Unraveling the Phylogenetic Relationships and Taxonomic Status of a Puzzling Freshwater Mussel Genus <i>Inversidens</i> (Bivalvia, Unionidae) through Multilocus Phylogeny and Mitochondrial Phylogenomics

Wu, Ruiwen; Zhang, Liping; Liu, Lili; Jia, Junli; Liu, Xiongjun

doi:https://doi.org/10.1155/2024/1499508

Journal of Zoological Systematics and Evolutionary Research

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Abstract Introduction Materials and Methods Results Discussion Conclusion Data Availability Conflicts of Interest Acknowledgments Supplementary Materials References Copyright Related Articles

Research Article | Open Access

Volume 2024 | Article ID 1499508 | https://doi.org/10.1155/2024/1499508

Unraveling the Phylogenetic Relationships and Taxonomic Status of a Puzzling Freshwater Mussel Genus Inversidens (Bivalvia, Unionidae) through Multilocus Phylogeny and Mitochondrial Phylogenomics

Ruiwen Wu,¹Liping Zhang,¹Lili Liu,¹Junli Jia,¹and Xiongjun Liu²

Academic Editor: Savel Daniels

Received28 Nov 2023

Revised17 Dec 2023

Accepted22 Dec 2023

Published08 Jan 2024

Abstract

Accurate phylogenetic reconstructions are crucial for comprehending the evolutionary histories, reproductive traits, and ecological habits of organisms. The subfamily Gonideinae of freshwater mussels is currently thought to include eight tribes. However, due to inadequate taxon sampling and molecular data, the assignment of the freshwater mussel genus Inversidens at the tribe level has been unstable. Additionally, the lack of phylogenetic data has hindered efforts to understand the basic biology and implementation of conservation efforts of Inversidens rentianensis, an endemic species to China. Here, we first present the complete mitochondrial genome of Inversidens rentianensis and offer a detailed description of its anatomical morphology. Based on DNA sequence data from five genes (COI, ND1, 16S rRNA, 18S rRNA, and 28S rRNA) and complete mitochondrial genomes, we investigated the phylogenetic position of Inversidens using various analytical methods. Both the concatenated five-gene and mitogenome datasets strongly supported that Inversidens classified to the tribe Gonideini in Gonideinae and formed a basal clade within the tribe Gonideini. Molecular dating analysis suggested that Inversidens originated during the mid-Cretaceous era (102.73 Mya, 95% highest posterior density Mya) and underwent diversification in the Late Paleogene era (37.92 Mya, 95% Mya). Moreover, based on the Quantitative Assessment of Species for Conservation (QASCP), Inversidens rentianensis is ranked as second priority, providing valuable insights for its management and conservation efforts. Taken together, this study provides a comprehensive understanding of the systematic position and evolutionary history of Inversidens within the currently accepted subfamily Gonideinae classification framework. These findings establish a solid foundation for future investigations on the ecology, reproductive behavior patterns, and conservation biology of this taxonomic group.

1. Introduction

Freshwater mussels of the family Unionidae occur in freshwater habitats worldwide except Antarctica and South America, with the most diverse regions being East Asia and North America [1–3]. These bivalves are ecologically significant benthic organisms that play pivotal roles in ecosystem functioning and services [4–7]. Additionally, they also possess substantial economic value; for instance, soft body serves as a natural food source for poultry and livestock, while shell holds high ornamental and collection value [8]. Regrettably, anthropogenic activities and environmental changes have posed severe threats to the habitats of freshwater mussels, resulting in a significant decline in their biodiversity over recent decades [9–11]. In many countries, especially in North America and Europe, the number of conservation actions for these mussels has increased substantially over the latest years [12, 13]. China is a biodiversity hotspot for freshwater mussels, but limited understanding of their biology and lack of precise systematic classification hinder the implementation of conservation measures for certain taxa [14].

The freshwater mussel genus Inversidens Haas, 1911 was initially described as a subgenus of Nodularia Conrad, 1853, containing two species endemic to Japan, i.e., Unio brandtii Kobelt, 1879 and Nodularia parcedentata Haas, 1911 [15]. Later, Thiele [16] elevated it to the generic level and designated U. brandtii as the type species, classifying it under the subfamily Unioninae in Unionidae. Haas [17] also placed Inversidens in Unioninae based on shell morphology and described six species within the genus. The subsequent efforts of malacologists focused on refining the classification on the genus level and assessing the species validity [18–21].

Currently, there are three recognized species within this genus, i.e., Inversidens brandtii (Kobelt, 1879), Inversidens pantoensis (Neumayr, 1899), and Inversidens rentianensis Wu & Wu, 2021 [22, 23]. I. brandtii is restricted to Japan [21], while I. pantoensis and I. rentianensis are endemic to China [24, 25].

The classification of the upper genus for Inversidens based on shell morphology has been subject to uncertainty [26, 27]. With the recent advancement of molecular systematics, researchers have attempted to employ molecular fragments (COI, 16S rRNA, and 28S rRNA) from type species Inversidens brandtii for resolving phylogenetic positions [28–31]. At present, the taxonomic status of this genus under the subfamily Gonideinae in Unionidae has been recognized [22, 23]. However, the classification position at the tribe level has not been solved (Figure 1). Based on three-gene data, Lopes-Lima et al. [30] resolved the diversity, biogeography, and evolutionary relationships of the Far East Asian Unionidae and assigned Inversidens brandtii to the tribe Gonideini based on a phylogenetic context of five tribes within Gonideinae. Later, Dai et al. [32] expanded the taxonomic sampling and found that the results do not support the classification of Inversidens to Gonideini and considered Gonideini to be polyphyletic. Recently, a phylogenetic framework for eight tribes of Gonideinae proposed by Zieritz et al. [33] by incorporating new taxa and molecular markers (five genes) has been accepted. Therefore, the systematic status of the genus Inversidens in the context of the updated phylogeny of Gonideinae necessitates reassessment.

Additionally, the species Inversidens rentianensis Wu & Wu, 2021, which was discovered and described by our research team [25], only had limited molecular data (i.e., mitochondrial COI) and shell morphological description. This species has a limited distribution range and specific ecological requirements, exhibiting a preference for inhabiting smaller rivers and streams. The vulnerability of this distinctive habitat to both anthropogenic and natural influences makes the species vulnerable to threats [34, 35]. The progress of modern biodiversity and conservation research heavily relies on the phylogenetic information [36]. Yet the lack of phylogenetic data for many imperiled species hinders our understanding of their basic biology and the implementation of conservation efforts [37–39]. Thus, phylogenetic information on I. rentianensis must be gathered to ensure scientific assessment of the endangered status and development of conservation and management strategies.

To determine the phylogenetic position of Inversidens, we constructed a combined gene dataset (including COI-mtDNA, ND1-mtDNA, mt 16S-rRNA, 18S-rRNA, and 28S-rRNA) based on Zieritz et al.’s [33] comprehensive sampling of eight tribes within the Gonideinae and utilized various software programs with partitioning strategies to construct phylogenies. Moreover, mitochondrial genomes are known for providing significantly more informative characters for phylogenetics and have proven highly effective in resolving both shallow and deep relationships in freshwater mussels [40–43]. In order to further validate the systematic position of Inversidens obtained from the five-gene dataset, we also conducted phylogenetic analyses using mitochondrial genomes. Therefore, a new mitogenome from Inversidens rentianensis was acquired.

Herein, the purpose of this study is to (1) describe the anatomical characteristics of Inversidens rentianensis and decipher its female complete mitochondrial genome, (2) elucidate the phylogenetic position of Inversidens based on both multilocus dataset and mitogenomic dataset, and (3) infer the evolutionary history of the genus Inversidens by time-calibrated phylogeny of Unionoidea.

2. Materials and Methods

2.1. Sample Collection and Anatomical Feature Observation

From 2021 to 2022, we conducted multiple field expeditions to the type locality of Mianshui River in Ganzhou City, Jiangxi Province, China, for collecting Inversidens rentianensis. However, it was not until October 2022 that we fortuitously obtained two live specimens (Figure 2). Both voucher specimens were deposited at the Museum of Zoology, Shanxi Normal University (SXNU), China (SXNU22102201-SXNU22102202). Anatomical features of the soft body were observed by visual examination and stereoscopic microscopy.

2.2. DNA Extraction, PCR Sequencing, and Mitogenome Assembly

Based on five genes, i.e., the mitochondrial cytochrome c oxidase subunit I (COI), NADH dehydrogenase subunit 1 (ND1), and 16S rRNA, as well as the nuclear 18S rRNA and 28S rRNA gene fragments, we compiled three multilocus datasets: a mtDNA gene dataset (COI+ND1+16S), a nuDNA gene dataset (18S+28S), and a concatenated five-gene dataset (COI+ND1+16S+18S+28S). We then employed various phylogenetic methods to generate a set of phylogenetic hypotheses.

Genomic DNA from samples was extracted from dissected foot tissue using the TIANamp Marine Animals DNA Kit (Tiangen Biotech, Beijing, China) according to the manufacturer’s instructions. Five gene regions (COI, ND1, 16S rRNA, 18S rRNA, and 28S rRNA) were amplified and sequenced using the same primers from Zieritz et al. [33]. PCR conditions were performed according to the TaKaRa Ex manufacturer’s protocol, including an initial denaturation step at 98°C for 10 s, followed by 35 cycles of amplification consisting of a denaturation step at 94°C for 1 min, annealing at 50°C for 1 min, and extension at 72°C for 1 min. The final extension was performed at 72°C for seven minutes. Amplified PCR products were purified and sequenced by Sangon Biotech (Shanghai).

Based on the dataset provided by Zieritz et al. [33], we expanded the taxa, resulting in a five-gene dataset comprising 35 Gonideinae taxa, with Margaritifera dahurica and Margaritifera margaritifera included as outgroup. The sequences of certain gene fragments from Postolata guangxiensis, Ptychorhynchus pfisteri, Koreosolenaia sitgyensis, and Parvasolenaia rivularis are newly obtained in this study (Table 1).

To further confirm the results of the multilocus datasets and clarify the phylogenetic relationship between Inversidens and other close relative, we sequenced the mitochondrial genome of Inversidens rentianensis. Qualified samples were sent to Novogene Co., Ltd. (China) for library construction and sequencing. The complete mitogenome was concatenated by Geneious (ver. 11.0, see https://www.geneious.com) [44]. Protein-coding genes were confirmed by the NCBI ORF Finder (https://www.ncbi.nlm.nih.gov/orffinder/) and BLAST search (http://blast.ncbi.nlm. http://nih.gov/) analysis. Subsequently, the mitogenome genomic sequence was submitted to GenBank under accession number OR823224.

The mitogenome sequence was annotated using MITOS WebServer (http://mitos.bioinf.uni-leipzig.de/index.py) [45]. Sequence length, base composition, and AT content were calculated using the built-in EditSeq program in DNAstar. Strand asymmetry was determined by calculating the and . The mitochondrial genome was visualized using GenomeVx (GenomeVx (ucd.ie)) [46] with manual modifications.

We combined the new genome with 92 Unionida mitogenomes from GenBank to construct the complete mitogenome dataset (Table 2). The nucleotide sequences of 12 protein-coding genes (12 PCGs, excluding ATP8 due to high sequence variation) and two rRNA genes were used for subsequent phylogenetic analyses.

2.3. Phylogenetic Analyses

The molecular phylogenetic reconstruction is consistent with the methods used in our previous studies [47]. In the datasets constructed above, protein-coding genes (PCGs) were aligned by built-in MUSCLE [48] with default settings implemented in PhyloSuite [49]. rRNA genes were aligned using MAFFT v7.2 [50] with the L-INS-i algorithm. Ambiguous alignment areas were trimmed by Gblocks (ver. 0.91b; see http://molevol.cmima.csic.es/castresana/Gblocks.html) [51], the parameter ribosomal gene block with a minimum length was set to 2 base pairs (bp), and allowed gap position was selected with half; the minimum length of protein-coding gene block was set to 3 bp, and allowed gap position was also selected with half. For the five-gene dataset, the COI, ND1, 16S, 18S, and 28S sequences were aligned and trimmed to lengths of 657 bp, 882 bp, 469 bp, 642 bp, and 743 bp, respectively. The sequences of various multigene datasets were concatenated using PhyloSuite v1.2.3. The mitogenomic data consisted of concatenated twelve protein-coding genes (12 PCGs) and two ribosomal RNA genes with a total length of 12,751 bp.

All built datasets were performed with partition schemes based on genes and codons. PartitionFinder (ver. 2.1.1; see http://www.robertlanfear.com/partitionfinder/) [52] was used to select Bayesian inference (BI) and BEAST analysis models for partitioning schemes. ModelFinder (ver. 1.4.2; see http://www.iqtree.org/ModelFinder/) [53] was used to select the maximum likelihood (ML) analysis models in IQ-TREE. The selection for best-fit models was based on the corrected Akaike information criterion (AICc). Substitution models assigned to each partition by PartitionFinder and ModelFinder are listed in Supplementary Table S1–S4.

ML analysis was performed in IQ-TREE [54] based on generated models in ModelFinder, using 1000 ultrafast bootstraps. BI analysis was used in MrBayes v2.01 [55] with generated models in PartitionFinder. Four independent Markov chain Monte Carlo (MCMC) were run simultaneously for ten million generations, and sampling was conducted every 1000 generations, with a burn-in of 25%. We terminated the process when the average standard deviation of splitting frequency falls below 0.01. The visualization and editing of phylogenetic trees were performed by using iTOL online software (http://itol.embl.de/itol.cgi) [56].

2.4. Divergence Time Estimation

The time-calibrated phylogenetic tree was generated using BEAST v1.8.4 [57] based on the concatenated five-gene dataset. We employed the uncorrelated lognormal clock model with a priori model of birth-death speciation process for tree construction. Three reliable fossil calibrations were selected according to our previous study [58]: (1) the minimum stem age of Margaritiferidae was set to 230 Mya (exponential prior, ), (2) the minimum age of Margaritifera dahurica and Margaritifera margaritifera for the most recent common ancestor (MRCA) was set to 34 Mya (exponential prior, ), and (3) the minimum age of the most recent common ancestor (MRCA) of Lamprotula leaii and Lamprotula caveata was also set to 34 Mya (exponential prior, ).

Four independent MCMC ran with 300 million generations each with sampling every 50,000 generations. Convergence and effective sample size (ESS) of parameters were checked using Tracer 1.7.1 [59]. LogCombiner v1.8.4 [57] was used to combine trees from these four runs with discarding the first 25% generations. A maximum clade credibility tree was summarized in TreeAnnotator v1.8.4 [57], with a burn-in of 25%.

3. Results

3.1. Soft-Body Anatomical Features

The fresh soft-body morphology of Inversidens rentianensis is depicted in Figure 3. The color of gills is milky white, and the inner gill is larger than the outer one (Figure 3(a)). The foot is butter yellow and darker than the gills (Figure 3(a)). The papillae of incurrent aperture are weakly developed, arranged in two rows, and almost stick together (Figure 3(b)); no papillae are observed in the excurrent aperture (Figure 3(c)). The pigmentation on both the incurrent and excurrent aperture is remarkable. The anal opening, located on the dorsal margin of the posterior adductor muscle, is connected to the excurrent aperture (Figure 3(d)). Labial palps are milky white and subtriangular (Figure 3(e)).

(a)

(b)

(c)

(d)

(e)

Figure 3

Anatomical features of Inversidens rentianensis with left valve remove. (a) Overall anatomical diagram of Inversidens rentianensis; (b–e) close-up of apertures, anal opening, and labial palps. Abbreviations: aam: anterior adductor muscle; pam: posterior adductor muscle; exa: excurrent aperture; ia: incurrent aperture; f: foot; ig: inner gill; og: outer gill; ao: anal opening; lp: labial palps; m: mantle; p ia: papillae in incurrent aperture; pg exa: pigmentation of excurrent aperture. Scale of (a) is 5 mm, and scale of (b–e) is 1 mm.

3.2. Mitochondrial Genome Structure

The mitogenome of Inversidens rentianensis is 15,987 bp in length (Figure 4). A+T bias has been observed in many unionid mitogenomes [60], and I. rentianensis genome is no exception: AT content of 60.91% (38.48% A, 22.42% T, 11.35% G, and 27.75% C). The AT skew and GC skew are 0.26% and -0.42%, respectively, indicating a bias towards A over T and C over G.

The genome contains all 37 genes commonly found in animal mtDNAs [61]: 13 protein-coding genes (PCGs), 22 tRNAs (including two trnL and two trnS), and 2 rRNAs (Figure 4). The gene arrangement pattern is consistent with species in the subfamily Gonideinae, except for Chamberlainia somsakpanhai [62, 63]. The gene distribution pattern in unionid mussels follows the typical arrangement, with 11 genes (trnH, cox2, cox1, cox3, atp6, trnD, atp8, nad4L, nad4, nad5, and nad3) located on the heavy strand and the remaining 26 genes encoded on the light strand (Figure 4). Additionally, a noncoding region of 1468 bp is observed which accounts for approximately 9.2% of the entire mitogenome.

The 13 PCGs have a total length of 10,995 bp, which accounts for 68.77% of the entire mitochondrial genome. The base composition is A, 21.44%; T, 38.73%; G, 24.09%; and C, 15.74%, with an AT content of 60.17%. The mitochondrial genome has four start codons: ATA, ATT, ATG, and TTG. Among 13 PCGs, cox1 starts with TTG while the other genes use standard codons ATN (ATA, ATG, and ATT). The mitogenome contains 22 tRNA genes, including two trnS and two trnL. Most of tRNA genes are located on the light strand, except for the trnH and trnD (Figure 4). The length of tRNA genes varies from 52 bp (trnA) to 68 bp (trnS1 and trnN). Both rrnS and rrnL are encoded on the light strand and are separated by trnK, trnT, and trnY (Figure 4), as in all the unionoid mt genomes studied so far [60, 63].

3.3. Multilocus Phylogenetic Analyses

Based on the mtDNA gene dataset, the phylogenetic trees obtained from Bayesian inference (BI) and maximum likelihood (ML) analyses show that all eight tribes in the subfamily Gonideinae are monophyletic groups, although most are weakly supported (, ). The sisterly group between Inversidens brandtii and Inversidens rentianensis is closely related to the tribe Lamprotulini, but with low support values ( and , Figure 5(a)). The BI and ML trees based on the nuDNA dataset do not form monophyly for all eight tribes, while taxa of Inversidens independently form distinct clades that do not belong to any tribe (Figures 5(b) and 5(c)).

(a) ML/BI-mtDNA

(b) BI-nuDNA

(c) ML-nuDNA

Figure 5

Phylogenetic trees inferred from Bayesian inference (BI) and maximum likelihood (ML) analyses. (a) Phylogram of BI and ML tree based on the mtDNA gene dataset (COI+ND1+16S rRNA). Numbers at nodes are support values for ML (bootstrap support (BS))/BI (posterior probability (PP)). (b) Phylogram of BI tree based on the nuDNA gene dataset (18S rRNA+28S rRNA). Numbers close to the nodes indicate for PP values. (c) Phylogram of ML tree based on nuDNA gene dataset (18S rRNA+28S rRNA). Numbers close to the nodes indicate BS values. Both Inversidens species are shown in red font. Colored clades represent eight tribes of the subfamily Gonideinae.

The ML and BI trees based on the concatenated five-gene dataset yield congruent topologies (Figure 6). The eight recognized tribes of the subfamily Gonideinae form monophyletic groups, with the following phylogenetic relationships: ((((Gonideini+Lamprotulini)+(Pseudodontini+Schepmaniini))+((Contradentini+Rectidentini)+Ctenodesmini))+Chamberlainiini). However, these clades are not well resolved in both ML and BI trees, as indicated by low supports at certain nodes (Figure 6; , ).

Both BI and ML phylogenetic analyses consistently support a sister group relationship between Inversidens brandtii and Inversidens rentianensis, with strong support values (, ; Figure 6). Furthermore, these two species are placed in a basal clade within the tribe Gonideini, as strongly supported by maximum likelihood bootstrap (BS) values of 97% and Bayesian posterior probabilities (PP) of 1.0 (Figure 6).

3.4. Fossil Calibrations and Molecular Dating

The tree topology obtained using the AICc partitioning scheme in BEAST does not align with our BI and ML phylogeny hypothesis at the tribe level; however, it consistently exhibits a phylogenetic relationship among taxa of Gonideini (Figures 6 and 7).

The genus Inversidens originates in the mid-Cretaceous (102.73 Mya, 95% Mya). The divergence time between I. rentianensis and I. brandtii is estimated to be 37.92 Mya (95% Mya), which occurs during the Late Paleogene (Figure 7).

3.5. Mitochondrial Phylogenomic Analyses

ML and BI trees based on the mitogenome dataset yield nearly identical topologies and are statistically well supported by 100% maximum likelihood bootstrap (BS) support values and Bayesian posterior probabilities (PP) in most nodes. Some discrepancies are observed only in specific branches, i.e., Anodonta nuttalliana, Anodonta anatina, and Pseudanodonta complanata (Figure S1-S2).

The phylogenetic results support the recognized four monophyletic groups of Unionidae, i.e., (Ambleminae+(Gonideinae+(Unioninae+Parreysiinae))) (Figure 8). Focusing on the subfamily Gonideinae, it is divided into six robust monophyletic clades, corresponding to six tribes with the following relationships: (Chamberlainiini+((Rectidentini+Contradentini)+(Lamprotulini+(Gonideini+Pseudodontini)))) by high support values (, ; Figure 8). In both trees, Inversidens rentianensis is positioned as a basal clade within the tribe Gonideini with strong support values ( and ; Figure 8).

Figure 8

Phylogenetic trees based on mitogenome dataset inferred from Bayesian inference (BI) and maximum likelihood (ML) analyses. (a) BI tree. (b) ML tree. Numbers close to the nodes for (a) indicate posterior probabilities (PP), and those for (b) indicate bootstrap support (BS). Colored backgrounds and colored clades represent six tribes of the subfamily Gonideinae. Red font indicates the species sequence from this study. Here, outgroups are not shown, and both Unioninae and Ambleminae taxa are collapsed in the phylogenetic tree, but complete trees can be found in supplementary Figure 1 and 2.

4. Discussion

4.1. Systematic Position of Inversidens from Multilocus and Mitogenomic Data

The classification of Gonideinae has undergone consistent revisions in recent years by the incorporation of additional taxa [33, 64–67]. Lopes-Lima et al. [30] classified Inversidens in Gonideini under the phylogenetic framework of the five tribes of Gonideinae based on the (COI+28S) dataset. However, Dai et al.’s [32] phylogenetic trees inferred from the three-gene (COI+16S+28S) dataset did not support this classification and suggested that “Inversidens was not within any tribe of Gonideinae.” Recently, Zieritz et al. [33] established and recognized a taxonomic system for eight tribes within Gonideinae based on five-gene phylogeny (COI+ND1+16S+18S+28S). Under this updated phylogenetic framework, we attempt to utilize multiple molecular datasets to address the phylogenetic location of Inversidens. The nuDNA trees were unable to effectively resolve the tribal relationship (Figures 5(b) and 5(c)), suggesting that nuclear genes 18S and 28S lack reliability in analyzing relationships at the tribe and lower levels within unionids. The phylogenetic relationships of all eight tribes were consistent between the mtDNA tree and the combined five-gene tree (Figures 5(a) and 6); however, the mtDNA tree exhibited limited confidence in accurately locating Inversidens. Therefore, we strongly support the genus Inversidens as a member of the tribe Gonideini and positioned in the basal clade based on the five-gene phylogeny (Figure 6, ). Additionally, our mitochondrial genome systematics further corroborate its taxonomic status (Figure 8, ).

Accurate systematics is essential for comprehending the evolutionary lineage of organisms. The previous divergence time estimation for the tribe Gonideini was 63.7 Mya, with Ptychorhynchus pfisteri as the basal clade [41]. In this study, the basal clade group of Gonideini was determined to be Inversidens. By reconstructing the species differentiation time, we revised the divergence time for Gonideini to be 102.73 Mya (95% Mya) (Figure 7). Despite we speculate that Inversidens rentianensis and Inversidens brandtii, which are endemic to China and Japan, respectively, originated in the Late Paleogene, their historical biogeography remains enigmatic due to a lack of comprehensive understanding on species diversity and distribution for this group. The recent discovery of new species in Southwest China [32, 68, 69] underscores the imperative for a more comprehensive examination of the overall diversity and distribution of freshwater mussels in other inadequately surveyed regions across China.

The accuracy of the phylogenetic framework is influenced by insufficient information loci and inadequate taxon selection [70, 71]. The phylogenetic reconstruction of eight tribes in Gonideinae that was conducted by Zieritz et al. [33] only included three taxa from the species-rich tribes Gonideini and Lamprotulini. The present study incorporated additional sampling taxa from Gonideini and Lamprotulini, which predictably revealed discrepancies in the phylogenetic relationships at the tribe level. The mitochondrial genomics in this study covered six out of the eight tribes within the subfamily Gonideinae, excluding Schepmaniini and Ctenodesmini due to unavailability of mitochondrial genome data. We found strong support for the monophyly of six tribes within Gonideinae, which is consistent with recent phylogenomic analyses utilizing anchored hybrid enrichment data [66] and mitogenomic data [32, 41, 63]. In this study, a novel phylogenetic hypothesis for the eight tribes of Gonideinae was proposed based on the five-locus dataset; however, further verification is required due to the low nodal support. To reconstruct and validate deep evolutionary relationships within Gonideinae, we suggest including genome-scale data such as mitochondrial genomes from Schepmaniini and Ctenodesmini taxa.

4.2. Threats and Conservation Implications

The genus Inversidens currently recognizes three species, with both I. pantoensis and I. brandtii classified as data deficient on the IUCN Red List, while the third species, I. rentianensis, remains unevaluated due to its recent discovery and description [25, 72, 73]. I. brandtii, an endemic species in Japan, was assessed as vulnerable based on extensive research conducted by Kondo [21] on its morphology, ecology, and reproductive biology. On the contrary, the understanding of the endemic species I. pantoensis and I. rentianensis in China is obviously insufficient. The knowledge about I. pantoensis is currently limited to its morphological shell description, which poses significant challenges in determining its validity due to the considerable shell morphological plasticity [74–76]. It is regrettable that the taxonomic status of this particular species has not been determined yet due to difficulties in collecting specimens for molecular examination. The species I. rentianensis is restricted to a narrow geographic range and primarily inhabits creeks and small streams characterized by substrata ranging from gravelly to sandy muddy compositions. Despite multiple visits to the type locality following the publication of this new species, only two live specimens were successfully collected. Through careful examination of these invaluable specimens, our understanding of the anatomy and molecular systematics of I. rentianensis has been significantly enriched. Since no brooding individuals were collected in this study, we could not determine the exact brooding period and examine the glochidia of this species. The accuracy of phylogenies is crucial for comprehending the evolutionary histories, reproductive traits, and ecological habits of organisms. The breeding period of the closely related species I. brandtii is from May to September [18]. The specimens of I. rentianensis were collected in October, and the gill exhibited characteristics indicative of postegg release. Therefore, we speculated that the breeding period of I. rentianensis likely occurs during August and September. Recently, Liu et al. [14] established a new conservation priority method for freshwater mussels, i.e., Quantitative Assessment of Species for Conservation (QASCP). According to this conservation assessment method, our preliminary evaluation ranks I. rentianensis as the second priority.

The impacts of urbanization and human activities have left river habitats containing endemic freshwater mussel species vulnerable to threats and require urgent attention and protection [38, 77]. Fortunately, from 2020, Chinese government has implemented comprehensive watershed protection measures (e.g., a decade-long fishing prohibition in crucial sections of the Yangtze River) and legislation (e.g., Yangtze River Protection Law) to effectively restore aquatic ecosystems, yielding remarkable outcomes [78, 79]. The recent discovery of new species, including Inversidens rentianensis, highlights the crucial role that small rivers and tributaries play as sanctuaries for freshwater mussels. However, the lack of comprehensive biodiversity surveys in these tributary basins hampers our understanding of aquatic biodiversity and the factors posing threats to it. Furthermore, the conservation efforts for unionid fauna are significantly hindered by a lack of knowledge regarding their distribution, population dynamics, and accurate taxonomic information pertaining to freshwater mussels. Therefore, we advocate researchers to pay more attention to the species diversity and population distribution of freshwater mussels in small tributaries. Simultaneously, we urge the government to formulate scientific protection measures aimed at safeguarding other endemic or endangered mussel species inhabiting small rivers and streams, in addition to those residing in larger rivers.

5. Conclusion

This study provides anatomical features and mitogenome data for Inversidens rentianensis. The mitochondrial phylogenomics and five-gene phylogeny clearly show that Inversidens rentianensis and Inversidens brandtii have a well-supported sister group relationship and form a basal clade in the tribe Gonideini, which should be classified to Gonideini in the subfamily Gonideinae. Molecular dating analysis suggests that Inversidens originated in the mid-Cretaceous era (102.73 Mya, 95% Mya) and diversified during the Late Paleogene era (37.92 Mya, 95% Mya). Additionally, preliminary assessment ranks Chinese endemic species Inversidens rentianensis as second priority based on the QASCP, which aids in managing and conserving this rare group. Taken together, this study shed light on the systematic position of Inversidens, while enriching information on the biology of Inversidens rentianensis, an endemic species to China. These findings pave the way for further comprehensive studies on the ecology, reproductive behavior patterns, and conservation biology of this fascinating group.

Data Availability

Sequences obtained in this study were uploaded to GenBank, and readers can find GenBank accession numbers in Tables 1 and 2.

Conflicts of Interest

The authors declare that there are no conflicts of interest regarding the publication of this paper.

Acknowledgments

This work was funded by the National Natural Science Foundation of China (No. 32200370) and the Basic Research Program of Shanxi Province, China (No. 20210302124253).

Supplementary Materials

Supplementary Figure 1: phylogenetic trees based on mitogenome dataset inferred from maximum likelihood (ML) analyses. Supplementary Figure 2: phylogenetic trees based on mitogenome dataset inferred from Bayesian inference (BI) analyses. Supplementary Table S1: partitioning strategies from ModerFinder and PartitionFinder for mtDNA dataset. Supplementary Table S2: partitioning strategies from ModerFinder and PartitionFinder for nuDNA dataset. Supplementary Table S3: partitioning strategies from ModerFinder and PartitionFinder for concatenated five-gene dataset. Supplementary Table S4: partitioning strategies from ModerFinder and PartitionFinder for mitogenome dataset. (Supplementary Materials)

References

A. E. Bogan, “Global diversity of freshwater mussels (Mollusca, Bivalvia) in freshwater,” Hydrobiologia, vol. 595, no. 1, pp. 139–147, 2008.
View at: Publisher Site | Google Scholar
D. L. Graf, “Patterns of freshwater bivalve global diversity and the state of phylogenetic studies on the Unionoida, Sphaeriidae, and Cyrenidae,” American Malacological Bulletin, vol. 31, no. 1, pp. 135–153, 2013.
View at: Publisher Site | Google Scholar
D. L. Graf and K. S. Cummings, “A ‘big data’ approach to global freshwater mussel diversity (Bivalvia: Unionoida), with an updated checklist of genera and species,” Journal of Molluscan Studies, vol. 87, no. 1, pp. 1–36, 2021.
View at: Publisher Site | Google Scholar
C. C. Vaughn and C. C. Hakenkamp, “The functional role of burrowing bivalves in freshwater ecosystems,” Freshwater Biology, vol. 46, no. 11, pp. 1431–1446, 2001.
View at: Publisher Site | Google Scholar
C. C. Vaughn, S. J. Nichols, and D. E. Spooner, “Community and foodweb ecology of freshwater mussels,” Journal of the North American Benthological Society, vol. 27, no. 2, pp. 409–423, 2008.
View at: Publisher Site | Google Scholar
C. C. Vaughn, “Ecosystem services provided by freshwater mussels,” Hydrobiologia, vol. 810, no. 1, pp. 15–27, 2018.
View at: Publisher Site | Google Scholar
J. K. Howard and K. M. Cuffeym, “The functional role of native freshwater mussels in the fluvial benthic environment,” Freshwater Biology, vol. 51, no. 3, pp. 460–474, 2006.
View at: Publisher Site | Google Scholar
Y. Y. Liu, W. Z. Zhang, Q. X. Wang, and E. Y. Wang, Economic Fauna of China: Freshwater Molluscs, Science Press, Beijing, 1979.
C. Lydeard, R. H. Cowie, W. F. Ponder et al., “The global decline of nonmarine mollusks,” Bioscience, vol. 54, no. 4, pp. 321–330, 2004.
View at: Publisher Site | Google Scholar
D. C. Aldridge, I. S. Ollard, Y. V. Bespalaya et al., “Freshwater mussel conservation: a global horizon scan of emerging threats and opportunities,” Global Change Biology, vol. 29, no. 3, pp. 575–589, 2023.
View at: Publisher Site | Google Scholar
X. J. Liu, Y. Y. Liu, R. W. Wu et al., “Systematics, distribution, biology, and conservation of freshwater mussels (Bivalvia: Unionida) in China,” Aquatic Conservation: Marine and Freshwater Ecosystems, vol. 32, no. 5, pp. 859–895, 2022.
View at: Publisher Site | Google Scholar
M. Böhm, N. I. Dewhurst-Richman, M. Seddon et al., “The conservation status of the world's freshwater molluscs,” Hydrobiologia, vol. 848, no. 12-13, pp. 3231–3254, 2021.
View at: Publisher Site | Google Scholar
M. Lopes-Lima, R. Sousa, J. Geist et al., “Conservation status of freshwater mussels in Europe: state of the art and future challenges,” Biological Reviews, vol. 92, no. 1, pp. 572–607, 2017.
View at: Publisher Site | Google Scholar
X. J. Liu, X. Yang, D. T. Zanatta et al., “Conservation status assessment and a new method for establishing conservation priorities for freshwater mussels (Bivalvia: Unionida) in the middle and lower reaches of the Yangtze River drainage,” Aquatic Conservation: Marine & Freshwater Ecosystems early online, vol. 30, no. 5, pp. 1000–1011, 2020.
View at: Publisher Site | Google Scholar
F. Haas, Die Unioniden, H. C. Küster, Ed., Systematisches Conchylien-Cabinet von Martini und Chemnitz, 1911, 9 (pt. 2, h. 45): 89-112, pls., 24-29.
J. Thiele, Handbuch der systematischen Weichtierkunde, Asher & Company, 1934.
F. Haas, “Superfamilia Unionacea,” Das Tierreich, vol. 88, pp. 1–663, 1969.
View at: Google Scholar
T. Kondo, “Taxonomic revision of Inversidens (Bivalvia: Unionidae),” Venus, vol. 41, no. 3, pp. 181–198, 1982.
View at: Google Scholar
T. Habe, “Bivalvia and Scaphopoda,” Systematics of Mollusca in Japan, vol. 373, 1977.
View at: Google Scholar
T. Kondo, “Revision of the genus Inversiunio (Bivalvia: Unionidae),” Venus, vol. 57, no. 2, pp. 85–93, 1998.
View at: Google Scholar
T. Kondo, “Monograph of Unionoida in Japan (Mollusca: Bivalvia),” Special Publication of the Malacological Society of Japan, vol. 3, pp. 32–34, 2008.
View at: Google Scholar
D. L. Graf and K. S. Cummings, “The Freshwater Mussels (Unionoida) of the World (and Other Less Consequential Bivalves),” MUSSEL Project Web Site, 2023. https://mussel-project.uwsp.edu/.
View at: Google Scholar
MolluscaBase eds, “MolluscaBase,” 2023, http://www.molluscabase.org.
View at: Google Scholar
J. He and Z. Zhuang, The Freshwater Bivalves of China, ConchBooks, 2013.
R. W. Wu, X. J. Liu, T. Kondo, S. Ouyang, and X. P. Wu, “New species of the genus Inversidens Haas, 1911 (Unionoida, Unionidae, Gonideinae) from Jiangxi Province, China,” Zookeys, vol. 1054, pp. 85–93, 2021.
View at: Publisher Site | Google Scholar
D. L. Graf and K. S. Cummings, “Review of the systematics and global diversity of freshwater mussel species (Bivalvia: Unionoida),” Journal of Molluscan Studies, vol. 73, no. 4, pp. 291–314, 2007.
View at: Publisher Site | Google Scholar
V. V. Bogatov and L. A. Prozorova, “Taxonomy and diversity of freshwater bivalve mollusks (Bivalvia) of China (based on analysis of the catalog by He and Zhuang, 2013),” Biology Bulletin, vol. 44, no. 8, pp. 922–940, 2017.
View at: Publisher Site | Google Scholar
I. Sano, A. Shirai, T. Kondo, and J.-I. Miyazaki, “Phylogenetic relationships of Japanese Unionoida (Mollusca: Bivalvia) based on mitochondrial 16S rDNA sequences,” Journal of Water Resources and Protection, vol. 9, no. 5, pp. 493–509, 2017.
View at: Publisher Site | Google Scholar
I. Sano, T. Saito, J.-I. Miyazaki et al., “Evolutionary history and diversity of unionoid mussels (Mollusca: Bivalvia) in the Japanese archipelago,” Plankton and Benthos Research, vol. 15, no. 2, pp. 97–111, 2020.
View at: Publisher Site | Google Scholar
M. Lopes-Lima, A. Hattori, T. Kondo et al., “Freshwater mussels (Bivalvia: Unionidae) from the rising sun (Far East Asia): phylogeny, systematics, and distribution,” Molecular Phylogenetics and Evolution, vol. 146, article 106755, 2020.
View at: Publisher Site | Google Scholar
M. Lopes-Lima, E. Froufe, V. T. Do et al., “Phylogeny of the most species-rich freshwater bivalve family (Bivalvia: Unionida: Unionidae): defining modern subfamilies and tribes,” Molecular Phylogenetics and Evolution, vol. 106, pp. 174–191, 2017.
View at: Publisher Site | Google Scholar
Y. T. Dai, X. C. Huang, C. X. Wu et al., “Multilocus and mitogenomic phylogenetic analyses reveal a new genus and species of freshwater mussel (Bivalvia: Unionidae) from Guangxi, China,” Invertebrate Systematics, vol. 37, no. 2, pp. 152–166, 2023.
View at: Publisher Site | Google Scholar
A. Zieritz, L. Jainih, J. Pfeiffer et al., “A new genus and two new, rare freshwater mussel (Bivalvia: Unionidae) species endemic to Borneo are threatened by ongoing habitat destruction,” Aquatic Conservation: Marine and Freshwater Ecosystems, vol. 31, no. 11, pp. 3169–3183, 2021.
View at: Publisher Site | Google Scholar
I. N. Bolotov, A. A. Makhrov, M. Y. Gofarov et al., “Climate warming as a possible trigger of keystone mussel population decline in oligotrophic rivers at the continental scale,” Scientific Reports, vol. 8, no. 1, pp. 1–9, 2018.
View at: Publisher Site | Google Scholar
D. Dudgeon, A. H. Arthington, M. O. Gessner et al., “Freshwater biodiversity: importance, threats, status and conservation challenges,” Biological Reviews, vol. 81, no. 2, pp. 163–182, 2006.
View at: Publisher Site | Google Scholar
M. Lopes-Lima, L. E. Burlakova, A. Y. Karatayev, K. Mehler, M. Seddon, and R. Sousa, “Conservation of freshwater bivalves at the global scale: diversity, threats and research needs,” Hydrobiologia, vol. 810, no. 1, pp. 1–14, 2018.
View at: Publisher Site | Google Scholar
P. D. Johnson, A. E. Bogan, K. M. Brown et al., “Conservation status of freshwater gastropods of Canada and the United States,” Fisheries, vol. 38, no. 6, pp. 247–282, 2013.
View at: Publisher Site | Google Scholar
W. R. Haag and J. D. Williams, “Biodiversity on the brink: an assessment of conservation strategies for North American freshwater mussels,” Hydrobiologia, vol. 735, no. 1, pp. 45–60, 2014.
View at: Publisher Site | Google Scholar
C. A. Taylor, R. J. DiStefano, E. R. Larson, and J. Stoeckel, “Towards a cohesive strategy for the conservation of the United States’ diverse and highly endemic crayfish fauna,” Hydrobiologia, vol. 846, no. 1, pp. 39–58, 2019.
View at: Publisher Site | Google Scholar
M. Lopes-Lima, M. M. Fonseca, D. C. Aldridge et al., “The first Margaritiferidae male (M-type) mitogenome: mitochondrial gene order as a potential character for determining higher-order phylogeny within Unionida (Bivalvia),” Journal of Molluscan Studies, vol. 83, no. 2, pp. 249–252, 2017.
View at: Publisher Site | Google Scholar
A. Zieritz, E. Froufe, I. Bolotov et al., “Mitogenomic phylogeny and fossil-calibrated mutation rates for all F- and M-type mtDNA genes of the largest freshwater mussel family, the Unionidae (Bivalvia),” Zoological Journal of the Linnean Society, vol. 193, no. 3, pp. 1088–1107, 2021.
View at: Publisher Site | Google Scholar
R. W. Wu, X. J. Liu, S. Wang, K. J. Roe, S. Ouyang, and X. P. Wu, “Analysis of mitochondrial genomes resolves the phylogenetic position of Chinese freshwater mussels (Bivalvia, Unionidae),” ZooKeys, vol. 812, no. 812, pp. 23–46, 2019.
View at: Publisher Site | Google Scholar
X. P. Wu, Y. T. Dai, N. Yin et al., “Mitogenomic phylogeny resolves Cuneopsis (Bivalvia: Unionidae) as polyphyletic: the description of two new genera and a new species,” Zoologica Scripta, vol. 51, no. 2, pp. 173–184, 2022.
View at: Publisher Site | Google Scholar
M. Kearse, R. Moir, A. Wilson et al., “Geneious basic: an integrated and extendable desktop software platform for the organization and analysis of sequence data,” Bioinformatics, vol. 28, no. 12, pp. 1647–1649, 2012.
View at: Publisher Site | Google Scholar
M. Bernt, A. Donath, F. Jühling et al., “MITOS: improved de novo metazoan mitochondrial genome annotation,” Molecular Phylogenetics and Evolution, vol. 69, no. 2, pp. 313–319, 2013.
View at: Publisher Site | Google Scholar
G. C. Conant and H. W. Kenneth, “GenomeVx: simple web-based creation of editable circular chromosome maps,” Bioinformatics, vol. 24, no. 6, pp. 861-862, 2008.
View at: Publisher Site | Google Scholar
R. W. Wu, L. L. Liu, L. P. Zhang et al., Molecular Phylogeny and Comparative Morphology Reveal the Species Validity and Systematic Position of Lepidodesma (Bivalvia: Unionidae), Organisms Diversity & Evolution, 2023.
View at: Publisher Site
R. C. Edgar, “MUSCLE: multiple sequence alignment with high accuracy and high throughput,” Nucleic Acids Research, vol. 32, no. 5, pp. 1792–1797, 2004.
View at: Publisher Site | Google Scholar
D. Zhang, F. L. Gao, I. Jakovlić et al., “PhyloSuite: an integrated and scalable desktop platform for streamlined molecular sequence data management and evolutionary phylogenetics studies,” Molecular Ecology Resources, vol. 20, no. 1, pp. 348–355, 2020.
View at: Publisher Site | Google Scholar
K. Katoh and D. M. Standley, “MAFFT multiple sequence alignment software version 7: improvements in performance and usability,” Molecular Biology and Evolution, vol. 30, no. 4, pp. 772–780, 2013.
View at: Publisher Site | Google Scholar
J. Castresana, “Selection of conserved blocks from multiple alignments for their use in phylogenetic analysis,” Molecular Biology and Evolution, vol. 17, no. 4, pp. 540–552, 2000.
View at: Publisher Site | Google Scholar
R. Lanfear, P. B. Frandsen, A. M. Wright, T. Senfeld, and B. Calcott, “PartitionFinder 2: new methods for selecting partitioned models of evolution for molecular and morphological phylogenetic analyses,” Molecular Biology and Evolution, vol. 34, no. 3, pp. 772-773, 2017.
View at: Publisher Site | Google Scholar
S. Kalyaanamoorthy, B. Q. Minh, T. K. F. Wong, A. von Haeseler, and L. S. Jermiin, “ModelFinder: fast model selection for accurate phylogenetic estimates,” Nature Methods, vol. 14, no. 6, pp. 587–589, 2017.
View at: Publisher Site | Google Scholar
B. Q. Minh, H. A. Schmidt, O. Chernomor et al., “IQTREE 2: new models and efficient methods for phylogenetic inference in the genomic era,” Molecular Biology and Evolution, vol. 37, no. 5, pp. 1530–1534, 2020.
View at: Publisher Site | Google Scholar
F. Ronquist, M. Teslenko, P. van der Mark et al., “MrBayes 3.2: efficient Bayesian phylogenetic inference and model choice across a large model space,” Systematic Biology, vol. 61, no. 3, pp. 539–542, 2012.
View at: Publisher Site | Google Scholar
I. Letunic and P. Bork, “Interactive tree of life (iTOL): an online tool for phylogenetic tree display and annotation,” Bioinformatics (Oxford, England), vol. 23, no. 1, pp. 127-128, 2007.
View at: Publisher Site | Google Scholar
A. J. Drummond, M. A. Suchard, D. Xie, and A. Rambaut, “Bayesian phylogenetics with BEAUti and the BEAST 1.7,” Molecular Biology and Evolution, vol. 29, no. 8, pp. 1969–1973, 2012.
View at: Publisher Site | Google Scholar
R. W. Wu, X. J. Liu, S. Ouyang, and X. P. Wu, “Comparative analyses of the complete mitochondrial genomes of three Lamprotula (Bivalvia: Unionidae) species: insight into the shortcomings of mitochondrial DNA for recently diverged species delimitation,” Malacologia, vol. 63, no. 1, pp. 51–66, 2020.
View at: Publisher Site | Google Scholar
A. Rambaut, A. J. Drummond, D. Xie, G. Baele, and M. A. Suchard, “Posterior summarization in Bayesian phylogenetics using Tracer 1.7,” Systematic Biology, vol. 67, no. 5, pp. 901–904, 2018.
View at: Publisher Site | Google Scholar
M. Soroka, “Characteristics of mitochondrial DNA of unionid bivalves (Mollusca: Bivalvia: Unionidae). I. Detection and characteristics of doubly uniparental inheritance (DUI) of unionid mitochondrial DNA,” Folia Malacologica, vol. 18, no. 4, pp. 147–188, 2010.
View at: Publisher Site | Google Scholar
D. R. Wolstenholme, “Animal mitochondrial DNA: structure and evolution,” International Review of Cytology, vol. 141, pp. 173–216, 1992.
View at: Publisher Site | Google Scholar
E. Froufe, I. Bolotov, D. C. Aldridge et al., “Mesozoic mitogenome rearrangements and freshwater mussel (Bivalvia: Unionoidea) macroevolution,” Heredity, vol. 124, no. 1, pp. 182–196, 2020.
View at: Publisher Site | Google Scholar
R. W. Wu, J. M. An, R. R. Chen, S. Ouyang, and X. P. Wu, “Taxonomy, phylogeny and evolution of freshwater mussels (Unionoida: Unionoidea) in China revealed by multilocus phylogenetic analyses and mitochondrial phylogenomics,” Journal of Lake Sciences, vol. 33, no. 6, pp. 1788–1804, 2021.
View at: Google Scholar
X. C. Huang, J. H. Su, J. X. Ouyang, S. Ouyang, C. H. Zhou, and X. P. Wu, “Towards a global phylogeny of freshwater mussels (Bivalvia: Unionida): species delimitation of Chinese taxa, mitochondrial phylogenomics, and diversification patterns,” Molecular Phylogenetics and Evolution, vol. 130, pp. 45–59, 2019.
View at: Publisher Site | Google Scholar
I. N. Bolotov, I. V. Vikhrev, A. V. Kondakov et al., “New taxa of freshwater mussels (Unionidae) from a species-rich but overlooked evolutionary hotspot in Southeast Asia,” Scientific Reports, vol. 7, no. 1, article 11573, 2017.
View at: Publisher Site | Google Scholar
J. M. Pfeiffer, J. W. Breinholt, and L. M. Page, “Unioverse: a phylogenomic resource for reconstructing the evolution of freshwater mussels (Bivalvia, Unionoida),” Molecular Phylogenetics and Evolution, vol. 137, pp. 114–126, 2019.
View at: Publisher Site | Google Scholar
J. M. Pfeiffer, D. L. Graf, K. S. Cummings, and L. M. Page, “Taxonomic revision of a radiation of South-east Asian freshwater mussels (Unionidae: Gonideinae: Contradentini+Rectidentini),” Invertebrate Systematics, vol. 35, no. 4, pp. 394–470, 2021.
View at: Publisher Site | Google Scholar
R. W. Wu, L. L. Liu, L. P. Zhang et al., “New species of the genus Pseudocuneopsis Huang, Dai, Chen & Wu, 2022 (Bivalvia, Unionidae) from Guangxi Province, China,” ZooKeys, vol. 1166, pp. 261–270, 2023.
View at: Publisher Site | Google Scholar
L. L. Liu, L. P. Zhang, D. D. Jin, H. T. Wang, X. J. Liu, and R. W. Wu, “Molecular and morphological evidence reveals a hidden new taxon in the endemic genus Pseudocuneopsis (Bivalvia, Unionidae) from China,” ZooKeys, vol. 1179, pp. 219–229, 2023.
View at: Publisher Site | Google Scholar
J. M. Pfeiffer and D. L. Graf, “Re-analysis confirms the polyphyly of Lamprotula Simpson, 1900 (Bivalvia: Unionidae),” Journal of Molluscan Studies, vol. 79, no. 3, pp. 249–256, 2013.
View at: Publisher Site | Google Scholar
I. N. Bolotov, A. V. Kondakov, I. V. Vikhrev et al., “Ancient river inference explains exceptional oriental freshwater mussel radiations,” Scientific Reports, vol. 7, no. 1, 2017.
View at: Publisher Site | Google Scholar
T. Rintelen and D. van Damme, Inversidens pantoensis, the IUCN red list of threatened species 2011, e.T189025A8679220, 2011.
View at: Publisher Site
K. Cummings, Inversidens brandtii, The IUCN red list of threatened species 2011, e.T173125A6962849, 2011.
View at: Publisher Site
K. Inoue, D. M. Hayes, J. L. Harris, and A. D. Christian, “Phylogenetic and morphometric analyses reveal ecophenotypic plasticity in freshwater mussels Obovaria jacksoniana and Villosa arkansasensis (Bivalvia: Unionidae),” Ecology and Evolution, vol. 3, no. 8, pp. 2670–2683, 2013.
View at: Publisher Site | Google Scholar
A. Zieritz, J. I. Hoffman, W. Amos, and D. C. Aldridge, “Phenotypic plasticity and genetic isolation-by-distance in the freshwater mussel Unio pictorum (Mollusca: Unionoida),” Evolutionary Ecology, vol. 24, no. 4, pp. 923–938, 2010.
View at: Publisher Site | Google Scholar
R. W. Wu, X. J. Liu, L. Guo, C. H. Zhou, S. Ouyang, and X. P. Wu, “DNA barcoding, multilocus phylogeny, and morphometry reveal phenotypic plasticity in the Chinese freshwater mussel Lamprotula caveata (Bivalvia: Unionidae),” Ecology and Evolution, vol. 12, no. 7, 2022.
View at: Publisher Site | Google Scholar
A. Zieritz, A. E. Bogan, E. Froufe et al., “Diversity, biogeography and conservation of freshwater mussels (Bivalvia: Unionida) in East and Southeast Asia,” Hydrobiologia, vol. 810, no. 1, pp. 29–44, 2018.
View at: Publisher Site | Google Scholar
L. Yang, M. Pan, J. R. Sun et al., “Short-term responses of macroinvertebrate assemblages to the "ten-year fishing ban" in the largest highland lake of the Yangtze basin,” Journal of Environmental Management, vol. 343, 2023.
View at: Publisher Site | Google Scholar
F. Liu, Z. X. Wang, Z. J. Xia, J. W. Wang, and H. Z. Liu, “Changes in fish resources 5 years after implementation of the 10-year fishing ban in the Chishui River, the first river with a complete fishing ban in the Yangtze River Basin,” Ecological Processes, vol. 12, no. 1, 2023.
View at: Publisher Site | Google Scholar

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