Mosaic evolution of grasping and genitalic traits in two sympatric scorpion species with reproductive interference

Mosaic evolution of grasping and genitalic traits in two sympatric scorpion species with reproductive interference | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Mosaic evolution of grasping and genitalic traits in two sympatric scorpion species with reproductive interference Mariela Oviedo-Diego, Camilo Mattoni, Fedra Bollatti, Eduardo M. Soto, and 1 more This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-2445373/v1 This work is licensed under a CC BY 4.0 License Status: Published Journal Publication published 27 Jan, 2024 Read the published version in Evolutionary Biology → Version 1 posted 8 You are reading this latest preprint version Abstract Reproductive interference (RI) can occur when two related species coexist in sympatry, involving sexual attraction, mating, and even hybridization between heterospecifics. Consequently, reproductive key characters of these species may suffer morphological shifts in sympatry to avoid the success of heterospecific sexual interactions, a phenomenon known as reproductive character displacement (RCD). RCD can be promoted by natural selection, although sexual selection pressures can act synergistically or agonistically so that phenotypic variation can respond in different directions and magnitudes to these forces. In turn, the size and shape of characters may respond differentially (mosaic evolution) to these pressures, so the analysis of multiple dimensions in traits is essential to understand the complexity of their phenotypic variability. To date, there are no studies evaluating this topic in scorpions, and two species ( Urophonius brachycentrus and U. achalensis ) sympatric and synchronous with RI represent an ideal model to evaluate the phenotypic variation and occurrence of RCD. In addition, the populations of these species are found in an altitudinal cline, so environmental factors may also be responsible for explaining their morphological variation. We compared the intra-specific variation, the size and shape of multiple characters involved in courtship, and sperm transfer in individuals from sympatric and allopatric populations using geometric morphometrics. We found asymmetric RCD of several sexual characters for courtship success (grasping structures) and sperm transfer (genital characters). This would evidence the action of natural selection pressures and the existence of a possible mechanism to avoid heterospecific mating success. In addition, we found a pattern of asymmetric morphological variation where one species in the sympatric zone suffered an increase in size in several characters due to environmental factors (pattern of morphological convergence). The convergence of characters combined with RI and a scramble competition mating system could intensify sexual selection pressures on specific characters, which was reflected in their high coefficients of variation. Our results suggest that in this sympatric zone, several selective regimes act differentially on various dimensions of the characters evaluated, which would support a possible mosaic evolution. This comprehensive study illuminates the complexity inherent in the evolution of multi-functional traits in a previously unexplored model, providing novel insights for evaluating traits under multiple selective pressures in animal systems experimenting RI. Reproductive character displacement Convergence Sexual selection Natural selection Geometric Morphometrics Scorpions. Figures Figure 1 Figure 2 Figure 3 Figure 4 1. Introduction Determining the factors underlying phenotypic variation in natural populations is important for comprehending the evolution of species and their biological diversity and is a fundamental task of evolutionary biology (Coyne & Orr, 2004 ). Morphological characters are shaped by multiple selective pressures, especially those involved in various components of the life history of organisms. Secondary sexual characters undergo relatively fast evolutionary divergence due to sexual and natural selection (Svensson & Gosden, 2007 ). Natural selection favors morphological traits linked to growth, reproduction, and survival resulting in greater reproductive success for certain environments. In contrast, sexual selection underlies morphological changes that favor reproductive success through intra-sexual competition, inter-sexual mate choice, or post-copulatory processes (Kraaijeveld et al., 2011 ; Maan & Seehausen, 2011 ; Safran et al., 2013 ). The study of interspecific interactions is crucial for understanding sex-linked ecological and evolutionary patterns (Cothran, 2015 ). Reproductive interference (henceforth referred as ‘RI’) is defined as any type of interspecific interaction between sympatric species associated with their mating systems caused by incomplete recognition between species (Gröning & Hochkirch, 2008 ; Burdfield-Steel & Shuker, 2011 ). This process may negatively affect the reproductive success of at least one species (Hochkirch et al., 2007 ). RI between species can lead to the displacement of key characters in reproductive interactions (i.e., reproductive character displacement - henceforth referred as ‘RCD’) (Howard, 1993 ), which generally results in a divergence of these characters alleviating RI and thus reinforcing reproductive isolation (Servedio & Noor, 2003 ; Coyne & Orr, 2004 ; Kyogoku, 2015 ). Characters of coexisting species should be more divergent in sympatry than in allopatry. The more similar the characters of interacting species are in sympatry, the greater the consequences of RI on reproductive success (Pfennig & Pfennig, 2010 ; Konuma & Chiba, 2012 ). In turn, the degree and direction of divergence of sexual characters may differ according to their function or the moment of the reproductive event in which RI occurs (Gröning & Hochkirch, 2008 ). In other cases, adaptive promiscuity may exist, and competition for exploitation (for females beyond their species) is propitiated, which may prevent divergence and even generate convergence of sexual characters (Grant, 1972 ; Grether et al., 2009 ; Tobias et al., 2014 ; Drury et al., 2015 ; Sobroza et al., 2021 ) with consequent maintenance or intensification of RI (Takakura et al., 2015 ; Wheatcroft, 2015 ; Yamaguchi & Iwasa, 2015 ). In sympatric areas, intraspecific sexual selection pressures may join interspecific interactions generating a mosaic of selective pressures with different outcomes in terms of morphological variation (Grether et al., 2009 ). Secondary sexual characters may play a role in specific recognition, so their divergence can be explained by natural selection (Mayr, 1963 ; Bennet-Clark & Ewing, 1970 ). However, it has been postulated that mate choice and specific recognition are part of a continuum and that sexual selection may also lead to reinforcement or RCD (Ryan & Rand, 1993 ; Boake et al., 1997 ; Liou & Price, 1994 ; Mendelson & Shaw, 2012 ). In cases where the female is the selective sex, it is hypothesized that female choice that promotes isolation will result in the divergence of male sexual characters to avoid RI or hybridization (Butlin, 1987 ; Gröning & Hochkirch, 2008 ; Hoskin & Higgie, 2010 ). RCD has been reported for body size (Ding et al., 2018 ; Sağlam et al., 2019 ), characters for grasping the female and genital characters (Kawano, 2002 ; Kameda et al., 2009 ; Anderson & Langerhans, 2015 ; Kosuda et al., 2016 ; Sağlam et al., 2019 ; Nishimura et al., 2022 ) or other types of characters (Marsteller et al., 2009 ; Kawakami & Tatsuta, 2010 ; Roth-Monzón et al., 2017 ). In many works where RCD is evaluated, one or a few characters linked to sexual reproduction are analyzed. However, phenotypic divergence can occur due to selective pressures along multiple phenotype axes simultaneously so that divergence can be multidimensional (Haines et al., 2021 ; White & Butlin, 2021 ; Vega-Sánchez et al., 2022 ). Animal genitalia, especially in the male, can exhibit relatively complex morphologies and show fast and divergent evolutionary changes compared to other body parts (Tuxen, 1970 ; Eberhard, 1985 ; Leonard & Córdoba-Aguilar, 2010 ). Sexual selection may play a key role in the evolution of genitalia (Eberhard, 1985 , 2010 ; Hosken & Stockley, 2004 ; Simmons, 2014 ). In turn, genital divergence can be explained by natural selection, as it contributes to reproductive isolation among species by promoting speciation (Eberhard, 1985 , 2010 ; Masly, 2012 ; Wojcieszek & Simmons, 2012 ; House et al., 2013 ). Phenomena such as RCD may contribute to differences in genitalia between species in sympatric zones, whereby mechanical or interlocking incompatibilities between male and female genitalia may be frequent (Masly, 2012 ). The relative importance of natural and sexual selection in genitalia evolution continues under discussion (Jennions & Kelly, 2002 ; Eberhard, 2010 ; Simmons, 2014 ; Brennan & Prum, 2015 ; Eberhard & Lehmann, 2019 ; Sloan & Simmons, 2019 ), although there is evidence that multiple selective pressures may be determinant in the morphological evolution of the genitalia (Langerhans et al., 2005 ; Song & Wenzel, 2008 ; McPeek et al., 2009 ; Simmons et al., 2009 ; House et al., 2013 ; Simmons, 2014 ; Frazee & Masly, 2015 ). This multiplicity of selective regimes can cause what is known as "mosaic evolution", where different portions of the same structure can respond in a mixed manner to concordant or antagonistic selective pressures (due to their multi-factorial nature) and where even shape and size of the same structure can diverge differentially (House & Simmons, 2005 ; Song & Wenzel, 2008 ; Werner & Simmons, 2008 ). Morphological variation in non-genital contact characters used during pre-copulatory or copulatory mating can be explained by some of the natural or sexual selection hypotheses that may generate genital morphological diversity (Robson & Richards, 1936 ; Eberhard 1985 , 2004 , 2010 ). These characters also possess a pattern of rapid evolutionary divergence and generally have the function of grasping or grasping the female during mating by resembling functionally genital ''claspers'' (Eberhard, 1985 , 2010 ). The intra-specific phenotypic variation can be considered as the raw material on which selection acts (West-Eberhard, 2005 ; Eberhard, 2009 ), and the patterns of variation are helpful in understanding the evolution of different morphological characters. In general, sexually selected display traits show high within-species phenotypic variation (Cuervo & Møller, 1999 ; Eberhard et al., 1998 ; Eberhard, 2009 ). High values of coefficient of variation (CVs) indicate directional selective forces, while low values of CVs are associated with stabilizing selective pressures (Eberhard et al., 1998 ; Eberhard, 2009 ). Phenotypic plasticity refers to the ability of organisms of a species to change their morphology, behavior, or physiology in response to environmental variation (Stearns, 1989 ; West-Eberhard, 2003 ; Whitman & Agrawal, 2009 ). When characters express some degree of phenotypic plasticity, environmentally based phenotypic differences among species and populations can underlie the patterns of morphological variation (Jennions & Kelly, 2002 ; Garnier et al., 2005 ; Song & Wenzel, 2008 ). The case of morphological divergence in environmental gradients deserves particular mention. In these cases, morphological differences between populations may be due to selective pressures for species differentiation and morphological changes linked to an environmental cline (Goldberg & Lande, 2006 ). Therefore, among the requirements for testing RCD, it is necessary to separate allopatric/sympatric context effects from other ecological effects. The environment can directly or indirectly influence genetic and phenotypic variation. Therefore, geographic variation among different populations is expected (Sota et al., 2000 ; Kosuda et al., 2016 ). Controlling for the effects of correlation between phenotype and environmental or geographic clines allows for finding patterns of divergence that might otherwise be undetectable (Goldberg & Lande, 2006 ). Variation in latitude or altitude is mainly linked to changes in temperature, an abiotic factor that affects animal growth, causing a substantial impact on the observed phenotypic variation results (Bergmann, 1847 ; Allen, 1877 ; Rensch, 1938 ; Atkinson, 1994 ). Examples of this RI exist in many animal and plant groups (e.g., Levin, 1970 ; Armbruster & Herzig, 1984 ; Hettyey & Pearman, 2003 ; Dame & Petren, 2006 ; Gröning & Hochkirch, 2008 ; Matsumoto et al., 2010 ), and among them, arthropods have been shown to provide interesting models for studying this phenomenon (Shuker & Burdfield-Steel, 2017 ). Although some cases of ecological character displacement have been described in insects and arachnids, there are fewer examples of RCD in these taxa due to the difficulty of empirically evidencing this process (Waage, 1979 ). However, in arthropods, evidence of RCD was found in pre-copulatory characters used during courtship (Marshall & Cooley, 2000 ; Jang & Gerhardt, 2006 ; Kronforst et al., 2007 ; Dyer et al., 2014 ; Rundle & Dyer, 2015 ; Yukilevich, 2021 ) and there are also examples of RCD in genital characters (Kawano, 2002 ; Kawakami & Tatsuta, 2010 ; Kosuda et al., 2016 ; Nishimura et al., 2022 ). In arachnids, there are some suggestions that RCD might be occurring between species in sympatry (Barth, 1990 ; Stratton, 1997 ; Agnarsson et al., 2016 ; Muster & Michalik, 2020 ), as is the case of genital characters between Paratrechalea spider species with RI (Costa-Schmidt & de Araújo, 2010 ). The study of phenotype variation and its causes may be complicated because adaptation can be viewed as a multivariate process acting on sets of characters (Lande & Arnold, 1983 ; Schluter & Nychka, 1994 ; Blows, 2007 ). Organisms can be interpreted as composite objects, with characters not necessarily independent of each other that respond in complex and different magnitudes and directions to different selective pressures (Klingenberg, 2009 ). Geometric morphometrics (GM) helps address the inherent complexity of characters separating their size and shape to evaluate the effect of selective pressures on these two dimensions of the phenotype (Bookstein, 1998; Adams et al., 2004 ; Zelditch et al., 2004 ). Indeed, shape metrics are better descriptors of genital morphology diversity, containing more information than size measures (Slice, 2007 ; Shen et al., 2009 ). These type of studies are ideally performed in species where the function of the characters to be assessed is well-known. Arachnids have proven to be exceptional models, although morphological quantification techniques have generally been applied mainly in systematic or ecomorphological studies (Costa-Schmidt & de Araújo, 2010 ; Kallal et al., 2019 ; Santibáñez-López et al., 2021 ; Wilson et al., 2021 ; Bellvert et al., 2022 ). Some studies have demonstrated the usefulness of these techniques in addressing sexual dimorphism (Fernández-Montraveta et al., 2017; Kallal et al., 2019 ), as well as the combination with other approaches such as the analysis of phenotypic variation (by the coefficient of variation -CV) of certain characters in some arachnids (Eberhard et al., 1998 ; Peretti et al., 2001 ; Calbacho-Rosa et al., 2019 ; Lai et al., 2021 ). Although studies applying fine morphological quantification methodologies in scorpions are scarce, these organisms appear to be excellent models for this type of analysis (Bechara & Liria, 2012 ; Santibáñez-López et al., 2017 ). It is known that different selective pressures act on specific scorpion characters (e.g., pedipalps, pectines, chelicerae) as demonstrated in studies that have evaluated their CV, allometric patterns, or where selection pressures behind dimorphic characters have been explored (Peretti et al., 2001 ; Carrera et al., 2009 ; Fox et al., 2015 ; Santibáñez-López et al., 2017 ; Visser & Geerts, 2021 ). Furthermore, during an elaborate courtship, both sexes displayed unique characters with functional roles such as stimulation or increased female receptivity with non-genital contact structures (e.g., the caudal gland in ‘rubbing with telson’, the sting in ‘sexual sting’) or grasping characters to overcome female resistance (e.g., apophyses in pedipalps, chelicerae) (Polis & Sissom, 1990 ; Carrera et al., 2009 ; Peretti, 2001). In particular, these characters were extensively studied in the family Bothriuridae in the evolutionary framework of sexual selection (Peretti et al., 2001 ; Carrera et al., 2009 ; Olivero et al., 2014 , 2019 ; Peretti, 2010 ). Lastly, scorpions present indirect sperm transfer via a sclerotized spermatophore deposited in the substrate (Weygoldt, 1990 ; Proctor, 1998 ). This spermatophore is regenerated each time the male mates from two chitinous halves (i.e., hemispermatophores) produced in internal glandular structures called paraxial organs (Polis & Sissom, 1990 ). These genital characters are incredibly complex and can be divided into subunits offering interesting opportunities for studying the evolution of genitalia (Peretti et al., 2001 ; Peretti, 2003 , 2010 ; Mattoni et al., 2012 ; Monod et al., 2017 ). For example, some characters follow a distinctive pattern of characters under sexual selection pressures (i.e., evolve rapidly and divergently), while others show only minor variations coinciding with what is predicted for characters under natural selection, such as structures with mechanical constraints or with key reproductive functions such as sperm passage (Peretti, 2010 ; Mattoni et al., 2012 ). The morphological diversity of sexual characters and spermatophores of scorpions responds to diverse (and not mutually exclusive) evolutionary hypotheses (Peretti, 2010 ). These mixed patterns result from complex synergistic or antagonistic interactions between different selective pressures (Peretti, 2010 ), so this genital structure could be found under mosaic evolution. This offers great possibilities for the morpho-functional study of diverse characters and contexts and allows different outcomes in a scenario of RCD. There are several records of interspecific mating in scorpions (Auber, 1963 ; Matthiesen, 1968 ; Probst, 1972 ; Le Pape & Goyffon, 1975 ; Peretti, 1993 ; Peretti et al., 2000 ). Although many scorpions use pheromones for sex encounter, males are vagrant in scenarios of indirect competition for females, and there are records of overlapping species distributions and coexistence of species, phenomena such as RI or RCD between closely related species have not yet been extensively assessed. Here, we explored the occurrence of RCD in two closely related scorpion species of the genus Urophonius Pocock, 1893 ( U. brachycentrus and U. achalensis , Bothriuridae) (Ojanguren-Affilastro et al., 2020 ) that have partially sympatric ranges with overlapping reproductive seasons and share the same habitat requirements and life-history traits (Maury, 1969 ; Acosta, 1985 ; Ojanguren-Affilastro et al., 2020 ). These scorpions have winter habits and adaptations for this lifestyle, which is rather peculiar among scorpions (Ojanguren-Affilastro et al., 2020 ; Garcia et al., 2021 ). These species do not possess specific recognition through chemical signals, which, together with a promiscuous mating system with scramble competition, leads to an asymmetric RI scenario with heterospecific mating (Oviedo-Diego et al., 2020 , 2021 ). The coexistence of these species raises the question of whether there are morphological, reproductive barriers, that may hinder or prevent the culmination of heterospecific mating, given the costs they may entail in terms of gamete loss, female plugging (Oviedo-Diego et al., 2019 , 2020 ; Romero-Lebrón et al., 2019 ) or potential hybridization. For these reasons, we evaluated the existence of RCD in the shape and size of somatic characters used in courtship (non-genital contact characters) and genital characters of hemispermatophores to observe whether these metrics responded concordantly or follow a mosaic pattern under specific recognition and sexual selection pressures. Additionally, we determined the phenotypic variation by analyzing the coefficient of variation of these characters in contexts of sympatry and allopatry of both species to complement the analysis of the selective regimes that could explain the morphological variability in these species. Complementarily, we consider the influence of environmental and geographic factors on the morphological patterns found. The results from multiple lines of evidence account for the inherent complexity of sexual characters in scorpions and provide clues about the possible selective pressures behind their evolution. 2. Material Amd Methods 2.1.1 Study Species and Sampling Urophonius brachycentrus has a wide geographic range distributed throughout central Argentina, while U. achalensis is endemic to the mountainous regions of Córdoba in central Argentina (Acosta, 1985 ; Ojanguren-Affilastro, 2020). The two species share partially sympatric distribution areas in the Sierras Grandes that are part of the Sierras Pampeanas Centrales (Acosta, 1985 ), part of a fundamental orographic system of extra-Andean mountain formations in Argentina, were formed in the Lower Paleozoic (about 300 and 350 million years ago). Adult scorpions of U. achalensis and U. brachycentrus were collected during the day during the mating season (May-August) (Acosta, 1985 ; Maury, 1969 ; Ojanguren-Affilastro et al., 2020 ) for three consecutive years (2018, 2019, 2020) by turning over rocks. We collected individuals in two allopatric populations of U. brachcyentrus (31°22'42.4"S 64°35'34.0"W, 876 m.a.s.l..; 31º31'46.3''S 64º51'52.7"W, 996 m.a.s.l.), two allopatric populations of U. achalensis (31°35'49.1"S 64°44'49.3"W, 2030 m.a.s.l., 31°21'17.3"S 64°48'21.3"W, 1927 m.a.s.l.), and in two sympatric populations (31°23'13.5"S 64°46'10.2"W, 1796 m.a.s.l.; 31°34'07.6"S 64°42'43.8"W, 1610 m.a.s.l.). 2.1.2 Processing of individuals and selected characters Individuals from field collections were identified and sexed (Ojanguren-Affilastro, 2005 ) with a Nikon SMZ 1500 stereo zoom microscope and preserved in 80% EtOH in glass containers for morphological studies. Classical and geometric morphometric studies were carried out, and measurements of characters were compared between sexes and study species in different contexts (sympatry vs. allopatry) (n = 25 per population context and per sex of each species) (Table 1 , Fig. 1 ). We selected characters under both natural and sexual selection pressures, used during feeding, defense, and courtship traits such as pedipalps, chelicerae and telson vesicle (Table 1 , Fig. 1 ). Also, we analyzed characters used only in a sexual context, such as characters for female stimulation (caudal gland) or characters for grasping the female pedipalps during courtship (pedipalp apophyses) (Table 1 , Fig. 1 ). Finally, we measured genital characters involved in sperm transfer that has also been shown to be under sexual selection pressures (Olivero et al., 2015 ; Peretti, 2010 ) (Table 1 , Fig. 1 ). To analyze the selected characters, individuals were dissected, and internal structures were extracted with fine tweezers for photographic treatment. The individuals were measured using images taken under the stereo zoom microscope with a digital coupled camera (Nikon Digital Sight DS-FI1-U2). Because the internal female genitalia consist of flexible structures that vary in size and shape according to the female reproductive status (Peretti, 2010 ), morphometric analysis was not performed. In subsequent analyses, individuals and characters with damaged or incomplete portions were not considered. Table 1 Morphological characters selected in Urophonius species analyzed. The type of character (somatic or genital), the corresponding sex, the functional role, and the measurement technique used are indicated. Abbreviations: AL, absolute length; RL, relative length; NS, Natural selection; SS, Sexual selection. See landmark positions in Fig. 1 and descriptions in Table S1. Morphological character Sex and type of character n Methodology Functional role Prosome Somatic in both sexes ♂ n = 122 Geometric morphometry (Landmarks = 8) Body size indicator (Polis & Sissom 1990 ; McLean et al., 2018 ). ♀ n = 112 Chelicera ♂ n = 113 Classic morphometry (AL, RL) Character used during feeding and courtship where the pair touch and rub chelicerae during ‘chelicera massage’ or ‘kiss’ (under NS and SS pressures) (Carrera et al., 2009 ). ♀ n = 114 Pectine ♂ n = 126 Classic morphometry (AL, RL) Character used for mechano-chemical-sensory recognition, foraing, mate searching and spermatophore deposition site in courtship (under NS and SS -slight- pressures) (Polis & Sissom, 1990 ; Peretti et al., 2001 ) ♀ n = 100 Pedipalp Grasping characters ♂ n = 128 Geometric morphometry (Landmarks = 5 + Semilandmarks = 21) Character used during defense, feeding and grasping of the other sex during courtship (under NS and SS pressures) (Polis & Sissom 1990 ; Peretti et al., 2001 ; Olivero et al., 2014 ). ♀ n = 121 Geometric morphometry (Landmarks = 4 + Semilandmarks = 21) Pedipalp apophysis Somatic in males ♂ n = 122 Geometric morphometry (Landmarks = 5 + Semilandmarks = 16) Character used for the correct grasping and locking of pedipalps during courtship (only under SS pressure) (Peretti et al., 2001 ). Telson vesicle Somatic in both sexes ♂ n = 122 Geometric morphometry (EFA = 8 harmonic) Character used during feeding and agonistic interactions, during courtship in sexual stinging of the female and gland rubbing (under NS and SS pressures) (Polis & Sissom, 1990 ; Peretti, 1993 ; Fox et al., 2015 ; Sentenská et al., 2017 ; Olivero et al., 2017, 2019 ). ♀ n = 122 Character used during feeding and agonistic interactions, sometimes during courtship movements indicative of receptivity (under NS and SS pressures) (Polis & Sissom 1990 ; Fox et al., 2015 ). Caudal gland Somatic in males Stimulation character ♂ n = 122 Geometric morphometry (EFA = 7 harmonic) External secretory gland on the dorsal side of the telson used during courtship where the male rubs the female to increase female receptivity (under SS pressures) (De la Serna de Esteban, 1978 ; Peretti, 1997 ; Olivero et al, 2017, 2019 ). Hemispermatophore lamella Genital in male Genital character ♂ n = 117 Geometric morphometry (Landmarks = 4 + Semilandmarks = 24) Genital character that will form the spermatophore involved in the copulatory mechanics for indirect sperm transfer, acting as a lever for sperm release (under NS and SS pressures) (Peretti et al., 2001 ). Hemispermatophore capsular lobe ♂ n = 108 Geometric morphometry (EFA = 6 harmonic) Genital character that will form the copulatory cone of the spermatophore that enters and evert inside the female genitalia, guiding the sperm during sperm transfer (under NS and SS pressures) (Peretti et al., 2001 ; Olivero et al., 2014 ). Table 2 Coefficients of variation (CVs) of multiple somatic and genital characters of male and female Urophonius achalensis and U. brachycentrus scorpions from sympatric and allopatric areas. Morphological character, sex, CVs value and statistical significance value (between species and contexts) are indicated (p-values < 0.05 indicated in bold). ♂: males, ♀: females. Letters indicate significant differences between character CVs (p-values < 0.05) Species U. achalensis U. brachycentrus Differences between spp. Differences between contexts Morphological character Sex/Context sympatry allopatry sympatry allopatry U. achalensis U. brachycentrus Chelicerae length ♀ 5.525 b 5.098 b 6.266 b 5.037 b 0.605 0.735 0.379 ♂ 4.220 b 5.605 b 5.09 b 5.831 b 0.134 0.169 0.354 Pecten length ♀ 7.263 b 6.141 b 6.824 b 6.955 b 0.150 0.195 0.251 ♂ 5.363 b 6.962 b 6.984 b 6.872 b 0.323 0.241 0.092 Pedipalp length ♀ 4.591 b 3.248 b 5.092 b 6.155 b 0.025 0.119 0.324 ♂ 3.581 b 4.439 b 5.416 b 5.643 b 0.010 0.274 0.506 Pedipalp apophysis length ♂ 10.741 a 11.267 a 12.329 a 16.227 a 0.018 0.301 0.191 Telson vesicle length ♀ 3.472 b 4.832 b 4.223 b 4.785 b 0.444 0.145 0.585 ♂ 4.471 b 5.315 b 5.413 b 6.331 b 0.215 0.371 0.169 Caudal gland length ♂ 10.359 a 12.653 a 16.111 a 13.020 a 0.249 0.320 0.307 Hemispermatophore lamella length ♂ 3.982 b 5.111 b 4.756 b 5.245 b 0.484 0.306 0.692 Hemispermatophore capsu l ar lobe length ♂ 4.447 b 5.925 b 4.951 b 5.451 b 0.677 0.285 0.730 Hemispermatophore frontal crest length ♂ 8.466 ab 8.539 ab 7.749 ab 7.842 ab 0.786 0.970 0.547 2.1.3 Morphometric studies 2.1.3.1 Classic morphometric and coefficient of variation analysis The chelicerae and the pectines were analyzed by linear measurements (due to methodological difficulties in applying geometrical morphometry) by analyzing absolute and relative lengths (prosome length as body size index - McLean et al., 2018 ) (Table 1 ). These measurements were taken from photographs obtained for each character with ImageJ software tools (Schneider et al., 2012 ). Measurements were taken three times by the same person, and the measurement error was calculated (Sokal & Rohlf, 1995 ). The coefficient of variation (CV) is widely used as indirect evidence to know the selective pressures that might be operating on morphological characters (Pomiankowski & Møller, 1995 ; Eberhard et al., 1998 ; Peretti et al., 2001 ). We compared this parameter across different types of characters in males and females from both contexts (sympatry versus allopatry). We used the modified formula: CV’ = [(sd y /mean y ) * (1 – r 2 ) 1/2 * 100], where sd y is the standard deviation of the character, mean y is the arithmetic mean of the character, r 2 is the determination coefficient between the character and a measure of body size (prosoma length) and * indicates multiplication symbol (Eberhard et al., 1998 ; Calbacho-Rosa et al., 2019 ). CVs were statistically compared using the 'asymptotic_test' function of the cvequality package (Marwick & Krishnamoorthy, 2019 ). 2.1.3.2 Geometric morphometric analysis We took digital images of selected characters in male and female scorpions with a scale close to the character, and the images were assembled with TPSutil software (Rohlf, 2015 ). Sets of anatomical Landmarks (Bookstein, 1991 ) and semilandmarks were established using TPSDig2 (Rohlf, 2004 ; Bookstein, 1997 ; Gunz & Mitteroecker, 2013 ). We used landmarks in the prosome, the hemispermatophore lamella, the pedipalp, and the apophysis of this structure (Table 1 , Fig. 1 , Table S1). Sliding landmarks or semilandmarks were used to enhance geometric information about curvatures between adjacent landmarks in the pedipalp, the pedipalp apophysis, and the hemispermatophore lamella (Fig. 1 ). In other characters (hemispermatophore capsular lobe, telson vesicle and caudal gland) we quantified shape using an elliptic Fourier analysis (EFA) (following Santibáñez-López et al., 2017 , 2021 ) that allowed us to explore small differences in defined shapes from contour characterization (Kuhl & Giardina, 1982 ; Ferson et al., 1985 ; Hammer & Harper, 2006 ) (Fig. 1 ). The shape coordinates of each character were subjected to a Generalized Procrustes Analysis (Gower, 1975 ) with the 'gpagen' function of the geomorph package (Schlager, 2017 ; Adams et al., 2017 ) in R software (R Core Team, 2021 ) to remove non-shape variables (translation, rotation, size) from the dataset to compare shape by contrasting with a mean generated from a consensus matrix (Rohlf & Slice, 1990 ; Adams et al., 2017 ). The size proxy of each character was retained from the GPA analysis (i.e., Centroid size) for subsequent analyses (Bookstein, 1991 ; Zelditch et al., 2004 ). To account for semilandmarks in the GPA calculation, we used the 'slider2d' function of the Morpho package (Schlager et al., 2021 ). EFA was performed using the momocs package (Iwata & Ukai, 2002 ; Bonhomme et al., 2014 ). A Principal Component Analysis (PCA) was performed to visualize and explore the general trends of the distribution of total morphological variation in morphospace from both the data yielded by the GPA as well as the data obtained from the EFA using the 'plotTangentSpace' function of the geomorph package. Principal components can be considered as reorganized and uncorrelated morphological features representing different aspects of the total shape variation. Additionally, vectors that reflected shape variation along x/y axes were used to visualize magnitudes and overall shape changes with the geomorph package (Bookstein, 1991 ). Multivariate analysis of variance (MANOVA) was performed with the function 'procD.lm' of the geomorph package with resampling permutations procedure to calculate the significance of shape variables. The variation in shape of the first two principal components (since they captured more than 70% of the morphological variation) was analyzed in detail. First, we checked the allometric component (influence of size on shape) of the characters with the functions 'procD.lm' and 'plotAllometry' of the geomorph package. If we found allometry in the sample, we calculated residual values of the shape variables for subsequent analyses (Outomuro & Johansson, 2017 ). 2.1.3.3 Statistical analysis Measurements obtained by classical and geometric morphometry were compared between species and contexts (sympatry versus allopatry) with linear mixed models (LMMs) in R. Separate models were performed for each character and sex (because in some characters the number of Landmarks was not equal for males and females) where we set as response variables the linear measurements, size variables (centroid size) or shape variables (PCs scores) and the fixed effects were species (levels: U. achalensis / U. brachycentrus ) and contexts (levels: sympatry / allopatry). The interaction between these explanatory variables was evaluated to corroborate RCD patterns. We added populations of origin as random effects to account for the variability contributed to this factor. Due to the influence of altitude on morphological variability, we added the altitude where individuals were collected as another random effect. Analyses were performed with the package lme4 (Bates et al., 2011 ) and lsmeans (Lenth, 2016 ) for a posteriori test (with Bonferroni correction) if necessary. Model validation was assessed graphically and by residual analysis. 2.1.4 Influence of environmental factors on morphological characters Complementarily, in a subset of data, we explored whether environmental factors might correlate with any of the phenotypic characters measured; because, for example, the clinal or geographic variation present in our study system may be influencing the patterns found (Goldberg & Lande, 2006 ). As altitude may be strongly associated with temperature and humidity, we considered the variation of these environmental factors in our analysis by obtaining the mean annual temperature and mean annual rainfall rasters from Geoportal IDESA ( http://geoportal.idesa.gob.ar/ , data from last year available: 2017). With the QGIS program 3.26 (QGIS Development Team, 2020), we mapped the distribution of the collected individuals (using the geo-referenced latitude and longitude data for each individual). We used the 'extractRandomClim' function of the raster package (Hijmans et al., 2015 ) in R to extract the mean annual temperature and mean annual rainfall values for each collection point. Subsequently, we explored the relationships between these environmental factors with size (centroid size, absolute length) and shape (PCs scores) previously calculated (see 2.1.3.2 ) with linear mixed models (LMMs). We acknowledge that other environmental factors (e.g., soil characteristics, atmospheric pressure, food availability) may sustain some of the phenotypic variation among species and populations. Still, the scoring of these factors was beyond the scope of this study, so our estimates of environmental effects on phenotype are prospective. 3. Results 3.1.1 Morphological variation across contexts We compared multiple sexual characters involved in courtship and sperm transfer in males' and females' scorpions from sympatric and allopatric contexts. We observed different patterns of phenotypic variation in different directions (convergences and divergences) in each species (Fig. 2 ), and the shape and size appear to respond independently to different selective pressures. The morphometric results for each character analyzed in both sexes are detailed below, first evaluating the size and then the variation in shape. 3.1.1.1 Chelicerae and pecten: asymmetric convergence in size only in females We observed an asymmetric convergence in the absolute length of both chelicerae (χ2 = 34.180, p < 0.001) and pectines (χ2 = 45.894, p < 0.001) in females ( U. brachycentrus more similar to U. achalensis in sympatry) (Fig. 2 ). Neither contexts nor species showed differences in the relative lengths of chelicerae or pectines. We only found interspecific differences in the relative cheliceral length in males, with U. brachycentrus males having larger chelicerae (χ2 = 64.348, p < 0.001). However, all the other variables did not differ between species or contexts. 3.1.1.2 Prosome and telson vesicle: size convergence Centroid size of the prosome showed symmetric convergence in females of both scorpion species, with species becoming more similar in sympatry than in allopatry (χ2 = 26.907, p < 0.001) and asymmetric convergence in males ( U. brachycentrus more similar in sympatry than in allopatry) (χ2 = 8.507, p = 0.004) (Fig. 2 ). In terms of shape, the Procrustes MANOVA showed no significant variation according to species and context. PC1 comprised almost half of the morphological variation (Females: 46.49%, Males: 45.85%), showing interspecific differences ( U. brachycentrus more compressed prosome than U. achalensis ) (Females: χ2 = 31.992, p < 0.001; Males: χ2 = 19.895, p < 0.001) (Fig. 2 ). PC2 explained an 18.44% of the variation in females and 13.82% in males and showed no differences between species or contexts in either sex. PC3 accounted for the 13.37% of the variability in females without differences between species or contexts. In contrast, PC3 in males representing the 12.52% of morphological variability was different between species (χ2 = 9.783, p = 0.002) and contexts (χ2 = 6.827, p = 0.006) but we found no significant interaction between these factors. Regarding the telson vesicle, in females, we found a pattern of symmetric convergence in the centroid size with both species becoming more similar in sympatry than in allopatry (χ2 = 32.176, p < 0.001) (Fig. 2 ). In males the convergence was asymmetric, as only males of U. brachycentrus presented a shift in the size of this character towards sympatry (χ2 = 6.118, p = 0.013). The Procrustes MANOVA showed significant shape variation according to species in both sexes (Females: F = 4.269, p = 0.001; Males: F = 4.404, p = 0.001), but the interaction between species and context was not significant (Fig. 2 ). In females, we found significant differences between species in telson vesicle shape reflected in PC1 (54%) (Females: χ2 = 22.441, p < 0.001) and PC2 (19.57%) (Females: χ2 = 21.034, p < 0.001). Also, in males, PC1 (67.48%) showed differences between species (χ2 = 36.965, p < 0.001) (Fig. 2 ), while in PC2 (12.21%) there were no significant differences between species or contexts. 3.1.1.3 Pedipalp in females: asymmetric convergence in size and divergence in shape We found asymmetric convergence in pedipalp centroid size, with species more similar in sympatry than in allopatry due to a shift of U. brachycentrus (χ2 = 19.812, p < 0.001) (Fig. 2 , 3 A). In terms of shape, the Procrustes MANOVA showed significant variation according to species and context (F = 7.788, p = 0.001). PC1 explained 38.10% of morphological variability, and we found asymmetric divergence in PC1, with U. brachycentrus females showing a shift relative to sympatric U. achalensis females and allopatric females (χ2 = 8.294, p = 0.004) (Fig. 3 B). PC2 explained 26.95% and PC3 10.60% of morphological variation although these shape variables showed no significant differences between species or contexts. 3.1.1.4 Pedipalp and apophysis in males: asymmetric divergence in shape Male pedipalp size showed only interspecific differences, with larger pedipalp and apophysis in U. achalensis than U. brachycentrus (χ2 = 84.839, p < 0.001) (Fig. 2 , 3 A). The Procrustes MANOVA showed significant variation by species and context (F = 3.321, p = 0.006). Regarding the pedipalp, the PC1 explained 45.25% of the morphological variability, and we found a pattern of asymmetric divergence in PC1 ( U. brachycentrus males with higher pedipalp and shorter fixed fingers than allopatric males and sympatric U. achalensis males) (χ2 = 10.069, p = 0.002) (Fig. 3 B, D-E). PC2 accounted for 20.21% and PC3 a 9.99% of the variability, and this component showed no differences between species or contexts (Fig. 3 D). For the pedipalp apophysis size, we found interspecific differences (χ2 = 38.651, p < 0.001), with apophysis of U. achalensis being larger than those of U. brachycentrus (Fig. 2 , 3 C). The Procrustes MANOVA showed significant variation in the interaction between species and context (F = 3.419, p = 0.014). PC1 (accounting for 31.11% of the variation) showed no significant differences between species or contexts. In contrast, PC2 explaining 21.07% of the morphological variation, showed significant differences between species in sympatry, and not in allopatry (χ2 = 10.221, p = 0.002) (Fig. 3 C, E). Moreover, the shape of the apophysis was different between sympatric and allopatric populations of U. brachycentrus so that this displacement pattern would be an asymmetric divergence. Morphological variability was also distributed between PC3 (9.34%) and PC4 (8.56%), although these morphological variables did not vary between contexts and only between species in PC4 (χ2 = 8.685, p = 0.003). 3.1.1.5 Caudal gland: asymmetrical convergence in size Caudal gland size showed a pattern of asymmetric convergence, with U. brachycentrus males more similar to U. achalensis males in sympatry and differing significantly from allopatric population males (with smaller gland) (χ2 = 10.087, p = 0.002) (Fig. 2 ). The Procrustes MANOVA showed significant variation only according to species (F = 155.064, p < 0.001), but the interaction between species and context was not significant. Regarding shape, PC1 almost completely comprised all morphological variability (92.81%), and we only found significant interspecific differences ( U. brachycentrus showing a more compressed and wider caudal gland than U. achalensis ) (χ2 = 155.774, p < 0.001). PC2, with an explanation of only 2.86% of the morphological variation, did not differ between species or contexts. 3.1.1.6 Hemispermatophore lamella: asymmetrical divergence in shape Hemispermatophore lamella size varied only at the interspecific level (χ2 = 86.714, p < 0.001), with lamella of U. achalensis males always being larger than those of U. brachycentrus (Fig. 2 , 4 A). In terms of shape, the Procrustes MANOVA showed significant variation according to species and context (F = 3.223, p = 0.006). Almost half of the lamella morphological variation was represented by PC1 (43.41%) (Fig. 4 B-C). This shape showed asymmetric divergence, as U. brachycentrus males differed from their allopatric conspecifics with a wider lamella, also differing from sympatric U. achalensis males (χ2 = 6.791, p = 0.009) (Fig. 4 C-D). PC2 comprised 15.33% and the PC3 14.02% of the morphological variation but these shape variables showed no differences between species or contexts (Fig. 4 C). 3.1.1.7 Hemispermatophore capsular lobes: asymmetrical divergence in size We found a pattern of asymmetric divergence in the hemispermatophore capsular lobe size, with males of U. brachycentrus in sympatry having larger lobes than the rest of the male groups (χ2 = 12.784, p < 0.001) (Fig. 2 ). We found no significant interaction between species and context in the Procrustes MANOVA, but there was variation in shape according to species (F = 4.847, p = 0.001). PC1 explained 31.96% and PC3 16.19% of the morphological variance, and none of the shape variables resulted in different between species or contexts. PC2 accounted for the 25.52% and differed between contexts (χ2 = 3.926, p = 0.048) and marginally between species (χ2 = 3.319, p = 0.068), but the interaction between context and species was not significant. 3.1.2 Coefficients of variation of morphological characters We found different values of CVs according to the type of character analyzed and sex (Table 3 ). The chelicerae, the pecten, the pedipalp, and the telson vesicle showed relatively low CVs values in both sexes and species, with no statistical differences in CVs between these characters, between species or between contexts. Only the pedipalp’ CVs differ between species, higher in U. brachycentrus than in U. achalensis in both sexes. In contrast, other male characters used exclusively during sexual interactions, such as the caudal gland and the pedipalp apophysis, showed high CVs, significantly different from the previously mentioned characters. In the case of the pedipalp apophysis for grasping during courtship, we found higher variation values in U. brachycentrus than in U. achalensis . Genital characters such as the length of the hemispermatophore lamella or the hemispermatophore capsular lobe showed low values of CVs with no differences between species or contexts. The only exception was the frontal crest of the hemispermatophore, which showed high CVs values compared to other genital characters in both species. Table 3 Influence of environmental factors on multiple somatic and genital characters of male and female Urophonius achalensis and U. brachycentrus scorpions from sympatric and allopatric areas. Character and compared parameter, sex, statistic value and statistical significance value are indicated (values < 0.05 indicated in bold). Abbreviations: AL, absolute length; cs, centroid size; hum, humidity (rainfall); hum:sp, interaction term between humidity and species fixed effect; PC, principal component 1–2; temp, temperature fixed effect; temp:sp, interaction between temperature and species fixed effects, ♂: males, ♀: females Morphological character Sex Fixed effect F p-value Sex Fixed effect F p-value Prosome cs ♂ temp:sp 12.102 0.001 ♀ temp:sp 68.449 < 0.005 PC1 ♂ temp 0.053 0.819 ♀ temp 3.324 0.072 PC2 ♂ temp 0.123 0.727 ♀ temp 0.589 0.445 PC3 ♂ temp 0.826 0.366 ♀ temp 0.165 0.686 cs ♂ hum 0.207 0.651 ♀ hum 5.424 0.022 PC1 ♂ hum 0.002 0.969 ♀ hum 0.021 0.885 PC2 ♂ hum 0.977 0.326 ♀ hum 3.929 0.051 PC3 ♂ hum 2.437 0.122 ♀ hum 0.231 0.632 Pedipalp cs ♂ temp:sp 5.129 0.026 ♀ temp:sp 8.876 0.004 PC1 ♂ temp 1.58 0.212 ♀ temp 2.715 0.103 PC2 ♂ temp 1.885 0.174 ♀ temp 0.205 0.652 PC3 ♂ temp 0.004 0.953 ♀ temp 0.015 0.904 cs ♂ hum 0.416 0.521 ♀ hum 1.505 0.223 PC1 ♂ hum 0.081 0.777 ♀ hum 0.069 0.793 PC2 ♂ hum 2.802 0.098 ♀ hum 0.987 0.323 PC3 ♂ hum 3.629 0.060 ♀ hum 0.818 0.365 Chelicerae AL ♂ temp:sp 12.904 0.001 ♀ temp:sp 15.457 0.0002 AL ♂ hum 0.001 0.973 ♀ hum 0.001 0.996 Pecten AL ♂ temp:sp 7.361 0.009 ♀ temp:sp 21.884 < 0.005 AL ♂ hum 0.421 0.653 ♀ hum 0.037 0.848 Telson vesicle cs ♂ temp:sp 4.957 0.029 ♀ temp:sp 8.371 0.005 PC1 ♂ temp 0.134 0.717 ♀ temp 1.783 0.185 PC2 ♂ temp 2.787 0.099 ♀ temp 0.897 0.348 cs ♂ hum 0.264 0.609 ♀ hum 2.614 0.109 PC1 ♂ hum 0.017 0.896 ♀ hum 0.476 0.492 PC2 ♂ hum 2.159 0.146 ♀ hum 0.753 0.389 Pedipalp apophysis cs ♂ temp 0.197 0.659 PC1 ♂ temp 0.325 0.570 PC2 ♂ temp 1.026 0.314 PC3 ♂ temp 0.136 0.713 PC4 ♂ temp 0.812 0.373 cs ♂ hum 0.019 0.888 PC1 ♂ hum 2.748 0.101 PC2 ♂ hum 1.796 0.184 PC3 ♂ hum 0.188 0.666 PC4 ♂ hum 1.107 0.298 Caudal gland cs ♂ temp:sp 8.485 0.003 PC1 ♂ temp 0.329 0.569 PC2 ♂ temp 2.068 0.154 cs ♂ hum 0.447 0.504 PC1 ♂ hum:sp 5.400 0.023 PC2 ♂ temp 0.764 0.385 Hemispermatophore Lamella cs ♂ temp:sp 13.602 0.0004 PC1 ♂ temp 2.648 0.108 PC2 ♂ temp 3.392 0.073 PC3 ♂ temp 2.144 0.147 cs ♂ hum 1.934 0.168 PC1 ♂ hum 0.015 0.902 PC2 ♂ hum 0.929 0.341 PC3 ♂ hum 0.159 0.691 Hemispermatophore capsular lobe cs ♂ temp:sp 4.152 0.046 PC1 ♂ temp 2.526 0.117 PC2 ♂ temp 0.005 0.945 PC3 ♂ temp 1.642 0.205 cs ♂ hum 0.725 0.398 PC1 ♂ hum 0.112 0.739 PC2 ♂ hum 3.025 0.087 PC3 ♂ hum 0.087 0.769 3.1.3 Influence of environmental factors on morphological characters We found that the size (centroid size and absolute length) of almost all the characters analyzed varied with temperature (Table 3 ). We found a significant statistical interaction between temperature and species in all cases, so temperature-dependent morphological variations were observed only in U . brachycentrus , with no relationship in U. achalensis . Generally, both sexes of this species had larger characters in colder areas (at higher altitudes) and smaller characters in warmer areas (at lower altitudes). This was observed for both sexes' prosome, pedipalp, chelicerae, pecten, and telson vesicle. In males, we also found this same pattern of variation in U. brachycentrus for the caudal gland and genital characters, although we did not observe it in the pedipalp apophysis. The pattern of variation found in the size of many characters coincides with the convergence asymmetrical in U. brachycentrus . The shape of none of the analyzed structures showed variation with temperature. As for humidity (rainfall), we found patterns of morphological variation of some characters regarding this environmental factor (Table 3 ). We observed that females of both species presented a larger prosoma in more humid areas. In addition, we found an interaction between humidity and species for caudal gland shape (PC1). That is, in U. brachycentrus , males presented a gland with negative PC1 values in more humid areas. This morphological change is associated with more slender and less rounded gland. The shape of no other character was affected by humidity. 4. Discussion We found great morphological variability between sympatric and allopatric contexts in the studied model species of scorpions. Our study revealed main novel insights about the evolution of shape and size of somatic and genital characters in an animal model so far understudied but with great potential for further research. We were able to observe complex patterns of phenotypic variation in different directions (convergences and divergences) in size and shape, which allows us to suggest a possible mosaic evolution in certain sexual characters in these scorpion species. The integration of the results allows us to infer an asymmetric RCD in the shape of certain sexual characters of both sexes key for courtship success (i.e., grasping characters) and sperm transfer (i.e., genital characters of the hemispermatophore). Intriguingly, although we found low phenotypic variation in some genital characters, others showed high variation which could reflect that some characters are under antagonistic selective pressures. The convergence patterns found in the size of many characters were due to environmental fluctuations linked to the altitude cline of the geographic system. In the following discussion, we analyze in depth the remarkable patterns of phenotypic variation, the possible selection pressures underlying this variability, and the consequences of the RCD in the mating system and coexistence for these scorpion species. 4.1 Reproductive character displacement in pedipalps We obtained evidence of phenotypic divergence in shape and size of multiple somatic characters used in courtship in U. brachycentrus , while U. achalensis showed no divergence in any character between sympatric and allopatric populations. Urophonius brachycentrus males of the sympatric zone differed from their conspecifics and U. achalensis males by having more globose pedipalps and apophyses with a lower crest deeper. U. brachycentrus females showed an RCD pattern also in the shape of their pedipalps, with the pedipalps being more globose in sympatric zones. Therefore, the pattern of divergence in pedipalp shape was complementary in males and females. RCD results in mechanical incompatibilities (due to mechanisms under natural selection such as the ''lock-and-key'' hypothesis) that can hinder the culmination of heterospecific matings, promoting reproductive isolation (Eberhard, 2004 ). We know that there is incompatibility at the behavioral level since, in heterospecific mating, females show more resistance events (Oviedo-Diego, M. pers. obs.), which sometimes causes the pedipalps of both sexes to be released, interrupting the mating. Multiple biomechanical variables may be involved in these events, such as the pedipalp muscles and grip strength, as well as probably the optimal fit given by the morphology of the apophysis. Analyzing these variables together could help better understand the determinants of ''pedipalp grasping'' success in scorpions (van der Meijden et al., 2012 ) and its relationship to species-specificity in heterospecific matings. Peretti et al. ( 2000 ) report that some of the intercrosses between Bothriurus flavidus and B. prospicus from areas of sympatry could progress to courtship but not to complete matings, but it is unclear which factors lead to mating interruption. Interestingly, it was noted that in intercrosses between B. cordubensis and B. noa , some matings were interrupted by female resistance events (Peretti et al., 2000 ). Although the latter species are allopatric, likely, mechanical incompatibilities in pedipalp grasping are also occurring in this pair of species. The pedipalp apophysis is a key character for the correct attachment of the pedipalps during the mating dance (Ábalos & Hominal, 1974 ; Maury, 1968 ; Peretti, 1993 ), although little is known about the mechanics of the coupling and adjustment with the female pedipalps (Peretti, 1993 ). The morphospace of this character was complex, and although its summary into a few dimensions allowed us to simplify this complexity, we believe that future studies should be carried out to complete the understanding of the selective forces underlying the evolution of this character. According to the hypotheses of morphological evolution, RCD could be expected under the hypotheses of structural or sensory "lock-and-key" natural selection. Under these hypotheses, species-specific morphological complementarity could exist with similar divergence in both sexes (Eberhard, 1985 ; Arnqvist, 1997 ), so it is necessary to include the morphological variability of female characters in future studies. At the same time, the male pedipalp apophysis showed a high coefficient of variation indicative of sexual selection pressures (directional selection) and not consistent with characters that are under stabilizing selection according to the "lock-and-key" hypothesis (Peretti et al., 2001 ). This could suggest that the pedipalp apophysis in U. brachycentrus is under different and maybe opposite selective pressures. On the one hand, the shape of the apophysis (which could be summarized as apophysis depth) showed divergence in sympatry to ensure mechanical isolation. Still, on the other hand, the size of the apophysis seems to be influenced by sexual selection pressures. 4.2 Reproductive character displacement in male genital characters As we observed in some somatic characters, we found evidence of RCD in characteristics of the hemispermatophores of U. brachycentrus . In addition, we found that these characters had low CVs, which would support some type of stabilizing selection on these characters (Peretti et al., 2001 ). Males of this species showed hemispermatophores with a more compressed lamella and larger capsular lobes than allopatric males and sympatric U. achalensis males. The size of larger capsular lobules in U. brachycentrus could be partly explained by the increase in size of females of this species towards the sympatric zone, as morphological complementarity is expected for mechanical isolation by the ''lock-and-key''. Although there are examples of these hypothesis in arthropods (Mikkola, 1992 , 2008 ; Sota & Kubota, 1998 ; Usami et al., 2006 ; Nagata et al., 2007 ; Takami et al., 2007 ; Tanabe & Sota, 2008 ; Sota & Tanabe, 2010 ; Wojcieszek, & Simmons, 2012 ; Kubota et al., 2013 ; Nishimura et al., 2022 ), it is a hypothesis that has been discarded in several species as in general, genitalia diverge much more in males than in females, and it is not so common to find morphological complementarity (Eberhard, 1985 ; Shapiro & Porter, 1989 ; Masly, 2012 ). Like to our conclusion for the pedipalp apophysis, it would be necessary to evaluate the female component to confirm this hypothesis in these species. However, although there may be rather cryptic differences, the female genital atrium is flexible and has a relatively ''uniform'' structure (Peretti, 2003 , 2010 ). Therefore, the female genitalia in these species does not mechanistically prevent the entry of heterospecific male genitalia, which would also not support the ''lock-and-key'' hypothesis. Some particular zones of the hemispermatophore (i.e., frontal crest area) had a very high phenotypic variation, suggesting that their variability is not so much restricted, which is not consistent with a stabilizing selection (Eberhard et al., 1998 ; Peretti et al., 2001 ). These results would indicate that the morphological variation of at least some areas of the genitalia of these species would be explained rather by sexual selection hypotheses (Peretti, 2003 , 2010 ; Monod et al., 2017 ). The frontal crest of the lamella fits into the inter-coxal space of the female, and there could be a 'passive' choice by 'mechanical adjustment' (Eberhard, 1985 ; Huber & Eberhard, 1997 ). Also, the capsular lobes possess micro-ornamentations contacting the female genital atrium wall that could have a stimulatory role, which could be contemplated in a female cryptic choice hypothesis (Peretti, 2003 , 2010 ). Larger capsular lobes could be related to a larger contact surface of ornamentations with the female genital atrium and, consequently, a greater stimulation that could be linked to cryptic female choice. An inevitable question at this point is: if some portions of the genitalia are under sexual selection pressures, why does RCD exist in others? An interesting option could be that the female may bias, by cryptic choice, the use of sperm or other variables (e.g., hardening of the genital plug) according to characteristics evaluated in the interaction of the genitalia, such as (a) greater stimulation by larger capsular lobes (would explain the RCD in the hemispermatophores capsule lobe), (b) by mechanical adjustment of coxae of the first pair of legs with hemispermatophore frontal crest (would explain the RCD in the lamella of hemispermatophore) or fit between male capsular lobe and female genital atrium. We could say that there would be a "combination" of the sensory/mechanical "lock-and-key" hypothesis, where females can recognize the species-specificity of the male genitalia (and thus RCD would be promoted) but where physiological changes would not occur immediately but at the post-copulatory level mediated by female cryptic choice processes. This interaction between sexual and natural selection hypotheses could be expected to explain the evolution of genitalia in these species, where there is intense competition between males at the intra and interspecific level and promiscuity in their mating and where females must not only exercise mate choice at the pre-copulatory level, but copulatory and post-copulatory mechanisms seem to be necessary to avoid hybridization. A similar example seems to occur in hybridizing Drosophila species, where the male genitalia differ in size and shape, and the external female genitalia shows no interspecific differences (Coyne, 1983 ). In interspecific mating, the intrusion of the male genitalia differentially contacts the female genitalia so that females can store and use sperm according to the specific identity of the male (Price et al., 2001 ). This is called "cryptic reproductive isolation" and maybe a by-product of multiple evolutionary forces acting at the intra- and interspecific level (Price et al., 2001 ). As we have emphasized above, it is now recognized that mate choice and specific recognition are part of a continuum and that the forces of sexual and natural selection may interact in multiple ways explaining patterns of sexual diversification across species (Ryan & Rand, 1993 ; Liou & Price, 1994 ; Boake et al., 1997 ; Mendelson & Shaw, 2012 ). Keeping these interactions in mind is critical for analyzing possible hypotheses of genital evolution (Simmons, 2014 ). The reinforcement model postulates the emergence of successive reproductive isolation barriers if these become ineffective (Howard, 1993 ; Coyne & Orr, 2004 ; Butlin & Smadja, 2018 ). If mechanical isolation existed in the past leading to an RCD pattern but subsequently the effectiveness of this barrier, weakened pre-copulatory barriers may have been generated (such as RCD in pedipalps and behavioral incompatibilities), and the RCD in the genitalia may have persisted rather than reverted to the previous morphological scenario. This, in turn, could have resulted in the genitalia (or some of its parts) being able to diversify under other pressures more "freely." This argument is supported by the fact that in this system, there is a pre-copulatory filter in heterospecific matings, and only 10 to 20% of these reach sperm transfer (Oviedo-Diego, M. per obs). Perhaps the existence of a percentage of matings that reach this point is sufficient for the maintenance of the RCD observed in genital characters. 4.3 Environmental variations promote size convergence of multiple characters The overall size, but no shape, of individuals converged in sympatry, i.e., individuals were more similar in size when the species were together, and this pattern was particularly strong for U. brachycentrus . This could be observed in males and females for the prosome and the telson vesicle. Convergence was also observed in females for cheliceral size and in males for caudal gland size and hemispermatophore lamella. The patterns of convergence found in size could follow the rule of Atkinson ( 1994 , 1995 ) that predicts larger body sizes at lower temperatures (Horne et al., 2015 ). Most ectotherms grow more slowly and mature with larger body sizes in colder environments (Angilletta et al., 2004). This increase in size may be adaptive when it allows for increased fecundity or higher survival or reproductive rates (Stearns, 1992 ). In scorpions, it is known that different numbers of molts or the period between molts can affect the final body size of individuals (Sarmento et al., 2008 ; Seiter et al., 2020 ), so through this mechanism, they could reach different sizes depending on environmental characteristics such as temperature or humidity, as well as variations in diet (Sarmento et al., 2008 ). The effect of altitude and temperature change was probably more drastic in U. brachycentrus due to the large altitudinal and temperature difference between the allopatric and sympatric populations compared. Temperature is predicted to affect the body size of individuals of both sexes similarly (Hirst et al., 2015 ), and in U. brachycentrus , we found that males and females increase in size. However, this increase could be seen reflected in different characters in each sex, which is perhaps related to sexual dimorphism due to different life habits or phenotypic plasticity in thermal gradients (Fairbairn, 2005 ; Blanckenhorn et al., 2006 ; Stillwell & Fox, 2007 ). Females had a general increase in size, including their chelicerae, a key character for digging and gestation chambers (Maury, 1968 , 1969 , 1977 ). Males increased in the body and caudal gland size, a character for sexual interactions (Peretti, 1997 ). The shape changes in the caudal gland related to humidity are intriguing, considering that this gland produces complex chemistry secretions with numerous compounds where geographic variation among different populations has been demonstrated previously (Olivero et al., 2015 ). Future studies will aim to determine whether the dynamics of secretion production or effectiveness of secretion rubbing may depend on these shape variations and whether this correlates with behavioral differences between species and allopatry and sympatry contexts. In scorpions, it is known that geographic variability may exist (Harington, 1983 ; Abdel-Nabi et al., 2004 ; Olivero et al., 2012 , 2015 ; Yamashita & Rhoads, 2013 ) and that the size of individuals may be affected by environmental gradients (Jochim et al., 2020 ; Lira et al., 2021 ). For example, Jochim et al. ( 2020 ), studying the morphology of a species complex of the family Vaejovidae, found a pattern of morphological convergence very similar to our results. In mountainous areas of Arizona, individuals at higher elevations were larger, resulting in individuals of different species being more similar in the middle areas of the gradient (Jochim et al., 2020 ). These authors argue that RCD does not occur in these species and that these scorpions probably follow Bergmann's rule, although they do not discuss these aspects further (Jochim et al., 2020 ). Because of this type of geographic variation, RCD studies must contemplate ecological factors as promoters of morphological variation (Goldberg & Lande, 2006 ; Kosuda et al., 2016 ). 4.4 Species asymmetry in morphological variability Asymmetric RI and RCD have been reported multiple times (Bordenstein et al., 2000 ; Pfennig & Simovich, 2002 ; Smadja & Ganem, 2005 ; Cooley et al., 2006 ; Cooley, 2007 ; Hochkirch et al., 2007 ; Costa-Schmidt & Machado, 2012 ) and it generally occurs when there are interspecific differences in the intensity of selective pressures to avoid heterospecific interactions because species suffer different costs from RI (Pfennig & Simovich, 2002 ; Cooley, 2007 ). Also, asymmetric outcomes in morphological variability between species may indicate interspecific differences in morphological plasticity. Divergent characters can also be plastic or can be expressed facultatively when individuals face competition with heterospecifics, so plasticity has been a proposed mechanism to explain character displacement (Robinson & Wilson, 1994 ; Pfennig & Murphy, 2002 ; Rice & Pfennig, 2007 ; Pfennig & Pfennig, 2010 ; Stuart et al., 2017 ). Species with broad distribution, exposed to a wide range of environmental conditions, and with ample genetic variation may exhibit more remarkable phenotypic plasticity (DeWitt & Scheiner, 2004 ; Lavergne et al., 2004 ; Pigliucci et al., 2006 ). For example, Crowder et al. ( 2010 ) found that the globally distributed whitefly Bemisia tabaco biotype exhibited greater plasticity in reproductive behavior, which could result in greater success in avoiding the costs of RI than other biotypes. Here, Urophonius species present asymmetries in the RI degree they may be undergoing since males of U. brachycentrus are more indiscriminate in their mating decisions than males of U. achalensis (Oviedo-Diego et al., 2021 ). Moreover, U. brachycentrus presented higher male-biased operational sex ratios than U. achalensis in the sympatric zone (Oviedo-Diego, M. pers. obs.), which could mean males under greater scramble competition to find females and that this species could suffer higher costs due to RI (Oviedo-Diego et al., 2020 ; 2021 ). In turn, U. brachycentrus showed the most remarkable morphological variations, being the most widely distributed species compared to U. achalensis , endemic to the highland area under analysis (Acosta 1985 , 1993 ; Ojanguren-Affilastro, 2005 ; Ojanguren-Affilastro et al., 2020 ). This complex social and geographic scenario could translate into strong selective pressures for interspecific recognition during mating or sperm transfer and the existence of RCD patterns in an asymmetric manner, being U. brachycentrus the species that suffers more RI costs and the most morphologically plastic to manifest changes under these pressures. 4.5 Mixed selective pressures on multiple characters in scorpions Our results reveal a strong variation in the size and shape of somatic and genital characters, which supports the notion that morphological traits are the result of multiple selective pressures and that different dimensions of the same character (e.g., shape, size) may be reflecting different evolutionary responses (mosaic evolution). This is most noticeable in characters used in multiple activities in the organism's life. We found evidence of the existence of RCD for the pedipalps shape of both sexes in sympatric populations, an evolutionary response to avoid crossbreeding and strengthen reproductive isolation among these species. In turn, other characters showed high geographic variability in size reflected in patterns of convergence towards the sympatry zone, which could affect the mating system of these species, promoting RI and explaining the high values of phenotypic variation found in characters used in sexual interactions (e.g., caudal gland, pedipalp apophysis). It is noteworthy the different selective pressures under which the genitalia would be, also under natural selection pressures showing an RCD pattern in shape, although manifesting in other portions of the hemispermatophore very high phenotypic variation which would indicate possible sexual selection pressures acting mainly in the crest zone. Peretti ( 2010 ) highlights the existence of mixed patterns in the genitalia of scorpions, were morphological complexity results from different selective regimes. This has also been observed in other arachnids (Huber, 1996 , 2004 ) and insects (Song & Wenzel, 2008 ; Simmons et al., 2009 ; Song & Bucheli, 2010 ; Rowe & Arnqvist, 2012 ; House et al., 2013 ; Frazee & Masly, 2015 ) where characters are under multiple, often contradictory or inconsistent pressures. Studies in water striders suggest that the non-intromittent genitalia have differing degrees of selection acting upon them (Danielsson & Askenmo, 1999 ; Bertin & Fairbairn, 2005 ; Rowe & Arnqvist, 2012 ). Another example was reported in the dung beetle Onthophagus taurus which has shown that different sections of male genital morphology may be under different selective regimes (Song & Wenzel, 2008 ; Simmons et al., 2009 ) as the shape of the aedeagus is subject to directional sexual selection, but genital sclerites that penetrate the female genitalia are subject to stabilizing and disruptive nonlinear selection (Simmons et al., 2009 ). In addition, in O. taurus , the genitalia shape diverges rapidly due to directional sexual selection, whereas size remains unaffected in the process (Simmons et al., 2009 ). Similarly, it has been reported for the millipede Antichiropus variabilis that genitalia shape responded to stabilizing pressures (supporting the occurrence of lock-and-key), although genitalia size did not follow this pattern and responded to environmental gradients (Wojcieszek & Simmons, 2012 ). This is like to our results, where the shape of certain structures responds to specific recognition variations with low phenotypic variation, and size shows patterns of variation linked to geographic and environmental differences. The size and shape of the same structure may respond in this mosaic manner, independently to different selective pressures, perhaps due to genetic or developmental decoupling (Macagno et al., 2011 ; Rowe & Arnqvist, 2012 ; Wojcieszek & Simmons, 2012 ; Richmond, 2014 ). Future studies will aim to assess the consistency of these results with allometric patterns between populations, and coevolution between female and male characters, as well as explore the morphological complexity of the traits by assessing the modularity of the subunits of the characters (e.g., Kuntner et al. 2009 ; Tatarnic & Cassis, 2010 ; Rowe & Arnqvist, 2012 ; Genevcius & Schwertner, 2017 ; Genevcius et al., 2020 ) Conclusions We found a remarkable morphological variability in both scorpion species that was determined in part by geographic and environmental variations, in part by sexual selection pressures at the intra- and interspecific level, and in part by natural selection pressures during species recognition. We report a pattern of asymmetric morphological variation where one of the scorpion species ( U. brachycentrus ) suffered an increase in size in several characters to the sympatric zone due to environmental factors (showing a pattern of morphological convergence). This increase in size and a scenario of promiscuity probably led to certain characters undergoing intense sexual selection pressures, which is reflected in the high phenotypic variation found. However, key characters for mating success, such as grasping or genital characters, experienced morphological divergence in males and females, implying a mechanical incompatibility that could function as a barrier promoting reproductive isolation. However, some characters that showed variation by RCD were also found to be under sexual selection pressures, suggesting a complex scenario of mixed selective regimes acting on these characters. On the other hand, the non-concordant results on the pressures on the size and shape of characters enlighten us on the complexity inherent in the evolution of multi-functional traits in scorpions. This comprehensive study presents novel results in an ancestral group that has not been studied from this perspective and provides interesting insights for evaluating characters under multiple selective pressures in animal systems with RI. Declarations Authors are required to disclose financial or non-financial interests that are directly or indirectly related to the work submitted for publication. Please refer to “Competing Interests and Funding” below for more information on how to complete this section. Competing interests: The authors declare no competing interests with regard to this manuscript and the material implicated. Ethical standards We declare that the experiments comply with the current laws of Argentina. 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Phenotypic plasticity of insects: Mechanisms and consequences , (pp. 1-63). Science Publishers, Enfield. Wilson, J. D., Zapata, L. V., Barone, M. L., Cotoras, D. D., Poy, D., & Ramírez, M. J. (2021). Geometric morphometrics reveal sister species in sympatry and a cline in genital morphology in a ghost spider genus. Zoologica Scripta , 50 (4), 485-499. https://doi.org/10.1111/zsc.12478 Wojcieszek, J. M., & Simmons, L. W. (2012). Evidence for stabilizing selection and slow divergent evolution of male genitalia in a millipede ( Antichiropus variabilis ). Evolution: International Journal of Organic Evolution , 66 (4), 1138-1153. https://doi.org/10.1111/j.1558-5646.2011.01509.x Yamaguchi, R., & Iwasa, Y. (2015). Reproductive interference can promote recurrent speciation. Population Ecology , 57 (2), 343-346. https://doi.org/10.1007/s10144-015-0485-2 Yamashita, T., & Rhoads, D. D. (2013). Species delimitation and morphological divergence in the scorpion Centruroides vittatus (Say, 1821): insights from phylogeography. PLoS One, 8 (7), e68282. https://doi.org/10.1371/journal.pone.0068282 Yukilevich, R. (2021). Reproductive character displacement drives diversification of male courtship songs in Drosophila . The American Naturalist , 197 (6), 690-707. https://doi.org/10.5061/dryad.m63xsj41k. Zelditch, M. L., Swiderski, D. L., & Sheets, H. D. (2004). Geometric morphometrics for biologists: a primer . Elsevier, San Diego. Additional Declarations No competing interests reported. Supplementary Files TableS1.xlsx Table S1 Description of morphological landmarks analyzed in each character in both sexes of Urophonius brachycentrus and achalensis . We used mainly Type II landmarks ( sensu Bookstein, 1991) (points of minimum and maximum curvature on an anatomical trait) and some Type I landmarks (points of intersection between anatomical features). Position indicated in Fig. 1. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-2445373","acceptedTermsAndConditions":true,"allowDirectSubmit":false,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":165190674,"identity":"65755a16-de8a-45c7-a3cf-81f3c9224ccd","order_by":0,"name":"Mariela Oviedo-Diego","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAAA/0lEQVRIiWNgGAWjYJCCAwwV/xkYmBkYJBgqgFxm5gYitJxhhmo5A9LCSFgLA2MbM5iWYGwDc/Fr0W0/+/DABzY2OYPjzAdvfJxXG83fDtTyo2IbTi1mZ9INDs7g4TE2OMyWbDlz2/HcGYcZGxh7ztzGreVAGsNhHgmJxJnNPGbSvNuO5TYAtTAztuHRcv4Zw+E/BgZALfzfpHnnHMudT1DLDaAtDAkJif3MPGzSvA01uRsIa3nGcLDnwAFjfmY2Y8sZxw7kbgRqOYjXL+fTmD/8/HdAjo3/8MMbH2rqcuedP3zwwY8K3FrQwWEweYBo9UBQR4riUTAKRsEoGCEAAJzdXPYY3nY8AAAAAElFTkSuQmCC","orcid":"","institution":"CONICET- UNC, Universidad Nacional de Córdoba","correspondingAuthor":true,"prefix":"","firstName":"Mariela","middleName":"","lastName":"Oviedo-Diego","suffix":""},{"id":165190675,"identity":"d79332c7-1519-4a93-9715-ca21634f3eb7","order_by":1,"name":"Camilo Mattoni","email":"","orcid":"","institution":"CONICET- UNC, Universidad Nacional de Córdoba","correspondingAuthor":false,"prefix":"","firstName":"Camilo","middleName":"","lastName":"Mattoni","suffix":""},{"id":165190676,"identity":"cff0bba9-1d78-43d4-82c1-3c51dd133d4a","order_by":2,"name":"Fedra Bollatti","email":"","orcid":"","institution":"CONICET- UNC, Universidad Nacional de Córdoba","correspondingAuthor":false,"prefix":"","firstName":"Fedra","middleName":"","lastName":"Bollatti","suffix":""},{"id":165190677,"identity":"15f550aa-82dc-4319-ab26-a426e51c4090","order_by":3,"name":"Eduardo M. Soto","email":"","orcid":"","institution":"IEGEBA (CONICET-UBA), Universidad de Buenos Aires","correspondingAuthor":false,"prefix":"","firstName":"Eduardo","middleName":"M.","lastName":"Soto","suffix":""},{"id":165190678,"identity":"cf2b6b07-9cd4-4745-984c-f3ea9c60d70f","order_by":4,"name":"Alfredo V. Peretti","email":"","orcid":"","institution":"CONICET- UNC, Universidad Nacional de Córdoba","correspondingAuthor":false,"prefix":"","firstName":"Alfredo","middleName":"V.","lastName":"Peretti","suffix":""}],"badges":[],"createdAt":"2023-01-05 06:59:25","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-2445373/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-2445373/v1","draftVersion":[],"editorialEvents":[{"content":"https://doi.org/10.1007/s11692-023-09623-2","type":"published","date":"2024-01-27T15:12:47+00:00"}],"editorialNote":"","failedWorkflow":false,"files":[{"id":31313125,"identity":"faf46a23-f7ed-4490-8d5f-fce7d1126079","added_by":"auto","created_at":"2023-01-09 15:47:13","extension":"png","order_by":1,"title":"Figure 1","display":"","copyAsset":false,"role":"figure","size":3687498,"visible":true,"origin":"","legend":"\u003cp\u003eSelected characters for morphological study in two scorpion species of \u003cem\u003eUrophonius achalensis\u003c/em\u003eand \u003cem\u003eU. brachycentrus\u003c/em\u003e. A. General diagrams of measured somatic and genital characters. B. Prosome. C. Lateral view of the male pedipalp. D. Apophysis of the male pedipalp. E. Lamella of the hemispermatophore with frontal crest. F. Capsular lobe of the hemispermatophore. G. Dorsal view of male telson. H. Ventral view of male telson with caudal gland. Abbreviations: ap, pedipalp apophysis; cg, caudal gland; ch, chelicerae; cl, hemispermatophore capsular lobe; fc, hemispermatophore frontal crest; ff, pedipalp fixed finger; la, hemispermatophore lamella; me, median eye; mf, pedipalp mobile finger; pe, pedipalp; pr, prosoma; tv, telson vesicle; st, sting. References: Red dots and numbers, Landmarks (descriptions in Table S1); dotted line with more separated stroke, positioning of semilandmarks; dotted line with a narrower stroke, character analyzed by elliptical Fourier analysis (EFA). Scales: A= 5 mm in scorpion, 0.5 mm in hemispermatophore, B-C, G-H=1 mm; D-F=0.5 mm\u003c/p\u003e","description":"","filename":"Figure1.png","url":"https://assets-eu.researchsquare.com/files/rs-2445373/v1/9d60287e167b78ffdd2a85af.png"},{"id":31315255,"identity":"f28241ac-6e6b-45b5-b5c8-76933aaa2085","added_by":"auto","created_at":"2023-01-09 16:03:12","extension":"png","order_by":2,"title":"Figure 2","display":"","copyAsset":false,"role":"figure","size":509862,"visible":true,"origin":"","legend":"\u003cp\u003eDiagrams showing the summary of morphological variation in size and shape of somatic and genitalia characters in scorpions in different contexts of sympatry and allopatry. Each character is scaled at the intrasexual level. Gray area in the middle of the plate indicates sympatric zone. Gray arrows, characters undergoing convergence; black arrows, characters undergoing divergence (RCD). ♂: males, ♀: females, ≠: Statistical differences between species in sympatry\u003c/p\u003e","description":"","filename":"Figure2.png","url":"https://assets-eu.researchsquare.com/files/rs-2445373/v1/7a4788b8ae7c96a506ad2c3d.png"},{"id":31314046,"identity":"42c1fc4c-dfc8-4061-9cd0-4c0e3b732200","added_by":"auto","created_at":"2023-01-09 15:55:12","extension":"png","order_by":3,"title":"Figure 3","display":"","copyAsset":false,"role":"figure","size":952481,"visible":true,"origin":"","legend":"\u003cp\u003eInterspecific and intraspecific morphological variation in pedipalp and male pedipalp apophysis in \u003cem\u003eUrophonius achalensis\u003c/em\u003e and \u003cem\u003eU. brachycentrus\u003c/em\u003e from sympatric and allopatric zones. A. Pedipalp size of males (first box) and females (second box) indicated by centroid size. B. Pedipalp shape (PC1) of males (first box) and females (second box) and differences between species and contexts C. Shape of pedipalp apophysis of males (PC2) and differences between species and contexts. Statistical differences indicated in each graph: continued line showed interspecific differences, dashed line: intraspecific differences (between allopatric and sympatric contexts), ♂: males, ♀: females. D. Male pedipalp morphospace indicating the morphological distribution of individuals along two principal components of variation. Numbers in parentheses on each axis showing percentage of variance explained by each principal component. Color reference following A-C. E. Summary of morphological changes in PC scores of extremes individuals (minimum in sympatric population and maximun in allopatric population) of \u003cem\u003eU. brachycentrus\u003c/em\u003e, Top: shape of male pedipalp (PC1 scores); Below: shape of male pedipalp’ apophysis (PC2 scores); black dots showing landmarks and semilandmarks showing consensus conformation, orientation of arrows (vectors) indicating direction of morphological change and arrow longitude indicating magnitude of change\u003c/p\u003e","description":"","filename":"Figure3.png","url":"https://assets-eu.researchsquare.com/files/rs-2445373/v1/301c4132384d49b9c996e758.png"},{"id":31313124,"identity":"cb15bc16-9cfa-4a5d-a4c2-ab5526404cb0","added_by":"auto","created_at":"2023-01-09 15:47:12","extension":"png","order_by":4,"title":"Figure 4","display":"","copyAsset":false,"role":"figure","size":868624,"visible":true,"origin":"","legend":"\u003cp\u003eInterspecific and intraspecific morphological variation in the hemispermatophore lamella of \u003cem\u003eUrophonius achalensis\u003c/em\u003e and \u003cem\u003eU. brachycentrus\u003c/em\u003emales from sympatric and allopatric zones. A. Size of hemispermatophore lamella indicated by centroid size. B. Hemispermatophore lamella shape (PC1) and differences between species and contexts. Statistical differences indicated in each graph: continued line showed interspecific differences, dashed line: intraspecific differences (between allopatric and sympatric contexts). C. Morphospace indicating the morphological distribution of individuals along two principal components of variation. Numbers in parentheses on each axis showing percentage of variance explained by each principal component. Color reference following A-B. D. Summary of morphological changes in PC1 scores of extremes individuals (maximum in sympatric population and minimum in allopatric population) of \u003cem\u003eU. brachycentrus\u003c/em\u003e, black dots showing landmarks and semilandmarks showing consensus conformation, orientation of arrows (vectors) indicating direction of morphological change and arrow longitude indicating magnitude of change\u003c/p\u003e","description":"","filename":"Figure4.png","url":"https://assets-eu.researchsquare.com/files/rs-2445373/v1/029ccca80b524aa34b8537e6.png"},{"id":50313911,"identity":"0243d155-9f1e-4b00-bd5c-16dd0607b41b","added_by":"auto","created_at":"2024-01-29 15:27:58","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":3370578,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-2445373/v1/d220325c-659c-4630-a79f-ad11b5396897.pdf"},{"id":31313121,"identity":"0db3ec72-b250-42a9-8ecc-644a62517199","added_by":"auto","created_at":"2023-01-09 15:47:12","extension":"xlsx","order_by":1,"title":"","display":"","copyAsset":false,"role":"supplement","size":11092,"visible":true,"origin":"","legend":"\u003cp\u003e\u003cstrong\u003eTable S1\u003c/strong\u003e Description of morphological landmarks analyzed in each character in both sexes of \u003cem\u003eUrophonius brachycentrus\u003c/em\u003e and \u003cem\u003eachalensis\u003c/em\u003e. We used mainly Type II landmarks (\u003cem\u003esensu \u003c/em\u003eBookstein, 1991) (points of minimum and maximum curvature on an anatomical trait) and some Type I landmarks (points of intersection between anatomical features). Position indicated in Fig. 1.\u003c/p\u003e","description":"","filename":"TableS1.xlsx","url":"https://assets-eu.researchsquare.com/files/rs-2445373/v1/1f548708cfd8e5d5706ad20b.xlsx"}],"financialInterests":"No competing interests reported.","formattedTitle":"Mosaic evolution of grasping and genitalic traits in two sympatric scorpion species with reproductive interference","fulltext":[{"header":"1. Introduction","content":"\u003cp\u003eDetermining the factors underlying phenotypic variation in natural populations is important for comprehending the evolution of species and their biological diversity and is a fundamental task of evolutionary biology (Coyne \u0026amp; Orr, \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). Morphological characters are shaped by multiple selective pressures, especially those involved in various components of the life history of organisms. Secondary sexual characters undergo relatively fast evolutionary divergence due to sexual and natural selection (Svensson \u0026amp; Gosden, \u003cspan citationid=\"CR210\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Natural selection favors morphological traits linked to growth, reproduction, and survival resulting in greater reproductive success for certain environments. In contrast, sexual selection underlies morphological changes that favor reproductive success through intra-sexual competition, inter-sexual mate choice, or post-copulatory processes (Kraaijeveld et al., \u003cspan citationid=\"CR100\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Maan \u0026amp; Seehausen, \u003cspan citationid=\"CR116\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Safran et al., \u003cspan citationid=\"CR178\" class=\"CitationRef\"\u003e2013\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe study of interspecific interactions is crucial for understanding sex-linked ecological and evolutionary patterns (Cothran, \u003cspan citationid=\"CR40\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Reproductive interference (henceforth referred as \u0026lsquo;RI\u0026rsquo;) is defined as any type of interspecific interaction between sympatric species associated with their mating systems caused by incomplete recognition between species (Gr\u0026ouml;ning \u0026amp; Hochkirch, \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Burdfield-Steel \u0026amp; Shuker, \u003cspan citationid=\"CR31\" class=\"CitationRef\"\u003e2011\u003c/span\u003e). This process may negatively affect the reproductive success of at least one species (Hochkirch et al., \u003cspan citationid=\"CR79\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). RI between species can lead to the displacement of key characters in reproductive interactions (i.e., reproductive character displacement - henceforth referred as \u0026lsquo;RCD\u0026rsquo;) (Howard, \u003cspan citationid=\"CR85\" class=\"CitationRef\"\u003e1993\u003c/span\u003e), which generally results in a divergence of these characters alleviating RI and thus reinforcing reproductive isolation (Servedio \u0026amp; Noor, \u003cspan citationid=\"CR189\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Coyne \u0026amp; Orr, \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Kyogoku, \u003cspan citationid=\"CR105\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Characters of coexisting species should be more divergent in sympatry than in allopatry. The more similar the characters of interacting species are in sympatry, the greater the consequences of RI on reproductive success (Pfennig \u0026amp; Pfennig, \u003cspan citationid=\"CR156\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Konuma \u0026amp; Chiba, \u003cspan citationid=\"CR98\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). In turn, the degree and direction of divergence of sexual characters may differ according to their function or the moment of the reproductive event in which RI occurs (Gr\u0026ouml;ning \u0026amp; Hochkirch, \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). In other cases, adaptive promiscuity may exist, and competition for exploitation (for females beyond their species) is propitiated, which may prevent divergence and even generate convergence of sexual characters (Grant, \u003cspan citationid=\"CR69\" class=\"CitationRef\"\u003e1972\u003c/span\u003e; Grether et al., \u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Tobias et al., \u003cspan citationid=\"CR215\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Drury et al., \u003cspan citationid=\"CR50\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Sobroza et al., \u003cspan citationid=\"CR198\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) with consequent maintenance or intensification of RI (Takakura et al., \u003cspan citationid=\"CR211\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Wheatcroft, \u003cspan citationid=\"CR226\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Yamaguchi \u0026amp; Iwasa, \u003cspan citationid=\"CR231\" class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eIn sympatric areas, intraspecific sexual selection pressures may join interspecific interactions generating a mosaic of selective pressures with different outcomes in terms of morphological variation (Grether et al., \u003cspan citationid=\"CR70\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Secondary sexual characters may play a role in specific recognition, so their divergence can be explained by natural selection (Mayr, \u003cspan citationid=\"CR128\" class=\"CitationRef\"\u003e1963\u003c/span\u003e; Bennet-Clark \u0026amp; Ewing, \u003cspan citationid=\"CR20\" class=\"CitationRef\"\u003e1970\u003c/span\u003e). However, it has been postulated that mate choice and specific recognition are part of a continuum and that sexual selection may also lead to reinforcement or RCD (Ryan \u0026amp; Rand, \u003cspan citationid=\"CR177\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Boake et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e1997\u003c/span\u003e; Liou \u0026amp; Price, \u003cspan citationid=\"CR114\" class=\"CitationRef\"\u003e1994\u003c/span\u003e; Mendelson \u0026amp; Shaw, \u003cspan citationid=\"CR131\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). In cases where the female is the selective sex, it is hypothesized that female choice that promotes isolation will result in the divergence of male sexual characters to avoid RI or hybridization (Butlin, \u003cspan citationid=\"CR32\" class=\"CitationRef\"\u003e1987\u003c/span\u003e; Gr\u0026ouml;ning \u0026amp; Hochkirch, \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Hoskin \u0026amp; Higgie, \u003cspan citationid=\"CR84\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). RCD has been reported for body size (Ding et al., \u003cspan citationid=\"CR49\" class=\"CitationRef\"\u003e2018\u003c/span\u003e; Sağlam et al., \u003cspan citationid=\"CR179\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), characters for grasping the female and genital characters (Kawano, \u003cspan citationid=\"CR96\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Kameda et al., \u003cspan citationid=\"CR94\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Anderson \u0026amp; Langerhans, \u003cspan citationid=\"CR9\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Kosuda et al., \u003cspan citationid=\"CR99\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Sağlam et al., \u003cspan citationid=\"CR179\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Nishimura et al., \u003cspan citationid=\"CR137\" class=\"CitationRef\"\u003e2022\u003c/span\u003e) or other types of characters (Marsteller et al., \u003cspan citationid=\"CR119\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Kawakami \u0026amp; Tatsuta, \u003cspan citationid=\"CR95\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Roth-Monz\u0026oacute;n et al., \u003cspan citationid=\"CR174\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). In many works where RCD is evaluated, one or a few characters linked to sexual reproduction are analyzed. However, phenotypic divergence can occur due to selective pressures along multiple phenotype axes simultaneously so that divergence can be multidimensional (Haines et al., \u003cspan citationid=\"CR73\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; White \u0026amp; Butlin, \u003cspan citationid=\"CR227\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Vega-S\u0026aacute;nchez et al., \u003cspan citationid=\"CR219\" class=\"CitationRef\"\u003e2022\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAnimal genitalia, especially in the male, can exhibit relatively complex morphologies and show fast and divergent evolutionary changes compared to other body parts (Tuxen, \u003cspan citationid=\"CR216\" class=\"CitationRef\"\u003e1970\u003c/span\u003e; Eberhard, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e1985\u003c/span\u003e; Leonard \u0026amp; C\u0026oacute;rdoba-Aguilar, \u003cspan citationid=\"CR112\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Sexual selection may play a key role in the evolution of genitalia (Eberhard, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e1985\u003c/span\u003e, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Hosken \u0026amp; Stockley, \u003cspan citationid=\"CR83\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Simmons, \u003cspan citationid=\"CR193\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). In turn, genital divergence can be explained by natural selection, as it contributes to reproductive isolation among species by promoting speciation (Eberhard, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e1985\u003c/span\u003e, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Masly, \u003cspan citationid=\"CR121\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Wojcieszek \u0026amp; Simmons, \u003cspan citationid=\"CR230\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; House et al., \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). Phenomena such as RCD may contribute to differences in genitalia between species in sympatric zones, whereby mechanical or interlocking incompatibilities between male and female genitalia may be frequent (Masly, \u003cspan citationid=\"CR121\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). The relative importance of natural and sexual selection in genitalia evolution continues under discussion (Jennions \u0026amp; Kelly, \u003cspan citationid=\"CR91\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Eberhard, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Simmons, \u003cspan citationid=\"CR193\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Brennan \u0026amp; Prum, \u003cspan citationid=\"CR30\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Eberhard \u0026amp; Lehmann, \u003cspan citationid=\"CR57\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Sloan \u0026amp; Simmons, \u003cspan citationid=\"CR196\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), although there is evidence that multiple selective pressures may be determinant in the morphological evolution of the genitalia (Langerhans et al., \u003cspan citationid=\"CR108\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Song \u0026amp; Wenzel, \u003cspan citationid=\"CR200\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; McPeek et al., \u003cspan citationid=\"CR130\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Simmons et al., \u003cspan citationid=\"CR194\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; House et al., \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Simmons, \u003cspan citationid=\"CR193\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Frazee \u0026amp; Masly, \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). This multiplicity of selective regimes can cause what is known as \"mosaic evolution\", where different portions of the same structure can respond in a mixed manner to concordant or antagonistic selective pressures (due to their multi-factorial nature) and where even shape and size of the same structure can diverge differentially (House \u0026amp; Simmons, \u003cspan citationid=\"CR81\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Song \u0026amp; Wenzel, \u003cspan citationid=\"CR200\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Werner \u0026amp; Simmons, \u003cspan citationid=\"CR222\" class=\"CitationRef\"\u003e2008\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eMorphological variation in non-genital contact characters used during pre-copulatory or copulatory mating can be explained by some of the natural or sexual selection hypotheses that may generate genital morphological diversity (Robson \u0026amp; Richards, \u003cspan citationid=\"CR169\" class=\"CitationRef\"\u003e1936\u003c/span\u003e; Eberhard \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e1985\u003c/span\u003e, \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2004\u003c/span\u003e, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). These characters also possess a pattern of rapid evolutionary divergence and generally have the function of grasping or grasping the female during mating by resembling functionally genital ''claspers'' (Eberhard, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e1985\u003c/span\u003e, \u003cspan citationid=\"CR56\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). The intra-specific phenotypic variation can be considered as the raw material on which selection acts (West-Eberhard, \u003cspan citationid=\"CR224\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Eberhard, \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2009\u003c/span\u003e), and the patterns of variation are helpful in understanding the evolution of different morphological characters. In general, sexually selected display traits show high within-species phenotypic variation (Cuervo \u0026amp; M\u0026oslash;ller, \u003cspan citationid=\"CR44\" class=\"CitationRef\"\u003e1999\u003c/span\u003e; Eberhard et al., \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Eberhard, \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). High values of coefficient of variation (CVs) indicate directional selective forces, while low values of CVs are associated with stabilizing selective pressures (Eberhard et al., \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Eberhard, \u003cspan citationid=\"CR55\" class=\"CitationRef\"\u003e2009\u003c/span\u003e).\u003c/p\u003e \u003cp\u003ePhenotypic plasticity refers to the ability of organisms of a species to change their morphology, behavior, or physiology in response to environmental variation (Stearns, \u003cspan citationid=\"CR205\" class=\"CitationRef\"\u003e1989\u003c/span\u003e; West-Eberhard, \u003cspan citationid=\"CR223\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Whitman \u0026amp; Agrawal, \u003cspan citationid=\"CR228\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). When characters express some degree of phenotypic plasticity, environmentally based phenotypic differences among species and populations can underlie the patterns of morphological variation (Jennions \u0026amp; Kelly, \u003cspan citationid=\"CR91\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Garnier et al., \u003cspan citationid=\"CR64\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Song \u0026amp; Wenzel, \u003cspan citationid=\"CR200\" class=\"CitationRef\"\u003e2008\u003c/span\u003e). The case of morphological divergence in environmental gradients deserves particular mention. In these cases, morphological differences between populations may be due to selective pressures for species differentiation and morphological changes linked to an environmental cline (Goldberg \u0026amp; Lande, \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Therefore, among the requirements for testing RCD, it is necessary to separate allopatric/sympatric context effects from other ecological effects. The environment can directly or indirectly influence genetic and phenotypic variation. Therefore, geographic variation among different populations is expected (Sota et al., \u003cspan citationid=\"CR203\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Kosuda et al., \u003cspan citationid=\"CR99\" class=\"CitationRef\"\u003e2016\u003c/span\u003e). Controlling for the effects of correlation between phenotype and environmental or geographic clines allows for finding patterns of divergence that might otherwise be undetectable (Goldberg \u0026amp; Lande, \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). Variation in latitude or altitude is mainly linked to changes in temperature, an abiotic factor that affects animal growth, causing a substantial impact on the observed phenotypic variation results (Bergmann, \u003cspan citationid=\"CR21\" class=\"CitationRef\"\u003e1847\u003c/span\u003e; Allen, \u003cspan citationid=\"CR8\" class=\"CitationRef\"\u003e1877\u003c/span\u003e; Rensch, \u003cspan citationid=\"CR165\" class=\"CitationRef\"\u003e1938\u003c/span\u003e; Atkinson, \u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1994\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eExamples of this RI exist in many animal and plant groups (e.g., Levin, \u003cspan citationid=\"CR113\" class=\"CitationRef\"\u003e1970\u003c/span\u003e; Armbruster \u0026amp; Herzig, \u003cspan citationid=\"CR11\" class=\"CitationRef\"\u003e1984\u003c/span\u003e; Hettyey \u0026amp; Pearman, \u003cspan citationid=\"CR76\" class=\"CitationRef\"\u003e2003\u003c/span\u003e; Dame \u0026amp; Petren, \u003cspan citationid=\"CR45\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Gr\u0026ouml;ning \u0026amp; Hochkirch, \u003cspan citationid=\"CR71\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Matsumoto et al., \u003cspan citationid=\"CR122\" class=\"CitationRef\"\u003e2010\u003c/span\u003e), and among them, arthropods have been shown to provide interesting models for studying this phenomenon (Shuker \u0026amp; Burdfield-Steel, \u003cspan citationid=\"CR192\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Although some cases of ecological character displacement have been described in insects and arachnids, there are fewer examples of RCD in these taxa due to the difficulty of empirically evidencing this process (Waage, \u003cspan citationid=\"CR221\" class=\"CitationRef\"\u003e1979\u003c/span\u003e). However, in arthropods, evidence of RCD was found in pre-copulatory characters used during courtship (Marshall \u0026amp; Cooley, \u003cspan citationid=\"CR118\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Jang \u0026amp; Gerhardt, \u003cspan citationid=\"CR90\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Kronforst et al., \u003cspan citationid=\"CR101\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Dyer et al., \u003cspan citationid=\"CR51\" class=\"CitationRef\"\u003e2014\u003c/span\u003e; Rundle \u0026amp; Dyer, \u003cspan citationid=\"CR176\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Yukilevich, \u003cspan citationid=\"CR233\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) and there are also examples of RCD in genital characters (Kawano, \u003cspan citationid=\"CR96\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Kawakami \u0026amp; Tatsuta, \u003cspan citationid=\"CR95\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Kosuda et al., \u003cspan citationid=\"CR99\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Nishimura et al., \u003cspan citationid=\"CR137\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). In arachnids, there are some suggestions that RCD might be occurring between species in sympatry (Barth, \u003cspan citationid=\"CR16\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; Stratton, \u003cspan citationid=\"CR208\" class=\"CitationRef\"\u003e1997\u003c/span\u003e; Agnarsson et al., \u003cspan citationid=\"CR7\" class=\"CitationRef\"\u003e2016\u003c/span\u003e; Muster \u0026amp; Michalik, \u003cspan citationid=\"CR135\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), as is the case of genital characters between \u003cem\u003eParatrechalea\u003c/em\u003e spider species with RI (Costa-Schmidt \u0026amp; de Ara\u0026uacute;jo, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2010\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe study of phenotype variation and its causes may be complicated because adaptation can be viewed as a multivariate process acting on sets of characters (Lande \u0026amp; Arnold, \u003cspan citationid=\"CR107\" class=\"CitationRef\"\u003e1983\u003c/span\u003e; Schluter \u0026amp; Nychka, \u003cspan citationid=\"CR185\" class=\"CitationRef\"\u003e1994\u003c/span\u003e; Blows, \u003cspan citationid=\"CR24\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Organisms can be interpreted as composite objects, with characters not necessarily independent of each other that respond in complex and different magnitudes and directions to different selective pressures (Klingenberg, \u003cspan citationid=\"CR97\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Geometric morphometrics (GM) helps address the inherent complexity of characters separating their size and shape to evaluate the effect of selective pressures on these two dimensions of the phenotype (Bookstein, 1998; Adams et al., \u003cspan citationid=\"CR5\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Zelditch et al., \u003cspan citationid=\"CR234\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). Indeed, shape metrics are better descriptors of genital morphology diversity, containing more information than size measures (Slice, \u003cspan citationid=\"CR195\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Shen et al., \u003cspan citationid=\"CR191\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). These type of studies are ideally performed in species where the function of the characters to be assessed is well-known. Arachnids have proven to be exceptional models, although morphological quantification techniques have generally been applied mainly in systematic or ecomorphological studies (Costa-Schmidt \u0026amp; de Ara\u0026uacute;jo, \u003cspan citationid=\"CR38\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Kallal et al., \u003cspan citationid=\"CR93\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Santib\u0026aacute;\u0026ntilde;ez-L\u0026oacute;pez et al., \u003cspan citationid=\"CR181\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Wilson et al., \u003cspan citationid=\"CR229\" class=\"CitationRef\"\u003e2021\u003c/span\u003e; Bellvert et al., \u003cspan citationid=\"CR19\" class=\"CitationRef\"\u003e2022\u003c/span\u003e). Some studies have demonstrated the usefulness of these techniques in addressing sexual dimorphism (Fern\u0026aacute;ndez-Montraveta et al., 2017; Kallal et al., \u003cspan citationid=\"CR93\" class=\"CitationRef\"\u003e2019\u003c/span\u003e), as well as the combination with other approaches such as the analysis of phenotypic variation (by the coefficient of variation -CV) of certain characters in some arachnids (Eberhard et al., \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Peretti et al., \u003cspan citationid=\"CR153\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Calbacho-Rosa et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Lai et al., \u003cspan citationid=\"CR106\" class=\"CitationRef\"\u003e2021\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eAlthough studies applying fine morphological quantification methodologies in scorpions are scarce, these organisms appear to be excellent models for this type of analysis (Bechara \u0026amp; Liria, \u003cspan citationid=\"CR18\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Santib\u0026aacute;\u0026ntilde;ez-L\u0026oacute;pez et al., \u003cspan citationid=\"CR180\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). It is known that different selective pressures act on specific scorpion characters (e.g., pedipalps, pectines, chelicerae) as demonstrated in studies that have evaluated their CV, allometric patterns, or where selection pressures behind dimorphic characters have been explored (Peretti et al., \u003cspan citationid=\"CR153\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Carrera et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Fox et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Santib\u0026aacute;\u0026ntilde;ez-L\u0026oacute;pez et al., \u003cspan citationid=\"CR180\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Visser \u0026amp; Geerts, \u003cspan citationid=\"CR220\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Furthermore, during an elaborate courtship, both sexes displayed unique characters with functional roles such as stimulation or increased female receptivity with non-genital contact structures (e.g., the caudal gland in \u0026lsquo;rubbing with telson\u0026rsquo;, the sting in \u0026lsquo;sexual sting\u0026rsquo;) or grasping characters to overcome female resistance (e.g., apophyses in pedipalps, chelicerae) (Polis \u0026amp; Sissom, \u003cspan citationid=\"CR158\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; Carrera et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Peretti, 2001). In particular, these characters were extensively studied in the family Bothriuridae in the evolutionary framework of sexual selection (Peretti et al., \u003cspan citationid=\"CR153\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Carrera et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Olivero et al., \u003cspan citationid=\"CR141\" class=\"CitationRef\"\u003e2014\u003c/span\u003e, \u003cspan citationid=\"CR143\" class=\"CitationRef\"\u003e2019\u003c/span\u003e; Peretti, \u003cspan citationid=\"CR151\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Lastly, scorpions present indirect sperm transfer via a sclerotized spermatophore deposited in the substrate (Weygoldt, \u003cspan citationid=\"CR225\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; Proctor, \u003cspan citationid=\"CR162\" class=\"CitationRef\"\u003e1998\u003c/span\u003e). This spermatophore is regenerated each time the male mates from two chitinous halves (i.e., hemispermatophores) produced in internal glandular structures called paraxial organs (Polis \u0026amp; Sissom, \u003cspan citationid=\"CR158\" class=\"CitationRef\"\u003e1990\u003c/span\u003e). These genital characters are incredibly complex and can be divided into subunits offering interesting opportunities for studying the evolution of genitalia (Peretti et al., \u003cspan citationid=\"CR153\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Peretti, \u003cspan citationid=\"CR150\" class=\"CitationRef\"\u003e2003\u003c/span\u003e, \u003cspan citationid=\"CR151\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Mattoni et al., \u003cspan citationid=\"CR124\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Monod et al., \u003cspan citationid=\"CR134\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). For example, some characters follow a distinctive pattern of characters under sexual selection pressures (i.e., evolve rapidly and divergently), while others show only minor variations coinciding with what is predicted for characters under natural selection, such as structures with mechanical constraints or with key reproductive functions such as sperm passage (Peretti, \u003cspan citationid=\"CR151\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Mattoni et al., \u003cspan citationid=\"CR124\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). The morphological diversity of sexual characters and spermatophores of scorpions responds to diverse (and not mutually exclusive) evolutionary hypotheses (Peretti, \u003cspan citationid=\"CR151\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). These mixed patterns result from complex synergistic or antagonistic interactions between different selective pressures (Peretti, \u003cspan citationid=\"CR151\" class=\"CitationRef\"\u003e2010\u003c/span\u003e), so this genital structure could be found under mosaic evolution. This offers great possibilities for the morpho-functional study of diverse characters and contexts and allows different outcomes in a scenario of RCD.\u003c/p\u003e \u003cp\u003eThere are several records of interspecific mating in scorpions (Auber, \u003cspan citationid=\"CR15\" class=\"CitationRef\"\u003e1963\u003c/span\u003e; Matthiesen, \u003cspan citationid=\"CR123\" class=\"CitationRef\"\u003e1968\u003c/span\u003e; Probst, \u003cspan citationid=\"CR161\" class=\"CitationRef\"\u003e1972\u003c/span\u003e; Le Pape \u0026amp; Goyffon, \u003cspan citationid=\"CR110\" class=\"CitationRef\"\u003e1975\u003c/span\u003e; Peretti, \u003cspan citationid=\"CR148\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Peretti et al., \u003cspan citationid=\"CR152\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). Although many scorpions use pheromones for sex encounter, males are vagrant in scenarios of indirect competition for females, and there are records of overlapping species distributions and coexistence of species, phenomena such as RI or RCD between closely related species have not yet been extensively assessed. Here, we explored the occurrence of RCD in two closely related scorpion species of the genus \u003cem\u003eUrophonius\u003c/em\u003e Pocock, 1893 (\u003cem\u003eU. brachycentrus\u003c/em\u003e and \u003cem\u003eU. achalensis\u003c/em\u003e, Bothriuridae) (Ojanguren-Affilastro et al., \u003cspan citationid=\"CR139\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) that have partially sympatric ranges with overlapping reproductive seasons and share the same habitat requirements and life-history traits (Maury, \u003cspan citationid=\"CR126\" class=\"CitationRef\"\u003e1969\u003c/span\u003e; Acosta, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1985\u003c/span\u003e; Ojanguren-Affilastro et al., \u003cspan citationid=\"CR139\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). These scorpions have winter habits and adaptations for this lifestyle, which is rather peculiar among scorpions (Ojanguren-Affilastro et al., \u003cspan citationid=\"CR139\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Garcia et al., \u003cspan citationid=\"CR63\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). These species do not possess specific recognition through chemical signals, which, together with a promiscuous mating system with scramble competition, leads to an asymmetric RI scenario with heterospecific mating (Oviedo-Diego et al., \u003cspan citationid=\"CR146\" class=\"CitationRef\"\u003e2020\u003c/span\u003e, \u003cspan citationid=\"CR147\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). The coexistence of these species raises the question of whether there are morphological, reproductive barriers, that may hinder or prevent the culmination of heterospecific mating, given the costs they may entail in terms of gamete loss, female plugging (Oviedo-Diego et al., \u003cspan citationid=\"CR145\" class=\"CitationRef\"\u003e2019\u003c/span\u003e, \u003cspan citationid=\"CR146\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Romero-Lebr\u0026oacute;n et al., \u003cspan citationid=\"CR173\" class=\"CitationRef\"\u003e2019\u003c/span\u003e) or potential hybridization. For these reasons, we evaluated the existence of RCD in the shape and size of somatic characters used in courtship (non-genital contact characters) and genital characters of hemispermatophores to observe whether these metrics responded concordantly or follow a mosaic pattern under specific recognition and sexual selection pressures. Additionally, we determined the phenotypic variation by analyzing the coefficient of variation of these characters in contexts of sympatry and allopatry of both species to complement the analysis of the selective regimes that could explain the morphological variability in these species. Complementarily, we consider the influence of environmental and geographic factors on the morphological patterns found. The results from multiple lines of evidence account for the inherent complexity of sexual characters in scorpions and provide clues about the possible selective pressures behind their evolution.\u003c/p\u003e"},{"header":"2. Material Amd Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003e2.1.1 Study Species and Sampling\u003c/h2\u003e \u003cp\u003e \u003cem\u003eUrophonius brachycentrus\u003c/em\u003e has a wide geographic range distributed throughout central Argentina, while \u003cem\u003eU. achalensis\u003c/em\u003e is endemic to the mountainous regions of C\u0026oacute;rdoba in central Argentina (Acosta, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1985\u003c/span\u003e; Ojanguren-Affilastro, 2020). The two species share partially sympatric distribution areas in the Sierras Grandes that are part of the Sierras Pampeanas Centrales (Acosta, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1985\u003c/span\u003e), part of a fundamental orographic system of extra-Andean mountain formations in Argentina, were formed in the Lower Paleozoic (about 300 and 350\u0026nbsp;million years ago). Adult scorpions of \u003cem\u003eU. achalensis\u003c/em\u003e and \u003cem\u003eU. brachycentrus\u003c/em\u003e were collected during the day during the mating season (May-August) (Acosta, \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1985\u003c/span\u003e; Maury, \u003cspan citationid=\"CR126\" class=\"CitationRef\"\u003e1969\u003c/span\u003e; Ojanguren-Affilastro et al., \u003cspan citationid=\"CR139\" class=\"CitationRef\"\u003e2020\u003c/span\u003e) for three consecutive years (2018, 2019, 2020) by turning over rocks. We collected individuals in two allopatric populations of \u003cem\u003eU. brachcyentrus\u003c/em\u003e (31\u0026deg;22'42.4\"S 64\u0026deg;35'34.0\"W, 876 m.a.s.l..; 31\u0026ordm;31'46.3''S 64\u0026ordm;51'52.7\"W, 996 m.a.s.l.), two allopatric populations of \u003cem\u003eU. achalensis\u003c/em\u003e (31\u0026deg;35'49.1\"S 64\u0026deg;44'49.3\"W, 2030 m.a.s.l., 31\u0026deg;21'17.3\"S 64\u0026deg;48'21.3\"W, 1927 m.a.s.l.), and in two sympatric populations (31\u0026deg;23'13.5\"S 64\u0026deg;46'10.2\"W, 1796 m.a.s.l.; 31\u0026deg;34'07.6\"S 64\u0026deg;42'43.8\"W, 1610 m.a.s.l.).\u003c/p\u003e \u003cdiv id=\"Sec4\" class=\"Section3\"\u003e \u003ch2\u003e2.1.2 Processing of individuals and selected characters\u003c/h2\u003e \u003cp\u003eIndividuals from field collections were identified and sexed (Ojanguren-Affilastro, \u003cspan citationid=\"CR138\" class=\"CitationRef\"\u003e2005\u003c/span\u003e) with a Nikon SMZ 1500 stereo zoom microscope and preserved in 80% EtOH in glass containers for morphological studies. Classical and geometric morphometric studies were carried out, and measurements of characters were compared between sexes and study species in different contexts (sympatry vs. allopatry) (n\u0026thinsp;=\u0026thinsp;25 per population context and per sex of each species) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). We selected characters under both natural and sexual selection pressures, used during feeding, defense, and courtship traits such as pedipalps, chelicerae and telson vesicle (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Also, we analyzed characters used only in a sexual context, such as characters for female stimulation (caudal gland) or characters for grasping the female pedipalps during courtship (pedipalp apophyses) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). Finally, we measured genital characters involved in sperm transfer that has also been shown to be under sexual selection pressures (Olivero et al., \u003cspan citationid=\"CR142\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Peretti, \u003cspan citationid=\"CR151\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). To analyze the selected characters, individuals were dissected, and internal structures were extracted with fine tweezers for photographic treatment. The individuals were measured using images taken under the stereo zoom microscope with a digital coupled camera (Nikon Digital Sight DS-FI1-U2). Because the internal female genitalia consist of flexible structures that vary in size and shape according to the female reproductive status (Peretti, \u003cspan citationid=\"CR151\" class=\"CitationRef\"\u003e2010\u003c/span\u003e), morphometric analysis was not performed. In subsequent analyses, individuals and characters with damaged or incomplete portions were not considered.\u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab1\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 1\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eMorphological characters selected in \u003cem\u003eUrophonius\u003c/em\u003e species analyzed. The type of character (somatic or genital), the corresponding sex, the functional role, and the measurement technique used are indicated. Abbreviations: AL, absolute length; RL, relative length; NS, Natural selection; SS, Sexual selection. See landmark positions in Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e and descriptions in Table S1.\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"6\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003eMorphological character\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c3\" namest=\"c2\"\u003e \u003cp\u003eSex and type of character\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003en\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003eMethodology\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eFunctional role\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eProsome\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"7\" rowspan=\"8\"\u003e \u003cp\u003eSomatic in both sexes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"5\" rowspan=\"6\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e♂ n\u0026thinsp;=\u0026thinsp;122\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGeometric morphometry (Landmarks\u0026thinsp;=\u0026thinsp;8)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eBody size indicator (Polis \u0026amp; Sissom \u003cspan citationid=\"CR158\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; McLean et al., \u003cspan citationid=\"CR129\" class=\"CitationRef\"\u003e2018\u003c/span\u003e).\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e♀ n\u0026thinsp;=\u0026thinsp;112\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eChelicera\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e♂ n\u0026thinsp;=\u0026thinsp;113\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eClassic morphometry (AL, RL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eCharacter used during feeding and courtship where the pair touch and rub chelicerae during \u0026lsquo;chelicera massage\u0026rsquo; or \u0026lsquo;kiss\u0026rsquo; (under NS and SS pressures) (Carrera et al., \u003cspan citationid=\"CR35\" class=\"CitationRef\"\u003e2009\u003c/span\u003e).\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e♀ n\u0026thinsp;=\u0026thinsp;114\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePectine\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e♂ n\u0026thinsp;=\u0026thinsp;126\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eClassic morphometry (AL, RL)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eCharacter used for mechano-chemical-sensory recognition, foraing, mate searching and spermatophore deposition site in courtship (under NS and SS -slight- pressures) (Polis \u0026amp; Sissom, \u003cspan citationid=\"CR158\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; Peretti et al., \u003cspan citationid=\"CR153\" class=\"CitationRef\"\u003e2001\u003c/span\u003e)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e♀ n\u0026thinsp;=\u0026thinsp;100\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePedipalp\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"2\" rowspan=\"3\"\u003e \u003cp\u003eGrasping characters\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e♂ n\u0026thinsp;=\u0026thinsp;128\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGeometric morphometry (Landmarks\u0026thinsp;=\u0026thinsp;5\u0026thinsp;+\u0026thinsp;Semilandmarks\u0026thinsp;=\u0026thinsp;21)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eCharacter used during defense, feeding and grasping of the other sex during courtship (under NS and SS pressures) (Polis \u0026amp; Sissom \u003cspan citationid=\"CR158\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; Peretti et al., \u003cspan citationid=\"CR153\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Olivero et al., \u003cspan citationid=\"CR141\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e♀ n\u0026thinsp;=\u0026thinsp;121\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGeometric morphometry (Landmarks\u0026thinsp;=\u0026thinsp;4\u0026thinsp;+\u0026thinsp;Semilandmarks\u0026thinsp;=\u0026thinsp;21)\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePedipalp apophysis\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSomatic in males\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e♂ n\u0026thinsp;=\u0026thinsp;122\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGeometric morphometry (Landmarks\u0026thinsp;=\u0026thinsp;5\u0026thinsp;+\u0026thinsp;Semilandmarks\u0026thinsp;=\u0026thinsp;16)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCharacter used for the correct grasping and locking of pedipalps during courtship (only under SS pressure) (Peretti et al., \u003cspan citationid=\"CR153\" class=\"CitationRef\"\u003e2001\u003c/span\u003e).\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTelson vesicle\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eSomatic in both sexes\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e\u0026nbsp;\u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e♂ n\u0026thinsp;=\u0026thinsp;122\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGeometric morphometry (EFA\u0026thinsp;=\u0026thinsp;8 harmonic)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCharacter used during feeding and agonistic interactions, during courtship in sexual stinging of the female and gland rubbing (under NS and SS pressures) (Polis \u0026amp; Sissom, \u003cspan citationid=\"CR158\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; Peretti, \u003cspan citationid=\"CR148\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Fox et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Sentensk\u0026aacute; et al., \u003cspan citationid=\"CR188\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Olivero et al., 2017, \u003cspan citationid=\"CR143\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e♀ n\u0026thinsp;=\u0026thinsp;122\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eCharacter used during feeding and agonistic interactions, sometimes during courtship movements indicative of receptivity (under NS and SS pressures) (Polis \u0026amp; Sissom \u003cspan citationid=\"CR158\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; Fox et al., \u003cspan citationid=\"CR61\" class=\"CitationRef\"\u003e2015\u003c/span\u003e).\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCaudal gland\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSomatic in males\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003eStimulation character\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e♂ n\u0026thinsp;=\u0026thinsp;122\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGeometric morphometry (EFA\u0026thinsp;=\u0026thinsp;7 harmonic)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eExternal secretory gland on the dorsal side of the telson used during courtship where the male rubs the female to increase female receptivity (under SS pressures) (De la Serna de Esteban, \u003cspan citationid=\"CR47\" class=\"CitationRef\"\u003e1978\u003c/span\u003e; Peretti, \u003cspan citationid=\"CR149\" class=\"CitationRef\"\u003e1997\u003c/span\u003e; Olivero et al, 2017, \u003cspan citationid=\"CR143\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHemispermatophore lamella\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGenital in male\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eGenital character\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e♂ n\u0026thinsp;=\u0026thinsp;117\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGeometric morphometry (Landmarks\u0026thinsp;=\u0026thinsp;4\u0026thinsp;+\u0026thinsp;Semilandmarks\u0026thinsp;=\u0026thinsp;24)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eGenital character that will form the spermatophore involved in the copulatory mechanics for indirect sperm transfer, acting as a lever for sperm release (under NS and SS pressures) (Peretti et al., \u003cspan citationid=\"CR153\" class=\"CitationRef\"\u003e2001\u003c/span\u003e).\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHemispermatophore capsular lobe\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e♂ n\u0026thinsp;=\u0026thinsp;108\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003eGeometric morphometry (EFA\u0026thinsp;=\u0026thinsp;6 harmonic)\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003eGenital character that will form the copulatory cone of the spermatophore that enters and evert inside the female genitalia, guiding the sperm during sperm transfer (under NS and SS pressures) (Peretti et al., \u003cspan citationid=\"CR153\" class=\"CitationRef\"\u003e2001\u003c/span\u003e; Olivero et al., \u003cspan citationid=\"CR141\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003cp\u003e \u003c/p\u003e \u003cp\u003e \u003cdiv class=\"gridtable\"\u003e\u003ctable float=\"Yes\" id=\"Tab2\" border=\"1\"\u003e \u003ccaption language=\"En\"\u003e \u003cdiv class=\"CaptionNumber\"\u003eTable 2\u003c/div\u003e \u003cdiv class=\"CaptionContent\"\u003e \u003cp\u003eCoefficients of variation (CVs) of multiple somatic and genital characters of male and female \u003cem\u003eUrophonius achalensis\u003c/em\u003e and \u003cem\u003eU. brachycentrus\u003c/em\u003e scorpions from sympatric and allopatric areas. Morphological character, sex, CVs value and statistical significance value (between species and contexts) are indicated (p-values\u0026thinsp;\u0026lt;\u0026thinsp;0.05 indicated in bold). ♂: males, ♀: females. Letters indicate significant differences between character CVs (p-values\u0026thinsp;\u0026lt;\u0026thinsp;0.05)\u003c/p\u003e \u003c/div\u003e \u003c/caption\u003e \u003ccolgroup cols=\"9\"\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c1\" colnum=\"1\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c2\" colnum=\"2\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c3\" colnum=\"3\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c4\" colnum=\"4\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c5\" colnum=\"5\"\u003e\u003c/div\u003e \u003cdiv align=\"left\" class=\"colspec\" colname=\"c6\" colnum=\"6\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c7\" colnum=\"7\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c8\" colnum=\"8\"\u003e\u003c/div\u003e \u003cdiv align=\"char\" char=\".\" class=\"colspec\" colname=\"c9\" colnum=\"9\"\u003e\u003c/div\u003e \u003cthead\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e\u0026nbsp;\u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003eSpecies\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c4\" namest=\"c3\"\u003e \u003cp\u003e\u003cem\u003eU. achalensis\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c6\" namest=\"c5\"\u003e \u003cp\u003e\u003cem\u003eU. brachycentrus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eDifferences between spp.\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colspan=\"2\" nameend=\"c9\" namest=\"c8\"\u003e \u003cp\u003eDifferences between contexts\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\"\u003e \u003cp\u003e\u003cb\u003eMorphological character\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003eSex/Context\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003esympatry\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003eallopatry\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003esympatry\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003eallopatry\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e\u003cem\u003eU. achalensis\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e\u003cem\u003eU. brachycentrus\u003c/em\u003e\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eChelicerae length\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e♀\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.525\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.098\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.266\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.037\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.605\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.735\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.379\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e♂\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.220\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.605\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.09\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.831\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.134\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.169\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.354\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePecten length\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e♀\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e7.263\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.141\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.824\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6.955\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.150\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.195\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.251\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e♂\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e5.363\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e6.962\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e6.984\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6.872\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e0.323\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.241\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.092\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003ePedipalp length\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e♀\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.591\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e3.248\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.092\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6.155\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.025\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.119\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.324\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003ctr\u003e \u003cth align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e♂\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.581\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.439\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.416\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.643\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.010\u003c/b\u003e\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c8\"\u003e \u003cp\u003e0.274\u003c/p\u003e \u003c/th\u003e \u003cth align=\"left\" colname=\"c9\"\u003e \u003cp\u003e0.506\u003c/p\u003e \u003c/th\u003e \u003c/tr\u003e \u003c/thead\u003e \u003ctbody\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003ePedipalp apophysis length\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e♂\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10.741\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e11.267\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e12.329\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e16.227\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e\u003cb\u003e0.018\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.301\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0.191\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\" morerows=\"1\" rowspan=\"2\"\u003e \u003cp\u003eTelson vesicle length\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e♀\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.472\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e4.832\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.223\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e4.785\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.444\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.145\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0.585\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e♂\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.471\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.315\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e5.413\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e6.331\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.215\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.371\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0.169\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eCaudal gland length\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e♂\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e10.359\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e12.653\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e16.111\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e13.020\u003csup\u003ea\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.249\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.320\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0.307\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHemispermatophore lamella length\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e♂\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e3.982\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.111\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.756\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.245\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.484\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.306\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0.692\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHemispermatophore capsu\u003cspan type=\"Underline\" class=\"Underline\" name=\"Emphasis\"\u003el\u003c/span\u003ear lobe length\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e♂\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e4.447\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e5.925\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e4.951\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e5.451\u003csup\u003eb\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.677\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.285\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0.730\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003ctr\u003e \u003ctd align=\"left\" colname=\"c1\"\u003e \u003cp\u003eHemispermatophore frontal crest length\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c2\"\u003e \u003cp\u003e\u003cb\u003e♂\u003c/b\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c3\"\u003e \u003cp\u003e8.466\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c4\"\u003e \u003cp\u003e8.539\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c5\"\u003e \u003cp\u003e7.749\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"left\" colname=\"c6\"\u003e \u003cp\u003e7.842\u003csup\u003eab\u003c/sup\u003e\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c7\"\u003e \u003cp\u003e0.786\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c8\"\u003e \u003cp\u003e0.970\u003c/p\u003e \u003c/td\u003e \u003ctd align=\"char\" char=\".\" colname=\"c9\"\u003e \u003cp\u003e0.547\u003c/p\u003e \u003c/td\u003e \u003c/tr\u003e \u003c/tbody\u003e \u003c/colgroup\u003e \u003c/table\u003e\u003c/div\u003e \u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec5\" class=\"Section3\"\u003e \u003ch2\u003e2.1.3 Morphometric studies\u003c/h2\u003e \u003cdiv id=\"Sec6\" class=\"Section4\"\u003e \u003ch2\u003e2.1.3.1 Classic morphometric and coefficient of variation analysis\u003c/h2\u003e \u003cp\u003eThe chelicerae and the pectines were analyzed by linear measurements (due to methodological difficulties in applying geometrical morphometry) by analyzing absolute and relative lengths (prosome length as body size index - McLean et al., \u003cspan citationid=\"CR129\" class=\"CitationRef\"\u003e2018\u003c/span\u003e) (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). These measurements were taken from photographs obtained for each character with ImageJ software tools (Schneider et al., \u003cspan citationid=\"CR186\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Measurements were taken three times by the same person, and the measurement error was calculated (Sokal \u0026amp; Rohlf, \u003cspan citationid=\"CR199\" class=\"CitationRef\"\u003e1995\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe coefficient of variation (CV) is widely used as indirect evidence to know the selective pressures that might be operating on morphological characters (Pomiankowski \u0026amp; M\u0026oslash;ller, \u003cspan citationid=\"CR159\" class=\"CitationRef\"\u003e1995\u003c/span\u003e; Eberhard et al., \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Peretti et al., \u003cspan citationid=\"CR153\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). We compared this parameter across different types of characters in males and females from both contexts (sympatry \u003cem\u003eversus\u003c/em\u003e allopatry). We used the modified formula: CV\u0026rsquo; = [(sd\u003csub\u003ey\u003c/sub\u003e/mean\u003csub\u003ey\u003c/sub\u003e) * (1 \u0026ndash; r\u003csup\u003e2\u003c/sup\u003e)\u003csup\u003e1/2\u003c/sup\u003e * 100], where sd\u003csub\u003ey\u003c/sub\u003e is the standard deviation of the character, mean\u003csub\u003ey\u003c/sub\u003e is the arithmetic mean of the character, r\u003csup\u003e2\u003c/sup\u003e is the determination coefficient between the character and a measure of body size (prosoma length) and * indicates multiplication symbol (Eberhard et al., \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Calbacho-Rosa et al., \u003cspan citationid=\"CR34\" class=\"CitationRef\"\u003e2019\u003c/span\u003e). CVs were statistically compared using the 'asymptotic_test' function of the \u003cem\u003ecvequality\u003c/em\u003e package (Marwick \u0026amp; Krishnamoorthy, \u003cspan citationid=\"CR120\" class=\"CitationRef\"\u003e2019\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec7\" class=\"Section4\"\u003e \u003ch2\u003e2.1.3.2 Geometric morphometric analysis\u003c/h2\u003e \u003cp\u003eWe took digital images of selected characters in male and female scorpions with a scale close to the character, and the images were assembled with TPSutil software (Rohlf, \u003cspan citationid=\"CR171\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Sets of anatomical Landmarks (Bookstein, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e1991\u003c/span\u003e) and semilandmarks were established using TPSDig2 (Rohlf, \u003cspan citationid=\"CR170\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Bookstein, \u003cspan citationid=\"CR28\" class=\"CitationRef\"\u003e1997\u003c/span\u003e; Gunz \u0026amp; Mitteroecker, \u003cspan citationid=\"CR72\" class=\"CitationRef\"\u003e2013\u003c/span\u003e). We used landmarks in the prosome, the hemispermatophore lamella, the pedipalp, and the apophysis of this structure (Table\u0026nbsp;\u003cspan refid=\"Tab1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e, Table S1). Sliding landmarks or semilandmarks were used to enhance geometric information about curvatures between adjacent landmarks in the pedipalp, the pedipalp apophysis, and the hemispermatophore lamella (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e). In other characters (hemispermatophore capsular lobe, telson vesicle and caudal gland) we quantified shape using an elliptic Fourier analysis (EFA) (following Santib\u0026aacute;\u0026ntilde;ez-L\u0026oacute;pez et al., \u003cspan citationid=\"CR180\" class=\"CitationRef\"\u003e2017\u003c/span\u003e, \u003cspan citationid=\"CR181\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) that allowed us to explore small differences in defined shapes from contour characterization (Kuhl \u0026amp; Giardina, \u003cspan citationid=\"CR103\" class=\"CitationRef\"\u003e1982\u003c/span\u003e; Ferson et al., \u003cspan citationid=\"CR60\" class=\"CitationRef\"\u003e1985\u003c/span\u003e; Hammer \u0026amp; Harper, \u003cspan citationid=\"CR74\" class=\"CitationRef\"\u003e2006\u003c/span\u003e) (Fig.\u0026nbsp;\u003cspan refid=\"Fig1\" class=\"InternalRef\"\u003e1\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe shape coordinates of each character were subjected to a Generalized Procrustes Analysis (Gower, \u003cspan citationid=\"CR68\" class=\"CitationRef\"\u003e1975\u003c/span\u003e) with the 'gpagen' function of the \u003cem\u003egeomorph\u003c/em\u003e package (Schlager, \u003cspan citationid=\"CR183\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Adams et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2017\u003c/span\u003e) in R software (R Core Team, \u003cspan citationid=\"CR164\" class=\"CitationRef\"\u003e2021\u003c/span\u003e) to remove non-shape variables (translation, rotation, size) from the dataset to compare shape by contrasting with a mean generated from a consensus matrix (Rohlf \u0026amp; Slice, \u003cspan citationid=\"CR172\" class=\"CitationRef\"\u003e1990\u003c/span\u003e; Adams et al., \u003cspan citationid=\"CR6\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The size proxy of each character was retained from the GPA analysis (i.e., Centroid size) for subsequent analyses (Bookstein, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e1991\u003c/span\u003e; Zelditch et al., \u003cspan citationid=\"CR234\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). To account for semilandmarks in the GPA calculation, we used the 'slider2d' function of the \u003cem\u003eMorpho\u003c/em\u003e package (Schlager et al., \u003cspan citationid=\"CR184\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). EFA was performed using the \u003cem\u003emomocs\u003c/em\u003e package (Iwata \u0026amp; Ukai, \u003cspan citationid=\"CR89\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Bonhomme et al., \u003cspan citationid=\"CR26\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eA Principal Component Analysis (PCA) was performed to visualize and explore the general trends of the distribution of total morphological variation in morphospace from both the data yielded by the GPA as well as the data obtained from the EFA using the 'plotTangentSpace' function of the \u003cem\u003egeomorph\u003c/em\u003e package. Principal components can be considered as reorganized and uncorrelated morphological features representing different aspects of the total shape variation. Additionally, vectors that reflected shape variation along x/y axes were used to visualize magnitudes and overall shape changes with the \u003cem\u003egeomorph\u003c/em\u003e package (Bookstein, \u003cspan citationid=\"CR27\" class=\"CitationRef\"\u003e1991\u003c/span\u003e). Multivariate analysis of variance (MANOVA) was performed with the function 'procD.lm' of the \u003cem\u003egeomorph\u003c/em\u003e package with resampling permutations procedure to calculate the significance of shape variables. The variation in shape of the first two principal components (since they captured more than 70% of the morphological variation) was analyzed in detail. First, we checked the allometric component (influence of size on shape) of the characters with the functions 'procD.lm' and 'plotAllometry' of the \u003cem\u003egeomorph\u003c/em\u003e package. If we found allometry in the sample, we calculated residual values of the shape variables for subsequent analyses (Outomuro \u0026amp; Johansson, \u003cspan citationid=\"CR144\" class=\"CitationRef\"\u003e2017\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section4\"\u003e \u003ch2\u003e2.1.3.3 Statistical analysis\u003c/h2\u003e \u003cp\u003eMeasurements obtained by classical and geometric morphometry were compared between species and contexts (sympatry \u003cem\u003eversus\u003c/em\u003e allopatry) with linear mixed models (LMMs) in R. Separate models were performed for each character and sex (because in some characters the number of Landmarks was not equal for males and females) where we set as response variables the linear measurements, size variables (centroid size) or shape variables (PCs scores) and the fixed effects were species (levels: \u003cem\u003eU. achalensis\u003c/em\u003e / \u003cem\u003eU. brachycentrus\u003c/em\u003e) and contexts (levels: sympatry / allopatry). The interaction between these explanatory variables was evaluated to corroborate RCD patterns. We added populations of origin as random effects to account for the variability contributed to this factor. Due to the influence of altitude on morphological variability, we added the altitude where individuals were collected as another random effect. Analyses were performed with the package \u003cem\u003elme4\u003c/em\u003e (Bates et al., \u003cspan citationid=\"CR17\" class=\"CitationRef\"\u003e2011\u003c/span\u003e) and \u003cem\u003elsmeans\u003c/em\u003e (Lenth, \u003cspan citationid=\"CR111\" class=\"CitationRef\"\u003e2016\u003c/span\u003e) for a posteriori test (with Bonferroni correction) if necessary. Model validation was assessed graphically and by residual analysis.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e \u003cdiv id=\"Sec9\" class=\"Section3\"\u003e \u003ch2\u003e2.1.4 Influence of environmental factors on morphological characters\u003c/h2\u003e \u003cp\u003eComplementarily, in a subset of data, we explored whether environmental factors might correlate with any of the phenotypic characters measured; because, for example, the clinal or geographic variation present in our study system may be influencing the patterns found (Goldberg \u0026amp; Lande, \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). As altitude may be strongly associated with temperature and humidity, we considered the variation of these environmental factors in our analysis by obtaining the mean annual temperature and mean annual rainfall rasters from Geoportal IDESA (\u003cspan class=\"ExternalRef\"\u003e\u003cspan class=\"RefSource\"\u003ehttp://geoportal.idesa.gob.ar/\u003c/span\u003e\u003cspan address=\"http://geoportal.idesa.gob.ar/\" targettype=\"URL\" class=\"RefTarget\"\u003e\u003c/span\u003e\u003c/span\u003e, data from last year available: 2017). With the QGIS program 3.26 (QGIS Development Team, 2020), we mapped the distribution of the collected individuals (using the geo-referenced latitude and longitude data for each individual). We used the 'extractRandomClim' function of the \u003cem\u003eraster\u003c/em\u003e package (Hijmans et al., \u003cspan citationid=\"CR77\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) in R to extract the mean annual temperature and mean annual rainfall values for each collection point. Subsequently, we explored the relationships between these environmental factors with size (centroid size, absolute length) and shape (PCs scores) previously calculated (see \u003cem\u003e2.1.3.2\u003c/em\u003e) with linear mixed models (LMMs). We acknowledge that other environmental factors (e.g., soil characteristics, atmospheric pressure, food availability) may sustain some of the phenotypic variation among species and populations. Still, the scoring of these factors was beyond the scope of this study, so our estimates of environmental effects on phenotype are prospective.\u003c/p\u003e \u003c/div\u003e \u003c/div\u003e"},{"header":"3. Results","content":"\u003cdiv class=\"Section2\" id=\"Sec11\"\u003e\n \u003ch2\u003e3.1.1 Morphological variation across contexts\u003c/h2\u003e\n \u003cp\u003eWe compared multiple sexual characters involved in courtship and sperm transfer in males\u0026apos; and females\u0026apos; scorpions from sympatric and allopatric contexts. We observed different patterns of phenotypic variation in different directions (convergences and divergences) in each species (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e), and the shape and size appear to respond independently to different selective pressures. The morphometric results for each character analyzed in both sexes are detailed below, first evaluating the size and then the variation in shape.\u003c/p\u003e\n \u003cdiv class=\"Section3\" id=\"Sec12\"\u003e\n \u003ch2\u003e3.1.1.1 Chelicerae and pecten: asymmetric convergence in size only in females\u003c/h2\u003e\n \u003cp\u003eWe observed an asymmetric convergence in the absolute length of both chelicerae (\u0026chi;2\u0026thinsp;=\u0026thinsp;34.180, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and pectines (\u0026chi;2\u0026thinsp;=\u0026thinsp;45.894, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) in females (\u003cem\u003eU. brachycentrus\u003c/em\u003e more similar to \u003cem\u003eU. achalensis\u003c/em\u003e in sympatry) (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). Neither contexts nor species showed differences in the relative lengths of chelicerae or pectines. We only found interspecific differences in the relative cheliceral length in males, with \u003cem\u003eU. brachycentrus\u003c/em\u003e males having larger chelicerae (\u0026chi;2\u0026thinsp;=\u0026thinsp;64.348, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). However, all the other variables did not differ between species or contexts.\u003c/p\u003e\n \u003cdiv class=\"Section4\" id=\"Sec13\"\u003e\n \u003ch2\u003e3.1.1.2 Prosome and telson vesicle: size convergence\u003c/h2\u003e\n \u003cp\u003eCentroid size of the prosome showed symmetric convergence in females of both scorpion species, with species becoming more similar in sympatry than in allopatry (\u0026chi;2\u0026thinsp;=\u0026thinsp;26.907, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and asymmetric convergence in males (\u003cem\u003eU. brachycentrus\u003c/em\u003e more similar in sympatry than in allopatry) (\u0026chi;2\u0026thinsp;=\u0026thinsp;8.507, p\u0026thinsp;=\u0026thinsp;0.004) (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). In terms of shape, the Procrustes MANOVA showed no significant variation according to species and context. PC1 comprised almost half of the morphological variation (Females: 46.49%, Males: 45.85%), showing interspecific differences (\u003cem\u003eU. brachycentrus\u003c/em\u003e more compressed prosome than \u003cem\u003eU. achalensis\u003c/em\u003e) (Females: \u0026chi;2\u0026thinsp;=\u0026thinsp;31.992, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001; Males: \u0026chi;2\u0026thinsp;=\u0026thinsp;19.895, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). PC2 explained an 18.44% of the variation in females and 13.82% in males and showed no differences between species or contexts in either sex. PC3 accounted for the 13.37% of the variability in females without differences between species or contexts. In contrast, PC3 in males representing the 12.52% of morphological variability was different between species (\u0026chi;2\u0026thinsp;=\u0026thinsp;9.783, p\u0026thinsp;=\u0026thinsp;0.002) and contexts (\u0026chi;2\u0026thinsp;=\u0026thinsp;6.827, p\u0026thinsp;=\u0026thinsp;0.006) but we found no significant interaction between these factors.\u003c/p\u003e\n \u003cp\u003eRegarding the telson vesicle, in females, we found a pattern of symmetric convergence in the centroid size with both species becoming more similar in sympatry than in allopatry (\u0026chi;2\u0026thinsp;=\u0026thinsp;32.176, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). In males the convergence was asymmetric, as only males of \u003cem\u003eU. brachycentrus\u003c/em\u003e presented a shift in the size of this character towards sympatry (\u0026chi;2\u0026thinsp;=\u0026thinsp;6.118, p\u0026thinsp;=\u0026thinsp;0.013). The Procrustes MANOVA showed significant shape variation according to species in both sexes (Females: F\u0026thinsp;=\u0026thinsp;4.269, p\u0026thinsp;=\u0026thinsp;0.001; Males: F\u0026thinsp;=\u0026thinsp;4.404, p\u0026thinsp;=\u0026thinsp;0.001), but the interaction between species and context was not significant (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). In females, we found significant differences between species in telson vesicle shape reflected in PC1 (54%) (Females: \u0026chi;2\u0026thinsp;=\u0026thinsp;22.441, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) and PC2 (19.57%) (Females: \u0026chi;2\u0026thinsp;=\u0026thinsp;21.034, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). Also, in males, PC1 (67.48%) showed differences between species (\u0026chi;2\u0026thinsp;=\u0026thinsp;36.965, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e), while in PC2 (12.21%) there were no significant differences between species or contexts.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv class=\"Section4\" id=\"Sec14\"\u003e\n \u003ch2\u003e3.1.1.3 Pedipalp in females: asymmetric convergence in size and divergence in shape\u003c/h2\u003e\n \u003cp\u003eWe found asymmetric convergence in pedipalp centroid size, with species more similar in sympatry than in allopatry due to a shift of \u003cem\u003eU. brachycentrus\u003c/em\u003e (\u0026chi;2\u0026thinsp;=\u0026thinsp;19.812, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eA). In terms of shape, the Procrustes MANOVA showed significant variation according to species and context (F\u0026thinsp;=\u0026thinsp;7.788, p\u0026thinsp;=\u0026thinsp;0.001). PC1 explained 38.10% of morphological variability, and we found asymmetric divergence in PC1, with \u003cem\u003eU. brachycentrus\u003c/em\u003e females showing a shift relative to sympatric \u003cem\u003eU. achalensis\u003c/em\u003e females and allopatric females (\u0026chi;2\u0026thinsp;=\u0026thinsp;8.294, p\u0026thinsp;=\u0026thinsp;0.004) (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eB). PC2 explained 26.95% and PC3 10.60% of morphological variation although these shape variables showed no significant differences between species or contexts.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv class=\"Section4\" id=\"Sec15\"\u003e\n \u003ch2\u003e3.1.1.4 Pedipalp and apophysis in males: asymmetric divergence in shape\u003c/h2\u003e\n \u003cp\u003eMale pedipalp size showed only interspecific differences, with larger pedipalp and apophysis in \u003cem\u003eU. achalensis\u003c/em\u003e than \u003cem\u003eU. brachycentrus\u003c/em\u003e (\u0026chi;2\u0026thinsp;=\u0026thinsp;84.839, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eA). The Procrustes MANOVA showed significant variation by species and context (F\u0026thinsp;=\u0026thinsp;3.321, p\u0026thinsp;=\u0026thinsp;0.006). Regarding the pedipalp, the PC1 explained 45.25% of the morphological variability, and we found a pattern of asymmetric divergence in PC1 (\u003cem\u003eU. brachycentrus\u003c/em\u003e males with higher pedipalp and shorter fixed fingers than allopatric males and sympatric \u003cem\u003eU. achalensis\u003c/em\u003e males) (\u0026chi;2\u0026thinsp;=\u0026thinsp;10.069, p\u0026thinsp;=\u0026thinsp;0.002) (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eB, D-E). PC2 accounted for 20.21% and PC3 a 9.99% of the variability, and this component showed no differences between species or contexts (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eD).\u003c/p\u003e\n \u003cp\u003eFor the pedipalp apophysis size, we found interspecific differences (\u0026chi;2\u0026thinsp;=\u0026thinsp;38.651, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), with apophysis of \u003cem\u003eU. achalensis\u003c/em\u003e being larger than those of \u003cem\u003eU. brachycentrus\u003c/em\u003e (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eC). The Procrustes MANOVA showed significant variation in the interaction between species and context (F\u0026thinsp;=\u0026thinsp;3.419, p\u0026thinsp;=\u0026thinsp;0.014). PC1 (accounting for 31.11% of the variation) showed no significant differences between species or contexts. In contrast, PC2 explaining 21.07% of the morphological variation, showed significant differences between species in sympatry, and not in allopatry (\u0026chi;2\u0026thinsp;=\u0026thinsp;10.221, p\u0026thinsp;=\u0026thinsp;0.002) (Fig. \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003eC, E). Moreover, the shape of the apophysis was different between sympatric and allopatric populations of \u003cem\u003eU. brachycentrus\u003c/em\u003e so that this displacement pattern would be an asymmetric divergence. Morphological variability was also distributed between PC3 (9.34%) and PC4 (8.56%), although these morphological variables did not vary between contexts and only between species in PC4 (\u0026chi;2\u0026thinsp;=\u0026thinsp;8.685, p\u0026thinsp;=\u0026thinsp;0.003).\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv class=\"Section4\" id=\"Sec16\"\u003e\n \u003ch2\u003e3.1.1.5 Caudal gland: asymmetrical convergence in size\u003c/h2\u003e\n \u003cp\u003eCaudal gland size showed a pattern of asymmetric convergence, with \u003cem\u003eU. brachycentrus\u003c/em\u003e males more similar to \u003cem\u003eU. achalensis\u003c/em\u003e males in sympatry and differing significantly from allopatric population males (with smaller gland) (\u0026chi;2\u0026thinsp;=\u0026thinsp;10.087, p\u0026thinsp;=\u0026thinsp;0.002) (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). The Procrustes MANOVA showed significant variation only according to species (F\u0026thinsp;=\u0026thinsp;155.064, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), but the interaction between species and context was not significant. Regarding shape, PC1 almost completely comprised all morphological variability (92.81%), and we only found significant interspecific differences (\u003cem\u003eU. brachycentrus\u003c/em\u003e showing a more compressed and wider caudal gland than \u003cem\u003eU. achalensis\u003c/em\u003e) (\u0026chi;2\u0026thinsp;=\u0026thinsp;155.774, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001). PC2, with an explanation of only 2.86% of the morphological variation, did not differ between species or contexts.\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv class=\"Section4\" id=\"Sec17\"\u003e\n \u003ch2\u003e3.1.1.6 Hemispermatophore lamella: asymmetrical divergence in shape\u003c/h2\u003e\n \u003cp\u003eHemispermatophore lamella size varied only at the interspecific level (\u0026chi;2\u0026thinsp;=\u0026thinsp;86.714, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001), with lamella of \u003cem\u003eU. achalensis\u003c/em\u003e males always being larger than those of \u003cem\u003eU. brachycentrus\u003c/em\u003e (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e, \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003eA). In terms of shape, the Procrustes MANOVA showed significant variation according to species and context (F\u0026thinsp;=\u0026thinsp;3.223, p\u0026thinsp;=\u0026thinsp;0.006). Almost half of the lamella morphological variation was represented by PC1 (43.41%) (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003eB-C). This shape showed asymmetric divergence, as \u003cem\u003eU. brachycentrus\u003c/em\u003e males differed from their allopatric conspecifics with a wider lamella, also differing from sympatric \u003cem\u003eU. achalensis\u003c/em\u003e males (\u0026chi;2\u0026thinsp;=\u0026thinsp;6.791, p\u0026thinsp;=\u0026thinsp;0.009) (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003eC-D). PC2 comprised 15.33% and the PC3 14.02% of the morphological variation but these shape variables showed no differences between species or contexts (Fig. \u003cspan class=\"InternalRef\"\u003e4\u003c/span\u003eC).\u003c/p\u003e\n \u003c/div\u003e\n \u003cdiv class=\"Section4\" id=\"Sec18\"\u003e\n \u003ch2\u003e3.1.1.7 Hemispermatophore capsular lobes: asymmetrical divergence in size\u003c/h2\u003e\n \u003cp\u003eWe found a pattern of asymmetric divergence in the hemispermatophore capsular lobe size, with males of \u003cem\u003eU. brachycentrus\u003c/em\u003e in sympatry having larger lobes than the rest of the male groups (\u0026chi;2\u0026thinsp;=\u0026thinsp;12.784, p\u0026thinsp;\u0026lt;\u0026thinsp;0.001) (Fig. \u003cspan class=\"InternalRef\"\u003e2\u003c/span\u003e). We found no significant interaction between species and context in the Procrustes MANOVA, but there was variation in shape according to species (F\u0026thinsp;=\u0026thinsp;4.847, p\u0026thinsp;=\u0026thinsp;0.001). PC1 explained 31.96% and PC3 16.19% of the morphological variance, and none of the shape variables resulted in different between species or contexts. PC2 accounted for the 25.52% and differed between contexts (\u0026chi;2\u0026thinsp;=\u0026thinsp;3.926, p\u0026thinsp;=\u0026thinsp;0.048) and marginally between species (\u0026chi;2\u0026thinsp;=\u0026thinsp;3.319, p\u0026thinsp;=\u0026thinsp;0.068), but the interaction between context and species was not significant.\u003c/p\u003e\n \u003c/div\u003e\n \u003c/div\u003e\n \u003cdiv class=\"Section3\" id=\"Sec19\"\u003e\n \u003ch2\u003e3.1.2 Coefficients of variation of morphological characters\u003c/h2\u003e\n \u003cp\u003eWe found different values of CVs according to the type of character analyzed and sex (Table \u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e). The chelicerae, the pecten, the pedipalp, and the telson vesicle showed relatively low CVs values in both sexes and species, with no statistical differences in CVs between these characters, between species or between contexts. Only the pedipalp\u0026rsquo; CVs differ between species, higher in \u003cem\u003eU. brachycentrus\u003c/em\u003e than in \u003cem\u003eU. achalensis\u003c/em\u003e in both sexes. In contrast, other male characters used exclusively during sexual interactions, such as the caudal gland and the pedipalp apophysis, showed high CVs, significantly different from the previously mentioned characters. In the case of the pedipalp apophysis for grasping during courtship, we found higher variation values in \u003cem\u003eU. brachycentrus\u003c/em\u003e than in \u003cem\u003eU. achalensis\u003c/em\u003e. Genital characters such as the length of the hemispermatophore lamella or the hemispermatophore capsular lobe showed low values of CVs with no differences between species or contexts. The only exception was the frontal crest of the hemispermatophore, which showed high CVs values compared to other genital characters in both species.\u003c/p\u003e\u0026nbsp;\u003ctable border=\"1\" id=\"Tab4\"\u003e\n \u003ccaption language=\"En\"\u003e\n \u003cdiv class=\"CaptionNumber\"\u003eTable 3\u003c/div\u003e\n \u003cdiv class=\"CaptionContent\"\u003e\n \u003cp\u003eInfluence of environmental factors on multiple somatic and genital characters of male and female \u003cem\u003eUrophonius achalensis\u003c/em\u003e and \u003cem\u003eU. brachycentrus\u003c/em\u003e scorpions from sympatric and allopatric areas. Character and compared parameter, sex, statistic value and statistical significance value are indicated (values\u0026thinsp;\u0026lt;\u0026thinsp;0.05 indicated in bold). Abbreviations: AL, absolute length; cs, centroid size; hum, humidity (rainfall); hum:sp, interaction term between humidity and species fixed effect; PC, principal component 1\u0026ndash;2; temp, temperature fixed effect; temp:sp, interaction between temperature and species fixed effects, ♂: males, ♀: females\u003c/p\u003e\n \u003c/div\u003e\n \u003c/caption\u003e\n \u003cthead\u003e\n \u003ctr\u003e\n \u003cth align=\"left\" colspan=\"2\"\u003e\n \u003cp\u003eMorphological character\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSex\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eFixed effect\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ep-value\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eSex\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eFixed effect\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003eF\u003c/p\u003e\n \u003c/th\u003e\n \u003cth align=\"left\"\u003e\n \u003cp\u003ep-value\u003c/p\u003e\n \u003c/th\u003e\n \u003c/tr\u003e\n \u003c/thead\u003e\n \u003ctbody\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"7\"\u003e\n \u003cp\u003eProsome\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp:sp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.102\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp:sp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e68.449\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt;\u0026thinsp;0.005\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.053\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.819\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.324\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.072\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.123\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.727\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.589\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.445\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.826\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.366\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.165\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.686\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.207\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.651\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.424\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.022\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.002\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.969\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.021\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.885\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.977\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.326\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.929\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.051\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.437\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.122\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.231\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.632\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"8\"\u003e\n \u003cp\u003ePedipalp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp:sp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.129\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.026\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp:sp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.876\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.004\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.58\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.212\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.715\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.103\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.885\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.174\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.205\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.652\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.004\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.953\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.015\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.904\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.416\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.521\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.505\u003c/p\u003e\n \u003c/td\u003e\n 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align=\"left\"\u003e\n \u003cp\u003ePC2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.802\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.098\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.987\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.323\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n 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\u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp:sp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e12.904\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.001\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp:sp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e15.457\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0002\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n 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\u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp:sp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e7.361\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.009\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp:sp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e21.884\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e\u0026lt;\u0026thinsp;0.005\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003eAL\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n 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\u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp:sp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.957\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.029\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp:sp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.371\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.005\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.134\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.717\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.783\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.185\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.787\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.099\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.897\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.348\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.264\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.609\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.614\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.109\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.017\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.896\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.476\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.492\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.159\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.146\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♀\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.753\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.389\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"10\"\u003e\n \u003cp\u003ePedipalp apophysis\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.197\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.659\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\" colspan=\"4\" rowspan=\"32\"\u003e\u0026nbsp;\u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.325\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.570\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.026\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.314\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.136\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.713\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.812\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.373\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.019\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.888\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.748\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.101\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.796\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.184\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.188\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.666\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC4\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.107\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.298\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"6\"\u003e\n \u003cp\u003eCaudal gland\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp:sp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e8.485\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.003\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.329\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.569\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.068\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.154\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.447\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.504\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum:sp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e5.400\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.023\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.764\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.385\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"8\"\u003e\n \u003cp\u003eHemispermatophore Lamella\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp:sp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e13.602\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.0004\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.648\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.108\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.392\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.073\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.144\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.147\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.934\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.168\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.015\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.902\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.929\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.341\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.159\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.691\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\" rowspan=\"8\"\u003e\n \u003cp\u003eHemispermatophore capsular lobe\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp:sp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e4.152\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e0.046\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e2.526\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.117\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.005\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.945\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003etemp\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e1.642\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.205\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ecs\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.725\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.398\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC1\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.112\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.739\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC2\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e3.025\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.087\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003ctr\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ePC3\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e\u003cstrong\u003e♂\u003c/strong\u003e\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003ehum\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.087\u003c/p\u003e\n \u003c/td\u003e\n \u003ctd align=\"left\"\u003e\n \u003cp\u003e0.769\u003c/p\u003e\n \u003c/td\u003e\n \u003c/tr\u003e\n \u003c/tbody\u003e\n \u003c/table\u003e\n \u003c/div\u003e\n \u003cdiv class=\"Section3\" id=\"Sec20\"\u003e\n \u003ch2\u003e3.1.3 Influence of environmental factors on morphological characters\u003c/h2\u003e\n \u003cp\u003eWe found that the size (centroid size and absolute length) of almost all the characters analyzed varied with temperature (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e). We found a significant statistical interaction between temperature and species in all cases, so temperature-dependent morphological variations were observed only in \u003cem\u003eU\u003c/em\u003e. \u003cem\u003ebrachycentrus\u003c/em\u003e, with no relationship in \u003cem\u003eU. achalensis\u003c/em\u003e. Generally, both sexes of this species had larger characters in colder areas (at higher altitudes) and smaller characters in warmer areas (at lower altitudes). This was observed for both sexes\u0026apos; prosome, pedipalp, chelicerae, pecten, and telson vesicle. In males, we also found this same pattern of variation in \u003cem\u003eU. brachycentrus\u003c/em\u003e for the caudal gland and genital characters, although we did not observe it in the pedipalp apophysis. The pattern of variation found in the size of many characters coincides with the convergence asymmetrical in \u003cem\u003eU. brachycentrus\u003c/em\u003e. The shape of none of the analyzed structures showed variation with temperature.\u003c/p\u003e\n \u003cp\u003eAs for humidity (rainfall), we found patterns of morphological variation of some characters regarding this environmental factor (Table\u0026nbsp;\u003cspan class=\"InternalRef\"\u003e3\u003c/span\u003e). We observed that females of both species presented a larger prosoma in more humid areas. In addition, we found an interaction between humidity and species for caudal gland shape (PC1). That is, in \u003cem\u003eU. brachycentrus\u003c/em\u003e, males presented a gland with negative PC1 values in more humid areas. This morphological change is associated with more slender and less rounded gland. The shape of no other character was affected by humidity.\u003c/p\u003e\n \u003c/div\u003e\n\u003c/div\u003e"},{"header":"4. Discussion","content":"\u003cp\u003eWe found great morphological variability between sympatric and allopatric contexts in the studied model species of scorpions. Our study revealed main novel insights about the evolution of shape and size of somatic and genital characters in an animal model so far understudied but with great potential for further research. We were able to observe complex patterns of phenotypic variation in different directions (convergences and divergences) in size and shape, which allows us to suggest a possible mosaic evolution in certain sexual characters in these scorpion species. The integration of the results allows us to infer an asymmetric RCD in the shape of certain sexual characters of both sexes key for courtship success (i.e., grasping characters) and sperm transfer (i.e., genital characters of the hemispermatophore). Intriguingly, although we found low phenotypic variation in some genital characters, others showed high variation which could reflect that some characters are under antagonistic selective pressures. The convergence patterns found in the size of many characters were due to environmental fluctuations linked to the altitude cline of the geographic system. In the following discussion, we analyze in depth the remarkable patterns of phenotypic variation, the possible selection pressures underlying this variability, and the consequences of the RCD in the mating system and coexistence for these scorpion species.\u003c/p\u003e \u003cdiv id=\"Sec22\" class=\"Section2\"\u003e \u003ch2\u003e4.1 Reproductive character displacement in pedipalps\u003c/h2\u003e \u003cp\u003eWe obtained evidence of phenotypic divergence in shape and size of multiple somatic characters used in courtship in \u003cem\u003eU. brachycentrus\u003c/em\u003e, while \u003cem\u003eU. achalensis\u003c/em\u003e showed no divergence in any character between sympatric and allopatric populations. \u003cem\u003eUrophonius brachycentrus\u003c/em\u003e males of the sympatric zone differed from their conspecifics and \u003cem\u003eU. achalensis\u003c/em\u003e males by having more globose pedipalps and apophyses with a lower crest deeper. \u003cem\u003eU. brachycentrus\u003c/em\u003e females showed an RCD pattern also in the shape of their pedipalps, with the pedipalps being more globose in sympatric zones. Therefore, the pattern of divergence in pedipalp shape was complementary in males and females. RCD results in mechanical incompatibilities (due to mechanisms under natural selection such as the ''lock-and-key'' hypothesis) that can hinder the culmination of heterospecific matings, promoting reproductive isolation (Eberhard, \u003cspan citationid=\"CR54\" class=\"CitationRef\"\u003e2004\u003c/span\u003e). We know that there is incompatibility at the behavioral level since, in heterospecific mating, females show more resistance events (Oviedo-Diego, M. pers. obs.), which sometimes causes the pedipalps of both sexes to be released, interrupting the mating. Multiple biomechanical variables may be involved in these events, such as the pedipalp muscles and grip strength, as well as probably the optimal fit given by the morphology of the apophysis. Analyzing these variables together could help better understand the determinants of ''pedipalp grasping'' success in scorpions (van der Meijden et al., \u003cspan citationid=\"CR218\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) and its relationship to species-specificity in heterospecific matings. Peretti et al. (\u003cspan citationid=\"CR152\" class=\"CitationRef\"\u003e2000\u003c/span\u003e) report that some of the intercrosses between \u003cem\u003eBothriurus flavidus\u003c/em\u003e and \u003cem\u003eB. prospicus\u003c/em\u003e from areas of sympatry could progress to courtship but not to complete matings, but it is unclear which factors lead to mating interruption. Interestingly, it was noted that in intercrosses between \u003cem\u003eB. cordubensis\u003c/em\u003e and \u003cem\u003eB. noa\u003c/em\u003e, some matings were interrupted by female resistance events (Peretti et al., \u003cspan citationid=\"CR152\" class=\"CitationRef\"\u003e2000\u003c/span\u003e). Although the latter species are allopatric, likely, mechanical incompatibilities in pedipalp grasping are also occurring in this pair of species.\u003c/p\u003e \u003cp\u003eThe pedipalp apophysis is a key character for the correct attachment of the pedipalps during the mating dance (Ábalos \u0026amp; Hominal, \u003cspan citationid=\"CR1\" class=\"CitationRef\"\u003e1974\u003c/span\u003e; Maury, \u003cspan citationid=\"CR125\" class=\"CitationRef\"\u003e1968\u003c/span\u003e; Peretti, \u003cspan citationid=\"CR148\" class=\"CitationRef\"\u003e1993\u003c/span\u003e), although little is known about the mechanics of the coupling and adjustment with the female pedipalps (Peretti, \u003cspan citationid=\"CR148\" class=\"CitationRef\"\u003e1993\u003c/span\u003e). The morphospace of this character was complex, and although its summary into a few dimensions allowed us to simplify this complexity, we believe that future studies should be carried out to complete the understanding of the selective forces underlying the evolution of this character. According to the hypotheses of morphological evolution, RCD could be expected under the hypotheses of structural or sensory \"lock-and-key\" natural selection. Under these hypotheses, species-specific morphological complementarity could exist with similar divergence in both sexes (Eberhard, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e1985\u003c/span\u003e; Arnqvist, \u003cspan citationid=\"CR12\" class=\"CitationRef\"\u003e1997\u003c/span\u003e), so it is necessary to include the morphological variability of female characters in future studies. At the same time, the male pedipalp apophysis showed a high coefficient of variation indicative of sexual selection pressures (directional selection) and not consistent with characters that are under stabilizing selection according to the \"lock-and-key\" hypothesis (Peretti et al., \u003cspan citationid=\"CR153\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). This could suggest that the pedipalp apophysis in \u003cem\u003eU. brachycentrus\u003c/em\u003e is under different and maybe opposite selective pressures. On the one hand, the shape of the apophysis (which could be summarized as apophysis depth) showed divergence in sympatry to ensure mechanical isolation. Still, on the other hand, the size of the apophysis seems to be influenced by sexual selection pressures.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec23\" class=\"Section2\"\u003e \u003ch2\u003e4.2 Reproductive character displacement in male genital characters\u003c/h2\u003e \u003cp\u003eAs we observed in some somatic characters, we found evidence of RCD in characteristics of the hemispermatophores of \u003cem\u003eU. brachycentrus\u003c/em\u003e. In addition, we found that these characters had low CVs, which would support some type of stabilizing selection on these characters (Peretti et al., \u003cspan citationid=\"CR153\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). Males of this species showed hemispermatophores with a more compressed lamella and larger capsular lobes than allopatric males and sympatric \u003cem\u003eU. achalensis\u003c/em\u003e males. The size of larger capsular lobules in \u003cem\u003eU. brachycentrus\u003c/em\u003e could be partly explained by the increase in size of females of this species towards the sympatric zone, as morphological complementarity is expected for mechanical isolation by the ''lock-and-key''. Although there are examples of these hypothesis in arthropods (Mikkola, \u003cspan citationid=\"CR132\" class=\"CitationRef\"\u003e1992\u003c/span\u003e, \u003cspan citationid=\"CR133\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Sota \u0026amp; Kubota, \u003cspan citationid=\"CR202\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Usami et al., \u003cspan citationid=\"CR217\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Nagata et al., \u003cspan citationid=\"CR136\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Takami et al., \u003cspan citationid=\"CR212\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Tanabe \u0026amp; Sota, \u003cspan citationid=\"CR213\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Sota \u0026amp; Tanabe, \u003cspan citationid=\"CR204\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Wojcieszek, \u0026amp; Simmons, \u003cspan citationid=\"CR230\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Kubota et al., \u003cspan citationid=\"CR102\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Nishimura et al., \u003cspan citationid=\"CR137\" class=\"CitationRef\"\u003e2022\u003c/span\u003e), it is a hypothesis that has been discarded in several species as in general, genitalia diverge much more in males than in females, and it is not so common to find morphological complementarity (Eberhard, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e1985\u003c/span\u003e; Shapiro \u0026amp; Porter, \u003cspan citationid=\"CR190\" class=\"CitationRef\"\u003e1989\u003c/span\u003e; Masly, \u003cspan citationid=\"CR121\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Like to our conclusion for the pedipalp apophysis, it would be necessary to evaluate the female component to confirm this hypothesis in these species. However, although there may be rather cryptic differences, the female genital atrium is flexible and has a relatively ''uniform'' structure (Peretti, \u003cspan citationid=\"CR150\" class=\"CitationRef\"\u003e2003\u003c/span\u003e, \u003cspan citationid=\"CR151\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Therefore, the female genitalia in these species does not mechanistically prevent the entry of heterospecific male genitalia, which would also not support the ''lock-and-key'' hypothesis.\u003c/p\u003e \u003cp\u003eSome particular zones of the hemispermatophore (i.e., frontal crest area) had a very high phenotypic variation, suggesting that their variability is not so much restricted, which is not consistent with a stabilizing selection (Eberhard et al., \u003cspan citationid=\"CR53\" class=\"CitationRef\"\u003e1998\u003c/span\u003e; Peretti et al., \u003cspan citationid=\"CR153\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). These results would indicate that the morphological variation of at least some areas of the genitalia of these species would be explained rather by sexual selection hypotheses (Peretti, \u003cspan citationid=\"CR150\" class=\"CitationRef\"\u003e2003\u003c/span\u003e, \u003cspan citationid=\"CR151\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Monod et al., \u003cspan citationid=\"CR134\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). The frontal crest of the lamella fits into the inter-coxal space of the female, and there could be a 'passive' choice by 'mechanical adjustment' (Eberhard, \u003cspan citationid=\"CR52\" class=\"CitationRef\"\u003e1985\u003c/span\u003e; Huber \u0026amp; Eberhard, \u003cspan citationid=\"CR88\" class=\"CitationRef\"\u003e1997\u003c/span\u003e). Also, the capsular lobes possess micro-ornamentations contacting the female genital atrium wall that could have a stimulatory role, which could be contemplated in a female cryptic choice hypothesis (Peretti, \u003cspan citationid=\"CR150\" class=\"CitationRef\"\u003e2003\u003c/span\u003e, \u003cspan citationid=\"CR151\" class=\"CitationRef\"\u003e2010\u003c/span\u003e). Larger capsular lobes could be related to a larger contact surface of ornamentations with the female genital atrium and, consequently, a greater stimulation that could be linked to cryptic female choice.\u003c/p\u003e \u003cp\u003eAn inevitable question at this point is: if some portions of the genitalia are under sexual selection pressures, why does RCD exist in others? An interesting option could be that the female may bias, by cryptic choice, the use of sperm or other variables (e.g., hardening of the genital plug) according to characteristics evaluated in the interaction of the genitalia, such as (a) greater stimulation by larger capsular lobes (would explain the RCD in the hemispermatophores capsule lobe), (b) by mechanical adjustment of coxae of the first pair of legs with hemispermatophore frontal crest (would explain the RCD in the lamella of hemispermatophore) or fit between male capsular lobe and female genital atrium. We could say that there would be a \"combination\" of the sensory/mechanical \"lock-and-key\" hypothesis, where females can recognize the species-specificity of the male genitalia (and thus RCD would be promoted) but where physiological changes would not occur immediately but at the post-copulatory level mediated by female cryptic choice processes.\u003c/p\u003e \u003cp\u003eThis interaction between sexual and natural selection hypotheses could be expected to explain the evolution of genitalia in these species, where there is intense competition between males at the intra and interspecific level and promiscuity in their mating and where females must not only exercise mate choice at the pre-copulatory level, but copulatory and post-copulatory mechanisms seem to be necessary to avoid hybridization. A similar example seems to occur in hybridizing \u003cem\u003eDrosophila\u003c/em\u003e species, where the male genitalia differ in size and shape, and the external female genitalia shows no interspecific differences (Coyne, \u003cspan citationid=\"CR41\" class=\"CitationRef\"\u003e1983\u003c/span\u003e). In interspecific mating, the intrusion of the male genitalia differentially contacts the female genitalia so that females can store and use sperm according to the specific identity of the male (Price et al., \u003cspan citationid=\"CR160\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). This is called \"cryptic reproductive isolation\" and maybe a by-product of multiple evolutionary forces acting at the intra- and interspecific level (Price et al., \u003cspan citationid=\"CR160\" class=\"CitationRef\"\u003e2001\u003c/span\u003e). As we have emphasized above, it is now recognized that mate choice and specific recognition are part of a continuum and that the forces of sexual and natural selection may interact in multiple ways explaining patterns of sexual diversification across species (Ryan \u0026amp; Rand, \u003cspan citationid=\"CR177\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Liou \u0026amp; Price, \u003cspan citationid=\"CR114\" class=\"CitationRef\"\u003e1994\u003c/span\u003e; Boake et al., \u003cspan citationid=\"CR25\" class=\"CitationRef\"\u003e1997\u003c/span\u003e; Mendelson \u0026amp; Shaw, \u003cspan citationid=\"CR131\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Keeping these interactions in mind is critical for analyzing possible hypotheses of genital evolution (Simmons, \u003cspan citationid=\"CR193\" class=\"CitationRef\"\u003e2014\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe reinforcement model postulates the emergence of successive reproductive isolation barriers if these become ineffective (Howard, \u003cspan citationid=\"CR85\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Coyne \u0026amp; Orr, \u003cspan citationid=\"CR42\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Butlin \u0026amp; Smadja, \u003cspan citationid=\"CR33\" class=\"CitationRef\"\u003e2018\u003c/span\u003e). If mechanical isolation existed in the past leading to an RCD pattern but subsequently the effectiveness of this barrier, weakened pre-copulatory barriers may have been generated (such as RCD in pedipalps and behavioral incompatibilities), and the RCD in the genitalia may have persisted rather than reverted to the previous morphological scenario. This, in turn, could have resulted in the genitalia (or some of its parts) being able to diversify under other pressures more \"freely.\" This argument is supported by the fact that in this system, there is a pre-copulatory filter in heterospecific matings, and only 10 to 20% of these reach sperm transfer (Oviedo-Diego, M. per obs). Perhaps the existence of a percentage of matings that reach this point is sufficient for the maintenance of the RCD observed in genital characters.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec24\" class=\"Section2\"\u003e \u003ch2\u003e4.3 Environmental variations promote size convergence of multiple characters\u003c/h2\u003e \u003cp\u003eThe overall size, but no shape, of individuals converged in sympatry, i.e., individuals were more similar in size when the species were together, and this pattern was particularly strong for \u003cem\u003eU. brachycentrus\u003c/em\u003e. This could be observed in males and females for the prosome and the telson vesicle. Convergence was also observed in females for cheliceral size and in males for caudal gland size and hemispermatophore lamella. The patterns of convergence found in size could follow the rule of Atkinson (\u003cspan citationid=\"CR13\" class=\"CitationRef\"\u003e1994\u003c/span\u003e, \u003cspan citationid=\"CR14\" class=\"CitationRef\"\u003e1995\u003c/span\u003e) that predicts larger body sizes at lower temperatures (Horne et al., \u003cspan citationid=\"CR82\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Most ectotherms grow more slowly and mature with larger body sizes in colder environments (Angilletta et al., 2004). This increase in size may be adaptive when it allows for increased fecundity or higher survival or reproductive rates (Stearns, \u003cspan citationid=\"CR206\" class=\"CitationRef\"\u003e1992\u003c/span\u003e). In scorpions, it is known that different numbers of molts or the period between molts can affect the final body size of individuals (Sarmento et al., \u003cspan citationid=\"CR182\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Seiter et al., \u003cspan citationid=\"CR187\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), so through this mechanism, they could reach different sizes depending on environmental characteristics such as temperature or humidity, as well as variations in diet (Sarmento et al., \u003cspan citationid=\"CR182\" class=\"CitationRef\"\u003e2008\u003c/span\u003e).\u003c/p\u003e \u003cp\u003eThe effect of altitude and temperature change was probably more drastic in \u003cem\u003eU. brachycentrus\u003c/em\u003e due to the large altitudinal and temperature difference between the allopatric and sympatric populations compared. Temperature is predicted to affect the body size of individuals of both sexes similarly (Hirst et al., \u003cspan citationid=\"CR78\" class=\"CitationRef\"\u003e2015\u003c/span\u003e), and in \u003cem\u003eU. brachycentrus\u003c/em\u003e, we found that males and females increase in size. However, this increase could be seen reflected in different characters in each sex, which is perhaps related to sexual dimorphism due to different life habits or phenotypic plasticity in thermal gradients (Fairbairn, \u003cspan citationid=\"CR58\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Blanckenhorn et al., \u003cspan citationid=\"CR23\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Stillwell \u0026amp; Fox, \u003cspan citationid=\"CR207\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Females had a general increase in size, including their chelicerae, a key character for digging and gestation chambers (Maury, \u003cspan citationid=\"CR125\" class=\"CitationRef\"\u003e1968\u003c/span\u003e, \u003cspan citationid=\"CR126\" class=\"CitationRef\"\u003e1969\u003c/span\u003e, \u003cspan citationid=\"CR127\" class=\"CitationRef\"\u003e1977\u003c/span\u003e). Males increased in the body and caudal gland size, a character for sexual interactions (Peretti, \u003cspan citationid=\"CR149\" class=\"CitationRef\"\u003e1997\u003c/span\u003e). The shape changes in the caudal gland related to humidity are intriguing, considering that this gland produces complex chemistry secretions with numerous compounds where geographic variation among different populations has been demonstrated previously (Olivero et al., \u003cspan citationid=\"CR142\" class=\"CitationRef\"\u003e2015\u003c/span\u003e). Future studies will aim to determine whether the dynamics of secretion production or effectiveness of secretion rubbing may depend on these shape variations and whether this correlates with behavioral differences between species and allopatry and sympatry contexts.\u003c/p\u003e \u003cp\u003eIn scorpions, it is known that geographic variability may exist (Harington, \u003cspan citationid=\"CR75\" class=\"CitationRef\"\u003e1983\u003c/span\u003e; Abdel-Nabi et al., \u003cspan citationid=\"CR2\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Olivero et al., \u003cspan citationid=\"CR140\" class=\"CitationRef\"\u003e2012\u003c/span\u003e, \u003cspan citationid=\"CR142\" class=\"CitationRef\"\u003e2015\u003c/span\u003e; Yamashita \u0026amp; Rhoads, \u003cspan citationid=\"CR232\" class=\"CitationRef\"\u003e2013\u003c/span\u003e) and that the size of individuals may be affected by environmental gradients (Jochim et al., \u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; Lira et al., \u003cspan citationid=\"CR115\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). For example, Jochim et al. (\u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e2020\u003c/span\u003e), studying the morphology of a species complex of the family Vaejovidae, found a pattern of morphological convergence very similar to our results. In mountainous areas of Arizona, individuals at higher elevations were larger, resulting in individuals of different species being more similar in the middle areas of the gradient (Jochim et al., \u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). These authors argue that RCD does not occur in these species and that these scorpions probably follow Bergmann's rule, although they do not discuss these aspects further (Jochim et al., \u003cspan citationid=\"CR92\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). Because of this type of geographic variation, RCD studies must contemplate ecological factors as promoters of morphological variation (Goldberg \u0026amp; Lande, \u003cspan citationid=\"CR67\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Kosuda et al., \u003cspan citationid=\"CR99\" class=\"CitationRef\"\u003e2016\u003c/span\u003e).\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec25\" class=\"Section2\"\u003e \u003ch2\u003e4.4 Species asymmetry in morphological variability\u003c/h2\u003e \u003cp\u003eAsymmetric RI and RCD have been reported multiple times (Bordenstein et al., \u003cspan citationid=\"CR29\" class=\"CitationRef\"\u003e2000\u003c/span\u003e; Pfennig \u0026amp; Simovich, \u003cspan citationid=\"CR155\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Smadja \u0026amp; Ganem, \u003cspan citationid=\"CR197\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Cooley et al., \u003cspan citationid=\"CR37\" class=\"CitationRef\"\u003e2006\u003c/span\u003e; Cooley, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Hochkirch et al., \u003cspan citationid=\"CR79\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Costa-Schmidt \u0026amp; Machado, \u003cspan citationid=\"CR39\" class=\"CitationRef\"\u003e2012\u003c/span\u003e) and it generally occurs when there are interspecific differences in the intensity of selective pressures to avoid heterospecific interactions because species suffer different costs from RI (Pfennig \u0026amp; Simovich, \u003cspan citationid=\"CR155\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Cooley, \u003cspan citationid=\"CR36\" class=\"CitationRef\"\u003e2007\u003c/span\u003e). Also, asymmetric outcomes in morphological variability between species may indicate interspecific differences in morphological plasticity. Divergent characters can also be plastic or can be expressed facultatively when individuals face competition with heterospecifics, so plasticity has been a proposed mechanism to explain character displacement (Robinson \u0026amp; Wilson, \u003cspan citationid=\"CR168\" class=\"CitationRef\"\u003e1994\u003c/span\u003e; Pfennig \u0026amp; Murphy, \u003cspan citationid=\"CR154\" class=\"CitationRef\"\u003e2002\u003c/span\u003e; Rice \u0026amp; Pfennig, \u003cspan citationid=\"CR166\" class=\"CitationRef\"\u003e2007\u003c/span\u003e; Pfennig \u0026amp; Pfennig, \u003cspan citationid=\"CR156\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Stuart et al., \u003cspan citationid=\"CR209\" class=\"CitationRef\"\u003e2017\u003c/span\u003e). Species with broad distribution, exposed to a wide range of environmental conditions, and with ample genetic variation may exhibit more remarkable phenotypic plasticity (DeWitt \u0026amp; Scheiner, \u003cspan citationid=\"CR48\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Lavergne et al., \u003cspan citationid=\"CR109\" class=\"CitationRef\"\u003e2004\u003c/span\u003e; Pigliucci et al., \u003cspan citationid=\"CR157\" class=\"CitationRef\"\u003e2006\u003c/span\u003e). For example, Crowder et al. (\u003cspan citationid=\"CR43\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) found that the globally distributed whitefly \u003cem\u003eBemisia tabaco\u003c/em\u003e biotype exhibited greater plasticity in reproductive behavior, which could result in greater success in avoiding the costs of RI than other biotypes. Here, \u003cem\u003eUrophonius\u003c/em\u003e species present asymmetries in the RI degree they may be undergoing since males of \u003cem\u003eU. brachycentrus\u003c/em\u003e are more indiscriminate in their mating decisions than males of \u003cem\u003eU. achalensis\u003c/em\u003e (Oviedo-Diego et al., \u003cspan citationid=\"CR147\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). Moreover, \u003cem\u003eU. brachycentrus\u003c/em\u003e presented higher male-biased operational sex ratios than \u003cem\u003eU. achalensis\u003c/em\u003e in the sympatric zone (Oviedo-Diego, M. pers. obs.), which could mean males under greater scramble competition to find females and that this species could suffer higher costs due to RI (Oviedo-Diego et al., \u003cspan citationid=\"CR146\" class=\"CitationRef\"\u003e2020\u003c/span\u003e; \u003cspan citationid=\"CR147\" class=\"CitationRef\"\u003e2021\u003c/span\u003e). In turn, \u003cem\u003eU. brachycentrus\u003c/em\u003e showed the most remarkable morphological variations, being the most widely distributed species compared to \u003cem\u003eU. achalensis\u003c/em\u003e, endemic to the highland area under analysis (Acosta \u003cspan citationid=\"CR3\" class=\"CitationRef\"\u003e1985\u003c/span\u003e, \u003cspan citationid=\"CR4\" class=\"CitationRef\"\u003e1993\u003c/span\u003e; Ojanguren-Affilastro, \u003cspan citationid=\"CR138\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Ojanguren-Affilastro et al., \u003cspan citationid=\"CR139\" class=\"CitationRef\"\u003e2020\u003c/span\u003e). This complex social and geographic scenario could translate into strong selective pressures for interspecific recognition during mating or sperm transfer and the existence of RCD patterns in an asymmetric manner, being \u003cem\u003eU. brachycentrus\u003c/em\u003e the species that suffers more RI costs and the most morphologically plastic to manifest changes under these pressures.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec26\" class=\"Section2\"\u003e \u003ch2\u003e4.5 Mixed selective pressures on multiple characters in scorpions\u003c/h2\u003e \u003cp\u003eOur results reveal a strong variation in the size and shape of somatic and genital characters, which supports the notion that morphological traits are the result of multiple selective pressures and that different dimensions of the same character (e.g., shape, size) may be reflecting different evolutionary responses (mosaic evolution). This is most noticeable in characters used in multiple activities in the organism's life. We found evidence of the existence of RCD for the pedipalps shape of both sexes in sympatric populations, an evolutionary response to avoid crossbreeding and strengthen reproductive isolation among these species. In turn, other characters showed high geographic variability in size reflected in patterns of convergence towards the sympatry zone, which could affect the mating system of these species, promoting RI and explaining the high values of phenotypic variation found in characters used in sexual interactions (e.g., caudal gland, pedipalp apophysis). It is noteworthy the different selective pressures under which the genitalia would be, also under natural selection pressures showing an RCD pattern in shape, although manifesting in other portions of the hemispermatophore very high phenotypic variation which would indicate possible sexual selection pressures acting mainly in the crest zone.\u003c/p\u003e \u003cp\u003ePeretti (\u003cspan citationid=\"CR151\" class=\"CitationRef\"\u003e2010\u003c/span\u003e) highlights the existence of mixed patterns in the genitalia of scorpions, were morphological complexity results from different selective regimes. This has also been observed in other arachnids (Huber, \u003cspan citationid=\"CR86\" class=\"CitationRef\"\u003e1996\u003c/span\u003e, \u003cspan citationid=\"CR87\" class=\"CitationRef\"\u003e2004\u003c/span\u003e) and insects (Song \u0026amp; Wenzel, \u003cspan citationid=\"CR200\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Simmons et al., \u003cspan citationid=\"CR194\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Song \u0026amp; Bucheli, \u003cspan citationid=\"CR201\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Rowe \u0026amp; Arnqvist, \u003cspan citationid=\"CR175\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; House et al., \u003cspan citationid=\"CR80\" class=\"CitationRef\"\u003e2013\u003c/span\u003e; Frazee \u0026amp; Masly, \u003cspan citationid=\"CR62\" class=\"CitationRef\"\u003e2015\u003c/span\u003e) where characters are under multiple, often contradictory or inconsistent pressures. Studies in water striders suggest that the non-intromittent genitalia have differing degrees of selection acting upon them (Danielsson \u0026amp; Askenmo, \u003cspan citationid=\"CR46\" class=\"CitationRef\"\u003e1999\u003c/span\u003e; Bertin \u0026amp; Fairbairn, \u003cspan citationid=\"CR22\" class=\"CitationRef\"\u003e2005\u003c/span\u003e; Rowe \u0026amp; Arnqvist, \u003cspan citationid=\"CR175\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). Another example was reported in the dung beetle \u003cem\u003eOnthophagus taurus\u003c/em\u003e which has shown that different sections of male genital morphology may be under different selective regimes (Song \u0026amp; Wenzel, \u003cspan citationid=\"CR200\" class=\"CitationRef\"\u003e2008\u003c/span\u003e; Simmons et al., \u003cspan citationid=\"CR194\" class=\"CitationRef\"\u003e2009\u003c/span\u003e) as the shape of the aedeagus is subject to directional sexual selection, but genital sclerites that penetrate the female genitalia are subject to stabilizing and disruptive nonlinear selection (Simmons et al., \u003cspan citationid=\"CR194\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). In addition, in \u003cem\u003eO. taurus\u003c/em\u003e, the genitalia shape diverges rapidly due to directional sexual selection, whereas size remains unaffected in the process (Simmons et al., \u003cspan citationid=\"CR194\" class=\"CitationRef\"\u003e2009\u003c/span\u003e). Similarly, it has been reported for the millipede \u003cem\u003eAntichiropus variabilis\u003c/em\u003e that genitalia shape responded to stabilizing pressures (supporting the occurrence of lock-and-key), although genitalia size did not follow this pattern and responded to environmental gradients (Wojcieszek \u0026amp; Simmons, \u003cspan citationid=\"CR230\" class=\"CitationRef\"\u003e2012\u003c/span\u003e). This is like to our results, where the shape of certain structures responds to specific recognition variations with low phenotypic variation, and size shows patterns of variation linked to geographic and environmental differences. The size and shape of the same structure may respond in this mosaic manner, independently to different selective pressures, perhaps due to genetic or developmental decoupling (Macagno et al., \u003cspan citationid=\"CR117\" class=\"CitationRef\"\u003e2011\u003c/span\u003e; Rowe \u0026amp; Arnqvist, \u003cspan citationid=\"CR175\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Wojcieszek \u0026amp; Simmons, \u003cspan citationid=\"CR230\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Richmond, \u003cspan citationid=\"CR167\" class=\"CitationRef\"\u003e2014\u003c/span\u003e). Future studies will aim to assess the consistency of these results with allometric patterns between populations, and coevolution between female and male characters, as well as explore the morphological complexity of the traits by assessing the modularity of the subunits of the characters (e.g., Kuntner et al. \u003cspan citationid=\"CR104\" class=\"CitationRef\"\u003e2009\u003c/span\u003e; Tatarnic \u0026amp; Cassis, \u003cspan citationid=\"CR214\" class=\"CitationRef\"\u003e2010\u003c/span\u003e; Rowe \u0026amp; Arnqvist, \u003cspan citationid=\"CR175\" class=\"CitationRef\"\u003e2012\u003c/span\u003e; Genevcius \u0026amp; Schwertner, \u003cspan citationid=\"CR65\" class=\"CitationRef\"\u003e2017\u003c/span\u003e; Genevcius et al., \u003cspan citationid=\"CR66\" class=\"CitationRef\"\u003e2020\u003c/span\u003e)\u003c/p\u003e \u003c/div\u003e"},{"header":"Conclusions","content":"\u003cp\u003eWe found a remarkable morphological variability in both scorpion species that was determined in part by geographic and environmental variations, in part by sexual selection pressures at the intra- and interspecific level, and in part by natural selection pressures during species recognition. We report a pattern of asymmetric morphological variation where one of the scorpion species (\u003cem\u003eU. brachycentrus\u003c/em\u003e) suffered an increase in size in several characters to the sympatric zone due to environmental factors (showing a pattern of morphological convergence). This increase in size and a scenario of promiscuity probably led to certain characters undergoing intense sexual selection pressures, which is reflected in the high phenotypic variation found. However, key characters for mating success, such as grasping or genital characters, experienced morphological divergence in males and females, implying a mechanical incompatibility that could function as a barrier promoting reproductive isolation. However, some characters that showed variation by RCD were also found to be under sexual selection pressures, suggesting a complex scenario of mixed selective regimes acting on these characters. On the other hand, the non-concordant results on the pressures on the size and shape of characters enlighten us on the complexity inherent in the evolution of multi-functional traits in scorpions. This comprehensive study presents novel results in an ancestral group that has not been studied from this perspective and provides interesting insights for evaluating characters under multiple selective pressures in animal systems with RI.\u003c/p\u003e"},{"header":"Declarations","content":"\u003cp\u003eAuthors are required to disclose financial or non-financial interests that are directly or indirectly related to the work submitted for publication. Please refer to \u0026ldquo;Competing Interests and Funding\u0026rdquo; below for more information on how to complete this section.\u003c/p\u003e\n\u003cp\u003e\u003cem\u003eCompeting interests:\u003c/em\u003e The authors declare no competing interests with regard to this manuscript and the material implicated.\u003c/p\u003e\n\u003cp\u003e\u003cstrong\u003eEthical standards\u003c/strong\u003e\u003c/p\u003e\n\u003cp\u003eWe declare that the experiments comply with the current laws of Argentina. This investigation adheres to the ASAB/ABS Guidelines for the Use of Animals in Research (Buchanan et al., 2012), and the use of animals was reviewed and approved by the animal care review committee at the Instituto de Diversidad y Ecolog\u0026iacute;a Animal (IDEA), CONICET-UNC, Argentina, where we performed the experiment.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003e\u0026Aacute;balos, J. W. y Hominal, C. 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Elsevier, San Diego. \u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":false,"hideJournal":false,"highlight":"","institution":"","isAcceptedByJournal":true,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"evolutionary-biology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"evol","sideBox":"Learn more about [Evolutionary Biology](http://link.springer.com/journal/11692)","snPcode":"11692","submissionUrl":"https://submission.nature.com/new-submission/11692/3","title":"Evolutionary Biology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false},"keywords":"Reproductive character displacement, Convergence, Sexual selection, Natural selection, Geometric Morphometrics, Scorpions. ","lastPublishedDoi":"10.21203/rs.3.rs-2445373/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-2445373/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eReproductive interference (RI) can occur when two related species coexist in sympatry, involving sexual attraction, mating, and even hybridization between heterospecifics. Consequently, reproductive key characters of these species may suffer morphological shifts in sympatry to avoid the success of heterospecific sexual interactions, a phenomenon known as reproductive character displacement (RCD). RCD can be promoted by natural selection, although sexual selection pressures can act synergistically or agonistically so that phenotypic variation can respond in different directions and magnitudes to these forces. In turn, the size and shape of characters may respond differentially (mosaic evolution) to these pressures, so the analysis of multiple dimensions in traits is essential to understand the complexity of their phenotypic variability. To date, there are no studies evaluating this topic in scorpions, and two species (\u003cem\u003eUrophonius brachycentrus\u003c/em\u003e and \u003cem\u003eU. achalensis\u003c/em\u003e) sympatric and synchronous with RI represent an ideal model to evaluate the phenotypic variation and occurrence of RCD. In addition, the populations of these species are found in an altitudinal cline, so environmental factors may also be responsible for explaining their morphological variation. We compared the intra-specific variation, the size and shape of multiple characters involved in courtship, and sperm transfer in individuals from sympatric and allopatric populations using geometric morphometrics. We found asymmetric RCD of several sexual characters for courtship success (grasping structures) and sperm transfer (genital characters). This would evidence the action of natural selection pressures and the existence of a possible mechanism to avoid heterospecific mating success. In addition, we found a pattern of asymmetric morphological variation where one species in the sympatric zone suffered an increase in size in several characters due to environmental factors (pattern of morphological convergence). The convergence of characters combined with RI and a scramble competition mating system could intensify sexual selection pressures on specific characters, which was reflected in their high coefficients of variation. Our results suggest that in this sympatric zone, several selective regimes act differentially on various dimensions of the characters evaluated, which would support a possible mosaic evolution. This comprehensive study illuminates the complexity inherent in the evolution of multi-functional traits in a previously unexplored model, providing novel insights for evaluating traits under multiple selective pressures in animal systems experimenting RI.\u003c/p\u003e","manuscriptTitle":"Mosaic evolution of grasping and genitalic traits in two sympatric scorpion species with reproductive interference","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2023-01-09 15:47:07","doi":"10.21203/rs.3.rs-2445373/v1","editorialEvents":[{"type":"communityComments","content":0},{"type":"decision","content":"Major revision","date":"2023-09-21T13:53:19+00:00","index":"","fulltext":""},{"type":"editorInvitedReview","content":"","date":"2023-09-04T17:10:51+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"ada0b2e9-eb92-495b-a418-e9855dbd7438","date":"2023-08-10T23:00:28+00:00","index":"hide","fulltext":""},{"type":"reviewerAgreed","content":"8976c158-ded2-4f85-a4eb-72ff874e9ad6","date":"2023-08-04T23:50:58+00:00","index":"hide","fulltext":""},{"type":"reviewersInvited","content":"","date":"2023-03-15T15:24:32+00:00","index":"","fulltext":""},{"type":"editorAssigned","content":"","date":"2023-01-05T07:17:31+00:00","index":"","fulltext":""},{"type":"checksComplete","content":"","date":"2023-01-05T07:17:29+00:00","index":"","fulltext":""},{"type":"submitted","content":"Evolutionary Biology","date":"2023-01-05T06:48:34+00:00","index":"","fulltext":""}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"evolutionary-biology","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":false,"externalIdentity":"evol","sideBox":"Learn more about [Evolutionary Biology](http://link.springer.com/journal/11692)","snPcode":"11692","submissionUrl":"https://submission.nature.com/new-submission/11692/3","title":"Evolutionary Biology","twitterHandle":"","acdcEnabled":true,"dfaEnabled":true,"editorialSystem":"em","reportingPortfolio":"Springer Hybrid","inReviewEnabled":true,"inReviewRevisionsEnabled":false}}],"origin":"","ownerIdentity":"ac61a841-d583-4a15-87aa-4b447818489b","owner":[],"postedDate":"January 9th, 2023","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"published-in-journal","subjectAreas":[],"tags":[],"updatedAt":"2024-01-29T15:23:57+00:00","versionOfRecord":{"articleIdentity":"rs-2445373","link":"https://doi.org/10.1007/s11692-023-09623-2","journal":{"identity":"evolutionary-biology","isVorOnly":false,"title":"Evolutionary Biology"},"publishedOn":"2024-01-27 15:12:47","publishedOnDateReadable":"January 27th, 2024"},"versionCreatedAt":"2023-01-09 15:47:07","video":"","vorDoi":"10.1007/s11692-023-09623-2","vorDoiUrl":"https://doi.org/10.1007/s11692-023-09623-2","workflowStages":[]},"version":"v1","identity":"rs-2445373","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-2445373","identity":"rs-2445373","version":["v1"]},"buildId":"J0_U0BvcaRcwD8yVFaRlm","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}