DOI: 10.5061/dryad.f1vhhmh3m
title: Data and code for: A supergene controlling social structure in Alpine ants also affects the dispersal ability and fecundity of each sex

description: Social organisation, dispersal and fecundity co-evolve, but whether they
 are genetically linked remains little known. Supergenes are prime
 candidates for coupling adaptive traits and mediating sex-specific
 trade-offs. Here, we test whether a supergene that controls social
 structure in Formica selysi also influences dispersal-related traits and
 fecundity within each sex. In this ant species, single-queen colonies
 contain only the ancestral supergene haplotype M and produce MM queens and
 M males, while multi-queen colonies contain the derived haplotype P and
 produce MP queens, PP queens, and P males. By combining multiple
 experiments, we show that the M haplotype induces phenotypes with higher
 dispersal potential and higher fecundity, for both sexes. Specifically, MM
 queens, MP queens, and M males are more aerodynamic and more fecund than
 PP queens and P males, respectively. Differences between MP and PP queens
 from the same colonies reveal a direct genetic effect of the supergene on
 dispersal-related traits and fecundity. The derived haplotype P,
 associated with multi-queen colonies, produces queens and males with
 reduced dispersal abilities and lower fecundity. More broadly,
 similarities between the Formica and Solenopsis systems reveal that
 supergenes play a major role in linking behavioural, morphological, and
 physiological traits associated with intraspecific social polymorphisms.
description: The Alpine silver ant, <em>F. selysi</em>, is a
 socially polymorphic species occupying riverine habitats in Alpine
 valleys. Colony social structure is controlled by a large (13.8 Mb long,
 ~745 genes) and ancient (~30-million-year-old) supergene. We collected
 winged queens, males, and workers of <em>F. selysi. </em>We
 genotyped the social supergene of three workers per colony, to determine
 colony social form. We genotyped the supergene of all queens from polygyne
 colonies, using DNA extracted from their wings or legs. We conducted the
 following experiments, comparing individuals with alternate
 genotypes: <em>Experiment one: supergene’s effect
 on queen and male morphology</em> We measured the
 fresh weight, estimated body size (distance between eyes), wing size
 (length, width, perimeter, and area); and estimated wing loading (fresh
 weight divided by the sum of the area of her four wings) of queens, and
 measured the size and wings of males.
 <em>Experiment two: supergene’s effect on queen flight
 tendencies and abilities</em> We evaluated the
 flight propensity, flight abilities, and costs of flying (weight lost) of
 virgin queens of all genotypes, using a custom-made mill that
 automatically records flying time, speed, and distance. See
 also:  De Gasperin, O. 2023 Queen flying in the flight
 mill. figshare. https://doi.org/10.6084/m9.figshare.23932791.v1
 <em>Experiment three:</em><em> supergene’s
 effect on male fecundity and mating success</em>
 a) We estimated the number of sperm cells produced by males and
 the number of sperm cells that males transfer to queens during their first
 and second mating event, respectively, using flow cytometry.
 b) We compared <em>MM </em>queen’s<em>
 </em>preference for the odour of <em>M </em>or
 <em>P </em>males. c) We evaluated the
 mating success of virgin males in experimental mating boxes, with a ratio
 close to 1:1 of <em>M:P </em>males, competing for a
 limiting<em> </em>number of <em>MM </em>queens, in
 artificial swarms. <em>Experiment four:
 supergene’s effect on queen fecundity and colony
 survival</em> We compared the survival and number
 of workers of 630 one-year old colonies established by queens of each
 supergene genotype and mated to M or P males (colonies formed by a single
 queen mated to a single male).
description: # Dispersing Supergenes In these files, you will find all the dataset
 supporting the paper "A supergene controlling social structure in
 Alpine ants also affects the dispersal ability and fecundity of each
 sex", and all scripts supporting the analyses. ## Description of the
 data and file structure **Experiment 1:** Data for queens:
 morphology_full_2022.csv Data for queens: final_mill.csv Data for males:
 MaleSize.csv Data for males: MaleWings.csv
 Script: DispersingSupergenesScript_Experiment1.Rmd **Experiment 2:**
 Data: final_mill.csv Script: DispersingSupergenesScript_Experiment2.Rmd
 **Experiment 3a:** Data: male_fertility_data_final.csv
 Script: DispersingSupergenesScript_Experiment3aSperm.Rmd **Experiment
 3b:** Data: QueenChoiceExperiment.csv
 Script: DispersingSupergenesScript_Experiment3b.Rmd **Experiment 3c:**
 Data: Mating_trials_males.csv
 Script: DispersingSupergenesScript_Experiment3c.Rmd **Experiment 4:**
 Data: HibernatingQueensMated.csv
 Script: DispersingSupergenesScript_Experiment4.Rmd ## Detailed Description
 of the data and file structure **Experiment 1:** **Data for queens:
 morphology_full_2022.csv** Variables used: Explanation of columns used in
 the analyses: size: distance between the eyes of the queen; units = mm
 mated: was the queen mated in artificial swarms or collected from her
 colony of origin, without placed in artificial swarms fem_fam: colony of
 origin of the queen population: population (De= high elevation population;
 Fi= low elevation population) genotype: Genotype of the queen year: year
 when the data was collected **Data for queens: morphology_full_2022.csv**
 Variables used: weightAv: weight of the queen (fresh weight), unit = mg
 colony_origin: colony of origin of the queen **Data for queens:
 final_mill.csv** Variables used: load: wing loading of the queen (weight
 of the queen (fresh weight; unit = mg), divided by the area of her wings;
 unit = mm squared) winglength_1: length of the longest wing of the
 queen; unit = mm wingarea_model1: area of the longest wing of the queen;
 unit = mm **Data for males: MaleSize.csv** Variables used: de: distance
 between the eyes of the male (size of the male); unit = mm pop: population
 (De= high elevation population; Fi= low elevation population)
 colony: colony of origin of the male fmss: Genotype of the male (or queen)
 sex: Sex of the individual **Data for males: MaleWings.csv** Variables
 used: ss: Genotype of the male winglength_1: length of the longest wing of
 the male; unit = mm pop: population (De= high elevation population; Fi=
 low elevation population) colony: colony of origin of the male
 **Experiment 2:** Data: final_mill.csv Variables used: channel: specific
 'box' where the queen was tested genotype: Genotype of the queen
 trial_duration: time in seconds the queen was allowed to fly for; unit =
 seconds flew: did the queen fly? y = yes, she flew population: population
 (De= high elevation population; Fi= low elevation population) weightAv:
 weight of the queen (fresh weight); unit = mg weight_diff: difference in
 the fresh weight of the queen before and after the trial (cost of flying),
 calculated as weight before the trial (weightAv) - weight after the trial
 (weightAp); unit = mg tot_flight.time: total time the queen spent flying;
 unit = in seconds colony_origin: colony of origin of the queen
 **Experiment 3a:** Data: male_fertility_data_final.csv Variables used:
 male_col: colony of origin of the male male_genotype: Genotype of the male
 absolute_count: absolute number of sperm cells collected from the male (or
 from the queen's spermatheca); unit = number of sperm cells
 de: distance between the eyes of the male (size of the male); units = mm
 pop: population of origin of the male (De= high elevation population; Fi=
 low elevation population) male_condition: virgin (the sperm sample came
 from the virgin male, otherwise = the sperm sample came from a
 queen's spermatheca, if 1stMating it came from a queen that mated to
 a virgin male, and 2ndMating, it came from a queen that mated to a
 previously male) **Experiment 3b:** Data: QueenChoiceExperiment.csv
 Variables used: ant: individual queen number zonesecond: seconds the queen
 spent in the 'chose' area controlorant: was the smell the
 control (solvent) or did it have a male's odour condition: did the
 odour came from a mated or virgin male zonetype: was the zone belonging to
 the odour of a P or of a M male? volume: quantity of liquid used (same
 within a replicate); unit = μL queenfullcolony: colony of origin of the
 queen malefullcolony: colony of origin of the male (sample for the odour)
 **Experiment 3c:** Data: Mating_trials_males.csv Variables used: Escaped:
 Did the male escape? 1= the male escaped (this data was excluded)
 Male_condition= was the male virgin or previously mated?
 Male_sf_New= Genotype of the male Box: Experimental box Male_col_New:
 Colony of origin of the male Population: population of origin of the male
 (De= high elevation population; Fi= low elevation population)
 QtoM_Ratio_end: Queen to male ratio in the experimental box Mating: Did
 the male mate? 1 = the male mated, 0 = the male failed to mate in the
 trial **Experiment 4:** Data: HibernatingQueensMated.csv Variables used:
 genotype: Genotype of the queen fail: Did the queen fail? 1= failed,
 because she died or because she did not produce a brood
 queen_genotype: Genotype of the queen male_genotype: Genotype of the male
 with whom the queen mated Population: population of origin of the queen
 (De= high elevation population; Fi= low elevation population) fem_fam:
 Colony of origin of the queen brood_size_end_experiment: Number of workers
 and pupae in the colony at the end of the experiment ## Code/Software
 **the MatLab code measures wing sizes.** --- – **MeasureWingSize.m ** is a
 script that generates a table for each image inside a folder. – It
 requires to add inside the folder an empty .mat file called
 AntWingList.mat .  – Make sure the file is empty before starting the
 script – The script (MeasureWingSize.m) runs on all the photos inside the
 current directory (MATLAB’s current folder). – Make sure that the folder
 contains ONLY photos that share a reference (as the script measures Pixels
 that are then converted into mm by a different script
 (GenerateTableInMM.m).