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Molecular Identification of Leporinus from the South Portion of South America

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Molecular Identification of Leporinus from the South Portion of South America DNA Barcodes 2015; Volume 3: 98–109 Research Article Open Access Gleisy S. Avelino*, Heraldo A. Britski, Fausto Foresti, Claudio Oliveira Molecular identification of Leporinus from the south portion of South America DOI 10.1515/dna-2015-0013 Received January 30, 2015; accepted July 6, 2015 1 Introduction Abstract: Anostomidae is a widely distributed group Anostomidae is a family of freshwater fishes from South that includes the species rich Leporinus. This genus America that ranges from the Atrato River, western is widespread in South America, including its south Colombia, down south to central portions of Argentina portion where they are mainly distributed in the São [1]. A hypothesis of phylogenetic relationships within Francisco and La Plata basins. Here we obtained the Anostomidae was recently proposed by Sidlauskas & sequences of mitochondrial and nuclear genes of 19 Vari [2], who recognize that the family includes fourteen species of Leporinus with the objective of testing our genera. Leporinus is the largest genus of Anostomidae methods of molecular species identification and to both in number of species and number of individuals test the similarities between species of Leporinus of in the rivers where they occur [1,3]; because of this, the São Francisco and La Plata basins. Results show several attempts were made trying to divide the species that two species pair have a genetic distance below of Leporinus in smaller groups. Fowler [4] created the 2% (L. paranensis x L. obtusidens and L. paranensis x subgenus Myocharax, based on the dentition presented L. aff. paranensis). Our results also show that a high by Leporinus desmotes, which shows a very long pair genetic distance occurs in L. obtusidens and L. striatus of symphyseal teeth. Borodin [5] attempted to divide suggesting a possible local differentiation of these the genus Leporinus in two subgenera and created species. An interesting finding of our study is that the subgenus Hypomasticus that included species the genetic similarities observed among species does with a lower mouth. Another subgenus, Leporinops, not corroborate the division of species of Leporinus in was created by Géry [6], based on tooth and cranial groups according to color patterns. Additionally, our differences. This subgenus was based on Leporinus results show that the species of Leporinus from the La moralesi [7]. All these proposals were not accept by Plata and São Francisco are not more similar among Garavello & Britski [1] in the catalog of species of the themselves than to species of other drainages which family but Hyposmastictus was recognized as valid by could be explained by the occurrence of recent fauna Sidlauskas & Vari [2]. exchanges between these two basins and adjacent Garavello & Britski [8], in a paper describing Leporinus basins. amblyrhynchus and L. paranensis from the upper Paraná Basin, showed that most species of the genus may be Keywords: Phylogeny, Molecular systematics, placed into three artificial groups based on the color Sequencing, Mitochondrial DNA, Fish. pattern of the trunk: 1) barred species, 2) striped species and 3) spotted species. The number of teeth is also a diagnostic feature for the species of the genus and is often *Corresponding author: Gleisy S. Avelino, Departamento de used in identification keys. With the exception of the Morfologia, Instituto de Biociências, Universidade Estadual Paulista recently described species Leporinus venerei [9], which “Júlio de Mesquita Filho” (UNESP), Caixa Postal 510, Rubião Júnior, has the dental formula 4/3, unique among species of s/n, 18618-970, Botucatu, SP, Brazil, E-mail: [email protected] the genus, all species are artificially placed into three Fausto Foresti, Claudio Oliveira, Departamento de Morfologia, Instituto de Biociências, Universidade Estadual Paulista “Júlio de groups according to the dental formula (number of Mesquita Filho” (UNESP), Caixa Postal 510, Rubião Júnior, s/n, tooth in premaxillary/dentary in each half of the jaw): 18618-970, Botucatu, SP, Brazil 3/3, 3/4 or 4/4 [9]. Heraldo A. Britski, Museu de Zoologia da Universidade de São Pau- Garavello & Britski [1] recognized nine species of lo, Caixa Postal 42494, 04299-970, São Paulo, SP, Brazil Leporinus in the La Plata Basin, four species in the São © 2015 Gleisy S. Avelino et al. licensee De Gruyter Open. This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivs 3.0 License. - 10.1515/dna-2015-0013 Downloaded from De Gruyter Online at 09/13/2016 04:12:13PM via free access Molecular identification of Leporinus from the south portion of South America 99 Francisco River Basin, and two species were reported for 2 Material and Methods both basins. Britski et al. [10] described a new species for the La Plata Basin (L. piavussu), recognized that L. Eighteen species of Leporinus (73 samples) were used, out elongatus is endemic to the Rio Jequitinhonha and Rio of which 40 samples were from the Parana River Basin, 9 Pardo (coastal rivers) and recognize L. aguapeiensis and L. samples from the Paraguay River Basin, 1 sample from the silvestrii as junior synonyms of L. obtusidens. Uruguay River Basin, 12 samples from the São Francisco Considering the previous hypothesis of closer River Basin and 8 samples from the coastal Brazilian rivers. relationships between the fish fauna of the São Francisco Additionally 1 sample from the Amazonas River Basin and and La Plata basins (including the Parana, Paraguay 2 samples from the Orinoco River Basin were analyzed and Uruguay rivers) [11], the widespread distribution of as comparative material. Species, collecting sites, and Leporinus in these basins [1,10] and in the south portion of museum collection numbers are shown in Table 1. Figure South America and the economical importance of species 1 shows the collecting sites of the samples obtained from of this genus, in the present study, partial sequences of the south part of South America. One species, from rio the mitochondrial genes 16S rRNA, Cytochrome B (CytB) Araguaia, could not be positively identified and was and Cytochrome c Oxidase I (COI) and of the nuclear gene named Leporinus sp. and one species was identified as α-Tropomyosin (Trop) from 19 species of Leporinus were L. aff. paranensis because it differs from L. paranensis in used to test the hypothesis that the current morphological the number of series of peduncule scales (12 vs. 16). recognized species in south portion of South America Tissue samples were taken from all specimens basins can be correctly identified using DNA markers and investigated for DNA isolation (preserved in 95% ethanol) to test the similarities between species from São Francisco and the specimens were fixed in 10% formaldehyde and La Plata basins. and preserved in 70% ethanol. The animals studied are Table 1. Specimens sequenced in the present study and Genbank sequence numbers. Voucher specimens are deposited in the fish collec- tion of Laboratório de Biologia e Genética de Peixes (LBP), Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil. Species LBP Locality Code Specimen Genes number 16S Cyt b COI Trop Leporinus 3505 Rio Itararé/Rio Paranapanema/ PA2 20121 EU181625 EU183041 EU185541 EU181641 amblyrhynchus Fartura/SP/S 23°24’44.9’’ W 49°34’15.4’’ 3917 Reservatório de Chavantes/Rio PA2 21828 EU181626 EU183046 EU185542 EU181642 Paranapanema/Chavantes/SP/S PA2 21829 EU181627 - EU185543 EU181644 23°08’01.1’’ W 49°40’34.4’’ PA2 21830 EU181628 EU183045 EU185544 EU181643 Leporinus 45851 Rio Capivari/Rui Barbosa/BA/S BA bahiensis 12°10’45’’S 40°24’16’’W Leporinus 2380 Lagoa Feia/Rio Paraíba do Sul/ EC2 16074 EU181599 EU183015 EU185593 EU181688 copelandii Leste/Campos dos Goytacazes/ RJ/S 22°00’ W 41°20’ Leporinus cf. 3077 Rio Orinoco/Rio Orinoco/Caicara OR 19698 EU181613 EU183027 EU185598 - fasciatus del Orinoco/Bolívar/Venezuela/N OR 19699 EU181614 - EU185599 - 07°38’11.6’’ W 66°19’04.2’’ Leporinus 655 Região de Rombado, tributary PY1 8087 EU181574 EU182993 EU185563 EU181659 friderici of the rio Pirai/Poconé/MT/S 16°25’40.8’’ W 56°25’08.58’’ 3179 Reservatório de Jurumirim/Rio PA2 16873 EU181602 EU183019 EU185551 EU181677 Paranapanema/Itatinga/SP/S 23°20’ W 48°34’ Leporinus 2340 Rio Tietê/Rio do Peixe/Rio PA3 15940 EU181638 EU183013 - - lacustris Paraná/Bofete/SP/S 22°46’29.9’’ PA3 15941 EU181598 EU183014 EU185566 - W 48°08’43.5’’ 4058 Região de Rombado, tributary PY1 8086 EU181573 EU182992 EU185562 EU181658 of rio Pirai/Poconé/MT/S 16°25’40.8’ W 56°25’08.6’’ - 10.1515/dna-2015-0013 Downloaded from De Gruyter Online at 09/13/2016 04:12:13PM via free access 100 G.S. Avelino, et al. contiunedTable 1. Specimens sequenced in the present study and Genbank sequence numbers. Voucher specimens are deposited in the fish collection of Laboratório de Biologia e Genética de Peixes (LBP), Instituto de Biociências, Universidade Estadual Paulista, Botucatu, São Paulo, Brazil. Species LBP Locality Code Specimen Genes Leporinus 1422 Rio Taquari/Rio Paraguai/Coxim/ PY3 12505 EU181590 EU183005 EU185582 EU181664 macrocephalus MS PY3 19492 - EU183039 EU185583 EU181662 S 18°25’42.5’’ W 54°50’02.8’’ PY3 19493 EU181612 EU183040 EU185584 EU181663 Leporinus 2491 Rio Montividiu/Rio Paraná/ PA5 16370 EU181634 EU183016 EU185585 - microphthalmus Montividiu/GO/S 17°26’25.8’’ W 51°10’26.7’’ Leporinus 250 Rio São Francisco/Três SF1 4211 EU181567 EU182984 EU185545 EU181648 obtusidens Marias/MG/S 18°11’28.5’’ W SF1 4212 EU181568 EU182985 EU185546 EU181646 45°14’51.42’’ 8091 Rio São Francisco/Três Marias/
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