Ectomycorrhizal fungal assessment of South African Pinus patula seedlings and their biological control potential to enhance seedling growth

Abstract

The South Africa forestry industry, covering 1.3 million hectares, is dependent on exotic pine and eucalyptus species. Nursery seedlings are not inoculated with ectomycorrhizal (ECM) fungi or other beneficial microbes. Fusarium circinatum is an economically important pathogen affecting seedling survival. The purpose of this investigation was to assess levels of naturally occurring ECM colonisation in South African nurseries and to determine the effects of seedling inoculation with selected ECM and bacterial isolates on plant growth and resistance to the fungal pathogen F. circiantum. Pinus patula seedlings from 10 different South African nurseries were assessed for ECM colonisation using a grid line intersect method and molecularly identified using morphological and next-generation Illumina sequencing. Explants from ECM basidiocarps, collected from Pinus stands, were plated onto MMN medium to obtain isolates which were verified using molecular techniques. Mycorrhizal helper bacteria (MHB) were also isolated from these basiocarps, tested for MHB properties, siderophore production, phospahte solubilising and IAA production. ECM and associated bacterial isolates were used to inoculate seedlings and growth was assessed over a 5 month period. Colonisation of seedlings in production nurseries was low (2-21%). Morphologically the ECM fungi T. terrestris, Suillus sibiricus, and the genera Russula, Pseudotomentella were identified. Molecularly the ECM fungi T. terrestris, Inocybe jacobi and the genera Sphaerosporella and several other ECM containing families were identified along with many saprotrophic/endophytic fungi belonging to genera such as, Penicillium, Ramasonia and Talaromyces. Inoculated seedlings showed a significant increase in growth in comparison to the un-inoculated control seedlings. ECM fungal colonisation levels of these seedlings were significantly increased and colonisation was promoted by the Suillus isolate, Salmon Suillus. Seedling growth in the presence of the pathogen F. circinatum was significantly increased and promoted by the Lactarius isolate Lactarius quieticolor. Inoculation of seedlings in the nursery would ensure the production of stronger healthy plants which may be more tolerant to fusarial infection increasing survival in the plantation.