Towards resolving Nothophaeocryptopus and Rhizosphaera inhabitants of spruce needles

Our knowledge of the endophytic fungal community associated with needles of Sitka spruce (Picea sitchensis) is rather limited, which contrasts with the importance of this tree species for forestry in the UK. In our study, we focused on the detailed characterization of multiple isolates tentatively placed into the genera Nothophaeocryptopus and Rhizosphaera that were obtained from needles of Sitka spruce and co-occurring Norway spruce (Picea abies) and Douglas fir (Pseudotsuga menziesii) at four sites in Scotland and Wales. After a thorough analysis of their phenotype and analysis of two nuclear regions (ITS rDNA and gene for β-tubulin), we propose two species new to science and one new combination. Nothophaeocryptopus piceae was isolated from healthy green or brown needles from spruces and is currently known only as a sterile culture, but the taxonomic novelty is well supported by host affinity and results of molecular data analysis. Rhizosphaera minteri may also be distinguished based on the combination of host and molecular data, but morphologically, it is similar to R. pini. Finally, Hormonema merioides and R. pseudotsugae are found to be conspecific and a new combination R. merioides is proposed following the phylogenetic placement of this species. Our study highlights the importance of multiple approaches used in the identification of microfungi associated with coniferous needles. It is evident that morphologically identical fungi may represent distinct species differing in their host range and severity on the host. This study also provides a basis for future monitoring of these fungi associated with important coniferous forestry trees in the UK.