Pseudo but actually genuine: Rhodeus pseudosericeus provides insight into the phylogeographic history of the Amur bitterling

Figures & data

Figure 1. Two alternative scenarios regarding the geographic isolation and colonization of R. pseudosericeus (RP), R. sericeus (RS), and European bitterling lineages (RA: R. amarus; RM: R. meridionalis; and RC: R. colchicus) on the basis of Bohlen et al. (2006; the above part of the figure) and the structure of freshwater systems among and within the Korean Peninsula (the below part of the figure).

Figure 2. Statistical parsimony network model drawn based on the cyt b sequence data obtained from 60 R. pseudosericeus individuals. Each circle represents a different haplotype with its diameter proportional to the haplotype frequency. Each line between haplotypes represents a nucleotide substitution. The six sampling localities are indicated with a relative frequency in each haplotype circle.

Table 1. List of the six R. pseudosericeus populations used in the present study.

Drainage system	Population	N	Location
Han River	Jucheon	10	N37°, 25′, 53″/E128°, 10′, 50″
	Heuk	10	N37°, 30′, 43″/E127°, 39′, 04″
	Seom	10	N37°, 32′, 26″/E127°, 57′, 56″
	Jojong	10	N37°, 45′, 53″/E127°, 26′, 50″
Sapgyo stream	Muhan	10	N36°, 33′, 51″/E126°, 46′, 53″
Daecheon stream	Daecheon	10	N36°, 23′, 25″/E126°, 39′, 49″

Table 2. GenBank accession numbers and the references of the haplotypes used in the BEAST species-tree reconstruction based on fossil calibration at the stem of Acheilognathinae lineage and a substitution rate for the cyt b gene (Figure 2).

Species	Accession nos.	Reference
Kichulchoia brevifasciata	KC524521	Kim et al. (2013)
Tanichthys albonubes	EU241475	Fang et al. (2009)
Acheilognathus tabira nakamurae	AB620146	Kitamura et al. (2012)
Acheilognathus yamatsutae	NC013712	Unpublished
Acheilognathus cyanostigma	AB620134	Kitamura et al. (2012)
Acheilognathus melanogaster	AB620136	Kitamura et al. (2012)
Acheilognathus rhombeus	AB620135	Kitamura et al. (2012)
Acheilognathus macropterus	NC013711	Unpublished
Acheilognathus typus	NC008668	Saitoh et al. (2006)
Acheilognathus chankaensis	DQ396628	Bohlen et al. (2006)
Tanakia lanceolata	EF483933	Unpublished
Tanakia koreensis	NC013704	Hwang et al. (2013)
Tanakia somjinensis	FJ515921	Hwang, Byeon, et al. (2014)
Rhodeus ocellatus	HQ113265	Yang et al. (2011)
Rhodeus ocellatus kurumeus	NC008642	Saitoh et al. (2006)
Rhodeus notatus	NC013709	Hwang, Lee, et al. (2014)
Rhodeus sinensis	EF483937	Unpublished
Rhodeus amurensis	DQ396627	Bohlen et al. (2006)
Rhodeus colchicus	DQ396678	Bohlen et al. (2006)
Rhodeus sericeus	DQ396683–DQ396686	Bohlen et al. (2006)
Rhodeus meridionalis	DQ396679–DQ396682	Bohlen et al. (2006)
Rhodeus amarus	DQ396620DQ396630–DQ396635DQ396637–DQ396649DQ396651–DQ396677	Bohlen et al. (2006)
Rhodeus pseudosericeus H1	KJ028096	The present study
Rhodeus pseudosericeus H2	KJ028097	The present study
Rhodeus pseudosericeus H3	KJ028098	The present study
Rhodeus pseudosericeus H4	KJ028099	The present study
Rhodeus pseudosericeus H5	KJ028100	The present study
Rhodeus pseudosericeus H6	KJ028101	The present study
Rhodeus pseudosericeus H7	KJ028102	The present study
Rhodeus pseudosericeus H8	KJ028103	The present study

Table 3. Summary of the AMOVA results that partition genetic variation among six R. pseudosericeus populations from the Korean Peninsula to two different geographic levels (among groups: the Han River vs. Muhan-Daechon).

Source of variation	df	Sum of squares	Variance components	Percentage of variation
Among groups	1	30.458	1.11992	71.44*
Among populations	4	2.375	0.01623	1.04
Within populations	54	23.3	0.43148	27.52*
Total	59	56.133	1.56763	100

* P < 0.001.

Figure 3. BEAST species-tree reconstructed to infer the relationship among the species within the subfamily Acheilognathinae and divergence time estimates based on fossil calibration at the stem of Acheilognathinae lineage and a substitution rate for the cyt b gene. Bars and numbers on nodes represent 95% credibility intervals and node-supporting values (posterior probabilities), respectively. The information (GenBank accession nos. and references) of haplotypes and species used were listed in Table 2. Multiple haplotypes were used for R. sericeus, R. amarus, and R. meridionalis in the tree (Table 2), and a single haplotype of R. colchicus was recovered to be in the R. amarus clade.

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