Neonectria bordenii sp. nov., a potential symbiote of the alder bark beetle, and its detection by quantitative PCR

A taxonomically comprehensive perspective on the fungal associates of bark beetles (Coleoptera: Curculionidae: Scolytinae), and powerful molecular tools for detection of these fungi, are imperative to understanding bark beetle impacts on forest ecosystems. The most common filamentous fungi living alongside bark beetles in infested trees are ophiostomatoids (Ascomycota: Ophiostomatales and Microascales), yet an undescribed species of Neonectria (Neonectria sp. nov.; Ascomycota: Hypocreales) was recently identified cohabitating with the alder bark beetle, Alniphagus aspericollis, in red alder, Alnus rubra. The hardwood-infesting alder bark beetle is found throughout the range of its red alder host in the Pacific Coast region of North America and is associated with Neonectria sp. nov. in southwestern British Columbia, Canada. The aim of this study was to describe and name Neonectria sp. nov. and to develop a quantitative PCR (qPCR) assay to enable rapid detection of Neonectria sp. nov. from individual adult alder bark beetles and to define the distribution of the fungus. Neonectria sp. nov. was phylogenetically and morphologically determined to represent a distinct species closely related to N. ditissima and is described herein as Neonectria bordenii sp. nov. Neonectria bordenii was reliably detected from individual whole-beetle DNA extractions using a probe-based qPCR assay targeting multi-copy internal transcribed spacers (ITS) of nuclear ribosomal DNA. The qPCR assay amplified the fungus from 87.8 % (36/41) of individual alder bark beetle samples and was highly sensitive to N. bordenii, with a lower limit of detection of 1 × 10^-6 ng/μL of culture DNA (or ~262 genome copies). Application of the qPCR assay developed in this study will expedite future research evaluating N. bordenii as a potential symbiote of the alder bark beetle. Citation: Wertman DL, Tanney JB, Hamelin RC, Carroll AL (2024). Neonectria bordenii sp. nov., a potential symbiote of the alder bark beetle, and its detection by quantitative PCR. Fungal Systematics and Evolution 13: 15-28. doi: 10.3114/fuse.2024.13.02.