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Front Microbiol
2018 Jan 01;9:612. doi: 10.3389/fmicb.2018.00612.
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Fungal Diversity and Community Composition of Culturable Fungi in Stanhopea trigrina Cast Gibberellin Producers.
Salazar-Cerezo S
,
Martinez-Montiel N
,
Cruz-Lopez MDC
,
Martinez-Contreras RD
.
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Stanhopea tigrina is a Mexican endemic orchid reported as a threatened species. The naturally occurring microorganisms present in S. tigrina are unknown. In this work, we analyzed the diversity of endophytic and epiphytic culturable fungi in S. tigrina according to morphological and molecular identification. Using this combined approach, in this study we retrieved a total of 634 fungal isolates that presented filamentous growth, which were grouped in 134 morphotypes that were associated to 63 genera, showing that S. tigrina harbors a rich diversity of both endophytic and epiphytic fungi. Among these, the majority of the isolates corresponded to Ascomycetes, with Trichoderma and Penicillium as the most frequent genera followed by Fusarium and Aspergillus. Non-ascomycetes isolated were associated only to the genus Mucor (Mucoromycota) and Schizophyllum (Basidiomycota). Identified genera showed a differential distribution considering their epiphytic or endophytic origin, the tissue from which they were isolated, and the ability of the orchid to grow on different substrates. To our knowledge, this work constitutes the first study of the mycobiome of S. tigrina. Interestingly, 21 fungal isolates showed the ability to produce gibberellins. Almost half of the isolates were related to the gibberellin-producer genus Penicillium based on morphological and molecular identification. However, the rest of the isolates were related to the following genera, which have not been reported as gibberellin producers so far: Bionectria, Macrophoma, Nectria, Neopestalotiopsis, Talaromyces, Trichoderma, and Diplodia. Taken together, we found that S. tigrina possess a significant fungal diversity that could be a rich source of fungal metabolites with the potential to develop biotechnological approaches oriented to revert the threatened state of this orchid in the near future.
Figure 1. Location map of the study site. Sampling was performed in the Botanical Garden Xoxoctic (Cuetzalan del Progreso, Puebla, México), a natural area located to the Sierra Norte (Puebla) devoted to preserve species characteristic of this region.
Figure 2. Abundance of fungal isolates in different S. tigrina tissues. The number of endophytic (orange), epiphytic (green), and rhizospheric (gray) fungal isolates recovered from different tissues is shown.
Figure 3. Tissue distribution of the fungal genera isolated from S. tigrina. Epiphytic (A) and endophytic (B) isolates were recovered from the leaf, pseudobulb, root, and flower of S. tigrina and the relative frequency for the different genera isolated is shown. Genera recovered from the rhizosphere are also shown (C).
Figure 4. Distribution of culturable fungi in different S. tigrina plants. The relative isolation frequency was estimated for the epiphytic (A) and endophytic (B) isolates that were recovered from each of the six different plants. Plants 1 and 2 grew as epiphytes, plant 3 was growing as litophyte, while plants 4, 5, and 6 were growing on substratum.
Figure 5. Fungal community composition according to the different growth habits of S. tigrina. Isolation frequency of genera growing as epiphytes, as litophytes, or in substratum are presented for the epiphytic (A) and endophytic (B) fungal communities. The number of genera shared by plants with different growth habits is shown for the epiphytic (C) and endophytic (D) fungal communities.
Figure 6. Phylogenetic relationship among the 21 fungal isolates that showed the ability to produce gibberellin. The tree with the highest log likelihood (−8844.44) is shown. The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. The analysis involved 45 nucleotide sequences with a total of 100 replicates in the bootstrap analysis.
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