Screening of ectomycorrhizal and other associated fungi in South African forest nurseries

The South African forestry industry covers approximately 1.3 million hectares and is dependent on exotic pine and eucalypt species. Nursery seedlings are not inoculated with ectomycorrhizal (ECM) fungi. This investigation assessed levels of naturally occurring ECM colonisation of Pinus patula seedlings from 10 different South African forestry nurseries using a grid line intersect method. Fungi from colonised roots were identified using morphological characteristics and Illumina sequencing. Colonisation of seedlings in production nurseries was low (2-21%). Morphologically, the ECM fungi Thelophora terrestris, Suillus sibiricus, and the genera Russula and Pseudotomentella were identified. Molecularly, the ECM fungi T. terrestris, Inocybe jacobi and the genus Sphaerosporella, as well as several other ECM-containing families were identified, along with many saprotrophic/ endophytic fungi belonging to genera such as Penicillium, Ramasonia and Talaromyces. As can be seen, a combination of both molecular and morphological identification techniques are needed as neither is able to give a full picture of the species present in isolation. This study reveals an initial insight into the root microbiome community associated with Pinus patula seedlings, which should be taken into account when inoculation with beneficial microbes is considered. It determined that natural ECM fungal root colonisation levels are very low throughout the South African nurseries investigated, indicating the need for ECM fungal inoculation, which can increase seedling growth, viability and resistance to pathogens.