Appendix S1 Character statements for discrete characters of cynolebiasine killifishes. Character statements (characters between brackets, character states between parentheses) for discrete characters used to erect the phylogenetic hypothesis among the Cynolebiasini are listed below. The distribution of character states among terminal taxa is presented in the data matrix of Appendix S2. Colours and colour patterns in males [1] Flank, ground colour: (0) brownish light grey; (1) bluish dark grey; (2) red; (3) black; (4) golden; (5) metallic green or greenish blue; (6) dark brown; (7) dark purple. [2] Flank, anterior portion, three dark blue bars alternating with red bars: (0) absent; (1) present. [3] Flank, dark grey bars: (0) absent; (1) present. [4] Flank, dark grey bars, extent: (0) whole flank; (1) anterior portion of flank; (2) posterior portion of flank. [5] Flank, anterior portion, round dark grey blotches: (0) absent; (1) present. [6] Flank, central portion, distinctive rounded black spot: (0) absent; (1) present. [7] Flank, iridescent blue marks: (0) absent; (1) present. [8] Flank, iridescent blue marks, design: (0) dots; (1) bars on anterior portion, dots on posterior portion; (2) bars; (3) dorso-ventrally elongated spots. [9] Flank, iridescent blue marks, extent: (0) whole; (1) posterior portion; (2) dorsal portion; (3) anterior portion; (4) ventral portion. [10] Head, posterior dorsolateral portion, distinctive concentration of intense red pigmentation on scale margins: (0) absent; (1) present. [11] Head, supra-opercular region, dark reddish brown spots: (0) absent; (1) present. [12] Head, suborbital region, dark grey to dark reddish grey bar: (0) absent; (1) present. Remarks: a supraorbital mark, considered as a distinct character in previous studies (e. g., Costa, 2006b) was always associated with the present character 12, thus herein considered a dependent condition. [13] Head, cephalic neuromast series, dark pigmentation: (0) not distinctively concentrated; (1) concentrated on parietal series; (2) concentrated on most series above, below and behind orbit. [14] Head, opercular region, three red bars: (0) absent; (1) present. [15] Head, opercular region, iridescent colour pattern: (0) pale greenish golden; (1) intense blue. [16] Head, iris, dark grey to dark reddish grey bar through center of eye: (0) absent; (1) present. [17] Head, iris, dark grey to dark reddish grey through posterior margin of eye: (0) absent; (1) present. [18] Head, iris, colour: (0) light yellow; (1) green; (2) metallic blue; (3) brownish orange. [19] Dorsal fin, distal edge, dark grey to black stripe: (0) absent; (1) present. [20] Dorsal fin, basal portion, black blotches: (0) absent; (1) present. [21] Dorsal fin, anterior portion, black spot: (0) absent; (1) present. [22] Dorsal fin, posterior portion, black bar: (0) absent; (1) present. Remarks: specimens of all terminal taxa exhibiting the apomorphic character state, also have a similar bar on the posterior portion of the anal fin, a condition thus considered as a dependent. [23] Dorsal fin, iridescent colour pattern: (0) dots on whole fin; (1) dots just on basal portion; (2) bars on whole fin; (3) vertically elongated spots on anterobasal portion, dots on remaining portions; (4) reticulate; (5) transverse vermiculate stripes; (6) longitudinal lines on fin rays; (7) vertical lines on anterior portion, dots on posterior portion; (8) vertically elongate spots along fin base; (-) iridescent marks inconspicuous. [24] Dorsal fin, distal margin, narrow iridescent blue outline: (0) absent; (1) present. Remarks: specimens of all terminal taxa having the apomorphic character state, also have a similar outline on the anal and caudal fins, thus, considered as dependent conditions. [25] Dorsal fin, distal margin, row of vertically elongated iridescent blue spots: (0) absent; (1) present. [26] Dorsal fin, subdistal zone, iridescent blue stripe: (0) absent; (1) present. Remarks: specimens of about all terminal taxa exhibiting the apomorphic character state, also have a similar stripe on the anal fin, therefore, considered as a dependent condition. [27] Dorsal fin, distal margin, dark red stripe: (0) absent; (1) present. [28] Anal fin, dark oblique bars: (0) absent; (1) present. [29] Anal fin, distal margin, dark grey to black stripe: (0) absent; (1) present. [30] Anal fin, iridescent marks: (0) absent; (1) present. [31] Anal fin, iridescent colour pattern: (0) dots on whole basal portion; (1) dots restricted to posterior portion of fin; (2) oblique bars; (3) lines parallel to fin rays; (4) dots on basal portion, short lines parallel to fin rays on distal half of fin. [32] Anal fin, subdistal zone, bright orange stripe: (0) absent; (1) present. [33] Anal fin, distal zone, distinctive bright orangish red pigmentation: (0) absent; (1) present. [34] Caudal fin, pigmentation: (0) pigmented; (1) hyaline. [35] Caudal fin, posterior margin, dark grey to black bar: (0) absent; (1) present. [36] Caudal fin, iridescent colour pattern: (0) dots; (1) bars; (2) lines parallel to fin rays; (3) elongated spots radially arranged. [37] Caudal fin, posterior margin, iridescent blue bar: (0) absent; (1) present. [38] Caudal fin, whole margin, row of iridescent blue spots: (0) absent; (1) present. [39] Caudal fin, subdistal zone, iridescent blue line: (0) absent; (1) present. [40] Pectoral fin, ventral margin, black stripe: (0) absent; (1) present. [41] Pectoral fin, basal portion, iridescent blue spots: (0) absent; (1) present. [42] Pectoral fin, ground colour: (0) hyaline; (1) reddish brown to red; (2) bright blue. Colours and colour patterns in females [43] Flank, anterocentral portion, distinctive black spot: (0) absent; (1) present. [44] Flank, dark pigmentation pattern: (0) dots; (1) continuous bars; (2) broken bars or spots. [45] Flank, caudal peduncle, posterior portion, black spot: (0) absent; (1) present. [46] Flank, caudal peduncle, posterior portion, black spot, number and arrangement: (0) one or more black spots irregularly arranged; (1) two black spots vertically arranged, sometimes coalesced forming 8-shaped blotch. [47] Flank, central portion, iridescent blue marks: (0) absent; (1) present. [48] Anal fin, posterior portion, iridescent blue spot: (0) absent; (1) present. [49] Anal fin, ground colour: (0) hyaline; (1) pale pink or pale orangish pink. External morphology of fins [50] Dorsal fin in males, distal extremity, filaments: (0) absent; (1) present. Remarks: taxa having the apomorphic condition also exhibited similar filaments on the anal fin, thus considered a dependent condition. [51] Dorsal and anal fins in males, distal extremity, filaments, extent: (0) reaching to middle caudal fin; (1) surpassing posterior caudal-fin margin. Remarks: taxa exhibiting the derived character state also had long filaments on the anal fin, which was interpreted as a single evolutionary event. [52] Dorsal fin in males, distal margin, anterior portion, filaments: (0) absent; (1) present. [53] Anal fin in males, distal border profile: (0) rounded; (1) pointed. Remarks: specimens of most terminal taxa had a similar morphology in dorsal and anal fins (i. e., both rounded or both pointed); to avoid a double weight to a putative dependent condition, dorsal fin shape is not analyzed as a distinct character. [54] Anal fin in females, shape: (0) approximately semi-circular; (1) approximately spatula-shaped. [55] Anal fin in females, antero-distal portion, membrane, dense whitish thickening: (0) absent; (1) present. [56] Anal fin in males, basal region, scales: (0) absent; (1) present. [57] Anal fin in males, basal region, transverse rows of scales projecting between fin rays: (0) absent; (1) present. [58] Caudal fin in males, shape: (0) rounded; (1) subtruncate. [59] Caudal fin in males, scales, extent: (0) restricted to basal portion of fin; (1) over about anterior half of fin. [60] Pectoral fin in males, basal portion, scales: (0) absent; (1) present. [61] Pelvic fin in males, bases, medial proximity: (0) separated by interspace; (1) in contact or united. [62] Pelvic fin in males, medial margin, membranes fusion: (0) not fused or fused only near basal portion of fin; (1) fused along most part of medial margin. External morphology of head [63] Mouth, cleft position: (0) terminal; (1) superior. [64] Mouth, cleft, ventro-lateral extremity: (0) straight; (1) curved upwards. [65] Mouth, cleft, posterior extent: (0) reaching vertical through anterior portion of orbit; (1) reaching vertical through median portion of orbit. [66] Anterior nostril, orientation: (0) anteriorly directed; (1) dorso-laterally directed. [67] Eye, orbital rim, dorsal attachment: (0) free; (1) attached. [68] Eye, position in head: (0) dorsolateral; (1) lateral. [69] Head, ventral surface, gap between laterosensory series and isthmus, posterior extent: (0) reaching opercle; (1) reaching transverse line through corner region of preopercular series; (2) reaching transverse line through middle of eye; (-) gap absent. [70] Head, ventro-lateral surface, gap below corner of pre-opercular series: (0) present; (1) absent. [71] Head in adult males, dorsal profile above eye: (0) about straight, sometimes slightly concave or slightly convex; (1) with pronounced concavity; (2) approximately straight and steep, abruptly terminating in convex nape region. [72] Frontal scales, geometric arrangement: (0) transverse; (1) circular. [73] Frontal squamation, modal arrangement-pattern determined by scale with all margins free: (0) G; (1) E; (2) A; (3) F or some other scale anterior to E-scale with variable position; (?) undetermined (taxa with small scales without a clear pattern). [74] Supraorbital scales: (0) present; (1) absent. [75] Suborbital region, scales: (0) present; (1) absent. [76] Preopercular region, scales: (0) present; (1) absent. [77] Frontal region, anterior portion adjacent to rostral neuromasts, scales: (0) present; (1) absent. External morphology of head and trunk [78] Flank in females, region above anal-fin base, scales: (0) present; (1) absent. [79] Urogenital papilla in males, shape: (0) globular; (1) tubular. [80] Urogenital papilla in males, proximity to anal fin: (0) separated by interspace; (1) close. In all terminal taxa having the derived state, the urogenital papilla is distinctively widened in its basal portion. [81] Urogenital papilla in males, attachment to anal fin: (0) free; (1) attached to first anal-fin ray by thin membrane. [82] Urogenital opening in females, shape: (0) gap; (1) prominent pocket-like structure overlapping anterior anal-fin origin. Contact organs in males [83] Flank, scales: (0) absent; (1) present. Remarks: most taxa having the apomorphic condition also have contact organs on opercular scales. [84] Dorsal fin: (0) absent; (1) present. [85] Anal fin: (0) absent; (1) present; (?) taxa with variable occurrence of the apomorphic condition. [86] Caudal fin: (0) absent; (1) present. [87] Pectoral fin, inner surface: (0) absent; (1) present. [88] Pectoral fin, inner surface, distribution: (0) on uppermost rays; (1) on most fin rays. [89] Pectoral fin, outer surface: (0) absent; (1) present. Laterosensory system [90] Supra-orbital series of neuromasts, lateral epidermal trenches: (0) present; (1) rudimentary or absent. [91] Supra-orbital series of neuromasts, anterior and posterior sections, connection: (0) separated; (1) continuous. [92] Supra-orbital series of neuromasts, anterior section, distribution of neuromasts: (0) continuous; (1) two anteriormost neuromasts separated from posterior neuromasts by space, making series interrupted at level of posterior nostril. Remarks: in all terminal taxa exhibiting the derived character state, pre-orbital and supraorbital series are continuous, thus considered a dependent condition. [93] Infra-orbital and preopercular canals: (0) close; (1) open. [94] Infra-orbital series of neuromasts, lower portion, design: (0) near orbit; (1) displaced antero-ventrally. [95] Infra-orbital series of neuromasts, lower portion, arrangement: (0) circular; (1) forming angle on antero-ventral region. [96] Otic and post-otic series of neuromasts, relative position: (0) otic on horizontal above post-otic; (1) otic and post-otic on same horizontal. [97] Mandibular series of neuromasts, posterior section, canal: (0) close; (1) open. [98] Latero-mandibular series of neuromasts, number of rows: (0) single; (1) double. [99] Preopercular and mandibular series of neuromasts, connection: (0) separated; (1) continuous. [100] Caudal fin, vertical rows of neuromasts: (0) single; (1) multiple. [101] Caudal fin, vertical row, number of neuromasts: (0) two; (1) three. Jaws, jaw suspensorium and opercular apparatus [102] Autopalatine, ventral process: (0) present; (1) rudimentary or absent. [103] Autopalatine, ventral process, extent relative to quadrate: (0) reaching dorsal portion; (1) not reaching; (-) not applicable by process being rudimentary or absent. [104] Autopalatine, prominent dorsomedial process: (0) absent; (1) present. [105] Autopalatine, dorsal pointed process: (0) absent; (1) present. [106] Autopalatine, dorsal extremity, torsion: (0) absent; (1) present. [107] Autopalatine, medial crest ventrally projected: (0) absent; (1) present; (2) present, with prominent ventral expansion. [108] Autopalatine, general morphology: (0) wide, bottle-shaped; (1) narrow, knifeshaped. [109] Entopterygoid, anterior portion, extent relative to autopalatine: (0) overlapping; (1) not overlapping. [110] Entopterygoid, posterior portion, extent: (0) reaching metapterygoid; (1) reaching vertical through middle portion of quadrate. [111] Entopterygoid, ventral portion, extent relative to quadrate: (0) overlapping; (1) not overlapping. [112] Metapterygoid, shape: (0) about rectangular; (1) about triangular, widening ventrally; (2) rod-like. Remarks: all taxa exhibiting character state 1 have a shorted anterior flap of hyomandibula, considered a dependent feature related to a constriction on the region comprising both the upper portion of the metapterygoid and the adjacent portion of hyomandibula. [113] Hyomandibula, shape: (0) short and wide, depth equal or slightly longer than width; (1) long and narrow, depth about twice width. [114] Hyomandibula, posteroventral process, extent relative to articular condyle for sympletic: (0) short, reaching transverse line through middle of condyle; (1) long, reaching transverse line through distal portion of condyle. [115] Hyomandibula, posteroventral flap, development expressed by width relative to main condyle of hyomandibula: (0) well developed, equally broad; (1) narrower; (2) rudimentary. [116] Sympletic, general shape: (0) short and deep, about so deep as long; (1) slender and long, longer than deep. [117] Quadrate, posterior process, length relative to quadrate length without process: (0) equal or shorter; (1) longer. [118] Maxilla, main axis, torsion: (0) not twisted; (1) slightly twisted. [119] Maxilla, dorsal process, shape: (0) triangular; (1) cylindrical. [120] Maxilla, ventral process, anterior expansion: (0) absent; (1) present. [121] Jaws, length relative to jaw suspensorium: (0) longer; (1) shorter. [122] Premaxilla, subdistal portion of posterior margin, expansion: (0) present; (1) absent. [123] Premaxilla, ascending process, shape: (0) about triangular; (1) about rectangular. [124] Premaxilla, medial portion close to symphysis, concavity: (0) absent; (1) present. [125] Premaxilla, alveolar arm, anterior process: (0) well developed; (1) rudimentary. [126] Rostral cartilage, shape: (0) approximately triangular; (1) approximately circular; (2) approximately hexagonal to rectangular, always longer than wide. [127] Dentary, coronoid process, extent relative to dorsal portion of angulo-articular: (0) extending; (1) not extending. [128] Dentary, coronoid process, shape: (0) robust; (1) slender. [129] Dentary, posterior teeth of external row, orientation: (0) directed to inside mouth; (1) antero-medially directed. [130] Dentary, posteroventral process of dentary, shape: (0) narrow, pointed; (1) broad, truncate. [131] Angulo-articular, ventral process, width relative to dorsal process of anguloarticular: (0) distinctively wider; (1) about equal; (-) terminal taxa with rudimentary ventral process. [132] Angulo-articular, ventral process, extent: (0) long, projecting below ventral process of dentary; (1) median, at same level of ventral process of dentary and retroarticular; (2) short, retro-articular slightly projecting below ventral process of anguloarticular. [133] Preopercle, shape: (0) robust, L-shaped; (1) thin, C-shaped. [134] Preopercle, dorsal arm, shape: (0) blunt; (1) pointed. [135] Preopercle, anteromedian rim, development: (0) well developed; (1) rudimentary or absent. [136] Preopercle, ventral arm, length relative to dorsal arm: (0) equal or longer; (1) shorter. [137] Opercle, dorsal portion length relative to central portion: (0) conspicuously longer; (1) about equal or shorter. Hyoid and branchial arches [138] Opercle, antero-dorsal margin: (0) about straight or convex; (1) concave. [139] Interhyal, ossification: (0) ossified; (1) cartilaginous. [140] Basihyal, cartilage, lateral border: (0) straight; (1) laterally expanded. [141] Urohyal, shape: (0) slender; (1) deep; (2) slender anteriorly, deep posteriorly. [142] First hypobranchial, ossification: (0) well-ossified, posterior margin welldelimited; (1) poorly ossified, posterior margin ill-delimited with cartilaginous border. Remarks: in addition, in all species having the apomorphic state, the anterior tip of the second hypobranchial is directed to the first hypobranchial, instead of to the first ceratobranchial. [143] First hypobranchial, lateral edge, articular face extension relative to apical cartilage of first ceratobranchial: (0) restricted to articulation area; (1) extended beyond articulation. [144] Second hypobranchial, anterior process, orientation: (0) anteriorly directed; (1) antero-medially directed, often branched with distinct process directed to second basibranchial. [145] Second hypobranchial, shape: (0) about pentagonal or rectangular; (1) round. [146] First branchial arch, gill-rakers, shape: (0) narrow, with wide interspaces; (1) wide, without interspaces. [147] Fourth ceratobranchial, ventral process: (0) present; (1) absent. Remarks: in all terminal taxa having the character state 1, teeth are absent on the fourth ceratobranchial, possibly constituting a single character. [148] Fourth ceratobranchial, proximal tip, width relative to proximal tip of third ceratobranchial: (0) wider; (1) narrower. [149] Fifth ceratobranchial, anterior length (between anterior tip and postero-medial angle) relative to postero-lateral length (between postero-lateral tip and postero-medial angle): (0) shorter; (1) longer. [150] Fifth ceratobranchial, medial fusion: (0) not fused; (1) fused. Remarks: in all taxa exhibiting the derived condition the anterior tip of the fifth ceratobranchial is narrow and long, thus considered a single derived condition. [151] Cylindrical ossification below fourth basibranchial cartilage: (0) absent; (1) present. [152] Second pharyngobranchial, articular facet for second epibranchial, position: (0) on medial margin; (1) continuous to proximal margin. [153] Second pharyngobranchial, articular facet for second epibranchial, orientation, angle relative to distal condyle: (0) perpendicular; (1) sharp angle. [154] Second pharyngobranchial, distal condyle, orientation: (0) distally oriented; (1) laterodistally oriented; (2) laterally oriented. [155] Second pharyngobranchial, distal condyle, width relative to medial condyle width: (0) equal or slightly wider; (1) conspicuously wider. [156] Second pharyngobranchial, place for attachment of interarcual cartilage: (0) near distal pharyngobranchial cartilage; (1) on medial margin; (2) near proximal edge. [157] Second pharyngobranchial, depth relative to width: (0) about so wide as deep; (1) conspicuously deeper than wide; (2) conspicuously wider than deep. [158] Second pharyngobranchial, teeth: (0) present; (1) absent. [159] Epibranchials, length relative to third pharyngobranchial width: (0) shorter; (1) longer. [160] Second epibranchial, subdistal process: (0) present; (1) absent. [161] Third epibranchial, uncinate process, length relative to distal process: (0) about so long or slightly shorter; (1) conspicuously shorter. Superficial dermal bones and neurocranium [162] Lachrymal, curvature on dorso-ventral axis: (0) absent; (1) present. [163] Dermosphenotic: (0) present; (1) absent; (?) terminal taxa with variable occurrence of one or two minute ossifications on dermosphenotic region. [164] Vomerine teeth: (0) present; (1) absent. [165] Vomer, anterior margin: (0) conspicuously convex; (1) nearly straight or slightly concave. [166] Lateral ethmoid, medial extent expressed by cartilaginous space width between medial margin of bone and vomer and parasphenoid relative to anterior parasphenoid width: (0) wider; (1) narrower. [167] Lateral ethmoid, anterior retrorse process: (0) rudimentary; (1) well developed. [168] Parasphenoid, posterior process, posterior widening: (0) gradual; (1) abrupt. [169] Frontal, lateral border, shape: (0) approximately straight; (1) concave. [170] Sphenotic, lateral process, width relative to posterior portion of parasphenoid: (0) conspicuously narrower; (1) about equal or wider. [171] Sphenotic, lateral process, pointed anterior expansion: (0) absent; (1) present. [172] Autopterotic, postero-lateral process, tip, thickened pointed extension (Costa, 2001): (0) absent; (1) present. [173] Supraoccipital, posterior process, shape: (0) short, without a narrow posterior extension; (1) long, terminating in narrow posterior extension. Vertebrae and unpaired fin skeleton [174] First vertebra, latero-dorsal laminar process: (0) present; (1) absent. [175] First vertebra, neural spine, connection with neural prezygapophysis of second vertebra: (0) thin ligaments; (1) directly attached. [176] Second vertebra, prezygapophysis, development: (0) well developed; (1) rudimentary. [177] Third vertebra, neural spine, width relative to neural spines of second and fourth vertebrae: (0) about so wide as second neural spine, distinctively wider than fourth; (1) much narrower than second neural spine, about so wide as fourth. [178] Caudal vertebrae, neural prezygapophyses, development: (0) well developed; (1) rudimentary. [179] Preural centrum 2, hemal spine, width relative to hemal spines anterior to it: (0) distinctively wider; (1) approximately equal in width. [180] Epural and parhypural, proximal region, shape: (0) broad and approximately straight; (1) narrow and curved anteriorly. [181] Parahypural, proximal tip, paired dorsal processes overlapping preural centrum: (0) present; (1) absent. [182] Hypurals, gap between two dorsal plates: (0) present; (1) absent. [183] Hypurals, middle gap: (0) present; (1) absent. [184] Dorsal and anal fins, sexual dimorphism in number of rays: (0) not dimorphic; (1) more rays in males than in females. [185] Dorsal fin, rays, branching: (0) branched; (1) branched, except on fin tip; (2) all unbranched. [186] Dorsal fin, medial radials, ossification: (0) well-ossified; (1) poorly ossified; (2) cartilaginous. [187] Dorsal fin, median radials, shape: (0) about twice longer than wide; (1) about three or four times longer than wide. [188] Anal fin, rays, branching: (0) branched; (1) branched, except on fin tip; (2) all unbranched. [189] Anal fin, medial radials, ossification: (0) ossified; (1) cartilaginous, sometimes poorly ossified. [190] Anal fin, first proximal radials, shape relative to middle proximal radials: (0) equally long and narrow; (1) distinctively shorter and wider. Shoulder and pelvic girdles [191] Pectoral fin, insertion: (0) lateral; (1) ventrolateral. [192] Pectoral fin, posttemporal, ventral process: (0) present; (1) absent (sometimes rudimentary). [193] Pectoral fin, cleithrum, posterior flange: (0) present; (1) absent. [194] Pectoral fin, cleithrum, dorsal wide portion, length relative to vertical length between dorsal margin of scapula and ventral margin of coracoid: (0) about equal; (1) about one time and half; (2) about twice. [195] Pectoral fin, first postcleithrum: (0) present; (1) absent. [196] Pectoral fin, proximal radials, shape: (0) cubical; (1) discoid. [197] Pectoral fin, proximal radials, cartilaginous interspace: (0) minute; (1) broad. [198] Pectoral fin, dorsalmost proximal radial: (0) present; (1) absent. [199] Coracoid, anteroventral condyle, shape: (0) entirely narrow; (1) widening towards extremity. [200] Pelvic bone, thickness and processes, development: (0) thick, processes well developed; (1) thin, ischial and lateral processes rudimentary or absent. [201] Pelvic bones, relative medial position: (0) separated by interspace; (1) overlapped. [202] Pelvic bone, development: (0) well developed; (1) rudimentary.