Fungal cultures were grown on PDA at 28°C. When colonies nearly covered the entire Petri dish (90 mm diam.), fresh mycelia were scraped from the agar surface with sterilised scalpels. Genomic DNA was extracted using a BIOMIGA Fungus Genomic DNA Extraction Kit (GD2416) following the manufacturer’s protocol. DNA amplification was performed in a 25 μl reaction volume following Liang et al. (2018). Primers ITS1 and ITS4 (White et al. 1990) were used to amplify the internal transcribed spacer regions and intervening 5.8S rRNA region (ITS) and LR0R and LR5 for 28S rRNA (LSU) region (Vilgalys and Hester 1990, Rehner and Samuels 1994). Two protein-coding gene fragments, the β-tubulin (tub2) and translation elongation factor 1-alpha (TEF1-a) were amplified with primer pairs BT2A/BT2B (Glass and Donaldson 1995, O'Donnell and Cigelnik 1997) and EF1-688F/EF1-986R, respectively (Carbone and Kohn 1999, Alves et al. 2008). Purification and sequencing of the PCR amplicons were done by SinoGenoMax, Beijing. The DNA sequences are deposited in the GenBank and their accession numbers are provided in Table 1. The DNA base differences of the four loci amongst our strains and ex-type or representative strains of relative taxa are shown (Table (Table22).

DNA base pair differences between Lasiodiplodia syzygii and L. rubropurpurea in four separate loci. T = ex-type

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