Dna extraction, , PCR

DNA extraction, PCR amplification and sequencing

DNA was extracted from 5-10 mg tissue of dried specimen with the Broad-spectrum Plant Rapid Genomic DNA Kit (Biomed, Beijing, China) following the manufacturer’s instructions and preserved at −20℃.

This study used primers ITS4 and ITS5 ( White et al. 1990) for the internal transcribed spacer (ITS) region of the nuclear ribosomal DNA repeat, primers LROR and LR5 ( Moncalvo et al. 2000) for the Large Subunit (nrLSU). Genes were amplified by polymerase chain reaction (PCR) using the procedures mentioned in ( Li et al. 2022; Wang et al. 2024b; Yang et al. 2024a; Han et al. 2024; Li et al. 2024b). The PCR products were sent to a commercial biotech company (Baimaide Biotechnique Company, Beijing, China) for sequencing region.