific Primers.

Astraeus species are valuable edible ectomycorrhizal mushrooms, particularly in Asia. The partial hypogenous nature of Astraeus fruit bodies in soil present challenges for sample collection and studying species distribution. In this study, we developed a PCR-based approach for identifying Astraeus. Two universal primers (AUPF1/AUPR1 and AUPF3/AUPR3) and specific primers for A. asiaticus (AAF4/AAR4), A. odoratus (AOF4/AOR4), and A. sirindhorniae (ASF2/ASR4) were designed based on the alignment of internal transcribed spacer sequences from various Astraeus species. Primer verification was performed by generating amplicons from extracted DNA of Astraeus fruit bodies and 130 soil samples collected from beneath various host plants of Astraeus spp. These novel primers were efficient and precise in identifying Astraeus species. Our results have implications for multi-sample assays for Astraeus identification and investigations into species distribution through large-scale inventories.