DOI: 10.5061/dryad.266qs
title: Data from: Brood ball-mediated transmission of microbiome members in the dung beetle, Onthophagus taurus (Coleoptera: Scarabaeidae)

description: Insects feeding on plant sap, blood, and other nutritionally incomplete
 diets are typically associated with mutualistic bacteria that supplement
 missing nutrients. Herbivorous mammal dung contains more than 86%
 cellulose and lacks amino acids essential for insect development and
 reproduction. Yet one of the most ecologically necessary and
 evolutionarily successful groups of beetles, the dung beetles
 (Scarabaeinae) feeds primarily, or exclusively, on dung. These
 associations suggest that dung beetles may benefit from mutualistic
 bacteria that provide nutrients missing from dung. The nesting behaviors
 of the female parent and the feeding behaviors of the larvae suggest that
 a microbiome could be vertically transmitted from the parental female to
 her offspring through the brood ball. Using sterile rearing and a
 combination of molecular and culture-based techniques, we examine
 transmission of the microbiome in the bull-headed dung beetle, Onthophagus
 taurus. Beetles were reared on autoclaved dung and the microbiome was
 characterized across development. A ~1425 bp region of the 16S rRNA
 identified Pseudomonadaceae, Enterobacteriaceae, and Comamonadaceae as the
 most common bacterial families across all life stages and populations,
 including cultured isolates from the 3rd instar digestive system. Finer
 level phylotyping analyses based on lepA and gyrB amplicons of cultured
 isolates placed the isolates closest to Enterobacter cloacae, Providencia
 stuartii, Pusillimonas sp., Pedobacter heparinus, and Lysinibacillus
 sphaericus. Scanning electron micrographs of brood balls constructed from
 sterile dung reveals secretions and microbes only in the chamber the
 female prepares for the egg. The use of autoclaved dung for rearing, the
 presence of microbes in the brood ball and offspring, and identical 16S
 rRNA sequences in both parent and offspring suggests that the O. taurus
 female parent transmits specific microbiome members to her offspring
 through the brood chamber. The transmission of the dung beetle microbiome
 highlights the maintenance and likely importance of this
 newly-characterized bacterial community.
description: READMEThis file describes the
 contents of all the files submitted to Dryad.BacterialGenomesConcatenated.ConstraintTreeThis NEXUS formatted file provides two items. It provides the masked alignment of 264 loci for the 1095 taxa used to construct the constraint tree used later for the phylotyping analysis. It also provides the single maximum likelihood estimate of the bacterial phylogenetic relationships, as estimated by RAxML-Light (http://bioinformatics.oxfordjournals.org/content/early/2012/05/24/bioinformatics.bts309.abstract). This NEXUS file can be read by the program Mesquite (http://mesquiteproject.org/mesquite/mesquite.html).ConcatenatedAlignmentPositionsThis file provides the positions in the alignment in the file ‘BacterialGenomesConcatenated.ConstraintTree.nex’ for each of the 264 loci used for the constraint tree phylogenetic analyses.Concatenated_gyrB_lepA_PhylotypingTreeThis NEXUS formatted file provides two items. It provides the masked alignment of gyrB and lepA for the 1095 taxa and cultured isolates, and it provides the single maximum likelihood phylogeny that includes the cultured dung beetle microbiota samples and the 1095 reference bacteria. The RAxML analysis was topologically constrained by the phylogeny resulting from the full analysis of the 1095 bacterial genomes and 264 loci.ConcatenatedAlignmentPositions_gyrB_lepAThis file provides locus positions in the alignment in the file ‘Concatenated_gyrB_lepA_PhylotypingTree.nex’ for the gyrB and lepA loci used by the phylotyping analysis. This file includes the gyrB and lepA sequences of the cultured microbiota samples from Othnophagus taurus.AnalysisThis text file contains commands to carry out the RaxML analyses of the matrices contained in BacterialGenomesConcatenated.ConstraintTree.nex and Concatenated_gyrB_lepA_PhylotypingTree.nex. Please note that the data matrices of the NEXUS files must be converted to the Phylip format for use with RAxML. The matrices can be exported using the program Mesquite.