DOI: 10.5061/dryad.fqz612jxx
title: Data from: Sympatry and parapatry among rocky reef cichlids of Lake Victoria explained by female mating preferences

description: Work on the Lake Victoria cichlids Pundamilia nyererei (red dorsum males,
 deeper water), Pundamilia pundamilia (blue males, shallower water) and
 related species pairs has provided insights into processes of speciation.
 Here, we investigate female mating behaviour of five Pundamilia species
 and four of their F1-hybrids through mate choice trials and paternity
 testing. We discuss the results in the context of the geography of
 speciation and coexistence. Complete assortative mating was observed among
 all sympatric species. Parapatric species with similar depth habitat
 distributions interbred whereas other parapatric and allopatric species
 showed complete assortative mating. F1-hybrids mated exclusively with
 species accepted by females of the parental species. Although consistent
 with reinforcement in sympatry, a closer look at our results suggests
 otherwise and it is more likely that pre-existing female preferences
 influence which taxa can co-exist in sympatry. Regardless of the
 mechanism, mating preferences may influence species distribution in
 potentially hybridizing taxa, such as in the adaptive radiations of
 cichlid fish. We suggest that this at least partly explains why some
 species fail to establish breeding populations in locations where they are
 occasionally recorded. Our result support the notion that mating
 preferences of potentially cross-breeding species ought to be included in
 coexistence theory.
description: <em>Experimental
 setup</em> A 240 x 80 x 40 cm aquarium was divided by grids
 into ten male compartments with a flowerpot as a standardized spawning
 site in each. This ‘partial partition’ design allowed females to move
 freely whereas the larger males were confined to their compartments
 (Supporting Information Figure S1). We used standard daylight full light
 spectrum aquarium T5 fluorescent tubes to enable nuptial colours to be
 visible for all females. The combination of ten males, two from each
 species, was changed every second month the first year when most clutches
 were spawned and thereafter every 3-5 months when we collected the
 remaining clutches (total number of males used: 9 <em>P.
 azurea</em>, 10 <em>P. igneopinnis</em>, 8 <em>P.
 nyererei</em>, 8<em> P. pundamilia</em> and 8
 <em>P</em>. sp. ‘<em>red head</em>’, Electronic
 supplementary material Table S1-S2). All experimental fish were marked
 with PIT tags and a clip from the dorsal fin provided a DNA sample.
 Females were introduced when large enough to be PIT-tagged (size
 differences depended on age). For the first three months only wild-type
 fish were present; F1 hybrid females and more wild-type fish were added
 over the following year. Females were stripped of embryos/juveniles once a
 month and released back into the experimental tank for a second and third
 clutch and thereafter removed. Females with eggs were placed in a separate
 aquarium until the eggs hatched. Larvae/juveniles were euthanized using
 MS-222 (tricaine methanesulfonate) and stored in 95% ethanol prior to
 paternity analyses. Electronic supplementary material Table S1 shows when
 each female spawned i.e. were present in the aquarium. The size of males
 were similar among species and initially 99-109 mm TL although they grew
 during the experiment. Female sizes when spawning were 65-97 mm TL and we
 confirmed for each female that it could pass the grids of the
 dividers. <em>Paternity
 analyses</em> Where possible, four embryos from each of two
 (sometimes three) clutches per female were genotyped at two to four
 microsatellite loci, Ppun5, Ppun7, Pun17 and Ppun21. (Taylor et al., 2003)
 using the methods for DNA extraction and PCR reactions as in Svensson et
 al. (2017). The amplified DNA samples were genotyped on a Beckman Coulter
 CEQ 8000 capillary sequencer. Genotypes were received from the CEQ 8000
 Series Genetic Analyses System 8.0.52. Paternities were determined
 manually by inspection of the allele size estimates, and males that
 possessed two alleles at a microsatellite locus that were both not present
 in the offspring were excluded as possible fathers. In all analysed
 offspring, paternity could be assigned with 100 % certainty to one species
 only (Electronic supplementary material table S1-S3). If a clutch was
 confirmed to be fathered by more than one male; each sire was considered
 to represent one mate choice decision by the
 female.
description: # Data from: Sympatry and parapatry among rocky reef cichlids of Lake
 Victoria explained by female mating preferences Authors: Ola Svensson,
 Katie Woodhouse, Alan Smith, Ole Seehausen, George F. Turner Contact
 details: Ola Svensson, University of Bors, 
 Title of study: Sympatry and parapatry among rocky reef cichlids of Lake
 Victoria explained by female mating preferences Brief summary: Here, we
 investigate female mating behaviour of five Pundamilia species and four of
 their F1-hybrids through mate choice trials and paternity testing. We
 discuss the results in the context of the geography of speciation and
 coexistence. Responsible for collecting data: Ola Svensson ## Description
 of the Data and file structure Supplementary tables S1-S3 The
 supplementary tables contain all microsatellite paternity analyses of the
 wild type females and F1-females used in the present study. Table S1
 contains a summary of the spawning decisions of all females, including
 their IDs. Table S2 includes the microsatellite genotype of all males used
 in the experiment as well as in which experimental round they were used.
 The experimental rounds were approximately four weeks each. The experiment
 was carried out over two years, from August 2006 to July 2008. Table S3
 includes microsatellite raw data of the offspring of the females, as well
 as the paternity analyses. The first row with the ID shows the genotype of
 the females whereas the following rows with the same ID show the IDs of
 the offspring. Brood No is the ID of the brood and the rows below, until
 the next Brood number, are siblings from the same brood. When available,
 number of eggs in the brood, the size of the brooding female, the date
 when the female was stripped of eggs/fry and the number and starting date
 of the experimental round are given on the same row as Brood No. A slash
 between potential fathers of the same species show that it was not
 possible to dettermine parentage between the two conspecific males.
 Fathers within brackets show that the conspecific male is a potential
 father. However, it is a less likely father e.g. because short time in the
 experimental setup. Note that all offspring were 100% assigned to one
 species only. Empty cells are marked with n/a. Species included in the
 present study: P. azurea, Ruti Island P. igneopinnis, Igombe Island P.
 nyererei, Makobe Island P. pundamilia, Makobe Island P. sp. red head, Zue
 Island Table S1. Spawning decisions of wild type females and F1 hybrid
 females Table S2. Microsatellite genotypes and experimental round of males
 Table S3. Microsatellite genotypes and paternity analyses Supplementary
 figures S1-S2 Supplemental figure S2a. ‘melanic’ v. ‘red dorsum’ and
 ‘blue’ F1-crosses. In the P. azurea x P. nyererei cross (both directions)
 approximately three fourth were ‘red dorsum’ morphs and one fourth of the
 crosses were ‘blue’ morphs. The same was true for the P. igneopinnis x P.
 nyererei cross (both directions). In the latter we followed 26 randomly
 picked males to old age and the ratio did not change (20 ‘red dorsum’ v. 6
 ‘blue’). One male and one female produced the cross except where noted
 (mother 1, mother 2). We do not have a photo of the P. igneopinnis x P.
 nyererei, ‘blue’ morph. Photos: Katie Woodhouse. Supplemental figure S2a.
 ‘melanic’ v. ‘red dorsum’ and ‘blue’ F1-crosses. In the P. azurea x P.
 nyererei cross (both directions) approximately three fourth were ‘red
 dorsum’ morphs and one fourth of the crosses were ‘blue’ morphs. The same
 was true for the P. igneopinnis x P. nyererei cross (both directions). In
 the latter we followed 26 randomly picked males to old age and the ratio
 did not change (20 ‘red dorsum’ v. 6 ‘blue’). One male and one female
 produced the cross except where noted (mother 1, mother 2). We do not have
 a photo of the P. igneopinnis x P. nyererei, ‘blue’ morph. Photos: Katie
 Woodhouse. Supplemental figure S2b. ‘blue’ v. ‘red chest’  and ‘red
 dorsum’ F1-crosses. One male and one female produced the cross except
 where noted (mother 1, mother 2). We do not have photos of the  P.
 pundamilia x P. nyererei and P. sp. ‘red head’ x P. pundamilia crosses.
 Photos: Katie Woodhouse and Ola Svensson. Figures S1-2. Experimental
 set-up and photos of F1 hybrid males ## Sharing/access Information The
 data is published in Journal of Evolutionary Biology with the same title
 and authors.