Dna extraction

Fan, Xiang-Ping, Song, Chang-Ge, Xu, Yi-Hua, Sun, Yi-Fei & Cui, Bao-Kai, 2025, Morphology and molecular phylogeny reveal one new species of Hydnellum (Thelephorales) from Eastern China, Phytotaxa 711 (1), pp. 43-60 : 44

DNA extraction and sequencing

The CTAB rapid plant genome DNA extraction kit DN14 (Aidlab Biotechnologies, Beijing, China) was used to extract DNA from dried samples. The experimental steps referred to the instructions of the kit and the modified methods in Song et al. (2022). The primers used for the amplification of the ITS gene fragment were ITS5 and ITS4 ( White et al. 1990), and the Polymerase Chain Reaction (PCR) procedure was as follows: initial denaturation at 95℃ for 3 min, followed by 35 cycles at 94℃ for 40 s, 56℃ for 45 s, and 72℃ for 1 min, and a final extension of 72℃ for 10 min. The primers used for the amplification of the nLSU gene fragment were LR0R and LR7 ( Hopple & Vilgalys 1999), and NS1 and NS4 were used for the amplification of the nSSU gene fragment ( Hibbett, 1996). The PCR procedure for nLSU and nSSU were as follows: initial denaturation at 94℃ for 1 min, followed by 35 cycles at 94℃ for 30 s, 50℃ for 1 min, and 72℃ for 90 s, and a final extension of 72℃ for 10 min. The primers used for the amplification of the rpb2 gene fragment were 5F and 7Cr ( Liu et al. 1999), and the PCR procedure was as follows: initial denaturation at 94℃ for 2 min, 9 cycles at 94℃ for 45 s, 60℃ for 45 s, followed by 36 cycles at 94℃ for 45 s, 53℃ for 1 min, 72℃ for 90 s, and a final extension of 72℃ for 10 min. The PCR amplification products were sequenced at the Beijing Genomics Institute (BGI), China, with the same primers. The newly generated sequences in this study were uploaded to Genbank (the National Center for Biotechnology Information, USA), and all the sequences used in this study were shown in Table 1.