Molecular Characterization of Ectomycorrhizal Fungi Associated with Pinus wallichiana A B Jackson in Kashmir Himalaya

Abstract

Ectomycorrhizal (ECM) fungi are functionally important in temperate forest ecosystems for plant nutrient and water uptake and provide protection from root pathogens. Studies of ECM community structure have revealed remarkable below ground richness in relatively small areas. Identifying ectomycorrhizal species associated with roots of higher plants has always been a laborious work mostly relying upon generation of pure cultures and morphotying. However, such culture-based techniques have limitations to document the entire diversity associated with soil or root samples. Notwithstanding the fact that conifers, which are known to harbour ectomycoorhizal species for their growth and survival, dominate the Kashmir Himalayan forests, studies on ectomycorrhizal fungi associated with conifers, such as Pinus wallichiana a dominant and economically important conifer are lacking. It is in this context that the fine root-tips harboring ectomycorrhizae were pooled and then directly processed for molecular characterization under highly stringent conditions using overlapped rRNA gene specific primers to generate multiple bands, each band being representative of an individual fungal species. This was followed by RFLP analysis to generate individual clades. Representatives of each clade are then sequenced, assembled and subjected to global BLAST analysis for identification of ectomycorrhizal specie(s). A total of 56 species comprising 34 unique fungal taxa were identified based on sequence scores and blast similarity. The fungi characterized included 25 species with confirmed ectomycorrhizal status while the ectomycorrhizal status in respect of 9 species is yet to be confirmed. newline