Root-associated fungi of Pinus wallichiana in Kashmir Himalaya

An important factor in the performance of out-planted conifers is the association of plant roots with ectomycorrhizal (EcM) fungi. However, limited information is available about the diversity of root-associated EcM fungi of Pinus wallichiana A.B. Jackson, a coniferous species endemic to Himalayan forests that has hampered the reforestation programs in the area. The study was carried at three major forest areas of the Kashmir Himalaya believed to be pure stands of P. wallichiana. Fine root tips harbouring EcM fungi were collected and processed for extraction of fungal DNA, which was subsequently subjected to ITS rDNA targeted PCR–RFLP profiling. DNA sequencing analysis of the overlapping ITS amplifications followed by global nucleotide BLAST analyses of the assembled nuclear ribosomal DNA (rDNA) revealed a total of 33 fungal taxa associated with P. wallichiana of which 23 species were EcM fungi. Of the 10 non-EcM fungi, we found a peculiar saprophytic wood decaying fungus, Chalara microchona, associated with P. wallichiana for the first time. The study not only reveals the species richness of fungi associated with this conifer but also documents new fungal associations with it, which have not been reported so far. The results in the study set a baseline for the broad association of ectomycorrhizal fungi with P. wallichiana, which may serve as guiding cue to design reforestation programs in the Kashmir Himalaya.