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Adult color patterns of clownfish species. Pictures of adult clownfishes classified depending on their color patterns. a No vertical stripe, b one vertical stripe on the head, c two vertical stripes (one on the head, the other on the body), d three vertical stripes (one on the head, one on the body trunk, and the last one on the peduncle), e fishes having stripes polymorphism

Adult color patterns of clownfish species. Pictures of adult clownfishes classified depending on their color patterns. a No vertical stripe, b one vertical stripe on the head, c two vertical stripes (one on the head, the other on the body), d three vertical stripes (one on the head, one on the body trunk, and the last one on the peduncle), e fishes having stripes polymorphism

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Background: Biologists have long been fascinated by the striking diversity of complex color patterns in tropical reef fishes. However, the origins and evolution of this diversity are still poorly understood. Disentangling the evolution of simple color patterns offers the opportunity to dissect both ultimate and proximate causes underlying color di...

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... can be classified into four categories ac- cording to their striped pattern at the adult stage: spe- cies without vertical stripe (group A) or species having one white vertical stripe (on the head-group B), two vertical stripes (on the head and the trunk-group C), or three vertical stripes (head, trunk, and caudal ped- uncle-group D) ( Fig. 1 and Additional file 1: Table S1). Interestingly, there is no species with a single stripe on the trunk or on the peduncle (Fig. 1). White stripe on the trunk is always associated with a head stripe. The white stripe on the peduncle is always preceded by stripes on the head and the ...
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... stripe (group A) or species having one white vertical stripe (on the head-group B), two vertical stripes (on the head and the trunk-group C), or three vertical stripes (head, trunk, and caudal ped- uncle-group D) ( Fig. 1 and Additional file 1: Table S1). Interestingly, there is no species with a single stripe on the trunk or on the peduncle (Fig. 1). White stripe on the trunk is always associated with a head stripe. The white stripe on the peduncle is always preceded by stripes on the head and the ...
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... understand the evolution of color pattern in clown- fishes, we performed a stochastic mapping of striped patterns on the most complete time-calibrated phyl- ogeny of Amphiprionini [27]. The analysis highly sug- gests that the common ancestor of extended clownfishes exhibited three vertical white stripes (90-100% of pos- terior probabilities; Fig. 2), independent from the color pattern polymorphism of some species (Additional file 2: Figure S1). The state reconstruction for every internal node of the phylogeny illustrates successive losses of ver- tical stripes from the caudal to the rostral region (Fig. 2). ...
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... document 19 ko. (XLSX 18 kb) Additional file 2: Figure S1. Successive caudo-rostral loss of stripes during evolution is independent of clownfish color polymorphism. ...
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... can be classified into four categories ac- cording to their striped pattern at the adult stage: spe- cies without vertical stripe (group A) or species having one white vertical stripe (on the head-group B), two vertical stripes (on the head and the trunk-group C), or three vertical stripes (head, trunk, and caudal ped- uncle-group D) ( Fig. 1 and Additional file 1: Table S1). Interestingly, there is no species with a single stripe on the trunk or on the peduncle (Fig. 1). White stripe on the trunk is always associated with a head stripe. The white stripe on the peduncle is always preceded by stripes on the head and the ...
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... stripe (group A) or species having one white vertical stripe (on the head-group B), two vertical stripes (on the head and the trunk-group C), or three vertical stripes (head, trunk, and caudal ped- uncle-group D) ( Fig. 1 and Additional file 1: Table S1). Interestingly, there is no species with a single stripe on the trunk or on the peduncle (Fig. 1). White stripe on the trunk is always associated with a head stripe. The white stripe on the peduncle is always preceded by stripes on the head and the ...
Context 7
... understand the evolution of color pattern in clown- fishes, we performed a stochastic mapping of striped patterns on the most complete time-calibrated phyl- ogeny of Amphiprionini [27]. The analysis highly sug- gests that the common ancestor of extended clownfishes exhibited three vertical white stripes (90-100% of pos- terior probabilities; Fig. 2), independent from the color pattern polymorphism of some species (Additional file 2: Figure S1). The state reconstruction for every internal node of the phylogeny illustrates successive losses of ver- tical stripes from the caudal to the rostral region (Fig. 2). ...
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... document 19 ko. (XLSX 18 kb) Additional file 2: Figure S1. Successive caudo-rostral loss of stripes during evolution is independent of clownfish color polymorphism. ...

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... Our results indicate that the common ancestor of all clownfishes was likely a generalist. Previous studies demonstrated that melanin production and white stripes formation and development appears to only be lost across the phylogenetic tree as specialist lineages evolved from generalist ancestors [37,38]. This is concordant with the present broad phenotype of generalists characterized by black body color and multiple white stripes. ...
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... In aquarium, breeding couples spawn with high frequency (twice a month) and can easily be maintained in 60L tanks. Rearing techniques for anemonefishes have been well documented (Divya et al. 2011, Kumar et al. 2012, Madhu et al. 2006, the diversity of this family allows to explore several fields of scientific research, such as symbiotic relationship (Marcionetti et al. 2019, Roux et al. 2019c, sex change (reviewed in Casas et al. 2022), social control of size hierarchy (Buston 2003, Buston andCant 2006), and color pattern evolution (Salis et al. 2018. Furthermore, the establishment of methods to perform pharmacological treatments of larvae in low volumes (less than 1L) allowed to carry out pharmacological experiments . ...
... As the color patterns of anemonefish arise during metamorphosis, they are providing an excellent marker to detect and analyze signs of phenotypic plasticity . Clownfish acquire white stripes (zero to three depending on the species) formed by iridophores (white pigment cells; Salis et al. 2018Salis et al. , 2019b. Depending on environmental conditions, the developmental timing of the white bars can be delayed, as noticed by studying young recruits of Amphiprion percula hosted in different species of sea anemones, namely carpet sea anemone Stichodactyla gigantea and magnificent sea anemone (Heteractis magnifica) ). ...
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... The ecological function of color patterns in anemonefishes is poorly understood, but at least three adaptive hypotheses have been proposed. The first is the species recognition hypothesis, as different species tend to have different numbers of white bars, especially among species living in the same areas (Hayashi et al., 2022b;Salis et al., 2018). In addition, Mitchell et al. (2023) showed experimentally that the orange and white bars of anemonefishes are highly UV reflective, and that the contrast and intensity of this UV reflection affected the determination of the anemonefish's social status. ...
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... The number and shape of white bars differ among anemonefish species, and can be divided into five categories (Klann et al., 2021;Salis et al., 2018Salis et al., , 2022: (1) no vertical bars (Amphiprion sandaracinos, A. akallopisos, A. ephippium), (2) one bar on the head (e.g. A. frenatus, A. perideraion, A. leucokranos), (3) two bars on the head and trunk (e.g. A. chrysopterus, A. sebae, A. akyndinos), (4) three bars on the head, trunk and the caudal peduncle (e.g. A. ocellaris, A. percula, A. biaculeatus), and (5) polymorphic bars (e.g. A. clarkii, A. polymnus, A. melanopus). Also, Amphiprion akallopisos, A. sandaracinos, A. pacificus and A. perideraion have a pattern with horizontal stripe, with a vertical head bar in some but not all of these 'skunk' species Allen, 1992, 1997;Salis et al., 2018). ...
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... We then present a detailed synthesis of recent research that has provided important insights into the incredible adaptive radiation anemonefish have undergone, [7][8][9][10][11] and how their genomic architecture underlies the evolution of complex phenotypic traits such as sex change 12,13 and color patterning. [14][15][16] We further describe how researchers are using anemonefish as a model system to understand the genomic basis of symbiosis with giant sea anemones 10,[17][18][19] and environmental plasticity. 20,21 2. Anemonefish as a model system for evolutionary biology Before presenting the various contributions made in the field of anemonefish research, we feel it is essential to clarify the difference between the terms "anemonefish" and "clownfish" both of which are used throughout this review. ...
... 97 The clownfish is a conspicuously colored species (possessing a bright orange body with three iridescent white bars bordered with black), and understanding the molecular basis of pigmentation has also become a fundamental question of evolutionary biology. 2,14,16,98 Studies on A. ocellaris and A. percula have provided insights on how pigmentation patterns are phylogenetically conserved but also exhibit developmental and environmental plasticity. 15 79 Transcriptomic analysis of each of the seven developmental stages of A. ocellaris has revealed three distinct phases: larval development, a pivotal stage that marks the onset of metamorphosis, and metamorphosis per se that corresponds to the actual transformation. ...
... 178 Loss of white vertical bars during ontogeny has indeed been observed in multiple Amphiprion species. 14 Mitchell and colleagues (2022) further showed that UV reflectance in anemonefish (from their orange and white bars) has a functional role in modulating aggression and signaling submissiveness in family groups. 194 Salis and colleagues (2018) mapped the occurrence of these bars throughout the phylogenetic tree and showed that the diversification of color patterns in anemonefish is the result of successive (posterior to anterior) losses of bars during clownfish radiation. ...
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... We then present a detailed synthesis of recent research that has provided important insights into the incredible adaptive radiation anemonefish have undergone, [7][8][9][10][11] and how their genomic architecture underlies the evolution of complex phenotypic traits such as sex change 12,13 and color patterning. [14][15][16] We further describe how researchers are using anemonefish as a model system to understand the genomic basis of symbiosis with giant sea anemones 10,[17][18][19] and environmental plasticity. 20,21 2. Anemonefish as a model system for evolutionary biology There are 28 species of anemonefish 2 (Figure 2a), yet the two clownfish A. ocellaris (Figure 2b) and A. percula are perhaps the most recognizable ones, especially following the Disney movie "Finding Nemo". ...
... 95 The clownfish is a conspicuously colored species (possessing a bright orange body with three iridescent white bars bordered with black), and understanding the molecular basis of pigmentation has also become a fundamental question of evolutionary biology. 2,14,16,96 Studies on A. ocellaris and A. percula have provided insights on how pigmentation patterns are phylogenetically conserved but also exhibit developmental and environmental plasticity. 15,86 Other applications of transcriptomics in anemonefish research have provided in-depth characterization of visual opsins 59,77,87 and the rhythmic expression of internal clock genes. ...
... 175 Loss of white vertical bars during ontogeny has indeed been observed in multiple Amphiprion species. 14 Salis and colleagues (2018) mapped the occurrence of these bars throughout the phylogenetic tree and showed that the diversification of color patterns in anemonefish is the result of successive (posterior to anterior) losses of bars during clownfish radiation. The sequential appearance/disappearance of white bars during the development of distantly related species is remarkable as it suggests a highly conserved mechanism of pigmentation pattern ontogeny across anemonefish 14 (Figure 5a). ...
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... The distinctive appearance and conserved development of the orange and white bars of anemonefishes (Salis et al. 2018(Salis et al. , 2019 has given rise to a variety of suggested functions, including for intra-and/or inter-species recognition (Fricke 1973;Buston 2003;Salis et al. 2018), camouflage, and aposematism for sea anemones (Merilaita and Kelley 2018). However, in a void of empirical testing, the function of their bard skin has remained elusive. ...
... The distinctive appearance and conserved development of the orange and white bars of anemonefishes (Salis et al. 2018(Salis et al. , 2019 has given rise to a variety of suggested functions, including for intra-and/or inter-species recognition (Fricke 1973;Buston 2003;Salis et al. 2018), camouflage, and aposematism for sea anemones (Merilaita and Kelley 2018). However, in a void of empirical testing, the function of their bard skin has remained elusive. ...
... One possible explanation for this difference could be that UV reflectance is cheap to produce by anemonefish, hence its utility as a submissive signal by juveniles. The white skin of early-juvenile false clownfish, A. ocellaris, is comprised of iridophores, and to a lesser extent melanophores (Salis et al. 2018(Salis et al. , 2019. As opposed to the pigmentary melanophores, iridophores provide structural coloration known to confer iridescence and UV reflectance (Cott 1940). ...
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... Anemonefish (Amphiprioninae, Pomacentridae) have conspicuous white bars against a background colour of orange, red or black, and the number of white bars varies depending on the species [7,[23][24][25]. The evolutionary function of colour patterns in anemonefish is poorly understood, but at least three adaptive hypotheses have been proposed [7,[26][27][28][29]. The first hypothesis is that the number of white bars has a recognition function since anemonefish have species-specific numbers of white bars. ...
... The first hypothesis is that the number of white bars has a recognition function since anemonefish have species-specific numbers of white bars. This hypothesis is supported by the fact that the differences in the number of white bars between species inhabiting the same area are significantly greater than would be expected at random [27]. In addition, anemonefish patterns change during ontogeny in some species [7,[23][24][25]27], and the different patterns of juveniles from adults may be a dishonest signal to conceal their presence and reduce agonistic interactions [26,27,30]. ...
... This hypothesis is supported by the fact that the differences in the number of white bars between species inhabiting the same area are significantly greater than would be expected at random [27]. In addition, anemonefish patterns change during ontogeny in some species [7,[23][24][25]27], and the different patterns of juveniles from adults may be a dishonest signal to conceal their presence and reduce agonistic interactions [26,27,30]. The second hypothesis is that the contrast of the bright base colour and white stripes is disruptive and functions to hide the fish silhouette. ...
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Colour patterns in fish are often used as an important medium for communication. Anemonefish, characterized by specific patterns of white bars, inhabit host anemones and defend the area around an anemone as their territory. The host anemone is used not only by the anemonefish, but also by other fish species that use anemones as temporary shelters. Anemonefish may be able to identify potential competitors by their colour patterns. We first examined the colour patterns of fish using host anemones inhabited by Amphiprion ocellaris as shelter and compared them with the patterns of fish using surrounding scleractinian corals. There were no fish with bars sheltering in host anemones, although many fish with bars were found in surrounding corals. Next, two fish models, one with white bars and the other with white stripes on a black background, were presented to an A. ocellaris colony. The duration of aggressive behaviour towards the bar model was significantly longer than that towards the stripe model. We conclude that differences in aggressive behaviour by the anemonefish possibly select the colour patterns of cohabiting fish. This study indicates that colour patterns may influence not only intraspecific interactions but also interspecific interactions in coral reef ecosystems.
... Another approach for linking a phenotype of interest with its genetic basis is by the experimental disruption (i.e., knockdown or enhancement) of normal gene expression. In anemonefishes, this can be induced using various pharmacological treatments that often serve to increase or block hormonal pathway activity (e.g., Nakamura et al. 2015;Salis et al. 2018; Iwata and Suzuki 2020). Using drugs can help deduce the genetic basis of a phenotype or trait but require caution and careful consideration of controls due to their often wide-ranging/whole-organism effects. ...
... The production of mutant strains using genome editing technologies requires the precise DNA sequence information of the target gene and well-established breeding and transfection (microinjection) techniques. Among these requirements, breeding technologies and genome/transcriptome information have been studied quite extensively in anemonefishes (Buston and Elith 2011;Maytin et al. 2018;Roux et al. 2021;Salis et al. 2018Salis et al. , 2019Salis et al. , 2021Tan et al. 2018). However, only recently several research groups have started to generate genetic mutants of anemonefish using newly developed microinjection methods in combination with the CRISPR/ Cas9 technology Yamanaka et al. 2021). ...