In a 2023 survey evaluating conifers with Rosellinia infections, five Pestalotiopsis-like fungal endophytes were isolated from plant samples obtained from Maine, New Hampshire, and Ohio by The Mycology & Nematology Genetic Diversity & Biology Laboratory at the United States Department of Agriculture. The two data sets provided herein contain species-specific base pair substitutions for the partial translation elongation factor 1-alpha gene (TEF). In the alignments, the novel Pestalotiopsis fungi are compared to their most closely related species. The data set can be used as a diagnostic tool to differentiate between the closely related species.
DNA was extracted from fungal samples using the E.Z.N.A HP Plant & Fungal DNA Kit (OMEGA\u00ae Bio-Tek, Norcross, GA, USA) following manufacturer\u2019s protocol. The TEF locus was amplified, and reactions were conducted in 25 \u03bcL volumes with 12.5 \u03bcL of KAPA2G Robust Hotstart\u00ae (Kapa Biosystems, Inc., Wilmington, MA, USA), 1.25 \u03bcL of the forward and reverse primers at 10 \u03bcM, 1-1.75 \u03bcL of DNA at 10-20 ng, and 8 \u03bcL of molecular grade H2O. Amplifications were performed following the protocol described by Maharachchikumbura et al. (2014) in a C1000 Touch PCR Thermal Cycler (Bio-Rad, Hercules, CA). PCR products were analyzed through capillary electrophoresis with the QIAxcel Advanced System instrument and the QIAxcel ScreenGel software (Qiagen, Hilden, Germany). PCR products were then purified using ExoSAP-IT Cleanup (Affymetrix, Santa Clara, CA) following the manufacturer\u2019s protocol. The BigDye\u2122 3.1 Terminator Cycle sequencing kit was used to sequence amplicons bi-directionally with the Applied Biosystems SeqStudio Genetic Analyzer (Thermo Fisher Scientific, Waltham, MA, USA).
Resources in this dataset: