Abstract
To date, many plants explored for their endospheric microbiomes have revealed a substantial amount of sterile mycelia, a group of fungi with pseudo-taxonomic positions. While unravelling the endophytic fungal assemblages of a plant <italic>Ephedra gerardiana</italic> Wall. ex Stapf. <italic>via</italic> culture-dependent methods, 73.2% of the total isolates failed to sporulate on the commonly used artificial media. Therefore, the current study was designed to investigate the diverse methods, either individually or in combination to check their impact in inducing fungal sporulation. The maximum number of isolates (N <sub>max</sub>) sporulated after individual treatments with 12 h alternate light & dark cycles and cold shocks at 4−6 °C were 22.91% and 18%, respectively. However, the combination of cold treatment with alternate light and dark cycles showed the N <sub>max</sub> of <sub/> 27.84% and proved to be the best method of sporulation induction out of the methods investigated. To the best of our knowledge, this involves the pioneer attempt of using cold treatment as a triggering factor for sporulating endophytic fungi. Accentuating better methods for endophytic fungal sporulation would not only solve the taxonomic position of these mycosymbionts, but also lessen the time to make them sporulate along with unveiling their eco-physiological behavior inside the host plant.
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