Nitrogen (N) deposition in many areas of the world is over an order of magnitude greater than it would be in absence of human activity. We ask how abiotic (N) and biotic (plant host and neighborhood) effects interact to influence root-associated bacterial (RAB) community assembly. Using 454 pyrosequencing, we examined RAB communities from two dominant alpine tundra plants, Geum rossii and Deschampsia cespitosa, under control, N addition and D. cespitosa removal treatments, implemented in a factorial design. We hypothesized that host would have the strongest effect on RAB assembly, followed by N, then neighbor effects. The most dominant phyla were Proteobacteria (mostly Gammaproteobacteria), Actinobacteria, Bacteroidetes and Acidobacteria. We found RAB communities were host specific, with only 17% overlap in operational taxonomic units. Host effects on composition were over twice as strong as N effects. D. cespitosa RAB diversity declined with N, while G. rossii RAB did not. D. cespitosa removal did not influence G. rossii RAB community composition, but G. rossii RAB diversity declined with N only when D. cespitosa was absent. We conclude that RAB of both hosts are sensitive to N enrichment, and RAB response to N is influenced by host identity and plant neighborhood.
NiwotB_all_denoised
All 454 sequences in fasta format. These have already been denoised, chimera checked, and quality filtered using Ampliconnoise as implemented in MacQiime (v.1.6.0)
NiwotB_all_map2
Mapping file that corresponds with fasta file (NiwotB_all_denoised.fna)
nG6XN_nchl_filtered_sorted_host
OTU table built in QIIME from NiwotB_all_denoised.fna. This table has had chloroplast sequences removed. Also the most shallowly sequenced sample (G6CXN) has been removed. Columns sorted by host and treatment. Unrarefied. This table was used for dbRDA. 100 rarefactions tables (in which each sample was rarefied to the most shallowly sequenced sample = 202 sequences) were also built from this table, and alpha diversity measures were conducted on each of those 100 rarefaction tables.
raref_nG6XN_nchl_filtered_sorted_host_L7
This OTU table was built in QIIME from NiwotB_all_denoised.fna. Chloroplast sequences and the most shallowly sequenced sample (G6CXN) are removed, and all samples have been rarefied to the next most shallowly sequenced sample (202 sequences). Columns are sorted by host. This table was used for ANOVAs conducted on individual taxa.
raref_nG6XN_nchl_filtered_sorted_host.txt
read.qiime.table
This script is used to read any qiime table into R. It was run prior to the other R scripts, so that the files would be uploaded correctly.
raref_nG6XN_nchl_filtered_sorted_host_L6
This genus table was built in QIIME from raref_nG6XN_nchl_filtered_sorted_host_L7.txt. Chloroplast sequences and the most shallowly sequenced sample (G6CXN) are removed, and all samples have been rarefied to the next most shallowly sequenced sample (202 sequences). Columns are sorted by host. This table was used for ANOVAs conducted on individual taxa.
raref_nG6XN_nchl_filtered_sorted_host_L5
This family table was built in QIIME from raref_nG6XN_nchl_filtered_sorted_host_L7.txt. Chloroplast sequences and the most shallowly sequenced sample (G6CXN) are removed, and all samples have been rarefied to the next most shallowly sequenced sample (202 sequences). Columns are sorted by host. This table was used for ANOVAs conducted on individual taxa.
raref_nG6XN_nchl_filtered_sorted_host_L4
This order table was built in QIIME from raref_nG6XN_nchl_filtered_sorted_host_L7.txt. Chloroplast sequences and the most shallowly sequenced sample (G6CXN) are removed, and all samples have been rarefied to the next most shallowly sequenced sample (202 sequences). Columns are sorted by host. This table was used for ANOVAs conducted on individual taxa.
raref_nG6XN_nchl_filtered_sorted_host_L3
This class table was built in QIIME from raref_nG6XN_nchl_filtered_sorted_host_L7.txt. Chloroplast sequences and the most shallowly sequenced sample (G6CXN) are removed, and all samples have been rarefied to the next most shallowly sequenced sample (202 sequences). Columns are sorted by host. This table was used for ANOVAs conducted on individual taxa.
raref_nG6XN_nchl_filtered_sorted_host_L2
This phylum table was built in QIIME from raref_nG6XN_nchl_filtered_sorted_host_L7.txt. Chloroplast sequences and the most shallowly sequenced sample (G6CXN) are removed, and all samples have been rarefied to the next most shallowly sequenced sample (202 sequences). Columns are sorted by host. This table was used for ANOVAs conducted on individual taxa.
PD_whole_tree
phylogenetic diversity calculated for 100 rarefactions (202 sequences) of each sample.
observed_species_L7
OTU richness calculated for each of 100 rarefactions (to 202 sequences) performed on each sample.
observed_species_L6
genus richness calculated for each of 100 rarefactions (to 202 sequences) performed on each sample.
observed_species_L5
family richness calculated for each of 100 rarefactions (to 202 sequences) performed on each sample.
observed_species_L4
order richness calculated for each of 100 rarefactions (to 202 sequences) performed on each sample.
observed_species_L3
class richness calculated for each of 100 rarefactions (to 202 sequences) performed on each sample.
observed_species_L2
phylum richness calculated for each of 100 rarefactions (to 202 sequences) performed on each sample.
simpson_L7
OTU diversity calculated by Simpson's diversity index in QIIME.
simpson_L6
Genus diversity calculated by Simpson's diversity index in QIIME.
simpson_L5
Family diversity calculated by Simpson's diversity index in QIIME.
simpson_L4
Order diversity calculated using Simpson's Diversity index in QIIME
simpson_L3
Class diversity calculated using Simpson's Diversity index in QIIME
simpson_L2
Phylum diversity calculated using Simpson's Diversity index in QIIME
even_L2
Phylum evenness, calculated using Simpson's evenness, calculated for each of 100 rarefactions (202 sequences) for each sample.
even_L3
Class evenness, calculated using Simpson's Evenness, calculated for each of 100 rarefaction tables (202 sequences) performed on each sample.
even_L4
Order evenness, calculated using Simpson's Evenness, calculated for each of 100 rarefaction tables (202 sequences) performed on each sample.
even_L5
Class evenness, calculated using Simpson's Evenness, calculated for each of 100 rarefaction tables (202 sequences) performed on each sample.
even_L6
Class evenness, calculated using Simpson's Evenness, calculated for each of 100 rarefaction tables (202 sequences) performed on each sample.
even_L7
Class evenness, calculated using Simpson's Evenness, calculated for each of 100 rarefaction tables (202 sequences) performed on each sample.
weighted_unifrac_nchl_filtered_sorted_host
Weighted unifrac table built in QIIME from unrarefied OTU table (chloroplasts removed) to use for dbRDA.
otu_anova_G vs D_Neffect
R code for ANOVA on individual root associated bacterial taxa in Geum rossii and Deschampsia cespitosa. Test to determine effect of nitrogen addition on abundance of any bacterial taxa. Tested in each host seperately.
otu_anova_Nxhost
R code ANOVA on individual root associated bacterial taxa in Geum rossii and Deschapmsia cespitosa roots. Purpose to determine effect of host plant identity and nitrogen addition on abundance of individual bacterial taxa.
otu_anova_remxN
R code for ANOVA on individual root associated bacterial taxa in Geum rossii. Test to determine whether nitrogen addition or Deschampsia cespitosa removal (principle competitor removal) had an effect on abundance of any bacterial taxa.
pie_charts
R code: Build pie charts for each host x treatment combination to see relative abundance of each phylum
alpha
R code: statistical analysis (ANOVA) on collated alpha diversity metrics (phylogenetic diversity, richness, simpson's evenness, and simpson's diversity) calculated for 100 rarefaction tables.
dbRDA_bact
R code: dbRDA and ordination plots, performed on weighted unifrac table derived from unrarefied OTU table.
README
README file, explains all other files