The percentage of workers that lost their Dufour's gland esters in queenless groups and colonies.

<p>Workers with less than 1% esters per total secretion were defined as workers that had lost their ester-sterility signal. The workers were 5-day-old and were kept in queenless groups of 3, 5 and 10. Workers' age in colonies was distributed normally (for more details see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0018238#pone.0018238-Amsalem1" target="_blank">[24]</a>). The numbers in brackets denote the numbers of workers per colony and the size of the sample (number of groups or colonies). All the workers in each group were dissected, therefore the n describes the number of Dufour glands as well. Letters above the columns denote statistical differences at p<0.05. Data are presented as mean ± SE.</p

Ovarian development and amount of esters in Dufour's gland of α- and β-workers in 3 different group-sizes.

<p>The experiments were performed in 5-day-old workers that were kept in queenless groups of 3, 5 and 10 (12 groups for each group-size). Data are presented as mean ± SE for the α-workers and β-workers in each group. Different letters denote statistical differences using two-way ANOVA test.</p

Aggression level per worker in α-workers and β-workers in 5-day-old workers.

<p>Workers were kept in queenless groups of 3, 5 and 10 (12 groups for each group-size). Each group was observed for a total of 120 minutes. Data are presented as mean ± SE for the α-workers and β-workers in each group. Different letters denote statistical differences using two-way ANOVA test.</p

Percentage of aggression exhibited by the α-worker towards the other females in the group.

<p>Aggression level includes cumulative aggression (attack and darting) during 5 days. Humming was excluded because of inability to determine directionality with certainty. Each group was observed for 120 minutes. Workers were kept in queenless groups of 3, 5 and 10 (12 groups for each group-size). Workers are presented in accordance with the amount of aggression each has received from the α-worker (e.g. 2′ female received the highest amount of aggression).</p

Molecular adaptation in olfactory functions on the fire ant social chromosome - supplementary data

<br><b><u>sinvgnG-SocialChr-genes-cDNA.fna  & sinvgnG-SocialChr-genes.gff:</u></b><br><br>For reference genome of <i>Solenopsis invicta</i> (version Si_gnG, http://www.ncbi.nlm.nih.gov/assembly/GCF_000188075.1/),<br>we have uploaded  gene-annotations and cDNA sequences for the genes that reside in the non-recombining region of the social chromosome.<br><br><br><u><b>socialChrORclade.tree:</b></u><br><br>Subtree, in newick format, of the hymenopteran odorant -receptor (OR) gene tree. Showing the subfamily that includes a tandem cluster of 23 ORs located in the <i>Solenopsis invicta</i> social chromosome.<br><br><br><b><u>SiORs.cDNA.fna:</u></b><br><br>cDNA sequences of <i>Solenopsis invicta </i>odorant -receptor (OR) genes<br><br><br><br><br><br><br><br

Effect of Bombus impatiens queen-produced cues on worker behavior, physiology, and gene expression

This file contains all the data associated with the manuscript. We include data on Bombus impatiens worker oocyte activation, egg-laying, aggressive behavior, and expression of the candidate genes vitellogenin and kr-h1. The data set allows comparison of these parameters in queenless workers, queenright workers, and workers exposed to volatile and contact cues generated by the queen

Supplementary Figure 1 from Chemical communication is not sufficient to explain reproductive inhibition in the bumblebee <i>Bombus impatiens</i>

Reproductive division of labour is a hallmark of eusociality, but disentangling the underlying proximate mechanisms can be challenging. In bumblebees, workers isolated from the queen can activate their ovaries and lay haploid, male eggs. We investigated if volatile, contact, visual or behavioural cues produced by the queen or brood mediate reproductive dominance in <i>Bombus impatiens.</i> Exposure to queen-produced volatiles, brood-produced volatiles and direct contact with pupae did not reduce worker ovary activation: only direct contact with the queen could reduce ovary activation. We evaluated behaviour, physiology and gene expression patterns in workers that were reared in chambers with all stages of brood and a free queen, caged queen (where workers could contact the queen, but the queen was unable to initiate interactions) or no queen. Workers housed with a caged queen or no queen fully activated their ovaries, whereas ovary activation in workers housed with a free queen was completely inhibited. The caged queen marginally reduced worker aggression and expression of an aggression-associated gene relative to queenless workers. Thus, queen-initiated behavioural interactions appear necessary to establish reproductive dominance. Queen-produced chemical cues may function secondarily in a context-specific manner to augment behavioural cues, as reliable or honest signal

Supplementary Table 1. Primer sequences from Chemical communication is not sufficient to explain reproductive inhibition in the bumblebee <i>Bombus impatiens</i>

Reproductive division of labour is a hallmark of eusociality, but disentangling the underlying proximate mechanisms can be challenging. In bumblebees, workers isolated from the queen can activate their ovaries and lay haploid, male eggs. We investigated if volatile, contact, visual or behavioural cues produced by the queen or brood mediate reproductive dominance in <i>Bombus impatiens.</i> Exposure to queen-produced volatiles, brood-produced volatiles and direct contact with pupae did not reduce worker ovary activation: only direct contact with the queen could reduce ovary activation. We evaluated behaviour, physiology and gene expression patterns in workers that were reared in chambers with all stages of brood and a free queen, caged queen (where workers could contact the queen, but the queen was unable to initiate interactions) or no queen. Workers housed with a caged queen or no queen fully activated their ovaries, whereas ovary activation in workers housed with a free queen was completely inhibited. The caged queen marginally reduced worker aggression and expression of an aggression-associated gene relative to queenless workers. Thus, queen-initiated behavioural interactions appear necessary to establish reproductive dominance. Queen-produced chemical cues may function secondarily in a context-specific manner to augment behavioural cues, as reliable or honest signal

Normalized read counts for PCA

Normalized read counts for PC

Phenotype raw data

raw data for ovarian activation, juvenile hormone levels, lipid and glycogen content, queen weight and gene expression data (fold change) for 11 genes that were tested in the stud