Research Article |
Corresponding author: Erick Cristofore Guimarães ( erick.ictio@yahoo.com.br ) Academic editor: Nicolas Hubert
© 2019 Erick Cristofore Guimarães, Pâmella Silva de Brito, Leonardo Manir Feitosa, Luis Fernando Carvalho Costa, Felipe Polivanov Ottoni.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Guimarães EC, Silva de Brito P, Feitosa LM, Carvalho Costa LF, Ottoni FP (2019) A new cryptic species of Hyphessobrycon Durbin, 1908 (Characiformes, Characidae) from the Eastern Amazon, revealed by integrative taxonomy. Zoosystematics and Evolution 95(2): 345-360. https://doi.org/10.3897/zse.95.34069
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Hyphessobrycon caru sp. nov. is described based on five different and independent methods of species delimitation, making the hypothesis of this new species supported by an integrative taxonomy perspective. This new species has a restricted distribution, occurring just in the upper Pindaré river drainage, Mearim river basin, Brazil. It is a member of the rosy tetra clade, which is characterized mainly by the presence of a dark brown or black blotch on dorsal fin and absence of a midlateral stripe on the body. Hyphessobrycon caru sp. nov. is distinguished from the members of this clade mainly by the shape of its humeral spot, possessing few irregular inconspicuous vertically arranged chromatophores in the humeral region, or sometimes a very thin and inconspicuous humeral spot, and other characters related to teeth count, and color pattern. The phylogenetic position of the new species within the rosy tetra clade was based on molecular phylogenetic analysis using sequences of the mitochondrial gene cytochrome oxidase subunit 1. In addition, a new clade (here termed Hyphessobrycon micropterus clade) within the rosy tetra clade is proposed based on molecular data, comprising H. caru sp. nov., H. micropterus, H. piorskii, and H. simulatus, and with H. caru sp. nov. and H. piorskii recovered as sister species. Our results suggest cryptic speciation in the rosy tetra clade and, more specifically, in the H. micropterus clade. We recommend the use of integrative taxonomy for future taxonomic revisions and species descriptions when dealing with species complexes and groups containing possible cryptic species.
bPTP, DNA barcoding, rosy tetra clade, species complex, Stethaprioninae
Hyphessobrycon Durbin, 1908 is a species-rich characid genus comprising about 160 valid species (
Several genetic studies focusing on characoid fishes, such as Astyanax Baird & Girard, 1854 (e.g.
In this context of integrative taxonomy, the present study aims to investigate the diversity within the rosy tetra clade sensu
This group has had its composition and name changed over the last decades, and a detailed taxonomic history is presented by
Individuals collected for this study were euthanized with a buffered solution of MS-222 at a concentration of 250 mg L−1 for a period of 10 min or more until opercular movements completely ceased. Specimens selected for morphological analysis were fixed in formalin and left for 10 days, after which they were preserved in 70% ethanol. Molecular data were obtained from specimens that were euthanized, fixed, and preserved in absolute ethanol.
Specimens for morphological analysis are listed in type and comparative material lists. Specimens for molecular approaches are listed in Table
List of species, specimens and their respective catalogue numbers, Region/state/country, and BOLD Systems and GenBank sequence accession numbers. Sequences available in the current study are in Bold.
N° | Species | Catalogue number | Region/state/country | Accession no. |
---|---|---|---|---|
1 | Hyphessobrycon erythrostigma | INPA 37681-HERY1 | Tabatinga/Amazonas/Brazil | HYP076-13 |
2 | Hyphessobrycon erythrostigma | INPA 37681-HERY10 | Tabatinga/Amazonas/Brazil | HYP077-13 |
3 | Hyphessobrycon erythrostigma | INPA 37681-HERY2 | Tabatinga/Amazonas/Brazil | HYP078-13 |
4 | Hyphessobrycon erythrostigma | INPA 37681-HERY3 | Tabatinga/Amazonas/Brazil | HYP079-13 |
5 | Hyphessobrycon pyrrhonotus | INPA 37672-TRO10 | Santa Isabel do Rio Negro/Amazonas/Brazil | HYP040-13 |
6 | Hyphessobrycon pyrrhonotus | INPA 37672-TRO11 | Santa Isabel do Rio Negro/Amazonas/Brazil | HYP041-13 |
7 | Hyphessobrycon pyrrhonotus | – | Barcelos/Amazonas/Brazil | HYP157-13 |
8 | Hyphessobrycon pyrrhonotus | – | Barcelos/Amazonas/Brazil | HYP158-13 |
9 | Hyphessobrycon socolofi | INPA_39530-6152 | Barcelos/Amazonas/Brazil | HYP131-13 |
10 | Hyphessobrycon socolofi | INPA_39530-6155 | Barcelos/Amazonas/Brazil | HYP134-13 |
11 | Hyphessobrycon socolofi | INPA_39530-6178 | Barcelos/Amazonas/Brazil | HYP135-13 |
12 | Hyphessobrycon socolofi | INPA 39530-BCR8 | Barcelos/Amazonas/Brazil | HYP148-13 |
13 | Hyphessobrycon copelandi | INPA_37683-TU1 | Tabatinga/Amazonas/Brazil | HYP094-13 |
14 | Hyphessobrycon copelandi | INPA_37683-TU2 | Tabatinga/Amazonas/Brazil | HYP095-13 |
15 | Hyphessobrycon copelandi | INPA_37683-TU3 | Tabatinga/Amazonas/Brazil | HYP096-13 |
16 | Hyphessobrycon eques | INPA_37678-IC2 | Santarém/Pará/Brazil | HYP070-13 |
17 | Hyphessobrycon eques | INPA_37679-PE1 | Macapá/Amapá/Brazil | HYP071-13 |
18 | Hyphessobrycon eques | INPA_37680-AL1 | Parintins/Amazonas/Brazil | HYP072-13 |
19 | Hyphessobrycon eques | OL-0544 | Bonito/Mato Grosso do Sul/Brazil | DSMIS077-09 |
20 | Hyphessobrycon epicharis | INPA_37665-JUF1 | São Gabriel da Cachoeira/Amazonas/Brazil | HYP002-13 |
21 | Hyphessobrycon epicharis | INPA_37665-JUF3 | São Gabriel da Cachoeira/Amazonas/Brazil | HYP004-13 |
22 | Hyphessobrycon epicharis | INPA_37665-JUF4 | São Gabriel da Cachoeira/Amazonas/Brazil | HYP005-13 |
23 | Hyphessobrycon epicharis | INPA_37665-JUF8 | São Gabriel da Cachoeira/Amazonas/Brazil | HYP006-13 |
24 | Hyphessobrycon compressus | CINV-NEC7411 | Flores Magon/Campeche/México | FYPM054-10 |
25 | Hyphessobrycon compressus | ECOCH | Hatie ville/Belize/Belize | MXV765-15 |
26 | Hyphessobrycon compressus | ECOCH | Hatie ville/Belize/Belize | MXV766-15 |
27 | Hyphessobrycon compressus | ECOCH | Hatie ville/Belize/Belize | MXV767-15 |
28 | Hyphessobrycon bentosi | INPA_37684-5939 | Barcelos/Amazonas/Brazil | HYP097-13 |
29 | Hyphessobrycon bentosi | INPA_37684-5940 | Barcelos/Amazonas/Brazil | HYP098-13 |
30 | Hyphessobrycon bentosi | INPA_39527-BA1 | – | HYP116-13 |
31 | Hyphessobrycon bentosi | INPA_39527-BA2 | – | HYP117-13 |
32 | Hyphessobrycon bentosi | CICCAA02349 | Santarém/Pará/Brazil | MK240339 |
33 | Hyphessobrycon bentosi | CICCAA02350 | Santarém/Pará/Brazil | MK240340 |
34 | Hyphessobrycon bentosi | CICCAA02351 | Santarém/Pará/Brazil | MK240341 |
35 | Hyphessobrycon simulatus | MHNG 2743.087 | Pisiemoengo/Commewijne/Suriname | GBOL761-15 |
36 | Hyphessobrycon simulatus | MHNG 2743.087 | Pisiemoengo/Commewijne/Suriname | GBOL762-15 |
37 | Hyphessobrycon simulatus | – | Sinnamary/Cayenne/French Guiana | GBOL1771-17 |
38 | Hyphessobrycon simulatus | MHNG 2735.007 | Sinnamary/Cayenne/French Guiana | GBOL3296-18 |
39 | Hyphessobrycon simulatus | MHNG 2757.080 | Kourou/Cayenne/French Guiana | GBOL3298-18 |
40 | Hyphessobrycon simulatus | MHNG 2759.026 | Kaw/Cayenne/French Guiana | GBOL3300-18 |
41 | Hyphessobrycon simulatus | MHNG 2759.026 | Kaw/Cayenne/French Guiana | GBOL3301-18 |
42 | Hyphessobrycon simulatus | MHNG 2759.035 | Régina/ Cayenne/French Guiana | GBOL3302-18 |
43 | Hyphessobrycon cf. sweglesi | INPA_37668-JAR3 | São Gabriel da Cachoeira/Amazonas/Brazil | HYP026-13 |
44 | Hyphessobrycon cf. sweglesi | INPA_37668-JAR4 | São Gabriel da Cachoeira/Amazonas/Brazil | HYP027-13 |
45 | Hyphessobrycon cf. sweglesi | INPA_37668-JAR5 | São Gabriel da Cachoeira/Amazonas/Brazil | HYP028-13 |
46 | Hyphessobrycon cf. sweglesi | INPA_37668-JAR7 | São Gabriel da Cachoeira/Amazonas/Brazil | HYP030-13 |
47 | Hyphessobrycon micropterus | – | Várzea da Palma/Minas Gerais/Brazil | BSB287-10 |
48 | Hyphessobrycon micropterus | – | Várzea da Palma/Minas Gerais/Brazil | BSB288-10 |
49 | Hyphessobrycon micropterus | – | Várzea da Palma/Minas Gerais/Brazil | BSB289-10 |
50 | Hyphessobrycon micropterus | – | Várzea da Palma/Minas Gerais/Brazil | BSB290-10 |
51 | Hyphessobrycon piorskii | CICCAA00725-1 | Chapadinha/Maranhão/Brazil | MF765796 |
52 | Hyphessobrycon piorskii | CICCAA00726-1 | Chapadinha/Maranhão/Brazil | MF765797 |
53 | Hyphessobrycon piorskii | CICCAA01650-1 | Barreirinhas/Maranhão/Brazil | MG791915 |
54 | Hyphessobrycon piorskii | CICCAA01651-1 | Barreirinhas/Maranhão/Brazil | MG791914 |
55 | Hyphessobrycon piorskii | CICCAA02164-1 | Codó/Maranhão/Brazil | MK240337 |
56 | Hyphessobrycon piorskii | CICCAA02164-4 | Codó/Maranhão/Brazil | MK240338 |
57 | Hyphessobrycon caru | CICCAA00748-1 | Buriticupu/Maranhão/Brazil | MH338230 |
58 | Hyphessobrycon caru | CICCAA00749-1 | Buriticupu/Maranhão/Brazil | MH338231 |
59 | Hyphessobrycon caru | CICCAA02300-1 | Buriticupu/Maranhão/Brazil | MH338232 |
60 | Hyphessobrycon caru | CICCAA02301-1 | Buriticupu/Maranhão/Brazil | MH338233 |
61 | Pristella maxillaris | – | – | KU568982.1 |
62 | Pristella maxillaris | – | – | KU568981.1 |
63 | Pristella maxillaris | – | Marlborough/Pomeroon-Supenaam/Guyana | TZGAA025-06 |
64 | Pristella maxillaris | – | Santa Cruz/Barima-Waini/Guyana | TZGAA178-06 |
65 | Moenkhausia hemigrammoides | INPA38532-PR1 | Guyana | HYP101-13 |
66 | Moenkhausia hemigrammoides | INPA_38532-PR2 | Guyana | HYP102-13 |
67 | Moenkhausia hemigrammoides | INPA_38532-PR3 | Guyana | HYP103-13 |
68 | Hyphessobrycon panamensis | STRI-05303 | Cocle/Panama | BSFFA760-07 |
69 | Hyphessobrycon flammeus | LBPV-40464 | Biritiba-Mirim/São Paulo/Brazil | FUPR988-09 |
Measurements and counts were made according to
DNA was extracted from fin clips using Wizard Genomic DNA Purification kit (Promega) according to the manufacturer’s protocol. Fragments of the cytochrome c oxidase subunit 1 gene (hereafter COI) from mitochondrial DNA were amplified, using the universal primers designed by
The dataset included the following gene: COI (680 Base pairs, BP). Sequences were aligned using ClustalW (
The unified species concept is herein adopted by expressing the conceptual definition shared by all traditional species concepts, “species are (segments of) separately evolving metapopulation lineages”, disentangling operational criterion elements to delimit taxa from species concepts (
Five distinct and independent operational criteria for species delimitation, based on morphological and molecular data, were implemented here: Population Aggregation Analysis (
The PAA (
We used the Kimura-2-parameters model (K2P) (
WP is based on the direct inspection of haplotype trees generated from the phylogenetic analysis having as terminals at least two individuals (haplotypes) of each focal species. In this method, the term “exclusive” is used instead of monophyletic, as the term monophyly is considered inapplicable below the species level (
The CBB is similar to the population aggregation analysis proposed by
The bPTP is a coalescent phylogeny-based species delimitation method aimed at delimiting species based on single locus molecular data (
CICCAA 02286, 22.2 mm SL, Brazil, Maranhão state, Buriticupu municipality, Buritizinho river, Pindaré river drainage, Mearim river basin, 04°22'52"S, 46°30'35"W, 24 Jan. 2017, Guimarães E. C., Brito P. S.
All from Brazil, Maranhão state: CICCAA 00706, 37, 15.9–25.4 mm SL; CICCAA 0709, 12 C&S, 15.1–20.6 mm SL; LIOP.UFAM 1009, 1, 16.2 mm SL collected with holotype. CICCAA00707, 3, 17.2–22.1 mm SL, Buriticupu municipality, Buritizinho river, Pindaré river drainage, Mearim river basin, 4°25'45"S, 46°29'41"W, 24 Jan. 2017, Guimarães E. C., Brito P. S. CICCAA00708, 2, 19.9–21.6 mm SL, Buriticupu municipality, Buritizinho river, Pindaré river drainage, Mearim river basin, 04°19'45"S, 46°29'46"W, 24 Jan. 2017, Guimarães E. C., Brito P. S. UFRJ11745, 1, 22.4 mm SL, Buriticupu municipality, Buritizinho river, Pindaré river drainage, Mearim river basin, 04°19'45"S, 46°29'46"W, 24 Jan. 2017, Guimarães E. C., Brito P. S.
The new species Hyphessobrycon caru sp. nov. differs from most of its congeners, except members of the rosy tetra clade, by the presence of a dark brown or black blotch on dorsal-fin (vs absence) and absence of a midlateral stripe on the body (vs presence).
The new species differs from most of its congeners in the rosy tetra clade by possessing few irregular inconspicuous vertically arranged chromatophores in the humeral region, or sometimes a very thin and inconspicuous humeral spot (Fig.
Furthermore, the new species differs from H. bentosi Durbin, 1908, H. erythrostigma, H. pyrrhonotus, H. rosaceus, and H. socolofi by presenting only one tooth in the outer row of premaxillary, and this unique tooth just slightly displaced from inner row [vs two or more teeth, displaced from the inner row]; from H. hasemani and H. micropterus by the dorsal-fin spot located approximately at the middle of the fin’s depth, not reaching its tip (vs spot located approximately at the middle of the fin’s depth, reaching its tip in adults); from H. hasemani by presenting tri to unicuspid teeth in the inner row of premaxillary and dentary (vs tricuspid or pentacuspid teeth); from H. piorskii by having the anal-fin profile usually nearly straight (vs anal-fin profile usually falcate). In addition, H. caru sp. nov. is easily distinguished from Pristella maxillaris (Ulrey, 1894), Moenkhausia hemigrammoides Géry, 1965, and Hemigrammus unilineatus (Gill, 1858) by the absence of a black oblique stripe or band on the anterior portion of the anal-fin (Fig.
Morphometric data of holotype and paratypes are presented in Table
Morphometric data (N = 45) of Hyphessobrycon caru sp. nov. SD: Standard deviation.
Holotype | Paratypes | Mean | SD | |
---|---|---|---|---|
Standard length | 22.2 | 14.8–25.4 | 18.9 | – |
Percentages of standard length | ||||
Depth at dorsal-fin origin (body depth) | 37.3 | 33.1–38.5 | 35.2 | 1.1 |
Snout to dorsal-fin origin | 53.7 | 49.4–55.0 | 51.7 | 1.2 |
Snout to pectoral-fin origin | 29.5 | 28.2–32.3 | 29.9 | 1.0 |
Snout to pelvic-fin origin | 46.0 | 43.6–48.8 | 45.6 | 1.0 |
Snout to anal-fin origin | 62.5 | 58.5–64.0 | 61.0 | 1.3 |
Caudal peduncle depth | 12.3 | 8.5–12.3 | 10.3 | 0.8 |
Caudal peduncle length | 11.7 | 9.5–12.7 | 11.2 | 0.8 |
Pectoral-fin length | 23.2 | 16.5–23.7 | 19.6 | 1.9 |
Pelvic-fin length | 20.6 | 14.1–20.5 | 17.4 | 1.4 |
Dorsal-fin base length | 15.2 | 12.9–15.7 | 14.3 | 0.8 |
Dorsal-fin height | 32.2 | 27.9–34.1 | 30.8 | 1.5 |
Anal-fin base length | 32.4 | 26.4–32.7 | 29.6 | 1.3 |
Eye to dorsal-fin origin | 37.5 | 34.4–38.8 | 37.3 | 0.9 |
Dorsal-fin origin to caudal-fin base | 55.1 | 50.6–56.1 | 53.4 | 1.1 |
Head length | 29.8 | 27.4–31.1 | 29.3 | 1.0 |
Percentages of head length | ||||
Horizontal eye diameter | 39.2 | 35.4–43.6 | 39.2 | 1.7 |
Snout length | 24.4 | 17.3–24.3 | 21.5 | 1.8 |
Least interorbital width | 29.1 | 22.4–30.7 | 27.2 | 1.8 |
Upper jaw length | 37.8 | 33.1–42.5 | 37.4 | 2.1 |
Premaxillary teeth in two rows. Outer row with one unicuspid or tricuspid tooth, just slightly displaced from inner row; inner row with 6(5), 7(6), or 8(1) tricuspid teeth and one unicuspid tooth. Maxilla with 3(2) tricuspid teeth and two unicuspid teeth, 4(3) tricuspid teeth and two unicuspid teeth or 5(7) tricuspid teeth. Dentary with five (10) or six (1) larger tricuspid teeth followed by one smaller tricuspid teeth 5(2), 6(2), 7(3), and 8(5) smaller unicuspid teeth (Fig.
Scales cycloid, three to eight radii strongly marked, circuli well-marked anteriorly, weakly marked posteriorly; lateral line incompletely pored, with 5(1), 6(2), 7(24), 8(14), or 9(4) perforated scales. Longitudinal scales series including lateral-line scales 31(1), 32(7), 33(14), 34(13), 35(3), or 36(7). Longitudinal scales rows between dorsal-fin origin and lateral line 5(3), 6(32), or 7(10). Horizontal scale rows between lateral line and pelvic-fin origin 4(43) or 5(2). Scales in median series between tip of supraoccipital spine and dorsal-fin origin 10(9), 11(12), 12(21), or 13(3). Circumpeduncular scales 11(6), 12(35), 13(2), or 14(2).
Dorsal-fin origin at midbody. Base of last dorsal-fin ray at vertical through first third of anal-fin. Dorsal-fin rays ii + 9(48), iii + 9(5), ii + 10(4). First dorsal-fin pterygiophore main body located behind neural spine of 4th vertebrae. Adipose-fin present. Anal-fin origin aligned with vertical line through middle of dorsal-fin, between 6th and 8th dorsal-fin rays base. Anteriormost anal-fin pterygiophore inserting posterior to haemal spine of 11th vertebrae. First anal-fin ray in vertical through the middle of dorsal-fin (with about 7th or 8th ray base). Anal-fin iii + 22(10) or iii + 23(47); anal-fin origin aligned with vertical line through middle of dorsal-fin (between base of 6th and 8th dorsal-fin rays); Anal-fin profile nearly straight; Anal-fin rays with a sexually dimorphic pattern, which is absent in females, described below. Pectoral-fin rays 12(57) total rays. Tip of pectoral-fin surpassing pelvic-fin base. Pelvic-fin rays 8(57) total rays, surpassing anal-fin origin. Pelvic-fin rays with a sexually dimorphic pattern, which are absent in females, described below. Caudal-fin forked, upper and lower lobes similar in size. Principal caudal-fin rays 11+10(50) or 10+9(7); dorsal procurrent rays 8(2), 9(8) or 11(2) and ventral procurrent rays 7(4) or 8(8).
Branchiostegal rays 4(12). First gill arch with 1(11), 2(1) hypobranchial, 11(1), 12(10), or 13(1) ceratobranchial, 1(12) on cartilage between ceratobranchial and epibranchial, and 5(1) or 6(11) epibranchial gill-rakers. Supraneurals 3(2), 4(9), or 5(1). Total vertebrae 28(2) or 29(10).
Ground coloration light yellowish brown. Humeral region with few irregular inconspicuous vertically arranged chromatophores, sometimes very thin and inconspicuous humeral spot. Flank with chromatophores homogeneously scattered, more concentrated on posterior region to humeral spot, posterior region of dorsal-fin base origin and below mid-portion of trunk, between anal-fin origin and caudal peduncle. Ventral region lacking dark-brown chromatophores. Dark-brown chromatophores present on head and more concentrated on dorsal portion, becoming sparser on cheek and preopercular regions.
Dorsal-fin ground coloration hyaline, with conspicuous black or dark-brown spot located on anterior portion of fin, reaching about 6th ray, approximately between one-half to two-thirds of fin depth. Anal and caudal-fins hyaline. Caudal-fin with a darker, usually dark brown, posterior margin and on its base. Adipose-fin hyaline to light brown, with dark-brown or black chromatophores more concentrated on its dorsal portion, depending on the specimen preservation state. Pectoral and pelvic-fins hyaline; pelvic-fin with variable amounts of dark-brown pigmentation remaining depending on the specimen preservation state.
Mature males with small bone hooks on anal and pelvic-fin rays. Bone hooks absent on females. Anal-fin presenting bone hooks from 3rd, 4th, or 5th rays to the last ray. Number of hooks variable, increasing from the first to the last rays. Pelvic-fin presenting 2nd, 3rd, 4th, or 5th rays with 5, 6, or 7 smaller hooks.
The specific epithet honors the term “Caru”. Caru is the name of an area (about 70.000 ha) inhabited by Brazilian native tribes from the ethnicities Guajá and Guajajara. People from this area use the Tupi language and have suffered consequences of European colonization and are under threat due to the pressure for exploration of the protected territory.
Hyphessobrycon caru sp. nov. has a restricted geographic distribution, being known only from the upper Pindaré river drainage, Mearim river basin, in the state of Maranhão, northeastern Brazil (Fig.
Hyphessobrycon caru sp. nov. belongs to the H. micropterus clade possessing 20 synapomorphic nucleotide substitutions: COI 73 (C→T), COI 88 (T→C), COI 217 (C→T), COI 274 (C→T), COI 298(C→T), COI 334 (C→G), COI 338 (T→C), COI 370 (A→G), COI 418 (A→G), COI 433 (C→T), COI 439 (C→A), COI 457 (A→G), COI 469 (T→C), COI 478 (A→T), COI 559 (A→G), COI 562 (T→A), COI 592(A→G), COI 631 (A→T), COI 655 (A→C), COI 673 (A→C). It shares nine synapomorphic nucleotide substitutions with H. piorskii, which separate them from H. simulatus and H. micropterus: COI 181 (A→C), COI 208 (A→G), COI 245 (C→T), COI 325 (T→C), COI 349 (T→C), COI 436 (A→T), COI 472 (A→G), COI 538 (C→T), COI 556 (T→C). In addition, it has six unique nucleotide substitutions within the H. micropterus clade: COI 148 (C→T), COI 154 (C→T), COI 175 (T→C), COI 364 (G→A), COI 487 (T→C), COI 517 (A→G) (Fig.
Phylogenetic tree based on Bayesian Inference (BI). Numbers above branches are posterior probability values. Posterior probability value supporting the Hyphessobrycon micropterus clade is indicated in green (haplotypes marked with a green bar); posterior probability value supporting the H. caru sp. nov. lineage under WP method is indicated in red (haplotypes marked with a red bar); and the other species (lineages) under WP method, within this clade, are indicated in black. b Strict consensus phylogenetic tree based on Maximum Parsimony (MP), obtained from the 38 most parsimonious trees, in which 587 characters were constant, 20 variable but parsimony-uninformative, and 248 parsimony-informative (total length 833, consistency index 0.489, retention index 0.901). The image is focusing on the Hyphessobrycon micropterus clade. Numbers above branch are bootstrap values and letters below branches correspond to nucleotide substitutions, listed in Suppl. material
COI sequences support the existence of a new species of Hyphessobrycon inhabiting the Pindaré river basin in Maranhão state. After trimming, the final alignment yielded 680 base pairs with 159 polymorphic sites and 26 haplotypes. Average genetic distances were 18.3%, with the highest values between H. epicharis and H. erythrostigma (23.4%), while the lowest value (0.7%) was between H. pyrrhonotus and H. erythrostigma (Table
1 | 2 | 3 | 4 | 5 | 6 | 7 | 8 | 9 | 10 | 11 | 12 | 13 | 14 | 15 | 16 | ||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
1 | H. erythrostigma | ||||||||||||||||
2 | H. pyrrhonotus | 0.007 | |||||||||||||||
3 | H. socolofi | 0.037 | 0.035 | ||||||||||||||
4 | H. simulatus | 0.192 | 0.194 | 0.179 | |||||||||||||
5 | H. micropterus | 0.205 | 0.202 | 0.198 | 0.041 | ||||||||||||
6 | H. piorskii | 0.188 | 0.185 | 0.178 | 0.063 | 0.064 | |||||||||||
7 | H. caru | 0.206 | 0.210 | 0.203 | 0.062 | 0.057 | 0.036 | ||||||||||
8 | H. eques | 0.175 | 0.178 | 0.163 | 0.154 | 0.157 | 0.160 | 0.160 | |||||||||
9 | H. copelandi | 0.192 | 0.189 | 0.186 | 0.170 | 0.176 | 0.158 | 0.168 | 0.102 | ||||||||
10 | H. epicharis | 0.234 | 0.230 | 0.220 | 0.170 | 0.175 | 0.189 | 0.190 | 0.187 | 0.197 | |||||||
11 | H. bentosi | 0.106 | 0.103 | 0.112 | 0.197 | 0.194 | 0.205 | 0.204 | 0.195 | 0.220 | 0.219 | ||||||
12 | H. cf. sweglesi | 0.210 | 0.207 | 0.197 | 0.182 | 0.187 | 0.201 | 0.209 | 0.181 | 0.197 | 0.030 | 0.222 | |||||
13 | P. maxillaris | 0.225 | 0.232 | 0.206 | 0.201 | 0.211 | 0.220 | 0.218 | 0.194 | 0.213 | 0.180 | 0.202 | 0.183 | ||||
14 | M. hemigrammoides | 0.219 | 0.219 | 0.209 | 0.179 | 0.185 | 0.178 | 0.179 | 0.203 | 0.211 | 0.196 | 0.221 | 0.199 | 0.166 | |||
15 | H. compressus | 0.214 | 0.218 | 0.215 | 0.202 | 0.208 | 0.198 | 0.208 | 0.198 | 0.201 | 0.212 | 0.212 | 0.212 | 0.203 | 0.215 | ||
16 | H. panamensis | 0.210 | 0.213 | 0.209 | 0.202 | 0.199 | 0.179 | 0.187 | 0.215 | 0.229 | 0.221 | 0.213 | 0.218 | 0.204 | 0.208 | 0.145 | |
17 | H. flammeus | 0.201 | 0.201 | 0.198 | 0.169 | 0.174 | 0.186 | 0.204 | 0.203 | 0.205 | 0.192 | 0.200 | 0.189 | 0.206 | 0.200 | 0.170 | 0.196 |
Both phylogenetic analysis based on BI and MP supported a clade comprising H. caru sp. nov., H. micropterus, H. piorskii, and H. simulatus, hereafter termed Hyphessobrycon micropterus clade, with maximum posterior probability value and 99% bootstrap value in BI and MP, respectively. Hyphessobrycon caru sp. nov. formed a single exclusive lineage with maximum posterior probability value (posterior probability = 1) and 99% bootstrap value in BI and MP, respectively.
These species delimitation analysis (WP and CBB) have identical results, delimitating four species within the Hyphessobrycon micropterus clade: H. caru sp. nov., H. micropterus, H. piorskii, and H. simulatus (Fig.
This species delimitation analysis also indicates four lineages (species) within the Hyphessobrycon micropterus clade: H. caru sp.n., H. micropterus, H. piorskii, and H. simulatus (Fig.
Currently molecular techniques are frequently useful for solve species complexes and discover cryptic species (e.g.
The large number of the described Hyphessobrycon species (about 160 spp.), with new species described every year, reveal an astonishing diversity within the genus. During the past 10 years, about 50 new species have been described (
Our results suggest a cryptic speciation in the rosy tetra clade, more specifically in a new clade here defined, the Hyphessobrycon micropterus clade, including H. caru sp. nov., H. micropterus, H. piorskii, and H. simulatus, so far only known from the Pindaré, Itapecuru, Munim, Preguiças, and São Francisco river drainages of Brazil and the coastal river basins of French Guiana and Suriname (
Hyphessobrycon caru sp. nov. is herein described within the Hyphessobrycon micropterus clade based on five different and independent methods of species delimitation (PAA, DBC, WP, CBB and bPTP), characterized by different criteria and assumptions. Hyphessobrycon caru sp. nov. is distinguished from all its congeners by a combination of unambiguous morphological character states [see Diagnosis (PAA)]. In our Bayesian phylogenetic analysis (Fig.
Hyphessobrycon amandae : UFRJ 1557, 5 spcms, Jussara municipality, Goiás state, Brazil. H. bentosi: MCZ 20842, 1 spcm (Syntype), Óbidos municipality, Pará state, Brazil. H. bifasciatus: UFRJ 0068, 6 spcms, Marataízes and Guarapari municipality, Espírito Santo state, Brazil. H. compressus: BMNH 1905.12.6.4-5, 2 spcms (Paratypes), Oaxaca state. México. H. copelandi: CAS 42683, 1 spcm (Syntype); MCZ 20771, 1 spcm (Syntype), Tabatinga municipality, Amazonas state, Brazil. H. eques: CICCAA 00715, 4 spcms (C&S); CICCAA 00710, 51 spcms, Tombos municipality Carangola river, Minas Gerais state, Brazil. H. erythrostigma: ANSP 70208, 1 spcm (Holotype), Peru and Brazil. H. epicharis: FMNH100609, 1 spcm (Paratype), Baria river, Amazonas, Venezuela. H. haraldschultzi: CICCAA 00873, 20 spcms, Ilha do Bananal municipality, Javaés river, Tocantins state, Brazil. H. hasemani: ANSP 39230, 1 spcm (Holotype), Guajaramirim municipality, Madeira river, Rondônia state, Brazil. H. micropterus: FMNHH 57916, 1 spcm (Holotype), São Francisco river at Lagoa de Porto, Minas Gerais state, Brazil. H. piorskii: CICCAA 00695, 1 spcm (Holotype); CICCAA 00430, 15 spcms (Paratype); CICCAA 00431, 21 spcms (Paratype); CICCAA 00696, 15 spcms (Paratype); CICCAA 00697, 16 spcms (C&S) (Paratype); CICCAA 00698, 6 spcms, 1 spcm (C&S) (Paratype); CICCAA 00750, 9 spcms (Paratype); CICCAA01654, 1 spcm (Paratype); CPUFMA 171664, 15 spcms (Paratype); UFRJ 11553, 6 spcms (Paratype), stream at the Anapurus municipality, Munim river, Maranhão state, Brazil. CICCAA 00089, 1 spcm (C&S) (Paratype); CICCAA 00881, 1 spcm (Paratype); CICCAA 01563, 1 spcm (Paratype); stream at Mata de Itamacaoca, Chapadinha municipality, Munim river, Maranhão state, Brazil. CICCAA 01382, 5 spcms (Paratype); CICCAA 02008, 12 (C&S) spcms (Paratype), stream at Mata Fome, Barreirinhas municipality, Preguiças river, Maranhão state, Brazil. H. pyrrhonotus: MZUSP 45714, 1 spcm (Holotype), Ereré river, Brazil. H. rosaceus: FMNH 52791, 1 spcm (Holotype), Gluck Island, Essequibo River, Guyana. H. werneri: MZUSP 42365, 1 spcm (Holotype), Santa Maria do Pará and São Miguel de Guamá municipality, Guamá river, Pará state, Brazil. CICCAA 00751, 1 spcm, Paragominas municipality, Candiru river, Pará state, Brazil. H. socolofi: MZUSP 13181, 1 spcm (Holotype), Barcelos municipality, Negro river, Amazonas state, Brazil.
We thank Axel Makay Katz, Axel Zarske, Flávio Lima, Ingo Schindler, Oscar Lasso-Alcalá and Ronald Fricke for providing useful literature; Vale S.A and Amplo Engenharia for the cession of part of the data analysed in this study; Raphael Covain and collaborators of the Project Gui-BOL Barcoding Guianese fishes for the cession of part of the data analysed in this study; Marcelo Rodrigues dos Anjos and Wilson Costa for the loan and donation of material; Mark Sabaj Pérez (CAS), James Maclaine (FMNH), Riedel Bettina (NMW) for providing photographs, x-ray images, and information on the type material of some species; FACEPE for providing the scholarship to LMF under the process IBPG-0089-2.05/17, CAPES and FAPEMA for providing the scholarship to PSB under the process 88887.159561/2017-00. This paper benefited from suggestions provided by P. Bragança and R. Collins. This study was supported by the project “PROCESSO UNIVERSAL-00724/17” from FAPEMA (Foundation for Scientific Research and Development of Maranhão). All material was collected with permits 51540-3/ from SISBIO (Brazilian Institute of Environment and Natural Resources).
Box 1
Data type: DOCX file
Explanation note: List of nucleotide substitutions (synapomorphies and autapomorphies) from each lineage (species) and some crucial points of the cladogram of the Fig.