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Article

DNA Barcode Reference Library and Undetected Diversity of Fish Species in the Yuanjiang River, China

by
Xian Shi
1,2,
Chunni Kou
1,
Chengdong He
1,3,
Hong Deng
1,3,
Hongfu Yang
4,
Xinhui Li
1,
Mingdian Liu
5,
Yaqiu Liu
1,6,7,*,
Jie Li
1,6,7 and
Weitao Chen
1,6,7,*
1
Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China
2
Ocean College, Fujian Agriculture and Forestry University, Fuzhou 350002, China
3
Fisheries and Life Science College, Shanghai Ocean University, Shanghai 201306, China
4
Fisheries Administration of Qiubei, Wenshan 663220, China
5
Yangtze River Fisheries Research Institute, Chinese Academy of Fishery Science, Wuhan 430223, China
6
Key Laboratory of Aquatic Animal Immune Technology of Guangdong Province, Guangzhou 510380, China
7
Guangzhou Scientific Observing and Experimental Station of National Fisheries Resources and Environment, Guangzhou 510380, China
*
Authors to whom correspondence should be addressed.
Fishes 2025, 10(8), 418; https://doi.org/10.3390/fishes10080418
Submission received: 15 July 2025 / Revised: 14 August 2025 / Accepted: 14 August 2025 / Published: 20 August 2025
(This article belongs to the Special Issue Mapping and Monitoring Aquatic Biodiversity in Hotspot Habitat Areas Using Environmental DNA (eDNA))
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Abstract

The Yuanjiang River, situated in the upper reaches of the Red River, is a crucial component of a biodiversity hotspot in the mountains of southwestern China, supporting a high diversity of fish species. Nevertheless, systematic research on fish diversity in the Yuanjiang River is scarce, scattered, and outdated. In our study, we produced 764 DNA barcodes belonging to 64 fish morphospecies to evaluate fish diversity in the Yuanjiang River. Barcoding gap analysis and DNA-based delimitation approaches achieved a high identification success rate (>93%), indicating that DNA barcoding is a practical approach for delimiting fish in the Yuanjiang River. However, four species were characterized by high levels of intraspecific divergence, generating multiple clades and/or molecular operational taxonomic units (MOTUs), suggesting that these species might comprise undetected species. Meanwhile, two closely related species within the genus Schistura, i.e., S. callichroma and S. caudofurca, cannot be delimited by the DNA barcoding technique, which is indicative of recent speciation. In summary, this study established a reliable DNA barcode reference library for fish species in the Yuanjiang River and revealed previously unknown fish diversity.
Keywords:
Yuanjiang River; DNA barcoding; fish species; species delimitation; cryptic diversity
Key Contribution: This study establishes the first comprehensive DNA barcode reference library for Yuanjiang River fish, comprising 764 barcodes from 64 species, enabling highly accurate (>93%) species identification. In addition, hidden diversity observed within four species suggests potential cryptic species.

1. Introduction

The Red River is an internationally important river that flows through China and Vietnam. The upper reach of the Red River—the Yuanjiang River—is an essential component of the biodiversity hotspot in the mountains of southwestern China, which harbors rich fish diversity and endemic fish species due to its complex geological history and dramatic topographical variations [1,2,3,4]. A total of 82 known fish species have been documented in the Yuanjiang River throughout history, and numerous species are endemic and even endangered, such as Bagarius rutilus and Poropuntius krempfi [1,2]. Furthermore, more and more invasive species such as Oreochromis niloticus, Cirrhinus mrigala, and Pterygoplichthys pardalis were documented in this river during our field surveys between 2023 and 2024. Meanwhile, several new species distributed in this river have been described over the last several years [5,6], indicating that unexplored fish diversity remains to be discovered. To better manage and protect fish resources in the Yuanjiang River, summarizing the current fish diversity is a necessary step.
Nevertheless, systematic research on fish diversity in the Yuanjiang River is rare, scattered, and outdated. For example, Zhou et al. (1999) compared β diversity among three branches within the Yuanjiang River based on field surveys conducted in the 1990s [7]. Liu et al. (2025) evaluated the length–weight and length–length relationships of twenty-three species of freshwater fish in the Yuanjiang River [8]. Other relevant reports were only found in ichthyography books and published research that focused on species descriptions [1,2]. Considering the timeliness and variability of the data on fish diversity in the Yuanjiang River, it is highly necessary to carry out investigations and research on fish diversity in this river.
Understanding detailed fish diversity in a river is crucial for utilization and conservation [9,10,11]. To date, most taxonomic descriptions of species have mainly been via morphological characterizations. Nevertheless, misidentifications often occurred due to phenotypic plasticity, cryptic speciation, and/or differing life stages [12,13,14,15,16,17,18]. DNA barcoding is a popular molecular approach for rapidly and accurately discriminating animal species, which has been shown to be highly successful in identifying fish, regardless of life stage [19,20,21,22,23,24]. Additionally, this technique is widely used for discovering cryptic species and novel species [25,26,27,28,29,30]. As a consequence, DNA barcoding enables accurate species identification and can detect morphologically cryptic species, as well as aid in the search for new species.
In this study, we employed DNA barcoding to investigate the fish diversity in the Yuanjiang River, using samples collected during four field surveys between 2023 and 2024. Our main goals were (1) to establish a DNA barcoding library for the ichthyofauna in the Yuanjiang River, (2) to shed new light on fish diversity via sequence-based delimitation methods, and (3) to reveal discrepancies in species observed between DNA barcodes and morphological features.

2. Materials and Methods

2.1. Sample Collection

During field surveys conducted in the Yuanjiang River between 2023 and 2024, fish specimens were randomly sampled using gill nets, throw nets, and fish cages. Species identification followed morphological criteria outlined in two monographs: “The Fishes of Yunnan, China” [1] and the “Checklist of Fishes of Yunnan” [2]. A total of 64 morphologically delimitated species were collected across 18 sampling sites spanning the mainstem and tributaries (Figure 1; Table S1). To construct a DNA barcoding library for Yuanjiang River fishes, 764 specimens representing all 64 species were selected for genetic analysis (Figure 1; Table S1). We created the sampling map using ArcGIS 10.2 and then modified it in Microsoft Office 2017. Fin clips or muscle tissues were clipped and conserved in absolute ethyl alcohol for DNA extraction in the laboratory. All the tissue samples used for genomic DNA extraction were deposited in the Freshwater Fish Museum at the Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences.

2.2. Genomic Extraction and Sequencing

Total genomic DNA was extracted using the Axygen DNA Extraction Kit, following the manufacturer’s instructions. We then checked DNA quality using 1% agarose gel electrophoresis. The standard barcoding fragment (~648 bp from the 5′ end of the mitochondrial COI gene) was selected as a DNA barcoding marker. We utilized the fish universal primers FishF1/FishF2 and FishR1/FishR2 to amplify and sequence the target locus [31].
The PCR reactions were performed in a total volume of 30 µL, containing the following components: 10 μL of 2 × PCR buffer, 2 μL of dNTPs (2.5 mM each), 0.2 µL of Taq DNA polymerase (2.0 U), 1 μL of template DNA, 1 μL of each primer (10 pmol), and 15 μL of ddH2O. The PCR conditions for the two fish universal primers contained initial denaturation at 95 °C for 5 min, 30 cycles at 94 °C for 30 s, 54 °C for 30 s, and 72 °C for 1 min, and a final extension at 72 °C for 10 min. The PCR products with high quality were sequenced bi-directionally on an ABI 3730XL DNA system (Perkin-Elmer Applied Biosystems, Foster City, CA, USA) to decrease the occurrence of sequencing mistakes. We checked and assembled the contigs using the SEQMEN in the Lasergene package (DNASTAR, Inc., Madison, WI, USA). The sequences were aligned and trimmed using the MEGA version X [32] and were submitted to the GenBank database under the accession numbers PV878687–PV879450 (Table S1).

2.3. Genetic Distance Calculations

Firstly, we calculated Kimura 2-parameter (K2P) [33] pairwise genetic distances within and between species. Secondly, we conducted a barcoding gap analysis by making species-level comparisons between maximum intraspecific genetic distances and minimum distances to the nearest neighbor [34]. Thirdly, we constructed a neighbor-joining (NJ) tree using K2P genetic distances with the “pairwise deletion” option and 1000 bootstrap replicates. All abovementioned analyses were performed using MEGA version X [32].

2.4. Sequence-Based Species Delimitation

Three species delimitation approaches were utilized to group specimens into molecular operational taxonomic units (MOTUs). Firstly, we performed automatic barcode gap discovery analysis (ABGD) using the default value for the relative gap width (X = 1.5) and K2P genetic distance [35] in ABGD program. Secondly, the version of the Poisson tree process (PTP) model in multiple rates was implemented on the web server (https://mcmc-mptp.h-its.org/; accessed on 5 May 2025 [36,37]) using a maximum likelihood (ML) tree constructed with the software RAxML-VI-HPC [38]. The GTR + I + R model selected in MrModelTEST [39] was employed for ML analyses. Thirdly, Generalized Mixed Yule Coalescent (GMYC) analyses [40] using single- and multiple-threshold values were also performed on a web server (http://species.h-its.org/gmyc/; accessed on 10 May 2025). A fully resolved ultrametric gene tree was inferred as input for the GMYC analysis using BEAST version 1.8.2 [41] as a Bayesian tree. A birth–death tree prior model and a GTR + I + R substitution model were employed in Bayesian reconstructions with haplotype sequences. We opted for an uncorrelated, relaxed lognormal clock model, estimating the rate from the data and ucld-mean parameters. A uniform prior with lower and upper boundaries of 0 and 10 was set. The remaining settings were set as the default parameters. Fifty million generations with 1000 sampling frequencies were used in the Monte Carlo Markov Chain (MCMC) run. Effective sample sizes (ESSs) for all parameters were assessed in the software Tracer v1.5 [42]. The final trees were summarized in a maximum credibility tree using TreeAnnotator version 1.8.2, with 50% of the total trees discarded [41]. A majority-rule consensus was adopted from the aforementioned three algorithms for the delimitation scheme.

2.5. Cryptic Diversity Analyses

Regarding morphological species that comprise multiple clades and/or MOTUs with higher genetic distances, the construction of an NJ tree for visual inspection was carried out using K2P distances, with 1000 bootstrap replicates. In addition, the mean K2P genetic divergence between observed clades was also calculated. The abovementioned analyses were performed using MEGA version X [32].

3. Results

3.1. Sequence Information

We successfully obtained 764 DNA barcodes that represent 64 morphospecies, 52 genera, 15 families, and 5 orders (Table S1). On average, 11.94 sequences were obtained per morphospecies, with a range of 1 to 42 (11 morphospecies were represented by singletons). The length of the barcoding sequences after aligning and trimming reached 632 base pairs (bp). The absence of any deletions, insertions, or stop codons in the amplified sequences suggests that all sequences constitute functional mitochondrial COI sequences.

3.2. Genetic Distance Calculation

A hierarchical increase in mean genetic distance was obtained, ranging from within species (0.47 ± 0.03%) to within congeners (4.48 ± 0.29%), as measured by the K2P distance (Table 1). Of the 53 species with more than two specimens, 49 had an intraspecific distance of less than 2%. Nevertheless, four species, i.e., Hemiculter leucisculus, Misgurnus anguillicaudatus, Paramisgurnus dabryanus, and Rhinogobius honghensis, exhibited relatively high intraspecific divergence (>2%). Among 53 species with more than two specimens, barcode gap analysis showed that 52 species had barcoding gaps with the minimum interspecific distance to the nearest neighbor larger than the maximum intraspecific distance (Figure 2). Spearman correlation analysis revealed that the intraspecific sequence divergence is not significantly correlated with sample size (R = 0.177, p = 0.206). The NJ trees (Figure 3) yielded 68 clades from 64 morphospecies (including the singleton species), suggesting that several morphospecies might form multiple clades. Four species, i.e., Hemiculter leucisculus, Opsariichthys bidens, Misgurnus anguillicaudatus, and Paramisgurnus dabryanus, yielded two clusters from the NJ trees. In addition, a visual inspection of the NJ tree revealed that 60 morphospecies formed concordant clusters of DNA barcodes.

3.3. Results of Sequence-Based Species Delimitation

Three delimitation approaches yielded discordant results (Figure 4). The ABGD analyses conducted with the K2P models resolved 65 MOTUs. Among the sixty-five MOTUs, we detected that Misgurnus anguillicaudatus and Opsariichthys bidens comprised two MOTUs. In addition, the ABGD analyses supported two Schistura species, namely S. callichroma and S. caudofurca, that shared a MOTU (Figure 4). The remaining MOTUs were found to be concordant with morphological identification (Figure 4).
The Poisson tree process (mPTP) obtained 52 MOTUs, which was fewer than the number of MOTUs yielded by ABGD analyses (Figure 4). Only 45 MOTUs were consistent with morphological results. mPTP analyses yielded conservative outcomes than other approaches. For example, mPTP clustered five morphologically distinct Schistura species into one MOTU. In addition, six species belonging to five genera, i.e., Pseudohemiculter dispar, Anabarilius sp., Chanodichthys dabryi, Culter alburnus, Toxabramis houdemeri, and Hemiculter leucisculus, were combined into one MOTU (Figure 4). A similar pattern was also seen in three species belonging to three genera, i.e., Discogobio yunnanensis, Ageneiogarra imberba, and Placocheilus caudofasciatus (Figure 4).
Regarding GMYC analyses, a total of 80 MOTUs were identified. GMYC analyses revealed that 11 morphological species had 2–4 MOTUs (Figure 4), resulting in more MOTUs than the other two approaches.
A majority-rule consensus established the final consensus, which obtained 65 MOTUs. In sum, sixty species were unambiguously identified by the delimitation analyses, two species (Opsariichthys bidens and Misgurnus anguillicaudatus) comprised two MOTUs, and two closely related species (Schistura callichroma and S. caudofurca) were found to display mixed genealogies with one MOTU (Figure 4).

3.4. Cryptic Diversity

Two clades were detected in four species, i.e., Hemiculter leucisculus, Opsariichthys bidens, Paramisgurnus dabryanus, and Misgurnus anguillicaudatus. Furthermore, higher genetic divergence between the two clades of each species was examined, with values ranging from 2.70% (Paramisgurnus dabryanus) to 10.08% (Misgurnus anguillicaudatus) (Figure 5).

4. Discussion

This is the first study to present a complete DNA barcode reference library for fish species in the Yuanjiang River in China. The library comprises 64 morphospecies and covers most of the river section including the main stem and tributaries (Figure 1). Of the 64 species, 54 are historically documented, accounting for approximately 65.85% of the reported species in this river [2]. In addition, our study collected six exotic species and five unnamed species. Barcoding analyses revealed that 60 morphospecies (93.75% of those analyzed) could be unambiguously identified, providing a simple identification system for the fish species in this river. However, the high intraspecific divergences observed for two species, as well as the failure to delimit two closely related species, demonstrate the need for further taxonomic research on these species. Our findings provide robust evidence that DNA barcoding is a reliable tool for refining extant taxonomic classifications and uncovering cryptic diversity within the ichthyofauna of the Yuanjiang River.

4.1. DNA Barcode Reference Library of Fish Species in the Yuanjiang River

The corrected mean K2P intraspecific distance of 0.47% (SE = 0.0003) calculated for the fish species in the Yuanjiang River is slightly greater than the values that reported in the several previous fish barcoding studies involving Chinese rivers, such as the Nujiang River of China section (0.41%), the middle of the Yangtze River (0.36%) and the middle and lower Pearl River (0.32%) and the Irtysh River (0.30%) [18,43,44,45]. The relatively higher level of intraspecific divergence observed in the Yuanjiang River can be explained by several species with high intraspecific genetic distance. We found that the mean intraspecific genetic distances of fifteen morphospecies (28.30% of the species analyzed, for which two specimens were available) exceeded 1.00%.
The barcoding gap analysis, which included 53 species with more than two samples, revealed that the mean nearest-neighbor distances (12.38%) were, on average, ~26-fold higher than intraspecific distances (0.47%), indicating that the present library is useful for identifying Yuanjiang River fish. Furthermore, we demonstrated the presence of a barcode gap in 52 species, with the exception of Schistura callichroma (Figure 2). This study also revealed that the interspecific divergence (3.77%) between S. callichroma and S. caudofurca was slightly lower than the maximum intraspecific divergence (3.94%). Additionally, while NJ tree analysis produced monophyletic clades for S. callichroma and S. caudofurca, Bayesian trees revealed that the two species were combined into a single phylogenetic clade (Figure 3 and Figure 4). These findings suggest that the two species diverged recently and share a close phylogenetic relationship. To clarify the relationships between the two species, future studies should conduct population-level analyses using larger sample sizes and nuclear loci.

4.2. Cryptic Diversity

The most salient finding was that four species (Hemiculter leucisculus, Opsariichthys bidens, Paramisgurnus dabryanus, and Misgurnus anguillicaudatus) generated two clades from the NJ topology, which suggested that these species have undetected genetic diversity. Hemiculter leucisculus is a widespread species in East Asia, with undetected diversity demonstrated in different drainages. For instance, Chen et al. (2017, 2021, 2022) found that H. leucisculus populations in the Pearl River generated two deep mitochondrial clades with higher genetic distance [18,46,47]. The upper Yuanjiang River is situated near two major rivers: the Pearl River and the Lancang River. Furthermore, the lower section of the Yuanjiang River in Vietnam is near the Hainan Island drainages. These can be connected due to the drop in sea level triggered by glaciation during the Late Pleistocene [48,49,50]. Collectively, the two H. leucisculus clades that occurred in the Yuanjiang River were considered a normal phenomenon. The same situation can be used to explain the pattern observed in Opsariichthys bidens populations. In addition, Opsariichthys bidens is a small, benthic taxon that was found to have an unexpected genetic diversity within the same drainage area, which could be because of limited dispersal ability, geographic separation, and geological events [51,52,53].
For Paramisgurnus dabryanus, two clades observed could be interpreted by their introductions from different sources. This species is an economically important taxon cultivated in many regions, including southwestern China. Introduction from various genetic resources can trigger multiple clades. Moreover, hybridization with other species may cause P. dabryanus to produce multiple clades, due to the introduction of mitochondrial genomes from other species [54]. Similar cases can be seen in other rivers in China, such as the Nujiang River and the lower Pearl River [18,43]. It was not unexpected that two deep clades were observed in the pond loach (Misgurnus anguillicaudatus) in an independent river. For example, Zhong et al. (2019) argued that Misgurnus anguillicaudatus populations both in the Pearl River and the Yangtze River had four mitochondrial clades with high genetic divergence among clades [55].

4.3. Implications for the Yuanjiang River Ichthyofauna

The present study provides the most comprehensive geographical, taxonomic, and molecular sampling of the Yuanjiang River to date, with several novel findings that contradict current taxonomic knowledge. The most challenging cases pertained to shallow interspecific divergence between two Schistura species, as well as the detection of multiple, highly divergent clades within four species. Given that close relationships between the two Schistura species may be the result of a recent divergence, using additional molecular loci with higher substitution rates—such as the control region or nuclear genome dataset—would certainly help to distinguish such cases more effectively. Furthermore, the presence of multiple clades/MOTUs in four species may be attributable to the presence of subtle, overlooked morphological differences, cryptic diversity, and unrecognized speciation events [46]. However, the taxonomic status of these cryptic clades, as delineated by the mitochondrial barcoding marker, is not robust. Further assessments of morphological divergence and phylogenetic placement should be performed using additional nuclear loci.
Five morphologically distinct species belonging to four genera—Anabarilius, Schistura, Rhinogobius, and Mastacembelus—cannot be assigned to exact species based on currently available publications. This observation suggests that many species in the Yuanjiang River remain undescribed because of a lack of systematic research on this river. Additionally, the ichthyofauna of the Yuanjiang River combines species from Vietnam and southwestern China, suggesting a complex composition of fish species. Collectively, to enhance our understanding of fish diversity in the Yuanjiang River, more field surveys and taxonomic research involving the ichthyofauna of the Yuanjiang River should be conducted in the future.

5. Conclusions

In the current study, a DNA barcode reference library was constructed, comprising 64 fish species from the Yuanjiang River in China, based on high-density sampling. The relatively high identification success rate (93.75%) demonstrates the reliability of DNA barcoding in automatically delimiting fish in the Yuanjiang River. The newly detected mitochondrial clades in four species necessitate a substantial effort to better determine the diversity of these groups. Nevertheless, we also confirmed the limitation of DNA barcoding when it comes to the automated identification of closely related species. Furthermore, five taxa cannot be delimited to the species level using morphological characteristics only, indicating that more taxonomic studies should be conducted for the fish in this river. Given that approximately 20% of species were not included in the current study, a more comprehensive barcode library containing unsampled species requires updating.

Supplementary Materials

The following supporting information can be downloaded at https://www.mdpi.com/article/10.3390/fishes10080418/s1, Table S1: Detailed information including collection data and GenBank accession numbers for sequences used in this study.

Author Contributions

W.C., J.L., Y.L., X.L. and M.L. conceived this study; X.S., C.K., C.H., H.D. and H.Y. conducted the experiments; X.S. and W.C. analyzed the data and drafted the manuscript. All authors have read and agreed to the published version of the manuscript.

Funding

This study was supported by the Finance Special Fund of the Chinese Ministry of Agriculture and Rural Affairs of the People’s Republic of China (fishery resources and environment survey in the key water areas of southwestern China).

Institutional Review Board Statement

The study was approved by the Laboratory Animal Ethics Committee of Pearl River Fisheries Research Institute (Nos. LAEC-PRFRI-2023-04-03).

Data Availability Statement

The sequences are available from the NCBI (https://dataview.ncbi.nlm.nih.gov; accessed on 1 July 2025) under the accession numbers of PV878687–PV879450.

Conflicts of Interest

The authors declare no competing interests.

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Fishes 10 00418 g001
Figure 1. Map of sampling locations in the present study. Green dots indicate sampling sites and the abbreviations of the sampling sites refer to Table S1.
Figure 1. Map of sampling locations in the present study. Green dots indicate sampling sites and the abbreviations of the sampling sites refer to Table S1.
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Figure 2. The maximum intraspecific distance is compared with the nearest-neighbor distance for fish in the Yuanjiang River. Species with more than two sequences are presented in Table S1. Green dots indicate maximum intraspecific distance and the nearest-neighbor distance for each species. The species fall above the 1:1 line, indicating the presence of a barcode gap.
Figure 2. The maximum intraspecific distance is compared with the nearest-neighbor distance for fish in the Yuanjiang River. Species with more than two sequences are presented in Table S1. Green dots indicate maximum intraspecific distance and the nearest-neighbor distance for each species. The species fall above the 1:1 line, indicating the presence of a barcode gap.
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Figure 3. Neighbor-joining tree of overall barcoding sequences based on the K2P model.
Figure 3. Neighbor-joining tree of overall barcoding sequences based on the K2P model.
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Figure 4. DNA-based species delimitation of 64 morphological species (green) in this study. MOTU delimitation schemes collected from the automatic barcode gap discovery analysis (ABGD), Poisson tree process (PTP) model in multiple rates (mPTP), and Generalized Mixed Yule Coalescent (GMYC) algorithms (orange), including the consensus delimitation scheme (light blue). A Bayesian tree conducted in BEAST was used to summarize delimitation results.
Figure 4. DNA-based species delimitation of 64 morphological species (green) in this study. MOTU delimitation schemes collected from the automatic barcode gap discovery analysis (ABGD), Poisson tree process (PTP) model in multiple rates (mPTP), and Generalized Mixed Yule Coalescent (GMYC) algorithms (orange), including the consensus delimitation scheme (light blue). A Bayesian tree conducted in BEAST was used to summarize delimitation results.
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Figure 5. Neighbor-joining trees with mean inter-clade distances of the four species based on the K2P model. (a) Hemiculter leucisculus, (b) Opsariichthys bidens, (c) Misgurnus anguillicaudatus, and (d) Paramisgurnus dabryanus.
Figure 5. Neighbor-joining trees with mean inter-clade distances of the four species based on the K2P model. (a) Hemiculter leucisculus, (b) Opsariichthys bidens, (c) Misgurnus anguillicaudatus, and (d) Paramisgurnus dabryanus.
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Table 1. Mean values, ranges, and standard deviations of genetic distances within and between species.
Table 1. Mean values, ranges, and standard deviations of genetic distances within and between species.
Mean%SEMin%Max%
Within species0.470.0003010.22
Within genus4.480.00323.3823.64
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MDPI and ACS Style

Shi, X.; Kou, C.; He, C.; Deng, H.; Yang, H.; Li, X.; Liu, M.; Liu, Y.; Li, J.; Chen, W. DNA Barcode Reference Library and Undetected Diversity of Fish Species in the Yuanjiang River, China. Fishes 2025, 10, 418. https://doi.org/10.3390/fishes10080418

AMA Style

Shi X, Kou C, He C, Deng H, Yang H, Li X, Liu M, Liu Y, Li J, Chen W. DNA Barcode Reference Library and Undetected Diversity of Fish Species in the Yuanjiang River, China. Fishes. 2025; 10(8):418. https://doi.org/10.3390/fishes10080418

Chicago/Turabian Style

Shi, Xian, Chunni Kou, Chengdong He, Hong Deng, Hongfu Yang, Xinhui Li, Mingdian Liu, Yaqiu Liu, Jie Li, and Weitao Chen. 2025. "DNA Barcode Reference Library and Undetected Diversity of Fish Species in the Yuanjiang River, China" Fishes 10, no. 8: 418. https://doi.org/10.3390/fishes10080418

APA Style

Shi, X., Kou, C., He, C., Deng, H., Yang, H., Li, X., Liu, M., Liu, Y., Li, J., & Chen, W. (2025). DNA Barcode Reference Library and Undetected Diversity of Fish Species in the Yuanjiang River, China. Fishes, 10(8), 418. https://doi.org/10.3390/fishes10080418

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