Fungal Diversity Associated with Sharp-dentated Bark Beetle Ips acuminatus (Coleoptera: Curculionidae) in Latvia

The dataset contains representative sequences obtained during year 2025 from fungi isolated from bark beetles Ips acuminatus in Latvia. Each sequence contains small subunit ribosomal RNA gene, partial sequence; internal transcribed spacer 1,5.8S ribosomal RNA gene, and internal transcribed spacer 2, complete sequence; and large subunit ribosomal RNA gene, partial sequence. For DNA extraction modified CTAB method were used followed by PCR amplification with fungi–specific primers ITS1F (5′-CTTGGTCATTTAGAGGAAGTAA-3′) and ITS4 (5′-TCCTCCGCTTATTGATATGC-3′). PCR reactions were performed in a volume of 10 µL containing approximately 50 ng DNA, 5× HOT FIREPol Blend Master Mix Ready to Load (Solis BioDyne, Tartu, Estonia) (containing 10 mM MgCl2), and 0.3 µM of each forward and reverse primers. PCR was carried out in a thermocycler (Eppendorf Master-cycler Epgradient; Eppendorf, Hamburg, Germany) using the following protocol: initial pre-denaturation step at 95 °C for 15 min, followed by 35 cycles of 95 °C for 30 s, annealing at 55 °C for 30 s, and 72 °C for 45 s, and a final extension step of 72 °C for 10 min. PCR fragments were analyzed by gel electrophoresis on 1.5% agarose gels stained with ethidium bromide and Sanger sequenced in two directions using ITS1F and ITS4 primers. The obtained nucleotide sequences were analyzed using the NCBI BLAST function (https://blast.ncbi.nlm.nih.gov/Blast.cgi) for identification of fungal species. The ITS sequence homology was set at 98-100% for fungal taxa and 94-97% for genus level. All work performed in Latvian State Forest Research Institute “Silava”.