Molecular phylogeny of the highly diversified catfish subfamily Loricariinae (Siluriformes, Loricariidae) reveals incongruences with morphological classification

Accepted Manuscript Molecular phylogeny of the highly diversified catfish subfamily Loricariinae (Siluriformes, Loricariidae) reveals incongruences with morphological classification Raphaël Covain, Sonia Fisch-Muller, Claudio Oliveira, Jan H. Mol, Juan I. Montoya-Burgos, Stéphane Dray PII: DOI: Reference:

S1055-7903(15)00321-8 http://dx.doi.org/10.1016/j.ympev.2015.10.018 YMPEV 5334

To appear in:

Molecular Phylogenetics and Evolution

Received Date: Revised Date: Accepted Date:

24 February 2015 15 September 2015 19 October 2015

Please cite this article as: Covain, R., Fisch-Muller, S., Oliveira, C., Mol, J.H., Montoya-Burgos, J.I., Dray, S., Molecular phylogeny of the highly diversified catfish subfamily Loricariinae (Siluriformes, Loricariidae) reveals incongruences with morphological classification, Molecular Phylogenetics and Evolution (2015), doi: http:// dx.doi.org/10.1016/j.ympev.2015.10.018

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Molecular phylogeny of the highly diversified catfish subfamily Loricariinae (Siluriformes, Loricariidae) reveals incongruences with morphological classification.

Raphaël Covain 1*, Sonia Fisch-Muller 1, Claudio Oliveira 2, Jan H. Mol 3, Juan I. MontoyaBurgos 4 & Stéphane Dray 5

1

Muséum d’histoire naturelle, Département d’herpétologie et d’ichtyologie, route de

Malagnou

1,

case

postale

6434,

CH-1211

Genève

6,

Switzerland;

e-mail:

[email protected]; [email protected] 2

Departamento de Morfologia, Universidade Estadual Paulista Júlio de Mesquita Filho,

Instituto de Biociências, Laboratório de Biologia e Genética de Peixes, Rubião Junior 18618970, Botucatu, SP, Brazil; e-mail: [email protected] 3

University of Suriname, Center for Agricultural Research in Suriname, CELOS and

Department of Biology, POB 9212, Paramaribo, Suriname; e-mail: [email protected] 4

Université de Genève, Département de Génétique et Evolution, Sciences III, quai E.

Ansermet 30, CH-1211 Genève 4, Switzerland; e-mail: [email protected] 5

Université de Lyon, F-69000, Lyon ; Université Lyon 1 ; CNRS, UMR5558, Laboratoire de

Biométrie

et

Biologie

Evolutive,

F-69622,

Villeurbanne,

France;

e-mail:

[email protected]

1

Abstract The Loricariinae belong to the Neotropical mailed catfish family Loricariidae, the most species-rich catfish family. Among loricariids, members of the Loricariinae are united by a long and flattened caudal peduncle and the absence of an adipose fin. Despite numerous studies of the Loricariidae, there is no comprehensive phylogeny of this morphologically highly diversified subfamily. To fill this gap, we present a molecular phylogeny of this group, including 350 representatives, based on the analysis of mitochondrial and nuclear genes (8426 positions). The resulting phylogeny indicates that Loricariinae are distributed into two sister tribes: Harttiini and Loricariini. The Harttiini tribe, as classically defined, constitutes a paraphyletic assemblage and is here restricted to the three genera Harttia, Cteniloricaria, and Harttiella. Two subtribes are distinguished within Loricariini: Farlowellina and Loricariina. Within Farlowellina, the nominal genus formed a paraphyletic group, as did Sturisoma and Sturisomatichthys. Within Loricariina, Loricaria, Crossoloricaria, and Apistoloricaria are also paraphyletic. To solve these issues, and given the lack of clear morphological diagnostic features, we propose here to synonymize several genera (Quiritixys with Harttia; East Andean members of Crossoloricaria, and Apistoloricaria with Rhadinoloricaria; Ixinandria, Hemiloricaria, Fonchiiichthys, and Leliella with Rineloricaria), to restrict others (Crossoloricaria, and Sturisomatichthys to the West Andean members, and Sturisoma to the East Andean species), and to revalidate the genus Proloricaria.

Keywords: Systematics, Neotropics, mitochondrial genes, nuclear genes, freshwaters, Amazon basin

1. Introduction The Loricariinae represent a highly diversified subfamily among the large Neotropical catfish family Loricariidae, or suckermouth armored catfish. Loricariids have undergone an evolutionary radiation at a subcontinental scale, from Costa Rica to Argentina, which has

2

been compared to that of the Cichlidae of the Great Lakes of the Rift Valley in Africa (Schaefer and Stewart, 1993). The species core flock Loricariidae (sensu Lecointre et al. (2013), represents the most species rich family of the Siluriformes with 898 valid species and an estimated 300 undescribed species distributed in more than 100 genera (Reis et al., 2003; Ferraris, 2007; Eschmeyer and Fong, 2015). Extremely variable color patterns and body shapes among loricariid taxa reflect their high degree of ecological specialization, and because of their highly specialized morphology loricariids were recognized as a monophyletic assemblage in the earliest classifications of the Siluriformes (de Pinna, 1998). The Loricariidae are characterized by a depressed body covered by bony plates, and above all, by the modification of the mouth into a sucker disk. Within the Loricariidae, members of the subfamily Loricariinae are diagnosed by a long and depressed caudal peduncle and by the absence of an adipose fin. They live stuck to the substrate and show marked variations in body shape according to the various habitats colonized, from lotic to lentic systems, on mineral or organic substrates. For example, members of Farlowella resemble a thin stick and blend remarkably among submerged wood and leafs, whereas members of Pseudohemiodon are large and flattened and bury themselves in sandy substrates like flatfishes. Some groups have numerous teeth, pedunculated, and organized in a comblike manner, while other groups have few teeth or even no teeth on the premaxillae. Teeth are often strongly differentiated, and can be bicuspid straight and thick, spoon-shaped, reduced in size or very long. An important diversity in lip characteristics, which can be strongly papillose, filamentous or smooth, also characterizes this subfamily (Isbrücker, 1979; Covain and Fisch-Muller, 2007). Different hypotheses have been proposed to classify the Loricariinae (summarized in Table 1). The first attempt was performed by Isbrücker (1979) who distributed them into four tribes and eight subtribes on the basis of external morphology, but without phylogenetic inferences. These included the Loricariini, comprising six subtribes (Loricariina,

3

Planiloricariina, Reganellina, Rineloricariina, Loricariichthyina, and Hemiodontichthyina), the Harttiini, including two subtribes (Harttiina and Metaloricariina), the Farlowellini, and the Acestridiini. The latter were subsequently placed in the subfamily Hypoptopomatinae by Schaefer (1991). In her PhD thesis, Rapp Py-Daniel (1997) proposed a phylogeny of the Loricariinae based on a phylogenetic analysis of morphological characters. She confirmed the monophyly of the subfamily, and split the Loricariinae into two tribes: Loricariini and Harttiini, the latter comprising Farlowellini (sensu Isbrücker, 1979). In a morphological phylogenetic analysis of the family Loricariidae, Armbruster (2004) also obtained a similar splitting based on a restricted sampling of the Loricariinae, with Harttiini (sensu Rapp PyDaniel, 1997, comprising Harttia, Lamontichthys, Sturisoma, and Sturisomatichthys), forming the sister group of Loricariini (sensu Rapp Py-Daniel, 1997, comprising Crossoloricaria, Loricaria, Loricariichthys, Ixinandria, and Rineloricaria). Covain and Fisch-Muller (2007), based on multivariate analyses of generic diagnostic characters, also split the subfamily into two tribes, the Harttiini and the Loricariini, and proposed four morphological groups within the Loricariini: (1) the Pseudohemiodon group, (2) the Loricaria group, (3) the Rineloricaria group, and (4) the Loricariichthys group. Montoya-Burgos et al. (1998) proposed a first molecular phylogeny of the family Loricariidae using mitochondrial genes. Although, their analysis included only nine representatives of the Loricariinae, they partially confirmed their subdivision into two main groups, but with Harttia (nominal genus of Harttiini) forming the sister genus of all other Loricariinae (comprising Farlowellini, part of Harttiini, and Loricariini). Covain et al. (2008) using mitochondrial genes, and Rodriguez et al. (2011) using mitochondrial and nuclear markers performed a molecular phylogeny on a small sampling of the Loricariinae. Both studies restricted Harttiini to Harttia, and included Farlowelliini as a subtribe of Loricariini. Within the latter, the Loricariichthys and LoricariaPseudohemiodon morphological groups (sensu Covain and Fisch-Muller, 2007) were

4

confirmed as natural groupings, whereas monophyly of the Rineloricaria group was rejected. Similar results were also obtained using different markers by Cramer et al. (2011) in a molecular phylogeny of the Hypoptopomatinae (including four Loricariinae in the outgroup), and by Lujan et al. (2015) in a molecular phylogeny of the Loricariidae (including 14 Loricariinae). In addition to the conflicting results obtained using either morphological or molecular data, the validity of several genera was regularly questioned by different authors rendering the taxonomy of the group confused. Isbrücker and Isbrücker & Michels (in Isbrücker et al. 2001) described four new genera: Fonchiiichthys, Leliella, Quiritixys and Proloricaria, and revalidated the genus Hemiloricaria Bleeker, 1862 on the basis of a very restricted number of characters. Rapp Py-Daniel and Oliveira (2001) considered Cteniloricaria a junior synonym of Harttia. Ferraris (2003) maintained the validity of Cteniloricaria, and considered junior synonyms of already described genera all the genera described by Isbrücker and Isbrücker & Michels (in Isbrücker et al. 2001). Covain and FischMuller (2007) followed Ferraris (2003) but maintained Cteniloricaria in synonymy with Harttia. Ferraris (2007) modified his previous statement and considered Fonchiiichthys, Proloricaria, and Hemiloricaria valid. Later on, Covain et al. (2012) revalidated Cteniloricaria. There are currently 239 species of Loricariinae considered valid, distributed in 32 genera (for a review see Covain and Fisch-Muller, 2007; also Ghazzi, 2008; Ingenito et al., 2008; Fichberg and Chamon, 2008; Rapp Py-Daniel and Fichberg, 2008; Rodriguez and Miquelarena, 2008; Rodriguez and Reis 2008; Rodriguez et al., 2008; Thomas and Rapp PyDaniel, 2008; de Carvalho Paixão and Toledo-Piza, 2009; Thomas and Sabaj Pérez, 2010; Rodriguez et al., 2011; Covain et al., 2012; Rodriguez et al., 2012; Vera-Alcaraz et al., 2012; Oyakawa et al., 2013; Thomas et al., 2013; Ballen and Mojica, 2014; Fichberg et al., 2014; Londoño-Burbano et al., 2014).

5

Given the confused systematics of the Loricariinae, we present a comprehensive and robust molecular phylogeny of this group based on mitochondrial and nuclear genes. The resulting phylogeny, in conjunction with morphological diagnostic characters, will be subsequently used to (1) redefine the tribal and subtribal ranks of the subfamily; (2) evaluate the validity and monophyly of the different genera, and (3) test alternative hypotheses of classification proposed in the literature.

2. Material and methods 2.1 Taxonomic sampling. The molecular phylogeny was reconstructed using the taxonomic sampling given in Covain et al. (2008), and Rodriguez et al. (2011) with the addition of 326 representatives of the Loricariinae and 16 outgroup species. The outgroup representatives were chosen in other subfamilies of the Loricariidae. The list of material used for this study is provided in Table 2. The analyzed samples came from the tissue collection of the Muséum d’histoire naturelle de la Ville de Genève (MHNG); Academy of Natural Sciences of Drexel University in Philadelphia (ANSP); Smithsonian Tropical Research Institute (STRI), Panama; Laboratório de Biologia de Peixes, Departamento de Morfologia, Universidade Estadual Paulista, Campus de Botucatu (LBP); Auburn University Museum, Montgomery (AUM); and Museu de Ciências e Tecnologia of the Pontifícia Universidade Católica do Rio Grande do Sul (MCP), Porto Alegre. The sequences were deposited in GenBank.

2.2 DNA extraction, choice of markers, amplification, and sequencing. Tissue samples were preserved in 80% ethanol and stored at -20°C. Total genomic DNA was extracted with the DNeasy Tissue Kit (Qiagen) following the instructions of the 6

manufacturer. The choice of markers was governed by their ability to resolve inter generic relationships at subfamilial ranks. We thus selected the mitonchondrial genes 12S and 16S for the resolution of phylogenetic relationships between close ralatives (between species to between genera relationships), and the nuclear Fish Reticulon-4 receptor (f-rtn4r) gene composed of two introns and three exons. Exons of this marker are rather conserved and provide information for deeper relationships (intra-familial to inter-ordinal relationships) whereas intronic regions are more variable and offer the possibility to investigate phylogenetic relationships between closely related species. Thus, the selected molecular makers were used to examine a range of taxomonic levels at subfamilial rank, from intraspecific to inter-tribal relationshisps. The PCR amplifications of mitochondrial 12S and 16S, and the nuclear f-rtn4r genes were carried out using the Taq PCR Core Kit (Qiagen). The methodology for PCR amplifications followed Chiachio et al. (2008) for f-rtn4r using the set of primers Freticul4-D, Freticul4-R, Freticul4 D2, and Freticul4 R2 (Roxo et al., 2014). For the complete sequencing of f-rtn4r, three internal primers were designed additionally to Freticul4-iR (Roxo et al., 2014): Freticul4-iD2 5’- CAA CAT CAC YTG GAT TGA GG -3’, Freticul4-LiD 5’- ATG ACC GTG AGC TGC CAG GC -3’, and Freticul4-LiR 5’- GCT CAG TAA TAC GGT TGT TCT GCA -3’. To amplify the almost complete 12S, tRNAval and 16S mitochondrial genes in a single 2,500 bp long fragment, a Nested PCR protocol was used. The external round of PCR was performed using the pair of primers Phe-L941 (Roxo et al., 2014) and H3059 (Alves-Gomes et al., 1995). The external amplifications were performed in a total volume of 50 μl, containing 5 μl of 10x reaction buffer, 1 μl of dNTP mix at 10mM each, 1 μl of each primer at 10 μM, 0.2 μl of Taq DNA Polymerase equivalent to 1 unit of Polymerase per tube, and 1 to 4 μl of DNA. Cycles of amplification were programmed with the following profile: (1) 3 min. at 94°C (initial denaturing), (2) 35 sec. at 94°C, (3) 30 sec. at 51°C, (4) 150 sec. at 72°C, and (5) 5 min. at 72°C (final elongation). Steps 2 to 4 were repeated 35 to 39

7

times according to the quality and concentration of DNA. The internal round of PCR was performed using 1 μl of DNA template sampled from external round PCR product, the pair of primers: An12S-1D: 5’- GTA TGA CAC TGA AGA TGT TAA G -3’ and iH3059: 5’- GAA CTC AGA TCA CGT AGG -3’, and the same protocol as above except for the annealing temperature that was set to 54°C. PCR products were sent to Macrogen Inc. (Seoul, Korea) for sequencing. For the complete sequencing of the 2,500 bp long mitochondrial fragment, two internal primers were used: Lor1D-1D: 5’- AGG AGC CTG TTC TAG AAC CG-3’ and Lor12S-3D (Covain et al. 2008) for a walking sequencing procedure with around 700 bp between each step.

2.3 Sequence alignment, phylogenetic reconstruction, and topological tests. The DNA sequences were edited and assembled using BioEdit 7.0.1 (Hall, 1999), aligned using ClustalW (Thompson et al., 1994) and final alignment optimized by eye. Regions with ambiguous alignments in loop regions of mitochondrial ribosomal genes were excluded from the analyses. The fragment of the f-rtn4r gene analyzed here contained relatively long introns, ranging from 489 bp in Rineloricaria altipinnis to 1385 bp in R. cf. latirostris for the first intron, and from 373 bp in R. pentamaculata to 656 bp in Harttiella lucifer for the second, with several species dependent indels that locally challenged the alignment process. To reduce the misleading effects of misaligned regions, we used the model alignment for this marker given by Rodriguez et al. (2011) that produced good assessment statistics according to SOAP 1.2a4 (Löytynoja and Milinkovitch, 2001), better than automatic alignments produced by CLUSTAL-X 1.83 under various parameter values (i.e. increase in mean nodal support, model adequacy, and tree balance). Moreover, and despite the presence of indels in its intronic regions, this same marker has proven to be efficient in solving the phylogenetic relationships among other catfish subfamilies (Chiachio et al., 2008; Cardoso

8

and Montoya-Burgos, 2009; Alexandrou et al., 2011; Cramer et al., 2011; Covain and FischMuller, 2012; Roxo et al., 2012; Costa Silva et al., 2014; Roxo et al., 2014). Additionally, Fisch-Muller et al. (2012) demonstrated that in Ancistrinae, the two introns of f-rtn4r were rather conserved, and displayed less variation than coding mitochondrial genes, making easier detection of homologous regions. Gaps were considered as missing data, and regions impossible to amplify or to sequence were coded as ambiguities (N). The final alignment of frtn4r marker is accessible on Dryad (http://datadryad.org/) using accession number XXXX. Since mitochondrial DNA is presumably transmitted through maternal lineage as a single non recombining genetic unit (Meyer, 1993), a first partition corresponding to the mitochondrial genes was created. With the mutational patterns in intronic and exonic regions of f-rtn4r being rather characterized by insertions/deletions and transitions/transversions respectively, two other partitions were created for introns and exons. Congruence in phylogenetic signals contained in mitochondrial and nuclear markers was secondarily assessed using the Congruence Among Distance Matrices (CADM) test (Legendre and Lapointe, 2004) as implemented in ape 2.6.2 (Paradis et al., 2004; Paradis, 2006) in R 2.12.1 (R Development Core Team, 2009). Patristic pairwise maximum likelihood (ML) (Felsenstein, 1981) distances were computed as estimates of tree topologies with Treefinder (Jobb et al., 2004) version of October 2008 for each partition using a likelihood model under which the distances are optimized. Appropriate substitution models corresponding to each potential partition were accordingly determined with the Akaike Information Criterion (Akaike, 1974) as implemented in Treefinder. The CADM test was computed using 9,999 permutations of the three ML distances matrices. Two phylogenetic reconstruction methods allowing the analysis of partitioned data were used. First, a ML reconstruction was performed with Treefinder, and robustness of the results was estimated by resampling the data set with the nonparametric bootstrap (Efron, 1979)

following

Felsenstein’s (1985)

methodology with 2,000 9

pseudoreplicates. Second, a Bayesian inference analysis was conducted in MrBayes 3.1.2 (Huelsenbeck and Ronquist, 2001; Ronquist and Huelsenbeck, 2003). Two runs of eight chains (one cold, seven heated) were conducted simultaneously for 2x107 generations, with the tree space sampled each 1000th generation. Convergence between chains occurred after 2x106 generations (average standard deviation of split frequencies <0.01). After a visual representation of the evolution of the likelihood scores, and checking for the stationarity of all model parameters using Tracer 1.5 (Rambaut and Drummond, 2007) (i.e.: potential scale reduction factor (PSRF), uncorrected roughly approached 1 as runs converged (Gelman and Rubin, 1992), and Effective Sample Size (ESS) of all parameters above 200), the 2x106 first generations were discarded as burn-in. The remaining trees were used to compute the consensus tree. Alternative hypotheses (i.e. topologies) were tested against the null hypothesis that all hypotheses provided equally good explanations of the data using the Approximately Unbiased (AU) (Shimodaira, 2002), the Bootstrap Probability (BP), and the ExpectedLikelihood Weights (ELW) of the alternative hypothesis (Strimmer and Rambaut, 2002) tests as implemented in Treefinder using 1x106 RELL replicates (Kishino et al., 1990). All alternative topologies were constructed in order to reflect, as much as possible given our taxonomic sampling, already proposed hypotheses (Isbrücker, 1979; Rapp Py-Daniel, 1997 and Armbruster 2004), or the monophyly of different groups.

3. Results 3.1 Phylogenetic analysis of the subfamily Loricariinae.

We sequenced the almost complete 12S and 16S mitochondrial genes, and the partial nuclear gene f-rtn4r for 326 specimens of 217 species of Loricariinae and 8 Loricariidae belonging to Hypostominae (7 species) and Neoplecostominae (1 species) as outgroup (Table 1). Other

10

sequences for 24 representatives of Loricariinae, and ten Loricariidae belonging to Rhinelepinae (1 species), and Hypostominae (9 species) were obtained from GenBank using the accession numbers provided in Covain et al. (2008), Chiachio et al. (2008), Rodriguez et al. (2011), Covain and Fisch-Muller (2012), and Fisch-Muller et al. (2012). The sequence alignment initially including 8,503 positions was restricted to 8,426 positions after removal of regions with ambiguous alignment. A subset of 2,545 positions corresponded to the mitochondrial genes (962 positions for the 12S rRNA gene, 74 for the tRNA Val gene, and 1,509 for the 16S rRNA gene), and 5,881 to the nuclear f-rtn4r gene (894 positions for the exonic regions, and 4,987 for the intronic regions). No significant conflicting phylogenetic signal was detected in the data set, as the global CADM test displayed a high coefficient of concordance between matrices and rejected the null hypothesis of incongruence between them (CADM: W = 0.7964, χ2ref = 163976.6, p(χ2ref≥χ2*) = 0.0001). The CADM a posteriori tests did not detect any conflicting matrix with the global phylogenetic signal ( rS mitochondrion = 0.6295, p( rS ref≥ rS *) = 0.0003; rS exons = 0.7239, p( rS ref≥ rS *) = 0.0003; rS introns = 0.7304, p( rS ref≥ rS *) = 0.0003). Thus, despite the presence of indels in the intronic regions, and of a lower contribution of the mitochondrial genes to the global phylogenetic signal, there was no indication to discard these regions from the phylogenetic analyses. The sequences were consequently concatenated, and three partitions corresponding to mitochondrial genes, exonic parts of f-rtn4r, and intronic parts of f-rtn4r were used to reconstruct the tree. The models GTR + G (Tavaré, 1986) for mitochondrial genes and intronic regions of f-rtn4r, and HKY + G (Hasegawa et al., 1985) for exonic regions of f-rtn4r displayed the smallest AIC and accordingly fitted our data the best as calculated with Treefinder. Maximun Likelihood and Bayesian phylogenetic reconstructions lead to equivalent tree topologies (Appendix A and B respectively), both comparable in broad outline to the one obtained by Covain et al. (2008), and Rodriguez et al. (2011). The ML tree (Fig1 and Appendix A; -lnL = 116343.2) and 11

Bayesian tree (Appendix B) both show a basal split within the Loricariinae [100 Bootstrap Probability (BP) and 1 Posterior Probability (PP)] resulting in two highly supported lineages: the Harttiini (clade 1; 95.65 BP and 1 PP) and the Loricariini (clade 2; 85.68 BP and 1 PP). The Loricariini was divided, in turn, into two strongly supported clades: the Farlowellina (clade A; 100 BP and 1 PP); and the Loricariina (clade B; 78.85 BP and 1 PP). Within the Loricariina, three main groups were resolved with high supports, one forming the Loricariichthys group (sensu Covain and Fisch-Muller, 2007; 85.95 BP and 1 PP), a second comprising Spatuloricaria in a sister position to the Loricaria plus Pseudohemiodon groups (sensu Covain and Fisch-Muller, 2007; 99.95 BP and 1 PP), and a third comprising all Rineloricaria representatives (99.6 BP and 1 PP). Metaloricaria, Dasyloricaria, and Fonchiiloricaria were not included in these morphological groups.

3.1.1 Harttiini

The Harttiini tribe formed a monophyletic group (Fig. 1, clade 1) and included the genera Harttia, Cteniloricaria, and Harttiella (Fig. 2). Cteniloricaria and Harttiella were resolved as monophyletic with high statistical supports (both with 100 BP and 1 PP; Appendices A and B). Cteniloricaria included two species, C. napova and C. platystoma (type species). Harttiella comprised six species, H. crassicauda (type species), H. parva, H. pilosa, H. longicauda, H. intermedia, and H. lucifer. Harttiella intermedia was nested within H. longicauda. Relationships among other Harttiini belonging to Harttia were partly unresolved. Guianese Harttia comprising H. guianensis, H. surinamensis, H. fluminensis, and H. tuna formed a highly supported monophyletic group (100 BP and 1 PP) but were weakly supported as members of Harttia (BP below 50). In the Bayesian inference they formed the sister group of Cteniloricaria with very low posterior probabilities (0.53). The only relationship better supported in Harttia was the clade including Amazonian representatives (H. punctata, H. 12

duriventris, H. dissidens, H. sp. Xingu1, H. sp. Xingu2, H. sp. Xingu3, H. sp. Tapajos, H. sp. Tocantins, and H. aff. punctata) plus the Guianese H. fowleri in a sister position to representatives from South East Brazil (including H. loricariformis, type species of the genus and H. leiopleura type species of Quiritixys) with a low bootstrap probability of 54.5 but a posterior probability of 1 (Appendices A and B respectively). Deeper relationships among genera were not statistically supported.

3.1.2 Loricariini, Farlowellina

The Loricariini tribe was resolved as monophyletic (Fig. 1, clade 2). Within Loricariini, the subtribe Farlowellina also constituted a monophyletic assemblage (Fig. 1, clade A), and comprised Lamontichthys, Pterosturisoma, Sturisoma, Farlowella, Aposturisoma, and Sturisomatichthys (Fig. 3). Interspecific relationships were congruent between both phylogenetic analyses. Lamontichthys (including L. filamentosus, type species) was monophyletic and formed, with high support (100 BP and 1 PP), the sister group of all other representatives of the subtribe. The second diverging genus was the monotypic Pterosturisoma microps that formed the sister group of the remaining Farlowellina. Then all cis-Andean (East of the Andes following the definition of Haffer, 1967; and Albert et al., 2006) representatives of Sturisoma included in this study, branched with high statistical support (100 BP and 1 PP) in a sister position to all other representatives of Sturisoma, Sturisomatichthys, Farlowella and Aposturisoma. The subsequent, also highly supported, group comprised a mix of representatives of Sturisomatichthys (including S. leightoni, type species) and of the trans-Andean (West of the Andes) Sturisoma rendering both genera paraphyletic. Within the last group of Farlowellina, a first group of Farlowella consisted of the stockiest forms of the genus (including F. platorynchus, F. amazona, F. aff. rugosa, F. taphorni and F. curtirostra) branched in a sister position to Aposturisoma myriodon forming 13

in turn and with high support (99.96 BP and 1 PP) the sister genus of a second group of Farlowella (including F. acus, type species) rendering Farlowella paraphyletic.

3.1.3 Loricariini, Loricariina

The subtribe Loricariina (Fig. 1, clade B) was also monophyletic and formed the sister group of Farlowellina (Fig. 1, clade A). The basal split (Fig. 4) gave rise to two strongly supported lineages, one comprising the representatives of Metaloricaria (including Metaloricaria paucidens, type species; 99.9 BP and 1 PP) and the second including all other Loricariina (Fig. 4; 99.8 BP and 1 PP). Then a second diverging group comprising Dasyloricaria representatives in a sister position to the monotypic Fonchiiloricaria nanodon, formed in turn the sister group of the remaining Loricariina. The sister relationship between Dasyloricaria and Fonchiiloricaria was however not supported by bootstrap values (BP<50) but displayed relatively high posterior probability (0.83). The sister group of these two genera split into two groups with on one side representatives of Rineloricaria and Ixinandria, and on the other side members of the Loricaria-Pseudohemiodon and Loricariichthys groups.

3.1.3.1 Loricariini, Loricariina, Rineloricaria

The genus Rineloricaria formed the most species rich group of the subfamily and constituted a monophyletic assemblage comprising members of Fonchiiichthys, Hemiloricaria, Leliella, and Ixinandria steinbachi, type species of Ixinandria, with high statistical support (Fig. 5; 99.6 BP and 1 PP). The first diverging group of Rineloricaria comprised the trans-Andean R. altipinnis in a sister relationship to the cis-Andean R. stewarti, R. fallax, R. formosa, R. melini, R. teffeana, R. morrowi, and several undescribed species (88.5 BP and 0.98 PP). The 14

second diverging group comprised different populations of R. lanceolata and R. hoehnei. The latter species was nested within R. lanceolata and all internal relationships were fully resolved and highly supported (82.4
3.1.3.2 Loricariini, Loricariina, Loricariichthys group

15

Within Loricariina, members of the Loricariichthys group formed a strongly supported natural group (85.95 BP and 1 PP) comprising Pseudoloricaria, Limatulichthys, Loricariichthys, and Hemiodontichthys (Fig. 6) and formed the sister clade of the Loricaria-Pseudohemiodon group. Loricariichthys (including L. maculatus, type species) was monophyletic and constituted the sister genus of all other members of its groups. The second diverging genus was the monotypic Hemiodontichthys acipenserinus with its different populations, and was the sister group of the genera Pseudoloricaria and Limatulichthys. All internal relationships within the Loricariichthys group were congruent in both reconstructions and fully resolved with high statistical supports (Appendices A and B).

3.1.1.3 Loricariini, Loricariina, Loricaria-Pseudohemiodon group

The Loricaria-Pseudohemiodon group formed a strongly supported clade (99.95 BP and 1 PP)

comprising

Brochiloricaria,

the

genera

Paraloricaria,

Spatuloricaria, Planiloricaria,

Loricaria

(including

Crossoloricaria,

Proloricaria),

Pseudohemiodon,

Apistoloricaria, and Rhadinoloricaria, and formed the most genera rich group (Fig. 7). Interspecific relationships were congruent between both reconstructions except for the species and populations closely related to L. cataphracta. Spatuloricaria was resolved as monophyletic and formed the sister genus of all other genera of the group. The remaining members of the Loricaria-Pseudohemidon group split into two strongly supported clades corresponding to the Loricaria group (sensu Covain and Fisch-Muller, 2007) on one side and the Pseudohemiodon group (sensu Covain and Fisch-Muller, 2007) on the other side. The Loricaria group was strongly supported (100 BP and 1 PP) and comprised Loricaria (including L. cataphracta type species), Brochiloricaria, and Paraloricaria. At the exclusion of L. prolixa (type species of Proloricaria) and L. apeltogaster, the remaining Loricaria

16

species formed a statistically highly supported monophyletic group (100 BP and 1 PP). Loricaria formed the sister genus of all other representatives of its group. The sister group of Loricaria comprised Loricaria prolixa in a sister position to Brochiloricaria representatives, both in turn forming the sister group of L. apeltogater as sister species of representatives of Paraloricaria. However, the positions of L. prolixa and L. apeltogaster were not statistically supported (50
3.2 Evaluation of alternative phylogenetic hypotheses Alternative hypotheses were evaluated using topological tests and results are summarized in Table 3. The hypothesis proposed by Isbrücker (1979) classifying the Loricariinae into three tribes: Harttiini, Loricariini, and Farlowellini without phylogenetic relationships between them (coded as a polytomy at origin of the three lineages) was significantly rejected by all testing procedures (Table 3, H1). The hypothesis of Rapp Py-Daniel (1997), partly confirmed by Armbruster (2004), and consisting in splitting the Loricariinae into two sister tribes:

17

Harttiini on one side (including Harttia, Cteniloricaria, Harttiella, Lamontichthys, and Pterosturisoma forming the subtribe Harttiina in a sister position to Farlowella, Aposturisoma, Sturisoma, and Sturisomatichthys forming the subtribe Farlowelliina), and Loricariini on the other side (comprising all other genera) was also significantly rejected (Table 3, H2). The hypothesis proposing, within Farlowellina (as defined by Covain et al., 2008; 2010), the monophyly of Farlowella as sister genus of Aposturisoma on one side, and of Surisoma as sister genus of Sturisomatichthys on the other side was also significantly rejected by all tests (Table 3, H3). The enforced monophyly of Crossoloricaria as sister genus of Apistoloricaria and Rhadinoloricaria (Table 3, H4), as well as the monophyly of Loricaria, comprising L. prolixa, and L. apeltogaster (Table 3, H5), were both significantly rejected. Within the Rineloricaria clade (Fig. 5), the validity of Ixinandria, Hemiloricaria, Rineloricaria, Leliella, and Fonchiiichthys was evaluated by assigning each species to the corresponding genus (as listed in Isbrücker, 2001) but without providing phylogenetic hypothesis of relationships between these five genera (coded as a polytomy at origin of all lineages). This hypothesis was also rejected (Table 3, H6). In the same way, the validity of Quiritixys was assessed by creating a polytomy at origin of Cteniloricaria, Harttia, Harttiella, and Quritixys lineages. This hypothesis was rejected by all testing procedures (Table 3, H7).

4. Discussion

The phylogenetic results confirmed the monophyly of the subfamily Loricariinae, and its splitting into two tribe-level clades, namely the Harttiini, and the Loricariini. Two subtribelevel clades were obtained for Loricariini, the Farlowellina and the Loricariina, the latter being the most diversified and further divided in three main clades. A reappraisal of these tribes, subtribes, and their respective genera is here proposed (summarized in Table 4). The

18

taxonomic status of some species will also be revised, with new synonymisations and generic reassignations.

4.1. Harttiini Corroborating previous results (Montoya-Burgos et al., 1998; Covain et al., 2008; Rodriguez et al., 2011; Lujan et al., 2015), the Harttiini are restricted to Harttia (type genus), Harttiella and Cteniloricaria. Moreover, the enlarged definition of Harttiini comprising Farlowellina (Rapp Py-Daniel, 1997; Armbruster, 2004) is here significantly rejected (Table 2). Harttiini were primarily diagnosed by 14 caudal-fin rays, no postorbital notches, no predorsal keels, a circular mouth with papillose lips, and numerous pedunculated teeth organized in a comblike manner (Isbrücker, 1979; Covain and Fisch-Muller, 2007). However, these features are shared by Harttiini, Farlowellina and partly by Fonchiiloricaria nanodon within Loricariini, rendering the definition of Harttiini sensu stricto invalid. We nevertheless note that in Harttiini, the abdominal cover made of small rhombic platelets can be present or absent, and when it is present, the abdominal cover never extends to the lower lip margin. The latter condition is, on the contrary, always observed in Farlowellina. If this criterion applies, then the recently described Harttia absaberi (Oyakawa et al., 2013) should be regarded as a putative member of Farlowellina. Deeper relationships within Harttiini are not resolved due to very short internal branches suggesting explosive radiation of the main lineages. The nested position of H. leiopleura, type species of Quiritixys, within the Southeastern species of Harttia which also included H. loricariformis, the type species of the genus, renders Harttia paraphyletic with the necessity to describe several new genera (considering our sampling, a total of four would be needed to render each lineage monophyletic). To prevent this taxonomic issue, a conservative approach consists in placing Quiritixys into synonymy with Harttia. This conclusion is reinforced by the significant rejection of the hypothesis evaluating

19

its validity (Table 2). A second problem concerns the position of Harttiella intermedia nested within H. longicauda. A rapid overview of this situation would probably lead to the placement of H. intermedia into the synonymy of H. longicauda. However, based on morphometric analyses, Covain et al. (2012) demonstrated that the former was perfectly distinct from the latter, and even belonged to another morphological group (the crassicauda group comprising all stockier species while H. longicauda belonged to the longicauda group comprising all slender species). In that study the barcode COI sequence of H. intermedia was also identical to that of H. longicauda, and the authors hypothesised introgressive hybridization or a recent founder effect in an isolated population to explain this phenomenon, both species being present in the same basin. The use of the nuclear f-rtn4r gene in the present study, and the topological result identical to that obtained using barcode sequences, invalidate the hypothesis of introgressive hybridization. Since the establishment of reciprocal monophyly between two sister taxa is a function of time (Hubert et al., 2008), when not enough time passed to accumulate mutations able to differentiate sister species, a paraphyletic grouping may be observed with one species nested within a second one [i.e. the coalescent of the first species is contained within the coalescent of the second (Meyer and Paulay, 2005)]. Harttiella intermedia thus represents a rather recent vicariant form of H. longicauda isolated in the Trinité Massif in French Guiana, and corroborates the alternative hypothesis of Covain et al. (2012) of a morphologically fast evolving species not yet genetically distinguishable from its ancestor following the example of the East African lacustrine cichlid species flock (e.g. Won et al., 2005).

4.2. Loricariini, Farlowellina Within Loricariini, the phylogeny of Farlowellina revealed unexpected results. All genera except Lamontichthys and Pterosturisoma appeared paraphyletic, and their enforced

20

monophyly was significantly rejected (Table 2). The nested position of Aposturisoma within Farlowella renders the latter polyphyletic. If one considers Aposturisoma a valid genus, based on its particular body shape, ecological habits and restricted distribution to the HuacamayoAguaytia drainage, members of the F. amazona species group (sensu Retzer and Page, 1997) should be placed in a new genus. However, the lack of significant distinctive features between the F. amazona group and other Farlowella, and the close relatedness of Aposturisoma and Farlowella, may imply that Aposturisoma corresponds to a local form of Farlowella adapted to rheophilic habits. This corroborates the hypothesis of Covain and Fisch-Muller (2007) who interpreted the morphological characteristics of Aposturisoma as adaptations to stream habitat rather than an intermediary shape between Farlowella and Sturisoma as supposed by Isbrücker et al. (1983). If this hypothesis is applied, Aposturisoma should be considered a junior synonym of Farlowella. Nevertheless, this question still deserves further investigation. The taxonomy of Farlowella is also confused and the group needs further revision. In the last revision of the genus, Retzer and Page (1997) described F. platorynchus without examining the holotype of F. amazona. However, examination of the holotypes of both species strongly suggests that F. platorynchus is a junior synonym of F. amazona. In addition, F. gladiolus was placed in synonymy with F. amazona, but should be regarded as a valid species. Moreover, identification at specific level remains difficult due to the very divergent morphology of the genus and the great similarity of its members. Consequently, species with a large geographic distribution may comprise species complexes (see e.g. F. oxyrryncha in Fig. 3). The second paraphyly highlighted here concerns the genera Sturisoma and Sturisomatichthys. Contrary to the preceding case, a strong geographical structure is represented in this result with one group of Sturisoma comprising all cis-Andean species, and a second group comprising all trans-Andean members of Sturisoma and Sturisomatichthys. Moreover, the type species of Sturisoma, S. rostrata, is described from Brazilian rivers,

21

whereas the type species of Sturisomatichthys, S. leightoni, is described from the Magdalena River in Colombia. For these reasons, Sturisoma is here restricted to the species occurring in the cis-Andean region (including S. barbatum, S. brevirostre, S. guentheri, S. lyra, S. monopelte, S. nigrirostrum, S. caquetae, S. robustum, S. rostratum, and S. tenuirostre) whereas Sturisomatichthys comprises all former trans-Andean species of Sturisoma and Sturisomatichthys (i.e. S. aureum, S. citurensis, S. dariense, S. festivum, S. frenatum, S. kneri, S. leightoni, S. panamense, and S. tamanae). The diagnostic feature provided by Isbrücker and Nijssen (in Isbrücker, 1979) to distinguish Sturisomatichthys from Sturisoma, i.e. the absence of a rostrum in Sturisomaticthys, is not phylogenetically informative (Covain et al., 2008), as it can be absent or present according to the species, and thus is not a valid criterion to diagnose the genus.

4.2. Loricariini, Loricariina

The Loricariina comprises some particular forms that can be seen as relictual species due to their particular morphological characteristics, restricted distributions, and long branches, rendering the phylogenetic signal noisy. Metaloricaria is indeed the first diverging group of the subtribe and it possesses a very particular morphology reminiscent to that of Harttia with which it shares the same habitat (stream waters in riffles). This resemblance probably resulted in the initial description of M. nijsseni as a member of Harttia (Boeseman, 1976), despite clear autapomorphic features such as a horse-shoe like mouth shape, teeth pedunculated yet reduced in size and number, and 13 caudal-fin rays, that indicate the future trends of the Loricariina (strong modifications in mouth, lips, and teeth characteristics, decrease of the number of caudal-fin rays…). Metaloricaria is restricted to the Guiana Shield in rivers flowing through Suriname and French Guiana. In the same way, Dasyloricaria which is restricted to the Pacific slope of the Andes (a unique pattern of distribution within the 22

subfamily), shares a mosaic of morphological characteristics with representatives of other Loricariina mainly distributed on the Atlantic slope. Along with members of Rineloricaria, it shares papillose lips and hypertrophied odontodes along the sides of the head in breeding males. With some representatives of the Loricariichthys group (sensu Covain and FischMuller, 2007), it shares deep postorbital notches, a strongly structured abdominal cover, and a similar mouth shape, including the hypertrophied lower lip of breeding males (Steindachner, 1878). Finally, with some representatives of the Loricaria group, it shares a triangular head, strong predorsal keels, and the upper caudal fin ray produced into a long whip. Finally, Fonchiiloricaria is restricted to the Upper Huallaga River in Peru. It possesses 14 caudal-fin rays, and no postorbital notches, two features characteristic for Harttiini and Farlowellina. In addition it also possesses autapomorphic features such as an extreme reduction in size and number of premaxillary teeth (when not missing) relative to dentary teeth (Rodriguez et al., 2011). All these relictual species exhibit features that will be successively lost or maintained in other Loricariina lineages. In this case the observed autapomorphic features would correspond to the retention of ancestral characters. Rineloricaria constitutes by far the most species rich genus of the Loricariinae, including 66 valid species and numerous undescribed ones. Several attempts have been made to split this genus. Isbrücker and Nijssen (1976) proposed the revalidation of Hemiloricaria Bleecker, 1862 (type species: Hemiloricaria caracasensis), but they finally left it in the synonymy of Rineloricaria because of the lack of obvious characters to split these two genera. Later on, in an aquarist hobbyist journal, Isbrücker (in Isbrücker et al., 2001) changed his mind and revalidated Hemiloricaria based on the disposition of breeding odontodes in males, and assigned 24 species to this genus (R. altipinnis, R. beni, R. cacerensis, R. caracasensis, R. castroi, R. eigenmanni, R. fallax, R. formosa, R. hasemani, R. hoehnei, R. jubata, R. konopickyi, R. lanceolata, R. magdalenae, R. melini, R. morrowi, R. nigricauda, R. parva, R.

23

phoxocephala, R. platyura, R. sneiderni, R. stewarti, R. teffeana, and R. wolfei), most of them belonging to different lineages according to the present results. Moreover, the breeding odontodes on the predorsal area of males are not always present in the species assigned to this group (e.g. R. platyura). In the same publication, Isbrücker and Michel described Fonchiiichthys (type species: Loricaria uracantha), and Isbrücker described Leliella (type species: Rineloricaria heteroptera) on the basis of subtle differences in sexual dimorphism. However, in our phylogenetic reconstruction R. uracantha, R. heteroptera and R. eigenmanni (a very close relative of R. caracasensis following the examination of type specimens) were nested within the same clade, and their enforced monophyly was significantly rejected (Table 3). For these reasons, Hemiloricaria, Fonchiiichthys, and Leliella are here placed in synonymy with Rineloricaria. In addition, the nested position of Ixinandria steinbachi in a sister position to R. misionera within Southeastern representatives of Rineloricaria, renders Rineloricaria paraphyletic. We therefore place Ixinandria in synonymy with Rineloricaria. The diagnostic feature given by Isbrücker and Nijssen (in Isbrücker, 1979) for Ixinandria, a naked belly and particular sexual dimorphism, should be considered as specific characters. This is reinforced by the appearance, in close relatives of R. steinbachi from Southeast Brazil or Argentina, of a gradual increase in the abdominal plating, rendering thus the belly partly covered (e.g. R. maquinensis, R. aequalicuspis or R. misionera). Finally, the nested position of R. hoehnei within R. lanceolata renders the latter paraphyletic. We confirm here results of Vera-Alcaraz et al. (2012) and also place R. hoehnei (Miranda Ribeiro, 1912) in synonymy with R. lanceolata (Günther, 1868). However, considering the branch lengths of the phylogenetic tree compared to other species of Rineloricaria, R. lanceolata may prove to host a species complex. The Loricariichthys group appears more structured, with all genera resolved as monophyletic and strongly supported. With the exception of the nominal genus, this group surprisingly

24

comprises mostly monotypic or poorly diversified genera (Limatulichthys, Pseudoloricaria, and Hemiodontichthys, with the addition of Furcodontichthys following results of Covain and Fisch-Muller, 2007). However, given their broad geographic range, and long branches among populations, Hemiodontichthys acipenserinus and Pseudoloricaria laeviuscula could comprise species complexes. Indeed, Isbrücker and Nijssen (1974) reported variations in morphometric features of H. acipenserinus, with populations from the Amazonian region tending to be more slender than those from the Paraguay and Guaporé Rivers. Within the Loricaria group, the nominal genus is resolved as paraphyletic, and its enforced monophyly is statistically rejected (Table 3). Loricaria prolixa connected in a sister position to representatives of Brochiloricaria, and L. apeltogater in a sister position to Paraloricaria. Loricaria prolixa was designated by Isbrücker (in Isbrücker et al., 2001) as type species of a new genus Proloricaria, based on a flattened and anteriorly broad body. The weakness of these supposed diagnostic features that are also present in other genera (e.g. Pyxiloricaria, Pseudohemiodon) lead several authors to consider Proloricaria as a junior synonym of Loricaria (Ferraris in Reis et al., 2003; Covain and Fisch-Muller, 2007). However, our results sustain the validity of Proloricaria which is here revalidated. The sister position of L. apeltogater to Paraloricaria needs further investigation. The specimen included in the present study was not preserved, and we can not be certain that it belonged to the species. However, in the description of P. agastor, Isbrücker (1979) had already noticed the close resemblance of both species (the smallest syntype of L. apeltogaster was even subsequently identified as P. agastor), distinguishing them on the basis of the dentition. Paraloricaria possesses small teeth on both jaws whereas L. apeltogater possesses the typical dentition for Loricaria with premaxillary teeth two times longer than dentary ones. Within the Pseudohemiodon group, the trans-Andean Crossoloricaria which includes C. variegata, type species, branches in a sister position to all other genera, whereas the cis-

25

Andean Crossoloricaria, are nested within the remaining members of the Pseudohemiodon group, rendering Crossoloricaria paraphyletic. Crossoloricaria is poorly diagnosed, its only distinctive character (incomplete abdominal cover consisting of a double median row of plates) being shared by Apistoloricaria and Rhadinoloricaria. Moreover, Crossoloricaria rhami possesses a complete abdominal plate development (Isbrücker and Nijssen, 1983), thus rendering the diagnostic feature of Crossoloricaria invalid. Apistoloricaria is also not well diagnosed and is distinguished from Rhadinoloricaria primarily by the presence or absence of the iris operculum (absent or vestigial in Apistoloricaria versus present in Rhadinoloricaria), a more conspicuous rostrum in Rhadinoloricaria, and by the number of fringed barbels (14 in Apistoloricaria versus 12 in Rhadinoloricaria). Based on the present phylogenetic results, the rejection

of

the

enforced

monophyly

of

Crossoloricaria,

Apistoloricaria,

and

Rhadinoloricaria (Table 3), and the weakness of these diagnostic features, Crossoloricaria is here restricted to the trans-Andean region (including C. variegata, C. venezuelae, and C. cephalaspis), whereas the cis-Andean species of Crossoloricaria (C. rhami, C. bahuaja) are transferred to Rhadinoloricaria. Apistoloricaria is also placed in synonymy with Rhadinoloricaria.

5. Conclusions

This work represents the first comprehensive phylogeny of the Loricariinae and provides considerable additional data to the evolutionary tree of one of the most diversified vertebrate families. The Loricariinae, the second species-rich subfamily of the Loricariidae, and hitherto phylogenetically underinvestigated, is analysed with 368 OTUs (350 Loricariinae). An important gap in the evolutionary tree of the Loricariidae is thus filled, and the present results complement other molecular works at familial and subfamilial ranks such as Lujan et al.

26

(2015) on the family Loricariidae with focus on Hypostominae (203 OTUs), Roxo et al. (2014) on Hypoptopomatinae, Neoplecostominae, and Otothyrinae (155 OTUs), Cramer et al. (2011) on the Loricariidae with focus on Neoplecostominae and Hypoptopomatinae (146 OTUs), Chiachio et al. (2008) on Hypoptopomatinae and Neoplecostominae (53 OTUs), and Montoya-Burgos et al. (1998) on the family Loricariidae with emphasis on Hypostominae (58 OTUs), providing thereby a more comprehensive view of the dramatic diversification of the Loricariidae in South America.

Acknowledgments

We are grateful to M. Sabaj Perez, and J. G. Lundberg, ANSP; E. Bermingham, and G. R. Reina, STRI; M. Lucena, MCP; J. Armbruster, AUM; O. T. Oyakawa, MZUSP; J. Casciota, and A. Almirón, MLP; Y.P. Cardoso, AUGM-UNLP; M. S. Rodriguez, LIRP; G. Costa Silva, LBP; P. Keith, R. Causse, and P. Pruvost, MNHN; M. van Oijen and K. van Egmond, RMNH; R. Vonk and H. Praagman, ZMA; J. Maclaine, BMNH; H. Wellendorf, NMW; S. Schaefer, AMNH; M. A. Rogers, FMNH; D. Catania, CAS; G. M. Gonzo, UNSA; L. Rapp Py-Daniel and M. Rocha, INPA; C. Dhlouy, San Lorenzo; and S. Rubin and L. Moissonier, France; for the loan of specimens and/or tissue samples. We acknowledge H. Ortega, M. Hidalgo and V. Meza Vargas, MUSM; T. Pequeño and collaborators from CIMA; L. Rodriguez (Conservación de Recursos Naturales); P. de Rham, Lausanne, P. Gaucher, CNRS Guyane; R. Vigouroux and P. Cerdan, Hydreco Guyane; C. Weber, and A. Merguin, MHNG; M. Dewynter, ONF Guyane; F. Melki, Biotope France; K. Wan Tong You and P. Ouboter, NZCS; C. Bernard, CSBD; the North Rupununi District Development Board; and the Iwokrama organization for their field and logistic assistance; the G. and A. Claraz Foundation for their financial support for the missions in Suriname in 2001, 2005, 2007 and 2008, and in

27

French Guiana in 2006; the Académie Suisse des Sciences Naturelles (ScNat) for their financial support for the missions in Guyana 2004 and French Guiana 2006; The C. Topali Fund for their financial contribution to the acquisition of field material for the mission Suriname 2007, and lab material in 2009; the All Catfish Species Inventory (NSF-DEB 0315963) for the financing of collection consultancies to RC (NMW and ANSP in 09/2007 and 12/2007), and open access in Zootaxa to Covain and Fisch-Muller (2007); and the A. Lombard Fund for data acquisition in 2010. We are also grateful to A. Huser, Sallmann-Fehr AG for the gift of gill nets for the mission in Suriname in 2005 and 2007; the Guyana Environmental Protection Agency, and Ministry of Amerindian affairs; the French Guiana Diren, and Préfecture; and the Surinamese Ministry of Agriculture, Animal Husbandry and Fisheries for the different necessary authorizations and collecting permits. Part of this project was supported by the Fond National Suisse de la Recherche Scientifique (JIMB 31003– 141233). CO researches are supported by CNPq and FAPESP in Brazil. The figures were finalized by Florence Marteau, MHNG. John Hollier, MHNG, improved language usage and style. Jan Pawlowski and José Fahrni, University of Geneva, are acknowledged for laboratory facilities.

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Table 1. Alternative classifications of the Loricariinae according to different authors.

Table 2. Taxa list, specimen and sequence data for the 350 Loricariinae and 18 outgroup representatives analyzed in this study. The acronyms of institutions follow Fricke and Eschmeyer (2015).

Table 3. Alternative phylogenetic relationships evaluated using the Approximately Unbiased (AU), Bootstrap Probability (BP), and the Expected-Likelihood Weights (ELW) of the alternative hypothesis tests using 1x106 RELL replicates. lnL: likelihood of the hypothesis; ΔlnL: difference in likelihood between the alternative hypothesis and the Maximum Likelihood (ML) tree as best explanation of the data. Reported results correspond to p-values.

Table 4. Loricariinae classification with list of valid genera following this study

Fig. 1. Maximum Likelihood tree of the Loricariinae (-lnL = 116343.2) inferred from the combined analysis of sequences of partial 12S and 16S mitochondrial genes, and partial frtn4r nuclear gene. The models GTR + G for mitochondrial genes and intronic regions of frtn4r, and HKY + G for exonic regions of f-rtn4r were applied for both ML and Bayesian reconstructions. Blackened branches in the ingroup indicate nodes with both bootstrap supports and posterior probabilities below 50 and 0.70 respectively. Stars indicate incongruence between ML and Bayesian reconstructions. 1: Harttiini; 2: Loricariini; A: Farlowellina, B: Loricariina. Circled numbers refer to subtrees figured in the text. Scale indicates the number of substitutions per site as expected by the model.

40

Fig. 2. Maximum Likelihood tree, labelled subtree of the Harttiini tribe. Numbers above branches indicate bootstrap supports above 50 followed by posterior probabilities above 0.70. Within species supports are provided in Appendices A and B. Dash (-) represent low supports. Blackened branches indicate nodes with both bootstrap supports and posterior probabilities below 50 and 0.70 respectively. Stars indicate incongruence between ML and Bayesian reconstructions and NAs indicate nodes absent in topologies of Appendices A and B. Bold type refers to type species of different genera. Scale indicates the number of substitutions per site as expected by the model.

Fig. 3. Maximum Likelihood tree, labelled subtree of the Loricariini tribe: Farlowellina subtribe. Numbers above branches indicate bootstrap supports above 50 followed by posterior probabilities above 0.70. Within species supports are provided in Appendices A and B. Dash (-) represent low supports. Blackened branches indicate nodes with both bootstrap supports and posterior probabilities below 50 and 0.70 respectively. Stars indicate incongruence between ML and Bayesian reconstructions and NAs indicate nodes absent in topologies of Appendices A and B. Bold type refers to type species of different genera. Scale indicates the number of substitutions per site as expected by the model.

Fig. 4. Maximum Likelihood tree, labelled subtree of the Loricariini tribe: Loricariina subtribe. Numbers above branches indicate bootstrap supports above 50 followed by posterior probabilities above 0.70. Within species supports are provided in Appendices A and B. Dash (-) represent low supports. Bold type refers to type species of different genera. Scale indicates the number of substitutions per site as expected by the model.

41

Fig. 5. Maximum Likelihood tree, labelled subtree of the Loricariini tribe: Loricariina subtribe, Rineloricaria group. Numbers above branches indicate bootstrap supports above 50 followed by posterior probabilities above 0.70. Within species supports are provided in Appendices A and B. Dash (-) represent low supports. Blackened branches indicate nodes with both bootstrap supports and posterior probabilities below 50 and 0.70 respectively. Stars indicate incongruence between ML and Bayesian reconstructions and NAs indicate nodes absent in topologies of Appendices A and B. Bold type refers to type species of different genera. Scale indicates the number of substitutions per site as expected by the model.

Fig. 6. Maximum Likelihood tree, labelled subtree of the Loricariini tribe: Loricariina subtribe, Loricariichthys group. Numbers above branches indicate bootstrap supports above 50 followed by posterior probabilities above 0.70. Within species supports are provided in Appendices A and B. Bold type refers to type species of different genera. Scale indicates the number of substitutions per site as expected by the model.

Fig. 7. Maximum Likelihood tree, labelled subtree of the Loricariini tribe: Loricariina subtribe, Loricaria-Pseudohemiodon group. Numbers above branches indicate bootstrap supports above 50 followed by posterior probabilities above 0.70. Within species supports are provided in Appendices A and B. Dash (-) represent low supports. Blackened branches indicate nodes with both bootstrap supports and posterior probabilities below 50 and 0.70 respectively. Stars indicate incongruence between ML and Bayesian reconstructions and NAs indicate nodes absent in topologies of Appendices A and B. Bold type refers to type species of different genera. Scale indicates the number of substitutions per site as expected by the model.

42

Appendix A. Bootstrap majority rule consensus tree (consensus level = 50) over 2,000 pseudoreplicates of the Maximum Likelihood tree of the Loricariinae inferred from the combined analysis of sequences of partial 12S and 16S mitochondrial genes, and partial frtn4r nuclear gene. Numbers above branches indicate bootstrap supports above 50. Scale indicates the number of substitutions per site as expected by the model.

Appendix B. Bayesian inference majority rule consensus tree (consensus level = 0.50) obtained from the posterior distributions of 18,000 trees of the Loricariinae inferred from the combined analysis of sequences of partial 12S and 16S mitochondrial genes, and partial frtn4r nuclear gene. Numbers above branches indicate posterior probabilities above 0.50. Scale indicates the number of substitutions per site as expected by the model.

43

Reference

Data

Classification of the Loricariinae Tribe Loricariini

Isbrücker, 1979

Morphological, diagnostic

Harttiini

Subtribe Loricariina Planiloricariina Reganellina Rineloricariina Loricariichthyina Hemiodontichthyina Harttiina Metaloricariina

Group

Farlowellini Acestridiini Loricariini Hemiodontichthyina Rapp Py-Daniel, 1997 Armbruster, 2004

Morphological, phylogenetic Planiloricariina

Harttiini

Harttiina Farlowellina

Loricariini Covain and Fisch-Muller, 2007

Morphological, phenetic Harttiini Loricariini

Covain et al ., 2008 Rodriguez et al ., 2011

Molecular, phylogenetic

Harttiini

Loricariina Loricariina Loricariina Loricariina Loricariina Farlowellina

Pseudohemiodon group Loricaria group Rineloricaria group Loricariichthy s group Loricaria -Pseudohemiodon group Loricariichthys group Rineloricaria group

Genus Brochiloricaria , Crossoloricaria , Loricaria , Paraloricaria , Pseudohemiodon , Rhadinoloricaria , Ricola Planiloricaria Reganella Dasyloricaria , Ixinandria , Rineloricaria , Spatuloricaria Limatulichthys , Loricariichthys , Pseudoloricaria Hemiodontichthys Cteniloricaria , Harttia , Harttiella , Lamontichthys , Pterosturisoma , Sturisoma , Sturisomatichthys Metaloricaria Farlowella Acestridium Furcodontichthys, Hemiodontichthys, Loricariichthys, Reganella Pseudoloricaria Limatulichthys Apistoloricaria, Crossoloricaria, Dentectus, Planiloricaria, Pseudohemiodon, Rhadinoloricaria Loricaria Spatuloricaria Rineloricaria Harttia, Lamontichthys Aposturisoma, Farlowella, Sturisoma Sturisomatichthys Apistoloricaria, Crossoloricaria, Dentectus, Planiloricaria, Pseudohemiodon, Pyxiloricaria, Rhadinoloricaria, Reganella Brochiloricaria, Loricaria, Paraloricaria, Ricola Dasyloricaria, Ixinandria, Rineloricaria, Spatuloricaria Furcodontichthys, Hemiodontichthys, Limatulichthys, Loricariichthys, Pseudoloricaria Aposturisoma, Farlowella, Harttia, Harttiella, Lamontichthys, Metaloricaria, Pterosturisoma, Sturisoma, Sturisomatichthys Crossoloricaria, Loricaria, Planiloricaria, Spatuloricaria Hemiodontichthys, Limatulichthys, Loricariichthys Rineloricaria Fonchiiloricaria Metaloricaria Farlowella, Lamontichthys, Sturisoma, Sturisomatichthys Harttia

Species

Catalog Number

Field Number

Locality

Harttia guianensis Loricaria parnahybae Crossoloricaria venezuelae Dasyloricaria tuyrensis Farlowella aff. oxyrryncha* Farlowella platorynchus Hemiodontichthys acipenserinus Lamontichthys stibaros

MHNG 2643.016 MHNG 2602.067 INHS 35467 MHNG 2674.052 MHNG 2588.064 MHNG 2588.093 MHNG 2651.012 MHNG 2677.039

GF00–351 BR98–274 VZ 049 PA00–012 PE96–022 PE96–071 GY04–015 MUS 208

Limatulichthys punctatus* Loricaria clavipinna Loricariichthys maculatus Loricariichthys microdon Metaloricaria paucidens Planiloricaria cryptodon

MHNG 2651.013 MHNG 2640.044 MHNG 2621.042 MHNG 2650.054 MHNG 2677.086 MHNG 2677.038

GY04–018 PE98–002 SU01–056 GY04–012 GF00–083 MUS 211

Rineloricaria lanceolata Rineloricaria osvaldoi* Rineloricaria platyura Sturisoma monopelte Sturisoma robustum* Sturisomatichthys citurensis Fonchiiloricaria nanodon Fonchiiloricaria nanodon Spatuloricaria aff. caquetae* Spatuloricaria sp. Nanay

MHNG 2588.059 UFRJ 6–EF4 MHNG 2651.009 MHNG 2651.033 MHNG 2588.055 MHNG 2676.004 MHNG 2710.048 MHNG 2710.060 MHNG 2710.050 MHNG 2677.071

PE96–011 BR 1114 GY04–083 GY04–187 PE96–001 PA97–032 PE08-199 PE08-336 PE08-230 PE05-014

Apistoloricaria ommation

ANSP 182331

P6265

French Guiana, Marouini River Brazil, Rio Parnahyba Venezuela, Rio Santa Rosa Panama, Rio Ipeti Peru, Rio Tambopata Peru, Rio Ucayali Guyana, Rupununi River Peru, aquarium trade, Rio Itaya2 Guyana, Rupununi River Peru, Rio Putumayo Surinam, Sarramacca River Guyana, Rupununi River French Guiana, Marouini River Peru, aquarium trade, Rio Itaya2 Peru, Rio Tambopata Brazil, Rio Maranhão Guyana, Rupununi River Guyana, Sawarab River Peru, Rio de las Piedras Panama, Rio Tuyra Peru, Rio Monzon Peru, Rio Aucayacu Peru, Rio Huallaga Peru, aquarium trade, Rio Nanay2 Peru, Rio Amazonas

Apistoloricaria ommation Aposturisoma myriodon Aposturisoma myriodon Brochiloricaria macrodon Brochiloricaria sp. Uruguay Crossoloricaria aff. bahuaja Crossoloricaria bahuaja Crossoloricaria cephalaspis Crossoloricaria cephalaspis Crossoloricaria rhami Crossoloricaria variegata Cteniloricaria napova Cteniloricaria platystoma Cteniloricaria platystoma Cteniloricaria platystoma Cteniloricaria platystoma Cteniloricaria platystoma Dasyloricaria latiura Dasyloricaria tuyrensis Farlowella schreitmuelleri Farlowella acus Farlowella aff. rugosa Farlowella amazona Farlowella curtirostra Farlowella hahni

MHNG 2708.086 MHNG 2710.035 MHNG 2710.043 LBP 5048 MCP 28414 MHNG 2710.072 ANSP 180793 Stri-1449 Stri-1577 MHNG 2710.041 Stri-6781 MHNG 2704.030 MHNG 2672.067 MHNG 2674.003 MHNG 2650.082 MHNG 2700.054 MHNG 2643.015 Stri-1559 Stri-4140 MHNG 2601.087 MER95T-22 ANSP 179 768 MHNG 2601.065 MER95T-13 MHNG 2678.022

MUS 437 PE08-004 PE08-131 LBPN 24033 MCP 28414 PE08-714 P4078 53 22 PE08-120 36 SU07-650 SU05-340 SU05-039 GY04-336 GF07-265 GF00-352 20 51 BR98-106 42 T2200 BR98-052 43 PR-029

Peru, aquarium trade, Rio Amazonas2 Peru, Rio Huacamayo Peru, Rio Huyhuantal Brazil, Rio Paraguay Brazil, Rio Ibicui-Mirim Peru, Rio Cushabatai Peru, Rio Madre de Dios Colombia, Rio San Juan Colombia, Rio Atrato Peru, Rio Aguaytia Panama, Rio Tuira Brazil, Paru de Oeste River Suriname, Corantijn River Suriname, Suriname River Guyana, Essequibo River French Guiana, Mana River French Guiana, Marouini River Panama, Rio Atrato Panama, Rio Tuira Brazil, Rio Guamá Venezuela, Valencia Lake Guyana, Simoni River Brazil, Rio Acara Venezuela, Rio Motatan Argentina, Santa Fé

mt 12S+16S bases + GenBank No. 2440 EU310447 2423 EU310452 2419 EU310444 2419 EU310445 2433 EU310443 2432 EU310446 2422 EU310448 2433 EU310449 2426 EU310450 2427 EU310451 2428 EU310453 2427 EU310454 2438 EU310455 2418 EU310456 2423 EU310457 2427 EU310459 2423 EU310458 2439 EU310461 2440 EU310460 2438 EU310462 2432 HM592626 2432 HM592627 2421 HM592624 2422 HM592625 2417 KR478088 2423 KR478089 2435 KR477910 2435 KR477911 2425 KR478052 2426 KR478053 2415 KR478091 2417 KR478092 2421 KR478077 2421 KR478076 2416 KR478083 2422 KR478075 2440 KR477882 2439 KR477888 2441 KR477889 2437 KR477881 2437 KR477902 2439 KR477887 2424 KR477966 2424 KR477965 2437 KR477943 2440 KR477936 2435 KR477948 2432 KR477937 2435 KR477938 2437 KR477941

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Covain et al. 2008 Covain et al. 2008 Covain et al. 2008 Covain et al. 2008 Covain et al. 2008 Covain et al. 2008 Covain et al. 2008 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study

F-RTN4 bases + GenBank No. 2092 FJ013232 1967 FJ013231 1994 HM623647 1996 HM623639 2228 HM623650 2292 HM623649 2231 HM623645 2029 HM623648 1950 HM623644 1964 HM623653 2212 HM623642 1934 HM623643 2064 HM623637 1991 HM623646 2211 HM623640 2008 HM623652 2204 HM623641 1965 HM623651 2547 HM623636 2259 HM623635 2006 HM623656 2006 HM623657 1971 HM623654 1980 HM623655 1981 KR478422 1982 KR478423 2289 KR478244 2287 KR478245 1972 KR478386 1986 KR478387 1983 KR478425 1967 KR478426 2001 KR478411 1997 KR478410 1984 KR478417 1986 KR478409 2086 KR478216 2091 KR478222 2090 KR478223 2090 KR478215 2089 KR478236 2089 KR478221 2017 KR478300 2018 KR478299 2241 KR478277 2311 KR477936 2298 KR478282 2299 KR478271 2301 KR478272 2235 KR478275

Ref. Chiachio et al . 2008 Chiachio et al . 2008 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 Rodriguez et al . 2011 This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study

Farlowella knerii Farlowella mariaelenae Farlowella martini Farlowella nattereri Farlowella nattereri Farlowella oxyrryncha Farlowella oxyrryncha Farlowella oxyrryncha Farlowella oxyrryncha Farlowella oxyrryncha Farlowella oxyrryncha Farlowella oxyrryncha Farlowella paraguayensis Farlowella paraguayensis Farlowella platorynchus Farlowella platorynchus Farlowella platorynchus Farlowella reticulata Farlowella reticulata Farlowella reticulata Farlowella smithi Farlowella taphorni Farlowella vittata Harttia aff. punctata Harttia carvalhoi Harttia carvalhoi Harttia dissidens Harttia dissidens Harttia duriventris Harttia fluminensis Harttia fowleri Harttia gracilis Harttia guianensis Harttia guianensis Harttia kronei Harttia kronei Harttia kronei Harttia kronei Harttia leiopleura Harttia leiopleura Harttia longipinna Harttia loricariformis Harttia novalimensis Harttia punctata Harttia punctata Harttia sp. 1 Xingu Harttia sp. 2 Xingu Harttia sp. 3 Xingu Harttia sp. Rio São Francisco Harttia sp. Serra do Cipó Harttia sp. Tapajos Harttia sp. Tocantins Harttia sp. Três Marias

MHNG 2710.052 VZ-59 VZ-126 MHNG 2650.099 MHNG 2654.067 MHNG 2710.034 MHNG 2613.035 MHNG 2601.095 LBP 2441 MHNG 2710.069 MHNG 2710.081 LBP4043 Stri-2205 LBP 5217 MHNG 2650.096 MHNG 2602.021 MHNG 2710.094 MHNG 2683.081 MHNG 2683.070 MHNG 2681.060 ANSP 180541 VZ-89 VZ-63 LBP 5839 MHNG 2587.027 LBP 2115 LBP 5859 LBP 5863 LBP 7505 MHNG 2690.013 MHNG 2643.022 LBP 6331 MHNG 2662.091 MHNG 2680.053 MHNG 2586.058 LBP 2661 LBP 2883 LBP 1269 LBP 6847 LBP 6492 DZSJRP 2819 LBP 2121 LBP 5836 MHNG 2645.059 MHNG 2645.053 LBP 5845 LBP 5860 LBP 5861 LBP 5838 LBP 6528 LBP 5857 LBP 5850 LBP 5838

PE08-259 45 49 GY04-291 GY04-306 PE08-051 CA 21 BR98-118 LBPN 16200 PE08-698 PE08-823 LBPN 22907 25 LBPN 26396 GY04-290 BR98-163 PE08-906 GF06-637 GF06-588 GF06-118 P4099 48 46 LBP 28353 BR 1236 LBP 21352 LBP 28331 LBP 28339 LBP 34804 SU01-445 GF99-202 LBP 29819 GF03-160 RV-21 BR 1166 LBP 17427 LBP 18609 LBP 11215 LBP 31528 LBP 31545 BR98-747 LBP 21362 LBP 28348 BR 995 BR 1051 LBP 28327 LBP 28333 LBP 28335 LBP 28352 LBP 31652 LBP 28329 LBP 28367 LBP 28351

Peru, Rio Aspuzana Venezuela, Rio Caipe Venezuela, Rio Aroa Guyana, Kurupukari cross Guyana, Kurupukari cross Peru, Rio Huacamayo Peru, Rio Ucayali Brazil, Rio Guamá Brazil, Rio Araguiaia Peru, Rio Neshua Peru, Rio Cushabatai Brazil, Rio Jurua Paraguay, Arroyo Curuguati Brazil, Rio Paraná Guyana, Kurupukari cross Brazil, Rio Peritoro Peru, Rio Ucayali French Guiana, Maroni River French Guiana, Mana River French Guiana, Oyapock River Peru, Rio Manuripe Venezuela, Rio Mayupa Venezuela, Rio Caipe Brazil, Rio Tocantins Brazil, Rio Paraíba do Sul Brazil, Rio Paraíba do Sul Brazil, Rio Tapajós Brazil, Rio Tapajós Brazil, Rio Tapajós Suriname, Coppename River French Guiana, Oyapock River Brazil, Rio Paraná French Guiana, Approuague River French Guiana, Sinnamary River Brazil, Rio Ribeira de Iguape Brazil, Rio Ribeira de Iguape Brazil, Rio Ribeira de Iguape Brazil, Rio Ribeira de Iguape Brazil, Rio São Francisco Brazil, Rio São Francisco Brazil, Rio São Francisco Brazil, Rio Paraíba do Sul Brazil, Rio São Francisco Brazil, Rio Tocantins Brazil, Rio Tocantins Brazil, Rio Xingu Brazil, Rio Xingu Brazil, Rio Xingu Brazil, Rio São Francisco Brazil, Rio São Francisco Brazil, Rio Tapajós Brazil, Rio Tocantins Brazil, Rio São Francisco

2437 KR477954 2439 KR477939 2436 KR477940 2439 KR477952 2438 KR477944 2437 KR477953 2437 KR477960 2440 KR477958 2436 KR477956 2437 KR477955 2437 KR477957 2437 KR477959 2437 KR477961 2434 KR477962 2435 KR477949 2435 KR477950 2432 KR477951 2439 KR477963 2438 KR477942 2437 KR477964 2436 KR477945 2433 KR477946 2438 KR477947 2433 KR477898 2432 KR477891 2433 KR477890 2435 KR477892 2435 KR477914 2432 KR477915 2435 KR477884 2442 KR477880 2433 KR477916 2438 KR477885 2443 KR477886 2424 KR477900 2426 KR477894 2425 KR477895 2426 KR477899 2435 KR477918 2436 KR477917 2429 KR477903 2435 KR477896 2429 KR477897 2431 KR477893 2430 KR477905 2433 KR477907 2432 KR478245 2436 KR477908 2429 KR477904 2431 KR477919 2436 KR477906 2433 KR477901 2429 KR477920

This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study

2234 KR478288 2309 KR478273 2307 KR478274 2305 KR478286 2306 KR478278 2239 KR478287 2241 KR478294 2240 KR477958 2183 KR478290 2242 KR478289 2242 KR478291 2228 KR478293 2236 KR478295 2238 KR478296 2296 KR478283 2302 KR478284 2301 KR478285 2243 KR478297 2242 KR478276 2242 KR478298 2236 KR478279 2168 KR478280 2303 KR478281 2084 KR478232 2046 KR478225 2046 KR478224 1955 KR478226 1954 KR478248 1951 KR478249 2092 KR478218 2086 KR478214 2041 KR478250 2092 KR478219 2092 KR478220 2081 KR478234 2083 KR478228 2080 KR478229 2080 KR478233 2068 KR478252 2068 KR478251 2072 KR478237 2041 KR478230 2060 KR478231 2084 KR478227 2084 KR478239 1971 KR478241 1973 KR478246 1973 KR478242 2061 KR478238 2061 KR477919 1968 KR478240 2085 KR478235 2061 KR478254

This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study

Harttia surinamensis Harttia torrenticola Harttia tuna Harttiella crassicauda Harttiella crassicauda Harttiella crassicauda Harttiella intermedia Harttiella intermedia Harttiella intermedia Harttiella longicauda Harttiella longicauda Harttiella longicauda Harttiella longicauda Harttiella longicauda Harttiella longicauda Harttiella lucifer Harttiella lucifer Harttiella lucifer Harttiella lucifer Harttiella lucifer Harttiella lucifer Harttiella lucifer Harttiella lucifer Harttiella lucifer Harttiella parva Harttiella parva Harttiella parva Harttiella pilosa Harttiella pilosa Harttiella pilosa Hemiodontichthys acipenserinus Hemiodontichthys acipenserinus Hemiodontichthys acipenserinus Hemiodontichthys acipenserinus Ixinandria steinbachi Lamontichthys filamentosus Lamontichthys filamentosus Lamontichthys llanero Lamontichthys stibaros Limatulichthys punctatus Limatulichthys punctatus Limatulichthys punctatus Limatulichthys punctatus Limatulichthys punctatus Limatulichthys punctatus Limatulichthys punctatus Loricaria sp. Guyana Loricaria sp. Guyana Loricaria aff. nickeriensis Loricaria aff. parnahybae Loricaria apeltogaster Loricaria cataphracta Loricaria cataphracta

MHNG 2674.042 LBP 5835 MHNG 2704.029 MHNG 2679.098 MHNG 2674.051 MHNG 2674.051 MHNG 2713.087 MHNG 2713.087 MHNG 2713.087 MHNG 2723.094 MHNG 2723.094 MHNG 2723.094 MHNG 2699.070 MHNG 2699.070 MHNG 2699.070 MHNG 2721.088 MHNG 2721.088 MHNG 2721.088 MHNG 2721.091 MHNG 2721.091 MHNG 2721.091 MHNG 2712.085 MHNG 2712.085 MHNG 2712.085 MHNG 2723.093 MHNG 2723.093 MHNG 2723.093 MHNG 2682.055 MHNG 2682.055 MHNG 2724.002 MHNG 2588.057 MHNG 2602.007 MCP 28819 LBP 5524 NA MHNG 2680.009 LBP 162 MHNG 2749.019 MHNG 2710.049 ANSP 182707 MHNG 2602.009 AUM 42223 LBP 5055 MHNG 2710.037 LBP 5285 LBP 5249 MHNG 2650.057 MHNG 2651.031 MHNG 2681.009 LBP 1615 NA MHNG 2749.022 MHNG 2683.061

SU05-001 LBP 28346 SU07-644 MUS 306 MUS 221 MUS 231 MUS 650 MUS 651 MUS 652 MUS 470 MUS 463 MUS 456 GF07-026 GF07-082 GF07-111 GF10-034 GF10-043 GF10-037 GF10-051 GF10-053 GF10-055 MUS 592 MUS 593 MUS 594 MUS 606 MUS 607 MUS 611 GF06-344 GF06-343 GF03-033 PE96-005 BR98-138 MCP 28819 LBPN 26640 IXS2 MUS 310 LBPN 4038 MUS 356 PE08-224 P6232 BR98-140 V5319 LBPN 23618 PE08-112 LBPN 26769 LBPN 26472 GY04-110 GY04-191 GF06-044 LBPN 11690 NA GF98-044 GF06-570

Suriname, Suriname River Brazil, Rio São Francisco Brazil, Paru de Oeste River Suriname, Nassau Mountains Suriname, Nassau Mountains Suriname, Nassau Mountains French Guiana, Trinité Mountains French Guiana, Trinité Mountains French Guiana, Trinité Mountains French Guiana, Balenfois Mountains French Guiana, Balenfois Mountains French Guiana, Balenfois Mountains French Guiana, Trinité Mountains French Guiana, Trinité Mountains French Guiana, Trinité Mountains French Guiana, Lucifer Mountains French Guiana, Lucifer Mountains French Guiana, Lucifer Mountains French Guiana, Lucifer Mountains French Guiana, Lucifer Mountains French Guiana, Lucifer Mountains French Guiana, Crique Limonade French Guiana, Crique Limonade French Guiana, Crique Limonade French Guiana, Atachi Bakka Mountains French Guiana, Atachi Bakka Mountains French Guiana, Atachi Bakka Mountains French Guiana, Tortue Mountains French Guiana, Tortue Mountains French Guiana, Tortue Mountains Peru, Madre de Dios Brazil, Rio Guamá Brazil, Rio Purus Brazil, Rio Jari Argentina, Salta Peru, aquarium trade Brazil, Rio Branco Colombia, aquarium trade Peru, Rio Huallaga Peru, Rio Itaya Brazil, Rio Guamá Venezuela, Rio Orinoco Brazil, Rio Jurua Peru, Rio Agaytia Brazil, Rio Jari Brazil, Rio Jari Guyana, Pirara River Guyana, Sawarab bridge French Guiana, Oyapock River Brazil, Rio Araguaia Argentina, Entre Rios French Guiana, Kourou River French Guiana, Mana River

2438 KR477883 2433 KR477913 2437 KR477909 2418 KR478145 2417 KR478146 2418 KR478131 2418 KR478164 2418 KR478165 2418 KR478135 2418 KR478136 2419 KR478159 2418 KR478133 2418 KR478144 2422 KR478134 2418 KR478139 2414 KR478153 2414 KR478154 2414 KR478155 2414 KR478156 2414 KR478158 2414 KR478157 2414 KR478478 2414 KR478151 2413 KR478152 2416 KR478147 2416 KR478148 2416 KR478477 2417 KR478132 2417 KR478137 2419 KR478138 2425 KR478140 2421 KR478141 2424 KR478142 2422 KR478143 2427 KR477986 2434 KR478262 2433 KR477930 2434 KR477928 2433 KR477931 2424 KR478095 2424 KR478094 2425 KR478097 2422 KR478096 2427 KR478093 2427 KR478183 2426 KR478182 2424 KR478068 2424 KR478069 2424 KR478061 2434 KR478059 2427 KR478056 2428 KR478057 2425 KR478062

This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study

2092 KR478217 2054 KR478247 2092 KR478243 2026 KR478474 2026 KR478475 2026 KR478460 2021 KR478490 2022 KR478491 2022 KR478464 2005 KR478465 2005 KR478485 2005 KR478462 2022 KR478473 2022 KR478463 2022 KR478468 2152 KR478479 2149 KR478480 2152 KR478481 2152 KR478482 2152 KR478484 2148 KR478483 NA NA NA 2026 KR478476 2026 KR478477 2026 KR478478 2026 KR478461 2026 KR478466 2026 KR478467 2259 KR478469 2217 KR478470 2238 KR478471 2283 KR478472 2513 KR478320 2116 KR478263 2115 KR478264 2089 KR478262 2040 KR478265 1957 KR478429 1963 KR478428 1960 KR478431 1959 KR478430 1959 KR478427 1956 KR478508 1958 KR478507 1979 KR478402 1983 KR478403 1944 KR478395 1977 KR478393 1973 KR478390 1943 KR478391 1933 KR478396

This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study

Loricaria cataphracta Loricaria cataphracta Loricaria cf. lata Loricaria nickeriensis Loricaria prolixa Loricaria prolixa

MHNG 2744.037 MHNG 2749.021 LBP 5053 MHNG 2672.080 LBP 7511 LBP 7511

SU08-042 SU08-943 LBPN 16148 SU05-334 LBPN 34925 LBPN 34924

Suriname, Suriname River Suriname, Commewijne River Brazil, Rio Araguaia Suriname, Corantijn River Brazil, Rio Paraná Brazil, Rio Paraná

Loricaria simillima Loricaria sp. Araguaia Loricaria sp. Branco Loricaria sp. Branco Loricaria sp. Mato Grosso Loricaria sp. Orinoco Loricaria sp. Paraguay Loricaria sp. Rupununi Loricaria tucumanensis Loricariichthys anus Loricariichthys anus Loricariichthys anus Loricariichthys castaneus Loricariichthys castaneus Loricariichthys castaneus Loricariichthys cf. ucayalensis Loricariichthys derbyi Loricariichthys derbyi Loricariichthys labialis Loricariichthys melanocheilus Loricariichthys platymetopon Loricariichthys sp. Amazonas Loricariichthys sp. Jari Loricariichthys sp. Jari Loricariichthys sp. Orinoco Loricariichthys sp. Rio Baia Metaloricaria nijsseni Metaloricaria nijsseni Metaloricaria nijsseni Metaloricaria paucidens Paraloricaria agastor Paraloricaria agastor Paraloricaria vetula

MHNG 2677.075 LBP 5049 LBP 169 LBP 223 MCP 36566 AUM 42224 MHNG 2677.003 MHNG 2651.036 NA MCP 28415 MCP 21317 LBP 578 MHNG 2583.068 LBP 7489 LBP 7490 ANSP 182668 MHNG 2602.061 LBP 5531 MHNG 2677.001 MCP 28915 MHNG 2677.004 Stri-531 LBP 5517 LBP 5420 AUM 42225 LBP 3094 MHNG 2674.025 MHNG 2672.053 MHNG 2690.016 MHNG 2681.042 NA NA NA

PE05-030 LBPN 11506 LBPN 4032 LBPN 4101 MCP 36566 V5315 PY9093 GY04-129 AG06-018 MCP 28415 MCP 21317 LBPN 7309 BR 162 LBPN 35548 LBPN 35549 P6046 BR98-250 LBPN 27214 PY9094 MCP 28915 PY9098 28 LBPN 26622 LBPN 27135 V5310 LBPN 19263 SU05-012 SU05-359 SU01-459 GF06-108 AG06-017 AG06-019 YC-008

Peru, aquarium trade, Rio Amazonas Brazil, Rio Araguaia Brazil, Rio Branco Brazil, Rio Branco Paraguay, Mato Grosso Venezuela, Rio Orinoco Paraguay, Rio Paraguay Guyana, Rupununi River Argentina, Ita-Ibate Brazil, Rio Grande do Sul Brazil, Rio Grande do Sul Brazil, Rio Guiaba Brazil, surroundings of Rio de Janeiro Brazil Brazil Peru, Rio Nanay Brazil, Rio Parnahyba Brazil, Rio Parnahyba Paraguay, Rio Paraguay Brazil, Rio Ibicui-Mirim Paraguay, Rio Paraguay Peru, Rio Amazonas Brazil, Rio Jari Brazil, Rio Jari Venezuela, Rio Orinoco Brazil, Rio Baia Suriname, Suriname River Suriname, Corantijn River Suriname, Coppename River French Guiana, Oyapock River Argentina, Ita-Ibate Argentina, Puerto Abra Argentina, Entre Rios

2

2424 KR478054 2424 KR478055 2423 KR478058 2424 KR478060 2429 KR478063 2428 KR478064 2424 KR478065

Pseudohemiodon aff. apithanos

MHNG 2677.070

PE05-009

Peru, aquarium trade, Rio Amazonas2

2429 KR478066 2424 KR478067 2424 KR478090 2429 KR478070 2426 KR478071 2427 KR478072 2427 KR478074 2423 KR478073 2430 KR478175 2430 KR478174 2429 KR478176 2430 KR478114 2429 KR478115 2429 KR478116 2424 KR478117 2428 KR478119 2425 KR478171 2429 KR478178 2426 KR478120 2427 KR478118 2426 KR478173 2428 KR478112 2427 KR478121 2426 KR478109 2429 KR478113 2435 KR477934 2437 KR477967 2441 KR477933 2439 KR477932 2427 KR478167 2426 KR478168 2425 KR478166 2416 KR478080

2

2418 KR478078 2411 KR478079 2414 KR478110

Pseudohemiodon aff. apithanos Pseudohemiodon aff. apithanos

ANSP 178115 MHNG 2710.086

P1759 PE08-852

Peru, aquarium trade, Rio Itaya Peru, Rio Cushabatai

Pseudohemiodon apithanos

MHNG 2677.073

PE05-020

Peru, aquarium trade, Rio Itaya2 2

Pseudohemiodon apithanos

MHNG 2677.074

PE05-026

Peru, aquarium trade, Rio Nanay

2414 KR478081

Pseudohemiodon laminus Pseudohemiodon laticeps Pseudohemiodon laticeps

MHNG 2677.077 LBP 4332 NA

PE05-035 LBPN 24034 NA

Peru, aquarium trade, Rio Amazonas2 Brazil, Rio Paraguay Argentina, Corrientes

2426 KR478082

Pseudohemiodon sp. Pseudoloricaria laeviuscula Pseudoloricaria laeviuscula Pseudoloricaria laeviuscula Pseudoloricaria laeviuscula

MHNG 2677.076 AUM 44646 AUM 44646 AUM 44646 INPA 28991

PE05-034 G5231 G5232 G5233 MUS 517

Peru, aquarium trade, Rio Amazonas2 Guyana, Takutu River Guyana, Takutu River Guyana, Takutu River Brazil, Rio Madeira

2424 KR478169 2424 KR478170 2406 KR478111 2427 KR478099 2427 KR478100 2427 KR478103 2430 KR478098

This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study

1932 KR478388 1932 KR478389 1961 KR478392 1932 KR478394 1975 KR478397 1975 KR478398 1950 KR478399 2006 KR478400 1969 KR478401 1967 KR478424 1949 KR478404 1961 KR478071 1930 KR478406 1967 KR478408 1976 KR478407 2280 KR478500 2276 KR478499 2272 KR478501 2230 KR478444 2275 KR478445 2279 KR478446 2197 KR478447 2240 KR478449 2243 KR478497 1956 KR478503 2271 KR478450 2232 KR478448 2231 KR478498 2228 KR478442 2232 KR478451 NA 2226 KR478443 2089 KR478268 2089 KR478301 2089 KR478267 2073 KR478266 1984 KR478493 1984 KR478494 1984 KR478492 2003 KR478414 2003 KR478412

This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study

This study

2004 KR478413 2012 KR478440

This study

2012 KR478415

This study

This study This study This study

2009 KR478416

This study This study This study

This study This study This study This study This study

2003 KR478495 2004 KR478496 1980 KR478441 1990 KR478433 1990 KR478434 1990 KR478436 1990 KR478432

This study

This study This study This study This study This study

Pterosturisoma microps Rhadinoloricaria aff. macromystax Rhadinoloricaria sp. Orinoco Rhadinoloricaria sp. Orinoco Rhadinoloricaria sp. Orinoco Rineloricaria aequalicuspis Rineloricaria aff. cadeae Rineloricaria aff. cadeae Rineloricaria aff. fallax Rineloricaria aff. langei Rineloricaria aff. latirostris Rineloricaria aff. phoxocephala Rineloricaria aff. phoxocephala Rineloricaria aff. phoxocephala Rineloricaria aff. phoxocephala Rineloricaria aff. phoxocephala Rineloricaria aff. phoxocephala Rineloricaria aff. stewarti Rineloricaria aff. stewarti Rineloricaria aff. stewarti Rineloricaria aff. stewarti Rineloricaria aff. stewarti Rineloricaria aff. stewarti Rineloricaria aff. stewarti Rineloricaria aff. strigilata Rineloricaria altipinnis Rineloricaria altipinnis Rineloricaria cadeae Rineloricaria catamarcensis Rineloricaria catamarcensis Rineloricaria cf. kronei Rineloricaria cf. latirostris Rineloricaria eigenmanni Rineloricaria fallax Rineloricaria fallax Rineloricaria fallax Rineloricaria fallax Rineloricaria fallax Rineloricaria fallax Rineloricaria formosa Rineloricaria formosa Rineloricaria heteroptera Rineloricaria heteroptera Rineloricaria heteroptera Rineloricaria heteroptera Rineloricaria hoehnei Rineloricaria jaraguensis Rineloricaria lanceolata Rineloricaria lanceolata Rineloricaria lanceolata Rineloricaria lanceolata Rineloricaria lanceolata Rineloricaria lanceolata

MHNG 2677.072 ANSP 182349 ANSP 185044 AUM 42094 AUM 42094 MCP 29282 LBP 901 LBP4772 LBP 4085 LBP 1189 MHNG 2749.014 MCP 28832 MCP 28832 MCP 28832 LBP 4123 MHNG 2749.013 ANSP 182368 MHNG 2663.003 MHNG 2681.019 MHNG 2682.091 MHNG 2683.049 MHNG 2617.015 MHNG 2704.040 MHNG 2749.011 LBP 2949 Stri-3589 MHNG 2709.086 MCP 21217 NA MHNG 2680.033 LBP 1248 LBP 771 NA MHNG 2651.054 MHNG 2672.015 MHNG 2650.072 MHNG 2651.034 MHNG 2650.067 LBP 4343 ANSP 185291 AUM 43885 AUM 43886 AUM 43886 AUM 43886 AUM 43928 MHNG 2678.018 LBP 729 MHNG 2613.029 MHNG 2651.029 MHNG 2651.059 MHNG 2680.008 Stri-2422 LBP 1557

PE05-016 T2364 T4029 V5507 V5508 MCP 29282 LBPN 7359 LBPN 25580 LBPN 23512 LBPN 10585 MUS 491 495 496 NA LBPN 23617 PI 720 T2101 GF03-196 GF06-077 GF06-428 GF06-538 MUS SUJM-064 SU08-945 LBPN 19534 40 PA97-045 MCP 21217 RC NA LBPN 11175 LBPN 8534 MUS 494 GY04-146 SU05-412 GY04-009 GY04-396 GY04-384 LBPN 24075 V5405 V5531 V5530 V5534 V5529 V5562 PR-018 LBPN 8268 CA-01 GY04-172 GY04-252 MUS 244 26 LBPN 11505

Peru, aquarium trade Guyana, Rupununi River Venezuela, Rio Orinoco Venezuela, Rio Orinoco Venezuela, Rio Orinoco Brazil, Arroio Molha Coco Brazil, Eldorado do Sul Brazil, Rio Guiaba Brazil, Rio Japim Brazil, Rio Iguaçu Brazil, aquarium trade Brazil, Rio Purus Brazil, Rio Purus Brazil, Rio Purus Brazil, Rio Jurua Peru, Rio Momon Guyana, Essequibo River French Guiana, Approuague River French Guiana, Oyapock River French Guiana, Maroni River French Guiana, Mana River French Guiana, Sinnamary River Suriname, Suriname River Suriname, Commewijne River Brazil, Córrego da batata Panama, Rio Chucunaque Panama, Rio Chucunaque Brazil, Rio Grande do Sul Argentina, Salta Argentina, Rio Sali Brazil, Rio Ribeira do Iguape Brazil, Rio Marumbi Colombia, aquarium trade Guyana, Takutu River Suriname, Corantijn River Guyana, Rupununi River Guyana, Berbice River Guyana, Demerara River Brazil, Boa Vista Venezuela, Rio Orinoco Venezuela, Rio Orinoco Venezuela, Rio Orinoco Venezuela, Rio Orinoco Venezuela, Rio Orinoco Venezuela, Rio Orinoco Paraguay, Paraguay River Brazil, Jaragua do Sul, Rio Itapocu Peru, Rio Ucayali Guyana, Takutu River Guyana, Mauishparu River Brazil, Purus River Argentine Rio Corrientes Brazil, Rio Araguaia

2439 KR477921 2414 KR478084 2415 KR478085 2414 KR478086 2415 KR478087 2427 KR478213 2424 KR477987 2424 KR477976 2420 KR477979 2425 KR478323 2425 KR478185 2427 KR478104 2426 KR478126 2427 KR478125 2431 KR478127 2431 KR478128 2431 KR478038 2425 KR478023 2422 KR478028 2425 KR478025 2425 KR478024 2425 KR478029 2429 KR478035 2426 KR478037 2417 KR477992 2425 KR478026 2423 KR478027 2427 KR477991 2429 KR477989 2426 KR477988 2429 KR478108 2429 KR478347 2428 KR477984 2429 KR477980 2427 KR477975 2427 KR477978 2427 KR477973 2427 KR477977 2429 KR477974 2429 KR477981 2429 KR477982 2429 KR477968 2429 KR477969 2429 KR477970 2428 KR477971 2424 KR477972 2426 KR477985 2423 KR478203 2424 KR477996 2424 KR477995 2424 KR477997 2425 KR478326 2424 KR477994

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2075 KR478255 1967 KR478418 1979 KR478419 1980 KR478420 1979 KR478421 2499 KR478534 2492 KR478321 2533 KR478310 1984 KR478313 2501 KR478324 2507 KR478510 2243 KR478437 2246 KR478455 2243 KR478454 2242 KR478456 2241 KR478457 2244 KR478372 2209 KR478357 2214 KR478362 2209 KR478359 2209 KR478358 2209 KR478363 2210 KR478369 2210 KR478371 2258 KR478326 1866 KR478360 1866 KR478361 2479 KR478325 2511 KR478323 2511 KR478322 2498 KR478439 2802 KR478348 2206 KR478318 2284 KR478314 2280 KR478309 2281 KR478312 2281 KR478307 2280 KR478311 2285 KR477974 2253 KR478315 2253 KR478316 2219 KR478302 2219 KR478303 2219 KR478304 2220 KR478305 2221 KR478306 2208 KR478319 2232 KR478528 2231 KR478330 2232 KR478329 2232 KR478331 2219 KR478327 2198 KR478328

This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study

Rineloricaria lanceolata Rineloricaria longicauda Rineloricaria melini Rineloricaria melini Rineloricaria microlepidogaster Rineloricaria misionera Rineloricaria morrowi Rineloricaria parva Rineloricaria parva Rineloricaria pentamaculata Rineloricaria platyura Rineloricaria platyura Rineloricaria platyura Rineloricaria platyura Rineloricaria platyura Rineloricaria platyura Rineloricaria platyura Rineloricaria platyura Rineloricaria quadrensis Rineloricaria sneiderni Rineloricaria sp. Agua Santa Rineloricaria sp. Ao Itai Rineloricaria sp. Araguaia Rineloricaria sp. Betari Rineloricaria sp. Betari Rineloricaria sp. Carombé Rineloricaria sp. Corantijn Rineloricaria sp. Corantijn Rineloricaria sp. Corantijn Rineloricaria sp. Corrego Seco Rineloricaria sp. Guama 4 Rineloricaria sp. Guama 5 Rineloricaria sp. Guama 5 Rineloricaria sp. Guama 5 Rineloricaria sp. Guama 6 Rineloricaria sp. Huacamayo Rineloricaria sp. Huacamayo Rineloricaria sp. Macacu Rineloricaria sp. Maroni 2 Rineloricaria sp. Maroni 2 Rineloricaria sp. Martinso Rineloricaria sp. Mongaguá Rineloricaria sp. Orinoco Rineloricaria sp. Orinoco Rineloricaria sp. Panama Rineloricaria sp. Paraiba do Sul Rineloricaria sp. Parguaza Rineloricaria sp. Piedade Rineloricaria sp. Previsto Rineloricaria sp. Puerto Ayacucho Rineloricaria sp. Ribeira Rineloricaria sp. Rio da Toca Rineloricaria sp. São João

MHNG 2710.033 MCP 38347 LBP 4409 MHNG 2680.011 MCP 21263 MHNG 2680.034 MHNG 2749.015 MHNG 2678.014 LBP 5 LBP 1319 LBP 1731 MHNG 2601.081 MHNG 2663.004 NA MHNG 2650.074 MHNG 2749.012 MHNG 2601.063 MCP 28832 MCP 21195 Stri-1399 MHNG 2587.054 MHNG 2587.015 LBP 1750 MHNG 2586.083 MHNG 2586.072 LBP 2654 MHNG 2671.085 MHNG 2704.035 MHNG 2722.048 MHNG 2586.065 MHNG 2601.091 MHNG 2601.044 MHNG 2601.042 MHNG 2602.025 MHNG 2601.063 MHNG 2613.032 MHNG 2710.032 MHNG 2587.082 MHNG 2749.018 MHNG 2749.018 MHNG 2586.089 LBP 2128 AUM 44067 AUM 44067 MHNG 2749.016 MHNG 2583.065 LBP 2308 MHNG 2586.055 MHNG 2710.047 MHNG 2749.017 MHNG 2586.088 MHNG 2587.078 LBP 802

PE08-053 MCP 38347 LBPN 24247 MUS 312 MCP 21263 MUS PI 719 PR-009 LBPN 3656 LBPN 11000 LBPN 12864 BR98-088 GF03-193 MUS 334 GY04-197 GF99-009 BR98-049 497 MCP 21195 52 BR1253 BR1215 LBPN 11818 BR1190 BR1184 LBPN 17410 SU05-450 SU07-017 SU01-417 BR1176 BR98-112 BR98-010 BR98-008 BR98-167 BR98-047 CA-33 PE08-057 BR1273 SU08-441 SU08-442 BR1196 LBPN 21378 V5435 V5437 PA00-011 BR 156 LBPN 15846 BR1163 PE08-186 MUS 489 BR1195 BR1269 LBPN 7954

Peru, Rio Huacamayo Brazil, Rio Grande do Sul Brazil, Rio Negro, Barcelos Brazil, aquarium trade Brazil, Rio Grande do Sul Argentina, Rio Cuna-Piru Peru, Rio Momon Argentina, Santa Fé Brazil, Rio Paraguay Brazil, Rio Tibagi Brazil, Rio Amazonas Brazil, Rio Guamá French Guiana, Approuague River Brazil, Rio Purus Guyana, Takutu River French Guiana, Kaw Brazil, Rio Acara Brazil, Rio Purus Brazil, Lagoa Fortaleza Colombia, Rio Baudo Brazil, Rio Paraíba do Sul Brazil, Rio Paraíba do Sul Brazil, Rio Araguaia Brazil, Rio Betari Brazil, Rio Betari Brazil, Rio Carombé Suriname, Corantijn River Suriname, Sipaliwini River Suriname, Nickerie River Brazil, Rio Ribeira do Iguape Brazil, Rio Guamá Brazil, Rio Gurupi Brazil, Rio Gurupi Brazil, Rio Piria Brazil, Rio Acara Peru, Rio Pisqui Peru, Rio Huacamayo Brazil, Rio da Toca Suriname, Paloemeu River Suriname, Paloemeu River Brazil, Rio Martinso Brazil, Riacho Sitio do Meio Venezuela, Rio Orinoco Venezuela, Rio Orinoco Panama, Rio Ipeti Brazil, Rio Paraíba do Sul Venezuela, Rio Parguaza Brazil, Rio Piedade Peru, Rio Previsto Venezuela, aquarium trade Brazil, Rio Ribeira do Iguape Brazil, Rio Macacu Brazil, Rio São João

2423 KR478202 2426 KR478018 2427 KR477983 2427 KR478013 2426 KR478212 2421 KR477999 2425 GBxxxxx 2433 KR478012 2433 KR478190 2432 KR478040 2421 KR478036 2428 KR478123 2427 KR478102 2426 KR478129 2424 KR478122 2427 KR478101 2428 KR478124 2426 KR478130 2427 KR478106 2426 KR478001 2426 KR478194 2424 KR478191 2428 KR478040 2429 KR478192 2429 KR478172 2427 KR478034 2431 KR478005 2430 KR478002 2433 KR478006 2429 KR478184 2426 KR478199 2425 KR478200 2425 KR478201 2417 KR478198 2427 KR478501 2431 KR478015 2431 KR478016 2425 KR478017 2430 KR478003 2430 KR478004 2429 KR478020 2429 KR478019 2428 KR478030 2428 KR478031 2427 KR478181 2427 KR478107 2427 KR478032 2429 KR478039 2433 KR478195 2427 KR478033 2429 KR478193 2423 KR478022 2429 KR478188

This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study

2230 KR478527 2804 KR478352 2204 KR478317 2203 KR478347 2504 KR478533 2214 KR478333 2280 KR478334 2522 KR478346 2506 KR478515 2021 KR478375 2220 KR478370 NA 953 KR478435 2229 KR478458 2220 KR478452 NA 949 KR478453 931 KR478459 2504 KR478438 2244 KR478335 2790 KR478519 2507 KR478516 2034 KR478040 2515 KR478517 NA 2208 KR478368 2188 KR478339 2188 KR478336 2194 KR478340 2716 KR478509 2474 KR478524 2508 KR478525 2510 KR478526 2508 KR478523 2285 KR478502 2453 KR478349 2528 KR478350 2506 KR478351 2249 KR478337 2249 KR478338 2802 KR478354 2786 KR478353 2197 KR478364 2197 KR478365 989 KR478506 NA 2217 KR478366 2226 KR478373 2528 KR478520 2279 KR478367 2493 KR478518 2573 KR478356 2793 KR478513

This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study

Rineloricaria sp. São João 2 Rineloricaria sp. São João 2 Rineloricaria sp. Ucayali Rineloricaria sp. Ucayali 2 Rineloricaria sp. Uruguay Rineloricaria sp. Von Humbolt Rineloricaria stewarti Rineloricaria stewarti Rineloricaria stewarti Rineloricaria stewarti Rineloricaria strigilata Rineloricaria teffeana Rineloricaria uracantha Rineloricaria uracantha Rineloricaria wolfei Spatuloricaria cf. evansii Spatuloricaria puganensis Spatuloricaria puganensis Spatuloricaria sp. Araguaia Spatuloricaria sp. Ireng Spatuloricaria sp. Magdalena 1 Spatuloricaria sp. Magdalena 2 Spatuloricaria sp. Magdalena 2 Spatuloricaria sp. Magdalena 2 Spatuloricaria sp. Orinoco Sturisoma aureum Sturisoma aureum Sturisoma cf. guentheri Sturisoma dariense Sturisoma festivum Sturisoma frenatum Sturisoma nigrirostrum Sturisoma panamense Sturisoma robustum Sturisoma sp. Rio Branco Sturisomatichthys leightoni Ancistrus cirrhosus 1

MHNG 2586.052 MHNG 2586.052 MHNG 2710.095 LBP 3273 MCP 21616 MHNG 2710.068 MHNG 2651.057 MHNG 2651.027 MHNG 2671.015 MHNG 2671.084 MCP 23751 NA Stri-1662 LBP 2762 ANSP 182695 LBP 2385 ANSP 180486 ANSP 180789 LBP 1556 ANSP 182372 MHNG 2722.096 IAvHP IAvHP IAvHP ANSP 185303 NA MHNG 2684.019 ANSP 182587 MHNG 2674.059 MER95T-20 Stri-872 ANSP 178322 MHNG 2674.058 MHNG 2677.002 LBP 1615 NA MHNG 2645.037

BR1155 BR1156 PE08-905 LBPN 20081 MCP 21616 PE08-697 GY04-257 GY04-183 SU05-592 SU05-457 MCP 23751 NA 23 LBPN 18551 P6236 LBPN 16145 P4743 P4747 LBPN 11507 T2361 MUS 353 6635 6637 6638 P4006 MUS 286 MUS 357 P6330 PA97-019 44 47 P1593 PA00-013 PY9091 LBPN 4044 MUS 327 MUS 202

Brazil, Rio Araraguara Brazil, Rio Araraguara Peru, Rio Ucayali Peru, Rio Huancabamba Brazil, Rio Grande do Sul Peru, bosque Von Humbolt Guyana, Mauishparu River Guyana, Takutu River Suriname, Coppename River Suriname, Corantijn River Brazil, Rio Grande do Sul SR, aquarium specimen Panama, Rio Mandinga Panama, Santa Rita Arriba Peru, Rio Itaya Brazil, Rio Araguaia Peru, Rio Yanatili Peru, Rio Urubamba Brazil,Rio Araguaia Guyana, Ireng River Colombia, aquarium trade Colombia, Rio Magdalena, Honda Colombia, Rio Magdalena, Honda Colombia, Rio Magdalena, Honda Venezuela, Rio Orinoco Colombia, aquarium trade Colombia, aquarium trade Peru, Rio Nanay Panama, Darien Venezuela, Maracaibo Lake Colombia, Rio San Juan Peru, Rio Amazonas Panama, Rio Ipeti Paraguay, Rio Paraguay Brazil, Rio Branco Colombia, aquarium specimen Argentina, Rio Uruguay

2428 KR478187 2425 KR478186 2426 KR478520 2433 KR478021 2432 KR478189 2430 KR478197 2424 KR478009 2429 KR478010 2427 KR478007 2426 KR478008 2426 KR477998 2427 KR478011 2426 KR478180 2424 KR478179 2424 KR478105 2427 KR478042 2421 KR478043 2421 KR478044 2427 KR478046 2423 KR478047 2426 KR478045 2418 KR478048 2418 KR478049 2419 KR478050 2427 KR478051 2438 KR477925 2442 KR478160 2446 KR477926 2440 KR477922 2443 KR477923 2440 KR477924 2444 KR478162 2443 KR478163 2443 KR478161 2442 KR477935 2440 KR477927 2425 EU310442

Covain et al . 2008

2505 KR478512 2501 KR478511 2481 KR478521 2536 KR478355 2506 KR478514 2532 KR478522 2289 KR478343 2286 KR478344 2289 KR478341 2270 KR478342 2780 KR478332 2281 KR478345 2245 KR478505 2230 KR478504 NA 1958 KR478376 1957 KR478377 1958 KR478378 1960 KR478380 1978 KR478381 1981 KR478379 1960 KR478382 1961 KR478383 1961 KR478384 1958 KR478385 2287 KR478259 2287 KR478486 1977 KR478260 2302 KR478256 2295 KR478257 2304 KR478258 2589 KR478488 2302 KR478489 2540 KR478487 2583 KR478269 2307 KR478261 1809 HM623638

Pseudorinelepis genibarbis 1

MHNG 2588.079

PE96-040

Peru, Rio Ucayali

2436 HM592623

Rodriguez et al . 2011

1925 HM623634

Rodriguez et al . 2011

Guyanancistrus brevispinis 1

MHNG 2725.099

GF00-103

French Guiana, Maroni River

2439 JN855735

Covain and Fisch-Muller 2012

1807 JN855772

Covain and Fisch-Muller 2012

Guyanancistrus longispinis 1

MHNG 2725.100

GF99-204

French Guiana, Oyapock River

2439 JN855720

Covain and Fisch-Muller 2012

1808 JN855757

Covain and Fisch-Muller 2012

French Guiana, Oyapock River

2438 JN855722

Covain and Fisch-Muller 2012

1809 JN855759

Covain and Fisch-Muller 2012

French Guiana, Approuague River

2442 JN855752

Covain and Fisch-Muller 2012

1818 JN855789

Covain and Fisch-Muller 2012

Peru, Rio Pauya

2432 JN855750

Covain and Fisch-Muller 2012

1790 JN855787

Covain and Fisch-Muller 2012

Guyana, Essequibo River

2428 JN855740

Covain and Fisch-Muller 2012

1715 JN855777

Covain and Fisch-Muller 2012 Covain and Fisch-Muller 2012

Guyanancistrus niger

1

MHNG 2722.089

Hypostomus gymnorhynchus Lasiancistrus heteracanthus Lithoxus lithoides

1

Hemiancistrus medians 1 Peckoltia sabaji

1

Peckoltia cavatica Panaqolus koko

1

1

MHNG 2621.098 MHNG 2613.037 MHNG 2651.087

Pseudancistrus barbatus Peckoltia oligospila

1

1

1

SU01-160 CA 13 GY04-136

This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study This study Rodriguez et al . 2011

French Guiana, Maroni River

Covain and Fisch-Muller 2012

1809 JN855761

MHNG 2664.078

GF00-074 GF00-084

2442 JN855724

Suriname, Tapanahony River

2437 JN855719

Covain and Fisch-Muller 2012

1820 JF747011

Fisch-Muller et al. 2012

MHNG 2602.017

BR98-154

Brazil, Rio Guamá

2439 KR478207

This study

1814 JF747020

Fisch-Muller et al . 2012

Guyana, Rupununi River

2437 KR478206

This study

1815 JF747019

Fisch-Muller et al . 2012

Guyana, Rupununi River

2441 KR478205

This study

1808 JF747017

Fisch-Muller et al . 2012

French Guiana, Maroni River

2435 KR478204

This study

1814 JF747016

Fisch-Muller et al . 2012

MHNG 2653.059

MHNG 2651.016 1

GF99-185

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MHNG 2651.020 MNHN 2011-0013

GY04-029 GY04-030 GF00-115

Scobinancistrus aureatus 1 Hypancistrus zebra

1

Megalancistrus cf. parananus Neoplecostomus microps 1 1 2

MHNG 2684.020 MHNG 2708.072 1

outgroup

according to the exporter * specimen reidentified after publication

MHNG 2711.048 MHNG 2588.002

MUS 358 MUS 420 MUS 332 BR 1283

Brazil, aquarium trade, Rio Xingu2

2442 KR478210

2

Brazil, aquarium trade, Rio Xingu

2435 KR478209

Brazil, aquarium trade

2442 KR478208

Brazil, Rio dos Frades

2442 KR478211

This study

1816 KR478531

This study

This study

1790 KR478530

This study

This study

1816 KR478529

This study

This study

1816 KR478532

This study

Hypothesis H0 Best ML tree H1 Three tribes: Farlowellini, Loricariini, Harttiini

Ref This study Isbrücker, 1979

H2 Two tribes: Harttiini (incl. Harttiina and Farlowellina) and Loricariini H3 Monophyly of Sturisoma , Sturisomatichthys , and Farlowella H4 Monophyly of Crossoloricaria , Apistoloricaria , and Rhadinoloricaria H5 Monophyly of Loricaria

Rapp Py-Daniel, 1997; Armbruster, 2004 This study

H6 Monophyly of Ixinandria, Rineloricaria , Hemiloricaria , Fonchiiichthys , and Leliella H7 Monophyly of Harttia , Cteniloricaria , Harttiella , and Quiritixys

as defined by Isbrücker, 2001

This study This study

as defined by Isbrücker, 2001

lnL -116343.2

Δ lnL -

AU -

BP -

ELW -

-116579.5

236.3

0

0

0

-116968.6

625.4

0

0

0

-116723.6

380.4

0

0

0

-116461.5

118.3

0

0

0

-116424.1

80.9

0

0

8.38 10-7

-117873.3

1530.1

0

0

0

-116532.8

189.6

0

0

0

Loricariidae Loricariinae Harttiini

Harttia (synonym: Quiritixys) Harttiella Cteniloricaria Loricariini Farlowellina

Lamontichthys Pterosturisoma Farlowella Aposturisoma (possibly a synonym of Farlowella ) Sturisoma (restricted to cis-Andean region) Sturisomatichthys (including all trans-Andean Sturisoma ) Loricariina Basal Loricariina group

Metaloricaria Dasyloricaria Fonchiiloricaria Rineloricaria group

Rineloricaria (synonyms: Fonchiiichthys , Hemiloricaria , Leliella , and Ixinandria) Loricariichthys group Pseudoloricaria Limatulichthys Loricariichthys Hemiodontichthys

Furcodontichthys (not available for this study; group assignment based on morphology) Loricaria-Pseudohemiodon group Spatuloricaria Loricaria Proloricaria (revalidated) Brochiloricaria Paraloricaria Crossoloricaria (restricted to trans-Andean region) Planiloricaria Pseudohemiodon Rhadinoloricaria (including all cis-Andean Crossoloricaria; synonym: Apistoloricaria ) Dentectus (not available for this study; group assignment based on morphology) Reganella (not available for this study; group assignment based on morphology) Pyxiloricaria (not available for this study; group assignment based on morphology) Ricola (not available for this study; group assignment based on morphology)

-

We performed a comprehensive molecular phylogeny of the species-rich subfamily Loricariinae. 350 loricariin OTUs were analyzed with mitochondrial and nuclear markers. Molecular results were incongruent with morphological classifications Tribal and subtribal ranks were redefined, and several genera were restricted, synonymized, or revalidated.

Graphical Abstract