Canopy height and epiphytic bryophytes shape fungal communities in a temperate rainforest
Data files
Sep 25, 2025 version files 2.01 GB
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EpiMicrMETAsqueaky.txt
16.45 KB
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FullITS_seqtab_newlabels.csv
3.40 MB
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FullITS_seqtab.csv
5.04 MB
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IR_data_loaded.csv
1.59 MB
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IR_map_loaded.csv
20.30 KB
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IR_taxonomy_loaded.csv
454.37 KB
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ITS_CleanTable_McToolsR.txt
3.92 MB
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ITS_FullTable_McToolsR.txt
3.98 MB
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ITS_Sequences.zip
1.99 GB
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ITS_taxonomy.xlsx
1.06 MB
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Laurel_Cleaned_NoUnassigned_Mctoolsr.txt
3.43 MB
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README.md
4.38 KB
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SamMap.csv
8.76 KB
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SamMap.txt
10.48 KB
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SamMapClean.txt
10.78 KB
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taxatab.csv
627.13 KB
Abstract
Fungal communities contribute to plant ecology and evolution in forested ecosystems. Their diverse interactions with associated host plants can vary along abiotic and biotic gradients, but these gradients are poorly understood in complex natural ecosystems. Given the high diversity of epiphytic plants in many ecosystems, forest canopies offer a unique and underexplored system for studying plant-associated microbial diversity and distribution. We explored both abiotic and biotic factors structuring arboreal fungal communities. Specifically, we hypothesized that bryophyte-associated fungal communities are structured by the vertical height gradient within host trees (from the ground to high in the canopy), vary across host plant species, and that living bryophytes host distinct fungal communities compared to dead bryophyte matter. To test these hypotheses, we sampled living and dead bryophytes (mosses and liverworts) across three different bryophyte host species and four heights, ranging from the forest floor to 18 meters above the ground. We characterized the fungal community composition in each sample using metabarcoding. Fungal communities showed significant variation across substrates: bryophytes collected from the ground exhibited 17% greater Shannon diversity and 34% higher taxonomic richness than epiphytic bryophytes, while living bryophytes had 15% higher diversity and 30% greater richness than dead tissues. This pattern suggests that the diverse microhabitats within living bryophytes may drive microbial diversity. Community analysis identified a core fungal community across living bryophyte samples, but rare taxa accounted for a majority of reads, driving differences in community composition between different heights and bryophyte species. Epiphytic bryophyte-associated fungal communities show high heterogeneity across different substrates and heights, which provides insight into the structuring of the forest microbiome and epiphyte ecology. These results demonstrate the importance of exploring canopy-associated microbes to better understand microbial diversity and function in forest ecosystems.
Dataset DOI: https://doi.org/10.5061/dryad.rr4xgxdjj
Description of the data and file structure
The data include fungal ITS sequencing results from bryophyte (moss and liverwort) samples, collected in order to determine how fungal diversity and community composition vary from the ground to the canopy, between bryophyte species, and between living and dead bryophytes.
Access information
Other publicly accessible locations of the data:
- None
Data was derived from the following sources:
- All data was generated by and for this project.
Code/software
All data processing and analysis were performed in R version 4.4.0. Packages are included within the code. All packages are free and easily downloadable.
BryoFung_ITS_Dada2.R
- Load in raw sequencing data
- Dereplication: combine identical sequences while retaining quality
BryoFung_MCToolsDecontamPhylo.R
- Use phyloseq to identify and remove contaminants
CanopyMicrobeAnalysis_ToShare.Rmd
- All analysis and figure generation.
- Analysis of microbial alpha/beta diversity, community composition, and functional diversity.
Files and variables
File: EpiMicrMETAsqueaky.txt
Description: Metadata for all samples remaining after data was decontaminated.
Variables
- SAMPLE: Name of sample. TreeID-Plot-SampleType (e.g. 01-V2-PORNAV)
- DateCollected
- Collector: Person who collected the sample.
- TreeID: Numerical ID of tree (1-12)
- Site: Site where sample was collected (Hoh Rainforest)
- Height: Categorical height within tree (T0=ground, V1/V2/V3 are ascending heights on trunk)
- MeasuredHeight: Measured height from ground (m)
- SampleType: Bryophyte species (ISOMYO = Isotheium myosuroides, PORNAV = Porella navicularis, METMEN = Metaneckera menziesii, COM = community, NECRO = dead bryophyte tissue)
- SampleControl: experimental sample or control
- ZYMO BATCH: batch number when extracting microbial DNA
- Extraction Date: date of extraction
- number in batch: tracking number within extraction batch
- Concentration_ngul: DNA concentration (ng/ul)
- CollNotes: notes from a collection of samples in the field
- LabNotes: notes from DNA extraction
File: Laurel_Cleaned_NoUnassigned_Mctoolsr.txt
Description: Community matrix of fungal ASVs in bryophyte samples, after cleaning, containing only fungal ASVs assigned at the phylum level
File: SamMap.txt
Description: Mapping file for DNA sequencing
Variables
- Same variables listed above
- Time: Height category in tree (T0/V1/V2/V3)
- Type: ISOMYO, PORNAV, METMEN, COM, or NECRO
File: SamMapClean.txt
Description: Sample Map after removing contaminants and insufficient samples
File: ITS_FullTable_McToolsR.txt
Description: Community matrix before cleaning/rarefaction
File: IR_taxonomy_loaded.csv
Description: Taxonomy of fungal ASVs present in samples
Variables
- taxonomy1: kingdom
- taxonomy2: phylum
- taxonomy3: class
- taxonomy4: order
- taxonomy5: family
- taxonomy6: genus
- taxonomy7: species
File: IR_map_loaded.csv
Description: Sample metadata
Variables
- SampleID
- See descriptions in EpiMicrMETAsqueaky.txt
File: FullITS_seqtab_newlabels.csv
Description: Community matrix with fungal ASVs as columns and samples as rows
File: IR_data_loaded.csv
Description: Community matrix after cleaning and rarefaction, with fungal ASVs as rows and samples as columns
File: ITS_CleanTable_McToolsR.txt
Description: Community matrix after cleaning
File: taxatab.csv
Description: Taxonomy data for fungal ASVs
Variables
- SeqID
- Taxonomy labels
File: FullITS_seqtab.csv
Description: Community matrix with fungal ASVs as columns and samples as rows
File: SamMap.csv
Description: Same as other mapping files
Variables:
- Same as other mapping files
File: ITS_taxonomy.xlsx
Description: Taxonomy data for fungal sequences present in samples.
File: ITS_Sequences.zip
Description: .zip file containing sequencing data from all samples (see below).
DNA SEQUENCING DATA
Format of file names:
SampleID_Read.fastq.gz
TreeID-Height-SampleType_Read#.fastq.gz
