DOI: 10.5061/dryad.cjsxksndz
title: Data from: Inter-clonal competition over queen succession imposes a cost of parthenogenesis on termite colonies

description: In social insect colonies, selfish behaviour due to intracolonial conflict
 among members can result in colony-level costs despite close relatedness.
 In certain termite species, queens use asexual reproduction for
 within-colony queen succession but rely on sexual reproduction for worker
 and alate production, resulting in multiple half-clones of a single
 primary queen competing for personal reproduction. Our study demonstrates
 that competition over asexual queen succession among different clone types
 leads to the overproduction of parthenogenetic offspring, resulting in the
 production of dysfunctional parthenogenetic alates. By genotyping the
 queens of 23 field colonies of Reticulitermes speratus, we found that
 clone variation in the queen population reduces as colonies develop. Field
 sampling of alates and primary reproductives of incipient colonies showed
 that overproduced parthenogenetic offspring develop into alates that have
 significantly smaller body sizes and much lower survivorship than
 sexually-produced alates. Our results indicate that while the production
 of earlier and more parthenogenetic eggs is advantageous for winning the
 competition for personal reproduction, it comes at a great cost to the
 colony. Thus, this study highlights the evolutionary interplay between
 individual-level and colony-level selection on parthenogenesis by queens.
description: <strong>(a) Clonal drive in queen
 population</strong> We collected 175 colonies
 with kings and queens of <em>R. speratus</em> in pine or
 Japanese cedar forests in Kyoto, Shiga, Wakayama, Nagano, and Chiba, Japan
 from May to September 2017–2019. All termites were extracted from the nest
 within 10 days of collection, and the phenotypes of kings and queens
 (primary or secondary) and the number of each were recorded. The data were
 included in sheets “colony information” and “no. of royals”. Of the 175
 total colonies, 28 were used for the genotyping analyses. Twenty secondary
 queens were randomly selected from each colony and stored at −80°C for
 genotyping. Primary queens were found in four of the colonies and were
 also stored for genotyping. The genotyping data were included in sheet
 “genotype of SQs”. Of the 28 colonies used for the genotyping analysis, 23
 were used for taking queen weight measurements. Termites in the 23
 colonies were extracted from the nest within 2 days of collection, and
 fresh weights of secondary queens were measured to the nearest 0.1 mg.
 Prior research has established that the total weight of queens, rather
 than the total number of queens, is a more accurate indicator of colony
 size. Therefore, the total weight of queens was used to analyse the
 correlation between the indicator of colony size and the degree of clonal
 drive. The data were included in sheet “no. of clones in SQ
 population”.<br>To investigate whether the frequency of certain
 alleles at specific loci increases in the secondary queen population from
 colonies dominated by a single clone type in queens, we conducted
 Fisher's exact test with Bonferroni correction. To compile the
 dataset for this analysis, we first defined a colony as a driven colony if
 over 90% of the genotyped secondary queens were composed of a single
 clone, which was then classified as the dominated clone type. Five
 colonies met this criterion (180626F, 180605I, 180721A, 180605L, 180609A).
 Colonies not meeting this criterion were categorized as non-driven
 colonies. Secondly, we listed the alleles found in each non-driven colony
 (sheet “alleles in non-driven colonies”), representing the alleles present
 in the primary queens of these colonies. Thirdly, to investigate the
 frequency of each allele present before the occurrence of clonal drive, we
 counted the number of each allele found in non-driven colonies for this
 analysis. If only one allele was detected at a locus, it means that the
 primary queens was homozygous for that allele, and therefore, that allele
 was double counted. Fourthly, we recorded the alleles of the dominated
 clone type in each driven colony (sheet "alleles in dominated
 clones"), and the count of each allele was used for the
 analysis. <strong>(b)
 Parthenogenetically-produced alates in the field</strong>
 (i) Sampling of dealates walking on the ground
 We collected a pre-foundation population of <em>R.
 speratus</em> by traps. Each trap consisted of a plastic board (210
 × 297 mm), a guiding wall (50 mm height), and four sticky traps (16 × 80 ×
 90 mm). The sticky trap, which has openings on all sides, was attached at
 each edge of the guiding wall. The guiding wall of this trap was set for
 leading dealates walking on the plastic board into the sticky traps. Fifty
 traps (a total of 200 sticky traps) were set haphazardly on the ground of
 an open forest of pine trees with grassy areas in Hieidaira, Kyoto, Japan,
 from 5 to 7 May 2015 during the swarming season of <em>R.
 speratus</em>. A massive synchronized flight was observed only on
 May 5th in this study site. On 7 May 2015, we brought all traps back to
 the laboratory and collected all the dealates from the sticky traps. The
 head width (the maximum distance across compound eyes) of each sample was
 measured under a stereoscope (Olympus, Tokyo, Japan) using a digital
 imaging system (FLVFS-LS; Flovel, Tokyo, Japan). Since the condition of
 the sticky trap samples varied among individuals, we measured head width
 as a stable body size indicator. The dealates were stored at −80°C for the
 following microsatellite analysis. Then, we analysed the genotypes of each
 individual for all eight microsatellite loci analysed. The data were
 included in sheet “genotype of alates”. (ii) Sampling
 of dealates after colony foundation We collected a
 post-foundation population of <em>R. speratus</em> from
 experimentally buried brown rotten pine wood pieces which were suitable
 for termite nesting material. The pine wood was cut into pieces of
 approximately 20W × 40D × 10H cm size, autoclaved, and half-buried in the
 ground at the same site on April 28, 2015. On May 14, 2015, which was 9
 days after the mating flight, all the buried pine wood was excavated and
 transported to the laboratory. The pine wood was meticulously dissected to
 extract founding units. We determined the sex of each individual by
 examining the morphology of the terminal abdominal sterna under a
 microscope. The individuals were placed separately in a collection tube
 (1.5 mL) and stored at −25ºC. Then, we analysed the genotypes of each
 individual for the eight microsatellite loci. The data were included in
 sheet “genotype of alates”.
 <strong> </strong><strong>(c) Survivorship of
 sexually- and parthenogenetically-produced alates</strong>
 Three colonies (colonies A−C) containing alates were collected
 from pine or Japanese cedar forests in Kyoto, Japan, just before the
 swarming season in 2016. These colonies were maintained at 20°C to keep
 alates from swarming until the experiment started. Just before the
 experiment, each colony was transferred to a room at 28°C, and alates
 stimulated by high temperature emerged from their chambers in wood. Alates
 were then separated by sex and maintained in Petri dishes lined with a
 moist unwoven cloth, and used for the experiments within a day of the
 flight. We used 40 males and 40 females randomly chosen from each of the
 three colonies. After removing the wings from the alates, we measured
 their fresh weight and used them in the following experiment. The wings of
 each individual were preserved in a test tube containing 99.5% ethanol.
 Then, we analysed the genotypes of each individual using the wings for
 four microsatellite loci. To compare viability and
 immunity levels between sexually-produced alates and
 parthenogenetically-produced alates, we investigated the survivorship of
 alates both with and without exposure to the entomopathogenic fungus
 <em>Metarhizium anisopliae</em> (source: National Institute of
 Technology Evaluation Biological Resource Centre, NBRC31961) that occurs
 with the termites in nature. Each termite was randomly assigned to one of
 the two treatments. See Text S1 for the methods of pathogen treatment. In
 the control treatment without the pathogen, the alates were individually
 exposed to a conidia-free 0.025% Tween 20 solution (without pathogen).
 After exposure to a conidia-free solution, termites were placed
 individually in a well of a 24-well plate (COSTAR®3526, Corning Inc. NY)
 lined with filter paper moistened with distilled water. The plates were
 maintained at 25ºC in darkness and checked every day for 30 days to
 investigate alate survival. The data were included in sheet “survivorship
 of alates”.
description: # Data from: Inter-clonal competition over queen succession imposes a cost
 of parthenogenesis on termite colonies
 [https://doi.org/10.5061/dryad.cjsxksndz](https://doi.org/10.5061/dryad.cjsxksndz) This dataset contains supplementary data associated with Yao *et al.* 2024 *Proceedings of the Royal Society B: Biological Sciences*. ## Description of the data and file structure This data contains dataset and R code. The "code.Rmd" file includes R code for statistical analyses. The "dataset.xlsx" file is divided into 8 sheets as detailed below: \- sheet "colony information" Contains data on the collection locations and collection dates of the colonies used in the study. Columns are described below: "colony" = data on colony IDs "location" = data on sites where colonies were collected "collection_date" = data on date colony was sampled (YYYY/MM/DD)   \- sheet "no. of royals" Contains the number of kings and queens in the colony. Columns are described below: "colony" = data on colony IDs "no_of_primary_king" = data on number of primary kings in the colony "no_of_secondary_king" = data on number of secondary kings in the colony "no_of_primary_queen" = data on number of primary queens in the colony "no_of_secondary_queen" = data on number of secondary queens in the colony   \- sheet "genotype of queens" Contains genotype data for queens. Columns are described below: "colony" = data on colony IDs "caste" = data on caste of genotyped individuals (PQ = primary queen, SQ = secondary queen) "Rf24_2" = genotypes of microsatellite locus *Rf24-2* "Rf6_1" = genotypes of microsatellite locus *Rf6-1* "Rf21_1" = genotypes of microsatellite locus *Rf21-1* "Rs15" = genotypes of microsatellite locus *Rs15* "Rs10" = genotypes of microsatellite locus *Rs10* "Rs78" = genotypes of microsatellite locus *Rs78* "Rs68" = genotypes of microsatellite locus *Rs68* "Rs02" = genotypes of microsatellite locus *Rs02*   \- sheet "alleles in non-driven colonies" Contains the list of alleles found in each non-driven colony. Columns are described below: "colony" = data on colony IDs "drive_level" = data on level of the clonal drive "no_secondary_queens_genotyped" = data on number of genotyped secondary queens "no_of_clone_types" = data on number of clone types found in the genotyped secondary queens "no_of_individuals_of_the_most_dominant_clone_type" = data on number of individuals of the most dominant clone type "Rf24_2" = list of alleles found in microsatellite locus *Rf24-2* "Rf6_1" = list of alleles found in microsatellite locus *Rf6-1* "Rf21_1" = list of alleles found in microsatellite locus *Rf21-1* "Rs15" = list of alleles found in microsatellite locus *Rs15* "Rs10" = list of alleles found in microsatellite locus *Rs10* "Rs78" = list of alleles found in microsatellite locus *Rs78* "Rs68" = list of alleles found in microsatellite locus *Rs68* "Rs02" = list of alleles found in microsatellite locus *Rs02*   \- sheet "alleles in dominant clones" Contains the alleles found in the dominant clone type in each driven colony. Columns are described below: "colony" = data on colony IDs "drive_level" = data on level of the clonal drive "no_secondary_queens_genotyped" = data on number of genotyped secondary queens "no_of_clone_types" = data on number of clone types found in the genotyped secondary queens "no_of_individuals_of_the_most_dominant_clone_type" = data on number of individuals of the most dominant clone type "Rf24_2" = genotypes of microsatellite locus *Rf24-2* of the dominant clone type "Rf6_1" = genotypes of microsatellite locus *Rf6-1* of the dominant clone type "Rf21_1" = genotypes of microsatellite locus *Rf21-1* of the dominant clone type "Rs15" = genotypes of microsatellite locus *Rs15* of the dominant clone type "Rs10" = genotypes of microsatellite locus *Rs10* of the dominant clone type "Rs78" = genotypes of microsatellite locus *Rs78* of the dominant clone type "Rs68" = genotypes of microsatellite locus *Rs68* of the dominant clone type "Rs02" = genotypes of microsatellite locus *Rs02* of the dominant clone type   \- sheet "no. of clones in SQ population" Contains the number of kings and queens in each colony, the total weight of the queens, the number of clone types found in the secondary queen, and the number of individuals of the most dominant clone type. Columns are described below: "colony" = data on colony IDs "no_of_primary_king" = data on number of primary kings in the colony "no_of_secondary_king" = data on number of secondary kings in the colony "no_of_primary_queen" = data on number of primary queens in the colony "no_of_secondary_queen" = data on number of secondary queens in the colony "total_weight_of_queen" = data on total fresh weight of the queens in the colony in milligrams "no_of_clone_types" = data on number of clone types found in the genotyped secondary queens in the colony "no_of_clone_types" = data on number of clone types found in the genotyped secondary queens "no_of_individuals_of_the_most_dominant_clone_type" = data on number of individuals of the most dominant clone type   \- sheet "genotype of alates" Contains genotype data for alates in pre- and post-founding population. Columns are described below: "ID" = data on IDs of the alate "sex" = data on sex of the alate "stage" = data on stage of the alate"sex" = data on sex of the alate (pre_founding = alate collected from pre-founding population, post_founding = alate collected from post-founding population) "Rf24_2" = genotypes of microsatellite locus *Rf24-2* "Rf6_1" = genotypes of microsatellite locus *Rf6-1* "Rf21_1" = genotypes of microsatellite locus *Rf21-1* "Rs15" = genotypes of microsatellite locus *Rs15* "Rs10" = genotypes of microsatellite locus *Rs10* "Rs78" = genotypes of microsatellite locus *Rs78* "Rs68" = genotypes of microsatellite locus *Rs68* "Rs02" = genotypes of microsatellite locus *Rs02* "heterozygosity" = data on the heterozygosity of the alate "parthenogenesis" = data indicating whether the alates are sexually or parthenogenetically produced (sexual = sexually-produced alate, asexual = parthenogenetically-produced alate)   \- sheet "survivorship of alates" Contains the survival time of individuals submitted to control and pathogen treatment. Columns are described below: "ID" = data on IDs of the alate "colony" = data on colony IDs "sex" = data on sex of the alate "treatment" = data on treatment group "weight" = data on fresh weight of the alate in milligrams "event_time" = data on the number of days from a starting point to the occurrence of death or censoring "status" = data indicating the occurrence of death or censoring (1 = death, 0 = censoring) "Rf24_2" = genotypes of microsatellite locus *Rf24-2* "Rf6_1" = genotypes of microsatellite locus *Rf6-1* "Rf21_1" = genotypes of microsatellite locus *Rf21-1* "Rs15" = genotypes of microsatellite locus *Rs15* "heterozygosity" = data on the heterozygosity of the alate "parthenogenesis" = data indicating whether the alates are sexually or parthenogenetically produced (sexual = sexually-produced alate, asexual = parthenogenetically-produced alate)   \- sheet "weight of young and old SQs" Contains the weight of the primary king and secondary queens. Columns are described below: "colony" = data on colony IDs "caste" = data on caste (PK = primary king, SQ = secondary queen) "body_weight" = data on fresh weight in milligrams   \- sheet "composition of founding units" Contains the data for composition of founding units. Columns are described below: "combination" = data on combination of founding units (M = single male, F = single female, MF = male-female pair, MM = male-male pair, MFF = single male with two females, MMF = single female with two males, MFFF = single male with three females, MMFF = two males and two females) "number_of_colony" = data on number of colonies found in the experiment ## Sharing/Access information This data is described in: * Yao *et al.* 2024 Inter-clonal competition over queen succession imposes a cost of parthenogenesis on termite colonies. *Proceedings of the Royal Society B: Biological Sciences* ## Code/Software All analyses were performed using R version 4.2.3.