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

Tharnrat Kaewgrajang; Leela Nakpong; Yatawee Foongchomchoi; Chatchai Ngernsaengsaruay; Runchida Khunkrai; Kantida Bunlerlerd; Sasitorn Hasin; Itsarapong Voraphab; Baramee Sakolrak; Penpitcha Choosa-nga; Cherdchai Phosri; Warong Suksavate; Mingkwan Nipitwattanaphon

doi:10.47371/mycosci.2024.09.002

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

Tharnrat Kaewgrajang, Leela Nakpong, Yatawee Foongchomchoi, Chatchai Ngernsaengsaruay, Runchida Khunkrai, Kantida Bunlerlerd, Sasitorn Hasin, Itsarapong Voraphab, Baramee Sakolrak, Penpitcha Choosa-nga, Cherdchai Phosri, Warong Suksavate, Mingkwan Nipitwattanaphon

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Tharnrat Kaewgrajang
Department of Forest Biology, Faculty of Forestry, Kasetsart University
Leela Nakpong
Department of Genetics, Faculty of Science, Kasetsart University
Yatawee Foongchomchoi
Department of Genetics, Faculty of Science, Kasetsart University
Chatchai Ngernsaengsaruay
Department of Botany, Faculty of Science, Kasetsart University
Biodiversity Center Kasetsart University (BDCKU)
Runchida Khunkrai
Department of Forest Biology, Faculty of Forestry, Kasetsart University
Kantida Bunlerlerd
Department of Forest Biology, Faculty of Forestry, Kasetsart University
Sasitorn Hasin
Innovation of Environmental Management, College of Innovative Management, Valaya Alongkorn Rajabhat University under the Royal Patronage
Itsarapong Voraphab
Department of National Park, Wildlife and Plant Conservation
Baramee Sakolrak
Department of National Park, Wildlife and Plant Conservation
Penpitcha Choosa-nga
Department of National Park, Wildlife and Plant Conservation
Cherdchai Phosri
Department of Biology, Faculty of Science, Nakhon Phanom University
Warong Suksavate
Department of Forest Biology, Faculty of Forestry, Kasetsart University
Mingkwan Nipitwattanaphon Corresponding author
Department of Genetics, Faculty of Science, Kasetsart University

Keywords: barometer earthstar, Diplocystaceae, ectomycorrhiza, molecular marker

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2024 Volume 65 Issue 6 Pages 288-297

DOI https://doi.org/10.47371/mycosci.2024.09.002

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Abstract

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.

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