DNA barcoding reveals hidden diversity in the Neotropical freshwater fish Piabina argentea (Characiformes: Characidae) from the Upper Paraná Basin of Brazil

Figures & data

Table I.  Specimen data.

					GPS			GenBank accession number
Cluster	Species name	Voucher number	Collection code	Locality state	Latitude	Longitude	BOLD process ID	COI	CytB
A	P. argentea	36233	LBP 7329	Paraná	− 25.092	− 52.495	FUPR120-09	HM144073	GU908189
A	P. argentea	34659	LBP 7111	Paraná	− 23.938	− 50.729	FUPR181-09	HM144065	GU908182
A	P. argentea	36235	LBP 7329	Paraná	− 25.092	− 52.495	FUPR122-09	HM144071	GU908187
A	P. argentea	36234	LBP 7329	Paraná	− 25.092	− 52.495	FUPR121-09	HM144072	GU908188
A	P. argentea	17229	LBP 2594	São Paulo	− 21.013	− 49.690	FUPR089-09	HM144104	GU908219
A	P. argentea	25091	LBP 6745	São Paulo	− 22.341	− 48.935	FUPR102-09	HM144091	GU908207
A	P. argentea	14154	LBP 1996	São Paulo	− 22.917	− 48.500	FUPR062-09	HM144107	GU908222
A	P. argentea	21305	LBP 3509	São Paulo	− 22.941	− 48.584	FUPR091-09	HM144102	GU908217
A	P. argentea	25050	LBP 6741	São Paulo	− 23.231	− 48.533	FUPR101-09	HM144092	GU908208
A	P. argentea	23459	LBP 6741	São Paulo	− 23.024	− 48.826	FUPR092-09	HM144101	GU908216
A	P. argentea	20764	LBP 6741	São Paulo	− 23.024	− 48.826	FUPR090-09	HM144103	GU908218
A	P. argentea	25410	LBP 6745	São Paulo	− 22.341	− 48.935	FUPR108-09	HM144085	GU908201
A	P. argentea	17228	LBP 2594	São Paulo	− 21.013	− 49.690	FUPR088-09	HM144105	GU908220
A	P. argentea	17227	LBP 2594	São Paulo	− 21.013	− 49.690	FUPR087-09	HM144106	GU908221
A	P. argentea	29280	LBP 6268	Minas Gerais	− 21.285	− 46.493	FUPR201-09	HM144056	GU908174
A	P. argentea	29282	LBP 6268	Minas Gerais	− 21.285	− 46.493	FUPR203-09	HM144054	GU908172
B	P. argentea	25214	LBP 6743	São Paulo	− 22.786	− 48.481	FUPR238-09	HM144052	GU908170
B	P. argentea	23222	LBP 6743	São Paulo	− 22.786	− 48.481	FUPR237-09	HM144053	GU908171
B	P. argentea	25219	LBP 6743	São Paulo	− 22.786	− 48.481	FUPR107-09	HM144086	GU908202
B	P. argentea	22884	LBP 4032	Distrito Federal	− 15.115	− 47.046	FUPR189-09	HM144063	GU908181
B	P. argentea	23220	LBP 6743	São Paulo	− 22.786	− 48.481	FUPR109-09	HM144084	GU908200
B	P. argentea	35870	LBP 7280	Goiás	− 17.120	− 48.740	FUPR192-09	HM144060	GU908178
B	P. argentea	35871	LBP 7280	Goiás	− 17.120	− 48.740	FUPR193-09	HM144059	GU908177
B	P. argentea	35849	LBP 7680	Goiás	− 17.099	− 48.762	FUPR191-09	HM144061	GU908179
B	P. argentea	35904	LBP 7292	Goiás	− 17.801	− 48.372	FUPR115-09	HM144078	GU908194
B	P. argentea	35906	LBP 7292	Goiás	− 17.801	− 48.372	FUPR117-09	HM144076	GU908192
B	P. argentea	35905	LBP 7292	Goiás	− 17.801	− 48.372	FUPR116-09	HM144077	GU908193
B	P. argentea	35848	LBP 7680	Goiás	− 17.099	− 48.762	FUPR190-09	HM144062	GU908180
C	P. argentea	31608	LBP 6513	Minas Gerais	− 19.385	− 43.659	FUPR113-09	HM144080	GU908196
C	P. argentea	31605	LBP 6513	Minas Gerais	− 19.385	− 43.659	FUPR110-09	HM144083	GU908199
C	P. argentea	31609	LBP 6513	Minas Gerais	− 19.385	− 43.659	FUPR114-09	HM144079	GU908195
C	P. argentea	31606	LBP 6513	Minas Gerais	− 19.385	− 43.659	FUPR111-09	HM144082	GU908198
C	P. argentea	31607	LBP 6513	Minas Gerais	− 19.385	− 43.659	FUPR112-09	HM144081	GU908197
D	P. argentea	25162	LBP 6744	São Paulo	− 23.231	− 48.533	FUPR103-09	HM144090	GU908206
D	P. argentea	25156	LBP 6744	São Paulo	− 23.231	− 48.533	FUPR097-09	HM144096	GU908211
D	P. argentea	25030	LBP 6744	São Paulo	− 23.231	− 48.533	FUPR093-09	HM144100	GU908215
D	P. argentea	25031	LBP 6744	São Paulo	− 23.231	− 48.533	FUPR094-09	HM144099	GU908214
D	P. argentea	25167	LBP 6744	São Paulo	− 23.231	− 48.533	FUPR106-09	HM144087	GU908203
D	P. argentea	25165	LBP 6744	São Paulo	− 23.231	− 48.533	FUPR104-09	HM144089	GU908205
D	P. argentea	25166	LBP 6744	São Paulo	− 23.231	− 48.533	FUPR105-09	HM144088	GU908204
D	P. argentea	25161	LBP 6744	São Paulo	− 23.231	− 48.533	FUPR099-09	HM144094	GU908209
D	P. argentea	25157	LBP 6744	São Paulo	− 23.231	− 48.533	FUPR098-09	HM144095	GU908210
D	P. argentea	25155	LBP 6744	São Paulo	− 23.231	− 48.533	FUPR096-09	HM144097	GU908212
E	P. argentea	29240	LBP 6226	Minas Gerais	− 21.321	− 46.511	FUPR200-09	HM144057	GU908175
E	P. argentea	29239	LBP 6226	Minas Gerais	− 21.321	− 46.511	FUPR199-09	HM144058	GU908176
E	P. argentea	29281	LBP 6268	Minas Gerais	− 21.285	− 46.493	FUPR202-09	HM144055	GU908173
F	P. argentea	35934	LBP 7301	Goiás	− 18.110	− 48.504	FUPR119-09	HM144074	GU908190
F	P. argentea	35933	LBP 7301	Goiás	− 18.110	− 48.504	FUPR118-09	HM144075	GU908191
F	P. argentea	28396	LBP 5994	Minas Gerais	− 21.732	− 46.423	FUPR129-09	HM144067	GU908184
F	P. argentea	28394	LBP 5994	Minas Gerais	− 21.732	− 46.423	FUPR127-09	HM144069	GU908185
F	P. argentea	28393	LBP 5994	Minas Gerais	− 21.732	− 46.423	FUPR126-09	HM144070	GU908186
–	P. anhembi	23373	LBP 6742	São Paulo	− 23.524	− 45.890	FUPR236-09	HM144047	GU908165
–	P. anhembi	23360	LBP 6742	São Paulo	− 23.524	− 45.890	FUPR235-09	HM144048	GU908166
–	P. anhembi	20821	LBP 6742	São Paulo	− 23.524	− 45.890	FUPR125-09	HM144049	GU908167
–	P. anhembi	23372	LBP 6742	São Paulo	− 23.524	− 45.890	FUPR124-09	HM144050	GU908168
–	C. meridionalis	35631	LBP 7557	Mato Grosso do Sul	− 20.343	− 55.726	FUPR188-09	HM144042	GU908160
–	C. paraguayensis	35628	LBP 7557	Mato Grosso do Sul	− 20.343	− 55.726	FUPR185-09	HM144045	GU908163

LBP, Laboratório de Biologia e Genética de Peixes.

Figure 1.  Map showing the distribution of samples of Piabina. Letters correspond to P. argentea clusters. Black square represents P. anhembi species.

Figure 2.  NJ tree of COI/CytB showing the seven major clusters obtained among Piabina specimens (A–F represent P. argentea). Node values represent statistic support: upper values, NJ bootstrap (1000 pseudo-replicates); lower values, maximum parsimony bootstrap (1000 pseudo-replicates). Numbers on fishes represent voucher number and size of photographed specimens (left and right, respectively).

Table II.  K2P genetic distance obtained among the seven major Piabina clusters.

	A	B	C	D	E	F	P. anhembi
A	0.009/0.010	0.047	0.047	0.042	0.054	0.052	0.048
B	0.042	0.007/0.010	0.023	0.030	0.047	0.050	0.027
C	0.056	0.025	0.003/0.004	0.031	0.053	0.053	0.030
D	0.035	0.021	0.029	0.002/0.002	0.041	0.049	0.025
E	0.048	0.025	0.031	0.020	0/0.005	0.049	0.049
F	0.047	0.039	0.045	0.032	0.036	0.003/0.003	0.051
P. anhembi	0.040	0.025	0.034	0.022	0.028	0.034	0.001/0.001

Note: COI below diagonal and CytB above diagonal. The average values of intra-K2P distance represented in bold on the diagonal (COI/CytB).

Table III.  Pairwise F_ST index obtained among the seven major Piabina clusters.

	A	B	C	D	E	F	P. anhembi
A	–	0.79254	0.84386	0.85336	0.85659	0.86067	0.86193
B	0.80038	–	0.65594	0.77328	0.78754	0.82325	0.70487
C	0.85826	0.76602	–	0.92187	0.91004	0.92957	0.91288
D	0.82645	0.78760	0.92876	–	0.94973	0.96113	0.95335
E	0.83305	0.77467	0.93190	0.93346	–	0.93750	0.95991
F	0.83610	0.84631	0.92541	0.92958	0.93352	–	0.96486
P. anhembi	0.81920	0.79240	0.93925	0.93510	0.97734	0.93127	–

Note: COI gene below diagonal, CytB gene above diagonal.

Figure 3.  Seven unconnected haplotype networks among Piabina specimens. P. anhembi is represented in gray. Numbers inside the figures represent specimens that share the same haplotype. Numbers on lines represent the mutational steps between haplotypes. Dashed lines represent the necessary steps to connect the independent networks.