An Insect Herbivore Microbiome with High Plant Biomass-Degrading Capacity Page: 3 of 14
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Leaf-Cutter Ant Fungus Garden Microbiome
2,794 near full-length bacterial 16S rDNA sequences were
generated for fungus garden top and bottom layers, respectively
(Table Si), and short-read pyrotag sequencing of the same samples
yielded 8,968 and 11,362 sequences, respectively. PCR using full-
length Archaea-specific primers failed to amplify Archaeal 16S
rDNA. Community metagenome sequencing of whole fungus
gardens using pyrosequencing [14] generated over 401 Mb of
sequence (Table S2), and assembly resulted in 155,000 contigs and
200,000 singletons, totaling 130 Mb.These DNA sequences indicate the presence of a diverse
community of bacteria in leaf-cutter ant fungus gardens (Figure 2,
Figure S1, Figure S2). Full-length 16S rDNA libraries contained
132 phylotypes (97% sequence identity) from 9 phyla in garden
tops (Figure 2A, Table S3), and 197 phylotypes from 8 phyla in
garden bottoms (Figure 2B, Table S3). Comparison of the phylo-
genetic diversity between top and bottom layer samples using
UniFrac [15] indicates that the top layer diversity is different
from bottom layer diversity (Figure S3). Both top and bottomB
Yproteobactera [2016]
p-proteobacteria [120]
BICP544
a-proteobacteria [329
8-proteobacteria [8Acidobacteria [8]
Firmicutes (20]
nActnobacterIa [51]
--Plarnctomycetes [4]
81G0396
BIGO39
BIG0637 TM7 [2]
Chloroflexr [2]
Bacteroidetes [15]
Verrucomicrobla [12]0.10
7
6
29
3
1
2
7
5Figure 2. Phylogenetic analysis of the leaf-cutter ant fungus garden. A phylogenetic analysis of near-full length 165 rDNA sequence libraries
from the top (A) and bottom (B) layers of leaf-cutter ant fungus gardens was performed. Identified phylotypes were tabulated and mapped to their
respective phyla as shown. Total numbers of phylotypes are shown to the right of each phylum, and the total number of clones for each phylum is
shown in square brackets. Comparison of top and bottom layers indicates that leaf-cutter ant fungus gardens are dominated by phylotypes
belonging to the c-proteobacteria, p-proteobacteria, y-proteobacteria, Actinobacteria, and the Bacteroidetes as highlighted. Phylotypes belonging to
specific phyla were found exclusive to top and bottom samples, including the Gemmatimonadetes and candidate phylum SPAM (blue lettering) in
the top, and the Chloroflexi and candidate phylum TM7 (red lettering) in the bottom of the garden.
doi:10.1371/journal.pgen.1001129.g002
. PLoS Genetics www.plosgenetics.org 3 September 2010 Volume 6 Issue 9 e1001129A
Y-proteobacteria [548] 56
p-proteobacteria [4] 4
TIGU1277
d-proteobacteria [3]11
a-proteobacterla [3] 2
TIGU1470
Acidobacleria [6] 6
Firm icutes [3] 3
Actinobacteria [40] 19
Planctomycetes [3] 3
Barteroidees [17] 13
Verrucomicrobia [3] 3
TIGU971
TIGU884
Gc~onn:nm cs 2 ] 2
TIBW100B
TIGU1362
SPArS - 2""
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73
15
385
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Suen, Garret; Barry, Kerrie; Goodwin, Lynne; Scott, Jarrod; Aylward, Frank; Adams, Sandra et al. An Insect Herbivore Microbiome with High Plant Biomass-Degrading Capacity, article, September 23, 2010; Berkeley, California. (https://digital.library.unt.edu/ark:/67531/metadc834301/m1/3/: accessed April 19, 2024), University of North Texas Libraries, UNT Digital Library, https://digital.library.unt.edu; crediting UNT Libraries Government Documents Department.