Research Article |
Corresponding author: Kyosuke Kitabatake ( 7621001k@st.toho-u.jp ) Academic editor: Célio Magalhães
© 2023 Kyosuke Kitabatake, Kentaro Izumi, Natsuko I. Kondo, Kenji Okoshi.
This is an open access article distributed under the terms of the Creative Commons Attribution License (CC BY 4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Citation:
Kitabatake K, Izumi K, Kondo NI, Okoshi K (2023) Phylogeography and genetic diversity of the Japanese mud shrimp Upogebia major (Crustacea, Decapoda, Upogebiidae): Natural or anthropogenic dispersal? ZooKeys 1182: 259-287. https://doi.org/10.3897/zookeys.1182.105030
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Upogebia major (De Haan, 1841) is known for forming huge burrows in sandy, intertidal areas that can extend to depths of over 2 m. Despite its widespread distribution in East Asia and Russia, the genetic relatedness of its regional populations remains uncertain, likely owing to difficulties in specimen collection. Therefore, to appraise the phylogeographic patterns, genetic diversity, and morphological variety of U. major, the mitochondrial DNA of specimens collected from Japan, Korea and China were subjected to molecular phylogenetic analyses of COI genes, alongside morphological assessment. As a result, we discovered four principal groups; of these, Group 1 consisted predominantly of Japanese specimens, while Groups 3 and 4 were interpreted as having originated from the continent. Group 2 exhibited genetic segregation from both continental and Japanese descent. Group 1 mostly comprising Japanese specimens implies that the planktonic larvae of U. major were disseminated north and south by ocean currents encompassing the Japanese archipelago. In contrast, individuals probably originating from the continent were discovered in Lake Notoro, Hokkaido and Matsukawa-ura, Fukushima in northeastern Japan, indicating possible introduction from the continent through ocean currents or unintentional introduction with other organisms imported. Additionally, one of the specimens collected from Matsukawa-ura exhibited significant genetic and morphological differences from other specimens, suggesting the possibility of being a subspecies. The outcomes of this study not only offer valuable insights into the origins of distribution of U. major but also introduce a novel challenge of assessing the coexistence of two routes: natural and anthropogenic dispersion.
Artificial introduction, Geographical distribution, Molecular phylogenetics, Morphological analysis, Ocean current, Topographic change, Upogebia major
The Upogebia genus constitutes a cluster of crustaceans, commonly referred to as mud shrimp, which inhabit all corners of the world and encompass approximately 280 species. Amongst these, the Japanese archipelago harbours 13 species (
The study of the phylogeography and genetic diversity of benthic organisms inhabiting coastal marine waters has been extensively researched on a global scale. For instance, Oratosquilla oratoria and Eriocheir japonica, crustaceans that inhabit the coast of China facing the East China Sea, display marked genetic divergence between the northern and southern regions of China, primarily because of topographic changes associated with the opening of the Sea of Japan (
In this study, molecular phylogenetic analyses were performed on U. major specimens collected from Japan, Korea and China. A phylogenetic tree and haplotype network, based on the cytochrome c oxidase subunit I (COI) of mitochondrial DNA, were constructed to reveal detailed interspecific relationships. Furthermore, the calculation of pairwise population differentiation (FST) and analysis of molecular variance (AMOVA) were performed to scrutinise the genetic variation amongst populations. Morphological measurements were conducted to examine the possible correlations between genetic variation and morphological characteristics. Through these examinations, the phylogeography and genetic diversity of U. major were evaluated and the physical and anthropogenic factors responsible for the genetic differentiation of the species in the vicinity of the Japanese archipelago were discussed. The findings of this study provide crucial insights into the geographical origins of U. major.
Samples of U. major were collected during the period of September 2021 to July 2022 from five sites in east and northern Japan (Fig.
(A) Provides a comprehensive representation of U. major, a Japanese species, that was obtained for this study. The depiction encompasses a multitude of details, including sample code, collection site, date of collection, isolation source and gene. (B) Depicts nucleotide sequence data obtained from GenBank, including crucial information such as sample code, species, accession number and elaborate information concerning collection location, literature and gene. The collected location of SK-03-05 was procured and has been verified by GBIF (Global Biodiversity Information Facility) (https://www.gbif.org/ja/species/2222995) on the grounds of the voucher number itemised in the registration information of the Accession Number.
A | Sample code | Collected location | Geographical coordinates | Collected date | Isolation source | Gene | Accession No. |
---|---|---|---|---|---|---|---|
NT-01 | Lake Notoro, Hokkaido | 44.0425°N, 144.1704°E | 29.july.22 | Abdomen tissue | COI | LC761102 | |
NT-02 | Lake Notoro, Hokkaido | 44.0425°N, 144.1704°E | 29.july.22 | Abdomen tissue | COI | LC761103 | |
NT-03 | Lake Notoro, Hokkaido | 44.0425°N, 144.1704°E | 29.july.22 | Abdomen tissue | COI | LC761104 | |
NT-04 | Lake Notoro, Hokkaido | 44.0425°N, 144.1704°E | 29.july.22 | Abdomen tissue | COI | LC761105 | |
NT-05 | Lake Notoro, Hokkaido | 44.0425°N, 144.1704°E | 29.july.22 | Abdomen tissue | COI | LC761106 | |
NT-06 | Lake Notoro, Hokkaido | 44.0425°N, 144.1704°E | 29.july.22 | Abdomen tissue | COI | LC761107 | |
NT-07 | Lake Notoro, Hokkaido | 44.0425°N, 144.1704°E | 29.july.22 | Abdomen tissue | COI | LC761108 | |
NT-08 | Lake Notoro, Hokkaido | 44.0425°N, 144.1704°E | 29.july.22 | Abdomen tissue | COI | LC761109 | |
NT-09 | Lake Notoro, Hokkaido | 44.0425°N, 144.1704°E | 29.july.22 | Abdomen tissue | COI | LC761110 | |
AK-01 | Lake Akkeshi, Hokkaido | 43.0256°N, 144.8792°E | 28.july.22 | Abdomen tissue | COI | LC761111 | |
AK-02 | Lake Akkeshi, Hokkaido | 43.0256°N, 144.8792°E | 28.july.22 | Abdomen tissue | COI | LC761112 | |
AK-03 | Lake Akkeshi, Hokkaido | 43.0256°N, 144.8792°E | 28.july.22 | Abdomen tissue | COI | LC761113 | |
MG-01 | Mangoku-ura, Miyagi | 38.4185°N, 141.3820°E | 22.March.22 | Abdomen tissue | COI | LC761114 | |
MG-02 | Mangoku-ura, Miyagi | 38.4185°N, 141.3820°E | 22.March.22 | Abdomen tissue | COI | LC761115 | |
MG-03 | Mangoku-ura, Miyagi | 38.4185°N, 141.3820°E | 22.March.22 | Abdomen tissue | COI | LC761116 | |
MG-04 | Mangoku-ura, Miyagi | 38.4185°N, 141.3820°E | 22.March.22 | Abdomen tissue | COI | LC761117 | |
MG-05 | Mangoku-ura, Miyagi | 38.4185°N, 141.3820°E | 19.May.22 | Abdomen tissue | COI | LC761118 | |
MG-06 | Mangoku-ura, Miyagi | 38.4185°N, 141.3820°E | 19.May.22 | Abdomen tissue | COI | LC761119 | |
MG-07 | Mangoku-ura, Miyagi | 38.4185°N, 141.3820°E | 19.May.22 | Abdomen tissue | COI | LC761120 | |
MT-01 | Matsukaura, Fukushima | 37.8128°N, 140.9723°E | 21.September.21 | Manus tissue | COI | LC761121 | |
MT-02 | Matsukaura, Fukushima | 37.8128°N, 140.9723°E | 21.September.21 | Manus tissue | COI | LC761122 | |
MT-03 | Matsukaura, Fukushima | 37.8128°N, 140.9723°E | 21.September.21 | Manus tissue | COI | LC761123 | |
MT-04 | Matsukaura, Fukushima | 37.8128°N, 140.9723°E | 20.May.22 | Abdomen tissue | COI | LC761124 | |
MT-05 | Matsukaura, Fukushima | 37.8128°N, 140.9723°E | 20.May.22 | Abdomen tissue | COI | LC761125 | |
MT-06 | Matsukaura, Fukushima | 37.8128°N, 140.9723°E | 20.May.22 | Abdomen tissue | COI | LC761126 | |
MT-07 | Matsukaura, Fukushima | 37.8128°N, 140.9723°E | 20.May.22 | Abdomen tissue | COI | LC761127 | |
SB-01 | Sanbanze, Chiba | 35.6709°N, 139.9689°E | 5.October.21 | Manus tissue | COI | LC761128 | |
SB-02 | Sanbanze, Chiba | 35.6709°N, 139.9689°E | 5.October.21 | Manus tissue | COI | LC761129 | |
SB-03 | Sanbanze, Chiba | 35.6709°N, 139.9689°E | 5.October.21 | Manus tissue | COI | LC761130 | |
SB-04 | Sanbanze, Chiba | 35.6709°N, 139.9689°E | 5.October.21 | Manus tissue | COI | LC761131 | |
SB-05 | Sanbanze, Chiba | 35.6709°N, 139.9689°E | 1.july.22 | Abdomen tissue | COI | LC761132 | |
MK-01 | Mikawa Bay, Aichi | Unknown | Purchased | Manus tissue | COI | LC761133 | |
MK-02 | Mikawa Bay, Aichi | Unknown | Purchased | Manus tissue | COI | LC761134 | |
MK-03 | Mikawa Bay, Aichi | Unknown | Purchased | Manus tissue | COI | LC761135 | |
MK-04 | Mikawa Bay, Aichi | Unknown | Purchased | Manus tissue | COI | LC761136 | |
MK-05 | Mikawa Bay, Aichi | Unknown | Purchased | Manus tissue | COI | LC761137 | |
MK-06 | Mikawa Bay, Aichi | Unknown | Purchased | Abdomen tissue | COI | LC761138 | |
MK-07 | Mikawa Bay, Aichi | Unknown | Purchased | Abdomen tissue | COI | LC761139 | |
KJ-01 | Kojima Bay, Okayama | Unknown | Purchased | Manus tissue | COI | LC761140 | |
KJ-02 | Kojima Bay, Okayama | Unknown | Purchased | Manus tissue | COI | LC761141 | |
KJ-03 | Kojima Bay, Okayama | Unknown | Purchased | Manus tissue | COI | LC761142 | |
KJ-04 | Kojima Bay, Okayama | Unknown | Purchased | Manus tissue | COI | LC761143 | |
KJ-05 | Kojima Bay, Okayama | Unknown | Purchased | Manus tissue | COI | LC761144 | |
KJ-06 | Kojima Bay, Okayama | Unknown | Purchased | Abdomen tissue | COI | LC761145 | |
KJ-07 | Kojima Bay, Okayama | Unknown | Purchased | Abdomen tissue | COI | LC761146 | |
KJ-08 | Kojima Bay, Okayama | Unknown | Purchased | Abdomen tissue | COI | LC761147 | |
KJ-09 | Kojima Bay, Okayama | Unknown | Purchased | Abdomen tissue | COI | LC761148 | |
KJ-10 | Kojima Bay, Okayama | Unknown | Purchased | Abdomen tissue | COI | LC761149 | |
KJ-11 | Kojima Bay, Okayama | Unknown | Purchased | Abdomen tissue | COI | LC761150 | |
AR-01 | Arao, Kumamoto | Unknown | Purchased | Manus tissue | COI | LC761151 | |
AR-02 | Arao, Kumamoto | Unknown | Purchased | Manus tissue | COI | LC761152 | |
AR-03 | Arao, Kumamoto | Unknown | Purchased | Manus tissue | COI | LC761153 | |
AR-04 | Arao, Kumamoto | Unknown | Purchased | Manus tissue | COI | LC761154 | |
B | Sample code, Species | Collected location | Gene | Reffrence | Accession No. | ||
SK-01 | Seosan, South Korea | COI | Kim et al. (2011) | JF793665.1 | |||
SK-02 | South Korea | COI | Kim et al. (Unpublished) | JX502989.1 | |||
SK-03 | South Korea | COI | Kim et al. (Unpublished) | JX502990.1 | |||
SK-04 | Gyeonggi-do, South Korea | COI | Kim, (Unpublished) | OL876961.1 | |||
SK-05 | Gyeonggi-do, South Korea | COI | Kim, (Unpublished) | OL876962.1 | |||
SK-06 | Gyeonggi-do, South Korea | COI | Kim, (Unpublished) | OL876963.1 | |||
CH-01 | Qingdao, China | COI | Liu et al. (2012) | JN897377.1 | |||
Upogebia yokoyai | Jeju-do, Korea | COI | Yang et al. (2014) | NC_025943.1 | |||
Wolffogebia heterocheir | India | COI | Rengaiyan et al. (2019) | MN579655.1 |
Less than 10 mg of abdominal tissue was removed from adult samples for DNA analysis. DNA was extracted using the ISOSPIN Tissue DNA Kit (NIPPON GENE, Tokyo, Japan) and stored at -25 °C until use.
Polymerase chain reaction (PCR) was used to amplify the COI region of the mitochondrial DNA. Up to 658 bp from the COI region was amplified using the universal primers LCO1490: 5′-GGTCAACAAATCATAAAGATATTG-3′ and HCO2198: 5′-TAAACTTCAGGGTGACCAAAAAATCA-3′ (
We conducted sequence analyses on 53 specimens of U. major for the COI gene, incorporating data obtained from GenBank for three species: U. major from Korea (Accession numbers: JF793665, JX502989, JX502990, OL876961, OL876962, OL876963) and China (JN897377), the closest relative species U. yokoyai (NC_025943) and Wolffogebia heterocheir (MN579655) as an outgroup, all with COI gene sequences (Table
The estimation of haplotypes, haplotype diversity (h) and nucleotide diversity (π) was performed through the utilisation of DnaSP v.6.12.03. The FST values between populations were executed using Arlequin v.3.5.2.2 and the K2P distance metric (
The parts for morphometric measurements are shown in Fig.
The data underpinning the analysis reported in this paper are deposited at GBIF, the Global Biodiversity Information Facility and are available at https://doi.org/10.15468/wmdf6k.
The final compilation of data comprised 637 bp of COI sequences from a total of 62 samples of U. major, outgroup species W. heterocheir and the closely-related outgroup species U. yokoyai.
Concatenate sequences are shown in Fig.
The disparity in base number between Group 2 and Group 3 was found to be minimal, ranging from 7 to 9 bp, while Group 1 and Group 4 showed a significant difference of 29 to 34 bp (Table
(A) The maximum, minimum, mean and standard deviation of the base number differences between groups. (B) The maximum, minimum, mean and standard deviation of the genetic distance between groups.
Between Group 1 and Group 2 | Between Group 1 and Group 3 | Between Group 1 and Group 4 | Between Group 2 and Group 3 | Between Group 2 and Group 4 | Between Group 3 and Group 4 | ||
---|---|---|---|---|---|---|---|
A | Maximum | 14 | 17 | 34 | 9 | 27 | 29 |
Minimum | 10 | 11 | 29 | 7 | 25 | 25 | |
Mean | 11.5 | 13.8 | 31.3 | 8.3 | 26.0 | 26.8 | |
Standard deviation | 0.862 | 1.114 | 1.271 | 0.699 | 1.000 | 1.115 | |
B | Maximum | 0.022 | 0.027 | 0.056 | 0.014 | 0.044 | 0.047 |
Minimum | 0.016 | 0.018 | 0.047 | 0.011 | 0.041 | 0.041 | |
Mean | 0.018 | 0.022 | 0.051 | 0.013 | 0.042 | 0.043 | |
Standard deviation | 0.001 | 0.002 | 0.002 | 0.001 | 0.002 | 0.002 |
Utilisation of the DnaSP software yielded 35 COI haplotypes (Table
Frequencies of selected haplotypes, haplotype diversity (h) and nucleotide diversity (π) in local samples from Japan, South Korea and China.
COI haplotype | Local populations | Total | ||||||||
---|---|---|---|---|---|---|---|---|---|---|
SK/CH | NT | AK | MG | MT | SB | MK | KJ | AR | ||
H_1 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 |
H_2 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 |
H_3 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 |
H_4 | 1 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 2 |
H_5 | 2 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 2 |
H_6 | 1 | 1 | 0 | 0 | 2 | 1 | 0 | 0 | 0 | 5 |
H_7 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 1 |
H_8 | 0 | 0 | 2 | 0 | 0 | 0 | 0 | 0 | 0 | 2 |
H_9 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 |
H_10 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 |
H_11 | 0 | 2 | 0 | 3 | 1 | 2 | 5 | 1 | 0 | 14 |
H_12 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 |
H_13 | 0 | 2 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 2 |
H_14 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 |
H_15 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 1 |
H_16 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 1 |
H_17 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 1 |
H_18 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 1 |
H_19 | 0 | 0 | 0 | 0 | 2 | 0 | 0 | 0 | 0 | 2 |
H_20 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 1 |
H_21 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | 1 |
H_22 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 0 | 1 |
H_23 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 1 |
H_24 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 1 | 0 | 2 |
H_25 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 1 |
H_26 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 1 |
H_27 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 1 |
H_28 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 2 | 0 | 3 |
H_29 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 1 |
H_30 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 1 |
H_31 | 0 | 0 | 0 | 0 | 0 | 0 | 0 | 1 | 0 | 1 |
H_32 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 1 |
H_33 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 1 |
H_34 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 1 |
H_35 | 0 | 0 | 0 | 1 | 0 | 0 | 0 | 0 | 0 | 1 |
h | 0.952 | 0.944 | 0.667 | 0.857 | 0.905 | 0.900 | 0.524 | 0.982 | 1.000 | 60 |
π | 0.02841 | 0.01151 | 0.00105 | 0.00344 | 0.02235 | 0.00188 | 0.00224 | 0.00320 | 0.00471 |
Pairwise FST revealed genetic differentiation in all groups (FST = 0.7931 to 0.9388, p < 0.05) (Table
Group 1 | Group 2 | Group 3 | Group 4 | |
---|---|---|---|---|
Group 1 | 0.0000 | |||
Group 2 | 0.8445 | 0.0000 | ||
Group 3 | 0.8608 | 0.7931 | 0.0000 | |
Group 4 | 0.9388 | 0.9256 | 0.9177 | 0.0000 |
A quantitative assessment of the genetic structural disparities between each group was conducted using AMOVA (Table
All measures of morphological traits of Japanese U. major are presented in Table
The size of each of the morphological parameters relative to carapace length (CL) A Carapace Width (CW) B Pleomere Total Length (PTL) C Telson Length (TL) D Telson Width (TW) E Left Propodus Length (PRL (L)), and F Right Propodus Length (PRL (R)). Shaded areas show the 95% confidence interval. Note that Group 4 was excluded from the analysis due to the carapace of the MT-03 being damaged, rendering the measurement of CL impossible.
Morphological traits observed in this study A projections located beneath the manus (a) and above the propodal finger (b) B ventral projections of pereopod 2 (arrow) C telson tip. (a) Linear shape, Female, MT-01 (b) Slightly concave shape, Male, MT-02 (c) Concave shape, MT-03 specimen from Matsukawa-ura D cross-sectional profile of the telson. (a) Linear shape, Female, MT-01 (b) Slightly arched shape, Male, MT-02 (c) Arched shape, MT-03
The morphometric measurements’ outcomes have been tabulated by group. The maximum and minimum values and the corresponding standard deviation, have been presented for each Group, except for group 4 (n = 1). The symbol “-” denotes that the observation was unattainable due to impairment.
Group | Sample Code | Sex | CL | CW | PL1 | PL2 | PL3 | PL4 | PL5 | PL6 | PTL | TL | TW | MW | PRL | ||
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
Left | Right | Left | Right | ||||||||||||||
1 | NT-01 | Male | 32.46 | 14.06 | 10.17 | - | 9.05 | 8.70 | 9.58 | 11.01 | - | - | 16.51 | 7.96 | 7.88 | 21.92 | 22.74 |
NT-02 | Male | 34.17 | 14.92 | 10.49 | 14.89 | 9.93 | - | 10.25 | 11.64 | - | 11.85 | 17.48 | 8.26 | - | 22.93 | - | |
NT-03 | Male | - | - | 11.94 | 8.71 | - | - | 8.06 | 11.13 | - | 10.48 | 14.13 | 8.33 | 8.31 | 23.40 | 8.31 | |
NT-07 | Female | 31.69 | 14.31 | 11.57 | 11.10 | 9.28 | 8.94 | 10.68 | 11.23 | 71.74 | 11.93 | 15.53 | 5.11 | 4.95 | 17.70 | 18.13 | |
NT-08 | Female | 32.97 | 13.79 | 10.60 | 12.61 | 10.13 | 9.44 | 9.05 | 11.39 | 72.66 | 11.68 | 15.60 | 5.79 | 5.22 | 18.07 | 17.71 | |
NT-09 | Female | 33.36 | 14.79 | 11.49 | 11.69 | 9.95 | 10.92 | 10.74 | - | - | - | - | 5.47 | 6.33 | 18.45 | 20.04 | |
AK-01 | Female | 33.46 | - | - | - | - | 8.63 | 7.20 | 10.25 | - | 10.44 | 12.17 | 5.56 | 5.24 | 17.23 | 16.85 | |
AK-02 | Female | - | - | - | - | - | - | - | - | - | - | - | 7.87 | - | 23.67 | - | |
AK-03 | Female | - | - | - | - | - | - | - | - | - | - | - | - | 8.24 | - | 24.48 | |
MG-01 | Female | 26.43 | 12.65 | 9.38 | 11.07 | - | 8.27 | 9.07 | 10.68 | - | 9.97 | 13.35 | 5.00 | - | 15.41 | - | |
MG-02 | Male | 31.03 | 21.49 | 10.21 | 11.17 | 8.19 | 7.68 | 8.70 | 9.61 | 63.24 | - | - | 6.06 | 6.22 | 18.22 | 18.03 | |
MG-03 | Female | - | - | 11.15 | 12.58 | 10.35 | 9.57 | - | 12.51 | 65.73 | 11.51 | 14.76 | - | - | - | - | |
MG-04 | Female | 19.26 | 8.57 | 6.10 | - | - | - | 5.86 | 7.49 | - | 7.02 | 8.60 | 3.24 | 3.12 | 10.81 | 10.77 | |
MG-05 | Female | 18.81 | 11.07 | 6.34 | - | 5.34 | 5.17 | 5.68 | 7.79 | - | 6.71 | 7.76 | 3.42 | 3.39 | 11.04 | 11.60 | |
MG-06 | Male | 21.95 | 8.73 | 6.87 | 8.09 | - | - | 5.97 | 7.51 | - | 6.71 | 9.83 | 4.10 | 3.72 | 13.23 | 12.52 | |
MG-07 | Male | 30.32 | 14.99 | 9.87 | - | 8.51 | 7.72 | 8.60 | 9.66 | - | 10.77 | 13.21 | 6.62 | 7.02 | 19.31 | 20.38 | |
MT-01 | Female | 32.44 | - | 10.13 | 12.63 | 9.13 | 7.58 | 7.46 | 11.49 | 66.00 | 11.38 | 15.51 | - | - | - | - | |
MT-02 | Male | 26.20 | 9.59 | 8.31 | 11.23 | 6.98 | 6.90 | 7.56 | 9.12 | 57.00 | 9.29 | 10.88 | 4.67 | 4.54 | 15.61 | 14.69 | |
MT-05 | Male | - | - | - | - | - | - | - | - | - | 11.49 | 15.33 | - | 8.89 | - | 23.64 | |
SB-01 | Male | 27.12 | - | 8.70 | 9.87 | 6.67 | 6.98 | 9.18 | 9.27 | 57.65 | 9.45 | 12.47 | - | 5.58 | - | 18.92 | |
SB-02 | Female | 27.08 | 13.28 | 9.87 | 11.50 | 8.76 | 6.82 | 8.81 | 9.47 | 62.05 | 10.07 | 13.64 | 4.67 | 4.97 | 15.75 | 16.30 | |
SB-03 | Male | 25.37 | 13.72 | 6.52 | 8.85 | - | 6.02 | 7.53 | 8.22 | - | 8.15 | 11.75 | 5.24 | 5.39 | 14.79 | 15.14 | |
SB-04 | Female | - | - | 9.18 | 10.43 | - | - | - | - | - | - | - | 5.14 | - | 16.09 | - | |
SB-05 | Male | 29.38 | 15.34 | - | - | - | - | - | 9.45 | - | 9.46 | 13.46 | 5.71 | 6.34 | 17.49 | 17.83 | |
MK-01 | Male | 25.89 | 12.36 | 8.68 | 10.14 | 6.85 | 5.99 | 6.40 | 8.57 | 52.62 | 8.98 | 12.24 | 5.09 | 4.81 | 15.93 | 15.78 | |
MK-02 | Male | 30.27 | 15.14 | 9.43 | 10.72 | 7.56 | 7.20 | 8.34 | 9.56 | 60.01 | 10.00 | 13.79 | - | 7.17 | - | 20.88 | |
MK-03 | Male | 30.64 | 13.03 | 9.29 | 10.38 | 7.60 | 6.96 | 7.53 | 8.66 | 57.38 | 9.19 | 12.85 | 6.08 | 6.00 | 17.44 | 17.36 | |
MK-04 | Male | 27.66 | 14.91 | 9.14 | 9.52 | 6.50 | 6.45 | 6.82 | 8.81 | 53.69 | 9.52 | 12.37 | 6.03 | 6.15 | 18.84 | 18.65 | |
MK-05 | Male | 23.85 | 11.81 | 8.01 | 8.49 | 5.82 | 6.68 | 7.30 | 7.40 | 50.38 | 8.37 | 11.08 | 4.74 | 4.82 | 14.47 | 15.39 | |
MK-06 | Female | 28.82 | 14.53 | 9.75 | 11.36 | 8.19 | 8.36 | 9.12 | 9.64 | 64.78 | 9.94 | 14.05 | 5.16 | 5.05 | 16.81 | 16.21 | |
MK-07 | Female | 29.97 | 15.79 | 9.58 | 10.91 | 7.30 | 7.39 | 8.79 | 9.78 | 61.14 | 10.05 | 13.46 | 4.91 | 5.38 | 16.28 | 16.30 | |
KJ-01 | Female | 23.99 | 12.76 | 8.30 | 9.51 | 6.25 | 6.10 | 6.43 | 8.13 | 50.82 | 8.52 | 11.46 | 5.57 | 5.66 | 16.09 | 17.35 | |
KJ-02 | Female | 22.16 | 10.00 | 6.93 | 6.84 | 5.69 | 4.72 | 5.63 | 7.05 | 41.58 | 7.19 | 9.76 | 3.98 | 4.36 | 12.05 | 13.10 | |
KJ-03 | Female | 23.34 | 10.59 | 7.42 | 8.63 | 6.18 | 6.13 | 6.46 | 7.77 | 48.72 | 7.65 | 11.01 | 4.36 | 4.33 | 13.25 | 13.30 | |
KJ-04 | Male | 25.86 | 12.17 | 8.03 | 9.56 | 6.94 | 6.67 | 7.31 | 8.03 | 53.21 | 8.68 | 12.39 | 5.32 | 5.03 | 16.06 | 15.21 | |
KJ-05 | Male | 20.88 | 11.04 | 7.08 | 8.21 | 5.63 | 5.38 | 6.02 | 7.08 | 44.78 | 6.96 | 9.65 | 4.62 | 4.61 | 13.99 | 13.53 | |
KJ-06 | Female | 19.61 | 10.56 | 6.93 | 8.06 | 5.07 | - | 5.82 | 6.78 | - | 6.98 | 9.61 | 4.13 | 4.05 | 12.62 | 12.66 | |
KJ-07 | Male | 27.12 | 12.82 | 8.53 | 10.12 | 6.65 | - | 7.13 | 8.05 | - | 8.55 | 10.83 | 5.28 | 5.89 | 15.75 | 16.82 | |
KJ-08 | Female | 22.82 | 11.21 | 7.36 | 8.63 | 6.04 | 4.67 | 7.00 | 7.57 | 45.94 | 7.66 | 10.99 | 4.71 | 4.65 | 13.98 | 13.70 | |
KJ-09 | Female | 22.42 | 10.01 | 7.91 | 9.01 | 6.20 | 6.03 | 6.50 | 7.58 | 49.26 | 7.92 | 10.27 | 4.90 | 4.78 | 14.16 | 14.33 | |
1 | KJ-10 | Male | 22.16 | 10.69 | 6.74 | 8.62 | 5.97 | 5.88 | 6.42 | 7.39 | 46.90 | 7.90 | 10.17 | 4.81 | 5.09 | 14.52 | 14.33 |
KJ-11 | Female | 23.85 | 11.17 | 7.41 | 9.41 | 6.15 | 6.94 | 7.10 | 7.37 | 51.32 | 7.82 | 10.18 | 4.61 | 4.71 | 15.31 | 14.67 | |
AR-01 | Male | 35.22 | 20.57 | 10.58 | 11.36 | 9.24 | 8.72 | 10.39 | 11.53 | 70.54 | 12.60 | 15.14 | - | 9.95 | - | 23.75 | |
AR-02 | Female | 32.18 | 17.08 | 11.50 | 12.10 | 10.03 | 8.84 | 10.35 | 10.86 | 72.52 | 12.12 | 15.88 | 6.73 | 6.41 | 20.51 | 20.85 | |
AR-03 | Female | 34.03 | 18.91 | 12.42 | 11.84 | 9.16 | 8.62 | 9.97 | 11.33 | 71.96 | 11.39 | 15.03 | 6.22 | 6.40 | 19.02 | 20.14 | |
AR-04 | Female | 34.66 | 19.36 | 12.19 | 12.52 | 9.80 | 9.83 | 10.29 | 11.82 | 76.28 | 11.86 | 14.65 | 11.48 | 6.64 | 24.97 | 20.17 | |
Maximum | 35.22 | 21.49 | 12.42 | 14.89 | 10.35 | 10.92 | 10.74 | 12.51 | 76.28 | 12.60 | 17.48 | 11.48 | 9.95 | 24.97 | 24.48 | ||
Minimum | 18.81 | 8.57 | 6.10 | 6.84 | 5.07 | 4.67 | 5.63 | 6.78 | 41.58 | 6.71 | 7.76 | 3.24 | 3.12 | 10.81 | 8.31 | ||
Mean | 27.5 | 13.5 | 9.1 | 10.3 | 7.6 | 7.3 | 7.9 | 9.3 | 58.6 | 9.5 | 12.7 | 5.6 | 5.7 | 16.7 | 16.8 | ||
Standard deviation | 4.717 | 3.093 | 1.739 | 1.673 | 1.608 | 1.493 | 1.554 | 1.620 | 9.675 | 1.722 | 2.316 | 1.523 | 1.464 | 3.362 | 3.638 | ||
2 | MT-04 | Male | 33.17 | - | 11.89 | 13.14 | 8.60 | 8.15 | - | 11.24 | - | 11.89 | 16.27 | - | 9.29 | - | 25.02 |
MT-07 | Male | 35.72 | 14.80 | 11.31 | 11.15 | 9.14 | 8.22 | 8.88 | 10.21 | 67.13 | 11.86 | 15.49 | 7.94 | - | 22.30 | - | |
Maximum | 35.72 | 14.80 | 11.89 | 13.14 | 9.14 | 8.22 | 8.88 | 11.24 | 67.13 | 11.89 | 16.27 | 7.94 | 9.29 | 22.30 | 25.02 | ||
Minimum | 33.17 | 14.80 | 11.31 | 11.15 | 8.60 | 8.15 | 8.88 | 10.21 | 67.13 | 11.86 | 15.49 | 7.94 | 9.29 | 22.30 | 25.02 | ||
Mean | 34.4 | 14.8 | 11.6 | 12.1 | 8.9 | 8.2 | 8.9 | 10.7 | 67.1 | 11.9 | 15.9 | 7.9 | 9.3 | 22.3 | 25.02 | ||
Standard deviation | 1.275 | 0.000 | 0.290 | 0.995 | 0.270 | 0.035 | 0.000 | 0.515 | 0.000 | 0.015 | 0.390 | 0.000 | 0.000 | 0.000 | 0.000 | ||
3 | NT-04 | Female | 33.89 | 16.03 | 11.71 | 11.12 | 10.43 | 9.56 | 11.70 | 11.69 | 75.77 | 11.53 | 16.61 | 5.92 | 5.58 | 17.94 | 17.52 |
NT-05 | Male | 31.46 | 12.69 | 11.36 | 11.03 | 8.42 | 9.18 | 9.76 | 11.70 | 70.63 | 11.01 | 15.49 | 7.34 | 7.14 | 21.38 | 20.40 | |
NT-06 | Female | 33.20 | 15.12 | - | 12.05 | 9.70 | 9.66 | 10.74 | 12.36 | - | 11.56 | 16.80 | 5.24 | 5.26 | 18.83 | 18.39 | |
MT-06 | Male | 34.13 | 17.16 | 10.19 | 12.12 | 8.82 | 8.26 | 9.87 | 10.70 | 68.22 | 11.07 | 15.49 | 8.16 | 7.96 | 24.26 | 24.10 | |
Maximum | 34.13 | 17.16 | 11.71 | 12.12 | 10.43 | 9.66 | 11.70 | 12.36 | 75.77 | 11.56 | 16.80 | 8.16 | 7.96 | 24.26 | 24.10 | ||
Minimum | 31.46 | 12.69 | 10.19 | 11.03 | 8.42 | 8.26 | 9.76 | 10.70 | 68.22 | 11.01 | 15.49 | 5.24 | 5.26 | 17.94 | 17.52 | ||
Mean | 33.2 | 15.3 | 11.1 | 11.6 | 9.3 | 9.2 | 10.5 | 11.6 | 71.5 | 11.3 | 16.1 | 6.7 | 6.5 | 20.6 | 20.1 | ||
Standard deviation | 1.045 | 1.645 | 0.650 | 0.507 | 0.780 | 0.552 | 0.781 | 0.593 | 3.149 | 0.254 | 0.611 | 1.149 | 1.110 | 2.460 | 2.533 | ||
4 | MT-03 | Male | - | - | - | 11.21 | 9.33 | 8.26 | 8.49 | 11.96 | - | 11.58 | 15.63 | - | 7.39 | - | 23.68 |
The COI phylogenetic tree analysis reveals that U. major is distinctly separated from its outgroups and closest species, forming its own clade comprising four groups (Fig.
The results of morphometric observations are summarised by group. NPLBM: Number of projections located beneath the manus, NPAPF: Number of projections above the propodal finger, NVPP2: Number of ventral projections of pereopod 2, MTT: Morphology of the telson tip, CST: Cross-sectional shape of the telson, L: Linear, SC: Slightly concave, C: Concave, SA: Slightly arched, A: Arched. The maximum and minimum values and the corresponding standard deviations are given for each group, except for Group 4, where the sample size is 1. Additionally, the percentage of MTT and CST traits are presented. The symbol “-” indicates an unattainable observation due to impairment.
Group | Sample code | Sex | NPLBM | NPAPF | NVPP2 | MTT | CST | ||||
---|---|---|---|---|---|---|---|---|---|---|---|
Left | Right | Left | Right | Left | Right | ||||||
1 | NT-01 | Male | 5 | 4 | 8 | 8 | – | 1 | SC | L | |
NT-02 | Male | 5 | – | 6 | – | 2 | 1 | SC | L | ||
NT-07 | Female | 4 | 5 | 4 | 7 | 2 | 2 | L | L | ||
NT-08 | Female | 6 | 5 | 7 | 4 | 1 | 1 | L | L | ||
NT-09 | Female | 5 | 4 | – | 2 | – | 1 | L | L | ||
AK-01 | Female | 5 | 6 | 5 | 4 | 1 | 1 | L | L | ||
MG-01 | Female | 4 | – | – | – | – | 2 | L | L | ||
MG-02 | Male | 3 | 4 | 8 | 9 | 3 | 2 | SC | L | ||
MG-07 | Male | 5 | 4 | 5 | 6 | 2 | 2 | L | L | ||
MT-01 | Female | – | – | – | – | – | 1 | L | L | ||
MT-02 | Male | 4 | 5 | 4 | 6 | 2 | 2 | SC | SA | ||
SB-01 | Male | – | 5 | – | 6 | 2 | – | SC | L | ||
SB-02 | Female | – | 3 | – | 8 | 2 | 1 | L | L | ||
SB-03 | Male | 5 | 6 | 4 | 5 | 2 | 1 | SC | L | ||
SB-05 | Male | 6 | 5 | 4 | 6 | 2 | 2 | SC | L | ||
MK-01 | Male | 7 | 6 | 4 | 3 | 2 | 2 | SC | L | ||
MK-02 | Male | – | 4 | – | 6 | 2 | 2 | SC | L | ||
MK-03 | Male | 6 | 6 | 6 | 5 | 2 | 2 | SC | L | ||
MK-04 | Male | 5 | 5 | 7 | 5 | 2 | 2 | SC | L | ||
MK-06 | Female | 4 | 4 | 6 | 4 | 2 | 2 | L | L | ||
MK-07 | Female | 5 | 6 | 4 | 6 | 3 | 2 | L | L | ||
KJ-04 | Male | 6 | 5 | 5 | 5 | – | – | SC | L | ||
KJ-05 | Male | 4 | 4 | 7 | 5 | 2 | 2 | SC | L | ||
AR-01 | Male | – | 5 | – | 6 | 2 | 1 | SC | L | ||
AR-02 | Female | 4 | 5 | 5 | 2 | 2 | 2 | L | L | ||
AR-03 | Female | 4 | 4 | 5 | 4 | 2 | 2 | L | L | ||
AR-04 | Female | 5 | 4 | 5 | 5 | 2 | 2 | L | L | ||
Maximum | 7 | 6 | 8 | 9 | 3 | 2 | % | L = 48.1 | L = 96.3 | ||
Minimum | 3 | 3 | 4 | 2 | 1 | 1 | SC = 51.9 | SA = 3.7 | |||
Mean | 4.9 | 4.8 | 5.5 | 5.3 | 2.0 | 1.6 | C = 0.0 | A = 0.0 | |||
Standard deviation | 0.919 | 0.829 | 1.322 | 1.695 | 0.426 | 0.480 | |||||
2 | MT-04 | Male | – | 6 | – | 8 | 1 | – | SC | L | |
MT-07 | Male | 4 | – | 8 | – | – | 2 | SC | L | ||
Maximum | 4 | 6 | 8 | 8 | 1 | 2 | % | L = 100 | L = 100 | ||
Minimum | 4 | 6 | 8 | 8 | 1 | 2 | SC = 0.0 | SA = 0.0 | |||
Mean | 4.0 | 6.0 | 8.0 | 8.0 | 1.0 | 2.0 | C = 0.0 | A = 0.0 | |||
Standard deviation | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | 0.000 | |||||
3 | NT-04 | Female | 7 | 6 | – | – | 1 | 2 | L | L | |
NT-05 | Male | 4 | 5 | 8 | 7 | 2 | 1 | SC | L | ||
NT-06 | Female | 5 | 4 | 6 | 6 | 2 | 2 | L | L | ||
MT-06 | Male | 5 | 4 | 8 | 9 | 2 | 1 | SC | L | ||
Maximum | 7 | 6 | 8 | 9 | 2 | 2 | % | L = 50.0 | L = 100 | ||
Minimum | 4 | 4 | 6 | 6 | 1 | 1 | SC = 50.0 | SA = 0.0 | |||
Mean | 5.3 | 4.8 | 7.3 | 7.3 | 1.8 | 1.5 | C = 0.0 | A = 0.0 | |||
Standard deviation | 1.090 | 0.829 | 0.943 | 1.247 | 0.433 | 0.500 | |||||
4 | MT-03 | Male | – | 4 | – | 9 | – | – | C | A |
Although the precise origin of U. major remains unknown, the oldest lineage traced in the inferred phylogenetic tree belongs to Group 4, with the South Korean sample (SK-03) being the first to diverge within this group. Thus, it is plausible that the coastal waters of Korea could serve as the putative origin of U. major. This study assumes that the origin of U. major is situated around South Korea and conducts an extensive evaluation of the genetic differentiation of this species throughout the entirety of the Japanese archipelago, taking into account various physical and anthropogenic factors.
Marine benthos possessing planktonic larval stages have the potential to increase their geographic range owing to their floating period and the hydrodynamic properties of ocean currents. Specifically, around the Japanese archipelago, three prominent oceanic currents exist: the warm Tsushima Current, the Kuroshio Current and the cold Oyashio Current (Fig.
Designations of the oceanic currents and their respective flow patterns in the Japanese archipelago. The Blue dotted arrow denotes the southern extremity of the First branch of the Oyashio current during its attenuation, while the Red dotted arrow demarcates the northern boundary of the Kuroshio Basin at that particular juncture. TC: Tsugaru Current, YSC: Yellow Sea Current, YSCC: Yellow Sea Counter Current, BP: Boso Peninsula.
Thus, ocean currents circulate around the Japanese archipelago. U. major has three larval stages and one decapod stage and its planktonic larval period is estimated to be approximately one month or more for individuals inhabiting Tokyo Bay (
The presence of U. major individuals belonging to Groups 3 and 4 in Lake Notoro and Matsukawa-ura, respectively, implies the potential for passive continental invasion of the species into Japan through the agency of oceanic currents. Included in Groups 3 and 4 were specimens from Gyeonggi-do, a coastal region situated in the western part of South Korea, as well as specimens from the Qingdao Peninsula in China and South Korean specimens with unspecified collection locations. To reach Lake Notoro, these individuals must follow the Yellow Sea Coastal Current and subsequently merge with the Tsushima Current before finally riding along the Soya Current (Fig.
Possible routes for U. major larvae from South Korea, China and Russia to Lake Notoro and Matsukawa-ura. The warm currents are denoted by red dotted arrows, while the cold currents are denoted by blue dotted arrows. The collection sites of specimens from Korea and China used in this study are indicated by green circles, while the localities of U. major in Russia are denoted by yellow circles (The exact collection site in Sakhalin Island is unknown). Furthermore, potential dispersal routes of U. major larvae from South Korean/Chinese specimens and Russian specimens are shown in green and yellow arrows, respectively. SC: Soya Current, TC: Tsugaru Current, YSC: Yellow Sea Current, YSCC: Yellow Sea Counter Current, VS: Vladivostok, VB: Vostok Bay, OB: Olga Bay, SH: Sakhalin Island.
While no prior research has documented the transfer of organisms from the Russian coastline to the Japanese archipelago via the Liman Current merging with the Tsushima Current, it is possible that U. major larvae from Russia may have travelled this route and ultimately arrived at Lake Notoro. Moreover, it is worth noting that although a greater distance must be traversed to reach Matsukawa-ura, two potential routes may be posited: one involves planktonic larvae of continental U. major inhabiting Lake Notoro riding the Soya Current into the Pacific Ocean and then utilising the Oyashio Current to reach Matsukawa-ura and the other involves reaching Matsukawa-ura via the Tsugaru Current from an intermediate relay point on the Sea of Japan (Fig.
While the sample size is not extensive, the evident genetic and morphological heterogeneity exhibited by the U. major population in Matsukawa-ura is noteworthy. This diversity may, to some extent, be influenced by changes in coastal topography. The Tohoku region, where Matsukawa-ura is situated, experiences significant earthquakes once every 500–800 years (
Additionally, an example of the spread of organisms of continental origin across the Japanese archipelago is evident in the “continental relict species”. During the Pleistocene glacial period, the southern coast of Korea and the present-day Ariake Sea were united by land, but regression of the Ariake Sea during the postglacial period resulted in their separation (
Group 1 individuals shared the haplotype H_11 at six different sites in Japan (Lake Notoro, Mangoku-ura, Matsukawa-ura, Sanbanze, Mikawa Bay and Kojima Bay) and displayed a trend of low genetic diversity, indicating that their planktonic larvae were dispersed over a broad north-south range by ocean currents surrounding the Japanese archipelago. Panulirus japonicus, a decapod crustacean resembling U. major, is broadly distributed in the Kuroshio region, but there is no identifiable genetic variation within populations. With a planktonic larval duration of about one year, it is speculated that P. japonicus with the same haplotype has moved long distances over an extended period through the Kuroshio region (
In contrast, two specimens (MT-04 and 07) from Matsukawa-ura, belonging to Group 2, exhibited a slight difference in PRL compared to Group 1 and no discernible morphological distinctions were noted between Group 2 and Group 3. Nevertheless, they exhibited a distinctive haplotype, which may suggest phenotypic plasticity. This occurrence is well established in marine snails, as demonstrated in previous studies (e.g.
In addition to physical factors, it is possible that species dispersion occurs because of anthropogenic activities. Japan began importing the Manila clam Ruditapes philippinarum, from China and North Korea in the 1980s. Jute bags of imported clams contain live organisms other than clams, which are released into domestic clam fisheries or added to aquaculture farms every year (
Conversely, in Lake Notoro, a specimen of continental origin was also detected, but there was no record of the introduction of R. philippinarum from China or Korea and none of the ten surrounding fishing tackle stores handled U. major as bait. Based on the current findings, it is highly probable that the introduction to Lake Notoro was natural and occurred via ocean currents.
In this study, we focused on various morphological traits, including spines and hairs and counted, measured and compared them amongst individuals. We identified several traits that were characteristic of the comparisons amongst individuals. The divergence exhibited by MT-03, which is affiliated with Group 4, is particularly noteworthy regarding the morphology of its telson tip and section. Additionally, genetic analysis revealed a significant genetic distance of up to 0.056 from the other Japanese specimens of Groups 1, 2 and 3, indicating that it might be considered a subspecies. However, there is a lack of available morphological data for SK-02, 03, and 05, which also belong to Group 4. Therefore, further investigation is required to confirm their classification as subspecies through continuous surveys in Matsukawa-ura and Korea.
Group 4 has been proposed as the most primordial lineage that diverged within the species U. major, with the potential occurrence of subspecies within this particular group.
We conducted morphological evaluations in conjunction with molecular phylogenetic analyses of COI genes, extracted from specimens collected in Japan, Korea, and China, to ascertain the phylogeographic patterns and genetic as well as morphological, diversity in Upogebia major. As a consequence of our analysis, U. major was classified into four primary groups: one with predominantly Japanese descent, two other groups inferred to have originated from the continent, and the other group genetically segregated from both Continental and Japanese descent. The group exclusively comprising Japanese specimens suggests that the planktonic larvae of this species are widely dispersed by ocean currents surrounding the Japanese Archipelago. In contrast, several Japanese specimens were included in the continental group, which may be due to the introduction of individuals from the continent via ocean currents, the possibility that this species being a continental relict or the unintentional introduction of biota imported from Korea and China. Matsukawa-ura demonstrated high genetic diversity, with specimens from all groups present. Moreover, one specimen sampled from Matsukawa-ura (MT-03) presented noteworthy genetic and morphological variances compared with the other specimens, indicating the possibility of its being a subspecies. A specimen from Gyeonggi-do, Korea, obtained from GenBank, was classified into the same group as MT-03, although information on its morphology was unavailable because the reference paper remains unpublished. To confirm these findings, further morphological and genetic investigations and analyses encompassing Matsukawa-ura and other sites are necessary.
The genetic dispersal of U. major suggests the existence of both natural and anthropogenic dissemination pathways, implying their intricate interplay in the shaping of regional populations. The outcomes of this study underscore the potential for analogous occurrences in all organisms, irrespective of intentional or unintentional introduction and release from neighboring regions, transcending the boundaries of this particular species. These insights not only contribute to a deeper understanding of the origins of distribution of U. major but also introduce a novel challenge of assessing the coexistence of these two dispersion routes.
We appreciate Mr. Satoru Takeyama, Director of Akkeshi Oyster Seeding Center, the Nishi-Abashiri Fisheries Cooperative Association in Hokkaido, the Ishinomaki Bay Branch of the Miyagi Prefecture Fisheries Cooperative Association and the Matsukawa-ura Branch of the Soma Futaba Fisheries Cooperative Association in Fukushima Prefecture for their understanding of the field survey and cooperation in transporting the samples. We also thank Ms. Rumiko Kaneko and Ms. Makoto Suzuki for their assistance during morphometric measurements. We would like to thank Editage (www.editage.com) for their English language editing. The comments received from Dr. Célio Magalhães, the subject editor, and the two anonymous reviewers have significantly contributed to the substantial improvement of our manuscript.
The authors have declared that no competing interests exist.
No ethical statement was reported.
This study was partly funded by JSPS KAKENHI Grant Number JP23H03519, the Sasakawa Scientific Research Grant from The Japan Science Society and The Research Institute of Marine Invertebrates.
KK conceived and designed the study, conducted the sampling, performed molecular and morphometric analyses and interpreted the data, as well as writing the manuscript. IK participated in the field survey and contributed to manuscript writing. KIN provided assistance with molecular analysis, as well as contributing to manuscript writing. OK oversaw the research, contributed to the sampling and reviewed and revised the manuscript. All authors have thoroughly reviewed and approved the final version of the manuscript.
Kyosuke Kitabatake https://orcid.org/0009-0006-4150-9622
Kentaro Izumi https://orcid.org/0000-0001-9802-071X
Natsuko I. Kondo https://orcid.org/0000-0002-0883-0506
Kenji Okoshi https://orcid.org/0000-0003-0421-2812
All of the data that support the findings of this study are available in the main text or Supplementary Information.
Molecular and morphometric analyses of Upogebia major
Data type: xlsx
Explanation note: The appendix contains certain outcomes of molecular analyses, such as variations in nucleotide numbers and genetic disparities, as well as the results and tests of morphometric analyses stratified by gender.