Diversity and Spatiotemporal Distribution of Fungal Endophytes Associated with Salvia multicaulis

Salvia multicaulis has been an important medicinal plant in Iran and several East Asian countries for hundreds of years. Because of growing demand, overharvesting of wild S. multicaulis has endangered its wild populations. Endophytes are well known for protecting wild plant populations against biotic and abiotic stresses, especially under harsh situations, as well as for their plant growth enhancement activities. Since no information was on endophyte biology in S. multicaulis, here we aimed at analyzing diversity and spatiotemporal distribution of fungal endophytes associating S. multicaulis in their main wild habitats in Iran, i.e., Qazvin, Alborz and Mazandaran provinces. A total of 153 fungal endophytes were isolated and identified according to their morphology and ribosomal ITS rDNA sequences. As results indicated Ascomycota dominated in colonizing S. multicaulis with a relative frequency (RF) of 96.77%, comprising of Eurotiomycetes (RF: 40.5%), Sordariomycetes (RF: 33.9%) and Dothideomycetes (RF: 20.5%). Mucoromycota, comprised the rest of endophytes (RF: 5.23%). The entire fungal microbiome was classified into nine genera including Fusarium (25.5%), Penicillium (21.5%), Aspergillus (17.0%), Alternaria (15.5%), Colletotrichum (5.2%), Rhizopus (5.2%), Macrophomina (4.5%), Trichoderma (3.25%) and Nodulisporium (2.0%). Analyses of different diversity indices indicated significant correlations with tissue type, sampling locations and season of recovery. Almost 43% of fungal endophytes were recovered at Mazandaran, Kojur; 35.4% at Qazvin, Barajin Forest Park; 30.1% at Alborz, Taleqan; and 21% at Alborz, Mahdasht. The highest overall endophyte recovery was in summer (36.8%), followed by spring (31.6%), autumn (21%), and winter (10.5%). In total, the number of endophytes recovered from roots (91) was higher than those of stems (32) and leaves (30), especially during autumn and winter. Accordingly, we conclude that Ascomycota are the major endophytic fungi colonizing S. multicaulis, and that sampling location, tissue type and season can affect the fungal endophyte composition of this medicinal plant. This knowledge could be further applied in protection and health management of this endangered species.