Research Article |
Corresponding author: Valiallah Khalaji-Pirbalouty ( vkhalaji@sci.sku.ac.ir ) Academic editor: Saskia Brix
© 2022 Valiallah Khalaji-Pirbalouty, Hamzeh Oraie, Carlos A. Santamaria, Johann Wolfgang Wägele.
This is an open access article distributed under the terms of the CC0 Public Domain Dedication.
Citation:
Khalaji-Pirbalouty V, Oraie H, Santamaria CA, Wägele JW (2022) Redescription of Tylos maindroni Giordani Soika, 1954 (Crustacea, Isopoda, Oniscidea) based on SEM and molecular data. ZooKeys 1087: 123-139. https://doi.org/10.3897/zookeys.1087.76668
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The woodlouse species Tylos maindroni Giordani Soika, 1954 (Crustacea, Isopoda, Oniscidea) is redescribed from the Persian Gulf based on light and scanning electron microscopy. This species differs from the closely related T. exiguus Stebbing, 1910, from the Red Sea (coasts of Sudan and Eritrea), and Socotra Island, by pereopod 1 superior margin without a prominent projection and pleopod 2 endopod 2.3 times as long as exopod, vs. 3.6 in T. exiguus. A distribution map for T. maindroni is provided. In addition, we studied the molecular differentiation of five populations of T. maindroni from the Persian Gulf, based on partial cytochrome c oxidase subunit I (COI) gene sequences. The results revealed low levels of population structuring between the analyzed populations.
DNA barcoding, haplotype network, Isopoda, Persian Gulf, Redescription, SEM
The isopod genus Tylos Audouin, 1826 has a worldwide distribution, with 21 species currently considered as valid (
Herein, we redescribe T. maindroni based on material from the Persian Gulf and provide new COI mtDNA sequence data.
Specimens used in this study were collected from four coastal sites in the Persian Gulf, Iran during field expeditions from 2006 to 2021 (Fig.
Samples of the Tylos maindroni from the Persian Gulf used in this study.
Museum number | Voucher numbers | Coordinates | Collection date | Locality | GenBank Accession number COI |
---|---|---|---|---|---|
ZSMU 1206 | 1043 | 26°16'20.48"N, 55°17'28.00"E | 03.01.2021 | Greater Tunb Island | OK513061 |
ZSMU 1206 | 1044 | 26°16'20.48"N, 55°17'28.00"E | 03.01.2021 | Greater Tunb Island | OK513060 |
ZMSU 1205 | 1051 | 26°6'59.99"N, 54°26'10.88"E | 30.12.2017 | Faroor Koochak Island | OK513062 |
ZMSU 1201 | 1098 | 26°42‘555"N, 54°14‘329"E | 05.12.2008 | Bandar-e-Charak | OK513063 |
ZMSU 1201 | C2 | 26°42‘555"N, 54°14‘329"E | 05.12.2008 | Bandar-e-Charak | OK513064 |
ZMSU 1202 | B1 | 27°07‘113"N, 53°01‘418"E | 30.01.2006 | Banda-e Bostaneh | OK513065 |
ZMSU 1202 | B2 | 27°07‘113"N, 53°01‘418"E | 30.01.2006 | Banda-e Bostaneh | OK513066 |
Specimens prepared for SEM were washed in a chilled 1% sodium acetate solution for 10 minutes, then cleaned for 10–20 seconds in an ultrasound cleaner in a weak solution of jewelry soap and distilled water to remove sediment and debris adhering to the cuticle. Specimens were dehydrated in an ethanol series (70, 75, 80, 85, 90, 95, 100%; 20 minutes per treatment). Specimens were transferred through ethanol and hexamethyldisilazane (HMDS) solutions (ethanol: HMDS ratios were 2:1, 1:1, 1:2) and finally into 100% HMDS (20 minutes per treatment). All samples were transferred to fresh HMDS, which evaporated overnight. Specimens were mounted on stubs using double adhesive carbon spots before being coated with gold in a sputter coater to 40 nm thickness. Micrographs were taken using a Hitachi S-2460N SEM at Zoologisches Forschungsmuseum Alexander Koenig in Bonn, Germany. Color images were taken using a Zeiss AxioCam ERc5s camera mounted on a Zeiss Stereomicroscope (Stemi 508).
We extracted genomic DNA from the legs of seven specimens, 1–2 individuals per locality, using the Aron-Gene Tissue DNA Extraction kit (Aron-Gene, Iran) following the manufacturer’s protocol. A 536 base pair fragment of the mitochondrial Cytochrome Oxidase I (COI) gene was PCR-amplified using the LCO-1490 and HCO-2198 primer pair under standard conditions (
Once assembled and edited, sequences produced in this study were combined with previously published COI sequences of T. maindroni as well as other Tylos species, provided that these sequences were > 500-bp long. Information for publicly available sequences included in this study can be found in Table
GenBank Accession information for sequences used in this study. Accession numbers of sequences produced in this study are in bold.
We used ASAP (
Lastly, we visualized relationships between T. maindroni COI haplotypes by reconstructing branch connections using the TCS network option (
Suborder Oniscidea Latreille, 1802
Tylos latreillii Audouin, 1826; from an unspecified location in Egypt (type locality); but current status is a nomen dubium (
A diagnosis for the genus was published by Schmalfuss (2000).
Tylos maindroni
Giordani Soika, 1954: 76, figs 8, 9, pl. 10, Oman Sea, Muscat (type locality); Ferrara and Taiti 1986: 94;
7 ♂♂ (5.1 to 9.8 mm), 3 ♀♀ (5.5, 8.5, 10 mm), the Persian Gulf, Bandar-e-Charak, sandy shore, under wood block and rubbish on sand, 05 Dec. 2008, 26°42'555"N, 54°14'329"E, coll. V. Khalaji (ZMSU 1201); 8 ♂♂ (5 to 8 mm), 6 ♀♀ (6 to 9.2 mm), Bandar-e-Bostaneh, sandy shore, 03 Jan. 2006, 27°07'113"N, 53°01'418"E, coll. R. Naderloo (ZMSU 1202); 1 ♀ (12.2 mm), Bandar-e-Lengeh, sandy beach, beneath wood, 03 May 2010, 26°34'10"N, 54°54'21"E, coll. V. Khalaji (ZMSU 1203); 2 ♀♀ (9 and 10 mm), Kish Island, northern coast, Derakht-e-Sabz, 24 Jun. 2006, 26°34'102"N, 53°58'098"E, coll. V. Khalaji (ZMSU 1204); 3 ♂♂ (9 to 11mm); 8 ♀♀ (8 to 10 mm), Faroor Koochak Island, rocky, sandy western coast, 30 Dec. 2017, 26°65'999"N, 54°26'108"E, coll. V. Khalaji (ZMSU 1205); 10 ♀♀ (7 to 11mm), 5 ♂♂ (7.7 to 11 mm), Greater Tunb Island, sandy beach, 03 Jan. 2021, 26°16'20.48"N, 55°17'28.00"E, coll. V. Khalaji and M. Majidi (ZSMU 1206).
(from the Persian Gulf). Color yellowish, or light brown dorsally with small, dark, pigmented dots of various densities (Fig.
Antennula
(Fig.
Antenna
(Fig.
Left mandible
(Fig.
Right mandible
(Fig.
Maxillule
(Fig.
Maxilla
(Fig.
Maxilliped
(Fig.
Pereopod 1–7
(Figs
Pleopod 2
(Fig.
Pleopod 3
(Fig.
Uropod
(Fig.
Female (Fig.
Oman, the Persian Gulf (Kuwait; Bandar-e-Charak, Bandar-e-Bostaneh, Bandar-e-Lengeh, Kish, Greater Tunb, and Faroor Koochak Islands, Iran)
We obtained seven 534-bp long COI sequences from T. maindroni individuals from four locations across the Persian Gulf coastline of Iran. These sequences were combined with a previously published COI sequence of T. maindroni from Kuwait (GenBank Acc. KJ468116; BIN: BOLD: ACQ3230). We identified four highly similar COI haplotypes as indicated by K2P divergences (0.0–0.4%, 1–3 nucleotide differences, Fig.
Average COI K2P divergences amongst Tylos species included in this study.
T. maindroni | T. punctatus | Tylos sp. BOLD:ACM2291 | Tylos sp. clade F* | Tylos sp. clade D* | Tylos sp. clade G* | Tylos sp. clade H* | Tylos sp. clade I* | Tylos sp. clade C* | Tylos sp. clade B* | Tylos sp. outgroup* | T. niveus | T. granulatus | T. capensis | T. marcuzzii | T. exiguus | T. opercularis | T. chilensis | T. spinulosus | Tylos sp. hachijoMN12 | T. granuliferus | T. wegeneri | |
---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|---|
T. maindroni | < 0.5 | |||||||||||||||||||||
T. punctatus | 20.5 | < 5.8 | ||||||||||||||||||||
Tylos sp. BOLD: ACM2291 | 20.5 | 0.1 | N/A | |||||||||||||||||||
Tylos sp. clade F* | 19.2 | 14.9 | 14.8 | 0.0 | ||||||||||||||||||
Tylos sp. clade D* | 17.7 | 12.5 | 12.4 | 11.0 | N/A | |||||||||||||||||
Tylos sp. clade G* | 18.2 | 13.6 | 13.6 | 13.2 | 12.5 | < 6.2 | ||||||||||||||||
Tylos sp. clade H* | 18.6 | 14.0 | 14.1 | 12.6 | 12.2 | 4.6 | 0.0 | |||||||||||||||
Tylos sp. clade I* | 18.9 | 14.2 | 14.3 | 13.6 | 13.0 | 5.6 | 4.5 | 0.0 | ||||||||||||||
Tylos sp. clade C* | 21.2 | 15.6 | 15.5 | 12.1 | 14.2 | 13.5 | 14.0 | 13.9 | N/A | |||||||||||||
Tylos sp. clade B* | 20.4 | 12.9 | 12.8 | 14.5 | 13.1 | 13.4 | 13.1 | 13.4 | 13.1 | N/A | ||||||||||||
Tylos sp. outgroup* | 19.9 | 16.0 | 15.9 | 16.7 | 18.2 | 15.8 | 15.9 | 16.3 | 18.4 | 14.7 | N/A | |||||||||||
T. niveus | 17.7 | 16.0 | 15.8 | 15.1 | 16.8 | 15.5 | 17.1 | 16.8 | 16.6 | 15.9 | 15.6 | N/A | ||||||||||
T. granulatus | 17.4 | 15.9 | 15.9 | 15.7 | 15.0 | 15.2 | 15.9 | 15.8 | 18.5 | 16.7 | 19.1 | 15.8 | < 13.2 | |||||||||
T. capensis | 19.5 | 16.3 | 16.3 | 17.9 | 17.7 | 17.5 | 18.4 | 17.7 | 20.8 | 17.8 | 19.0 | 19.8 | 12.2 | < 2.8 | ||||||||
T. marcuzzii | 21.5 | 17.8 | 17.8 | 21.6 | 22.7 | 20.2 | 20.9 | 21.3 | 19.9 | 17.3 | 18.0 | 20.6 | 19.4 | 19.6 | N/A | |||||||
T. exiguus | 20.7 | 21.3 | 21.2 | 20.5 | 19.2 | 18.7 | 18.8 | 19.2 | 21.9 | 20.5 | 21.0 | 22.9 | 18.9 | 19.1 | 23.1 | N/A | ||||||
T. opercularis | 25.4 | 25.5 | 25.5 | 21.9 | 25.2 | 25.1 | 25.2 | 25.2 | 27.3 | 25.8 | 23.6 | 19.2 | 23.3 | 23.3 | 26.0 | 23.0 | N/A | |||||
T. chilensis | 25.6 | 23.7 | 23.6 | 26.2 | 25.4 | 24.9 | 25.0 | 24.6 | 24.8 | 25.1 | 24.3 | 25.5 | 22.7 | 24.8 | 24.4 | 24.1 | 29.1 | N/A | ||||
T. spinulosus | 23.2 | 21.1 | 21.0 | 22.6 | 22.2 | 21.0 | 21.4 | 22.7 | 22.8 | 21.3 | 23.5 | 23.0 | 20.9 | 22.1 | 27.0 | 22.0 | 31.3 | 13.7 | N/A | |||
Tylos sp. hachijoMN12 | 23.9 | 27.3 | 27.3 | 25.1 | 26.6 | 25.2 | 27.4 | 27.0 | 25.7 | 24.9 | 26.2 | 21.2 | 25.2 | 25.8 | 29.7 | 28.2 | 24.1 | 33.5 | 28.3 | N/A | ||
T. granuliferus | 28.9 | 30.5 | 30.4 | 27.1 | 25.6 | 27.6 | 27.2 | 27.2 | 28.0 | 27.5 | 31.5 | 27.3 | 25.6 | 26.2 | 32.0 | 26.3 | 24.4 | 32.2 | 30.3 | 24.4 | < 25.2 | |
T. wegeneri | 26.2 | 28.7 | 28.7 | 27.3 | 27.2 | 26.3 | 25.4 | 27.7 | 26.2 | 26.0 | 27.7 | 27.4 | 25.9 | 26.0 | 25.7 | 27.1 | 26.4 | 27.8 | 27.3 | 26.5 | 29.2 | N/A |
Haplotype networks for the COI mitochondrial gene fragment of Tylos from the Persian Gulf. Colors correspond to locations as indicated in figure. Dashes along branches represent the number of nucleotide differences between haplotypes. Frequency of haplotype recovery represented through relative sizes of circles.
Combining the T. maindroni sequences with other previously published sequences of the genus Tylos resulted in a 517-bp long alignment containing 410 sequences. ASAP analyses of this dataset identified two partitioning schemes with nearly similar numbers of hypothetical species groups (23 and 24), threshold distances (0.068107 and 0.051440), and low ASAP scores (7 and 8). This last measure reflects both the p-value and the relative barcode gap width rank for a given partitioning scheme, with lower values reflecting stronger support for a given partitioning scheme. All COI haplotypes from T. maindroni individuals were placed in a single cluster that included no sequences from other Tylos species, regardless of the partitioning scheme.
Tylos maindroni was first described by Giordani Soika in 1954; however, the original description was brief and did not include a discussion or illustration of characters used in the taxonomy of this genus. A later work by
Our Persian Gulf specimens correspond morphologically quite well to the brief description and illustrations of T. maindroni from Oman by
Molecular data are in concordance with the above findings. All Tylos specimens that were morphologically identified as T. maindroni have highly similar COI haplotypes differing by a maximum of three positions (K2P distances amongst haplotypes < 0.5%). Furthermore, sequences recovered from T. maindroni individuals were highly divergent from all other COI sequences recovered from other Tylos species including T. exiguus (16.2–33.9% COI K2P). Not surprisingly, all T. maindroni haplotypes were assigned to a single species cluster in species delimitation analyses carried out in ASAP, regardless of the partitioning scheme.
The low level of diversification herein reported between individuals of T. maindroni collected at Persian Gulf locations stands in contrast with those reported for other coastal oniscid taxa (
The low levels of genetic divergence within T. maindroni in the Persian Gulf is likely a reflection of the young age of this marine waterbody. Although there is disagreement on the extent of the Persian Gulf coastline during the Holocene and Late Pleistocene (
We would like to thank Dr. Saskia Brix (Senckenberg Research Institute, Germany), Dr. Michael Raupach (Bavarian State Collection of Zoology, Munich, Germany) and one anonymous reviewer for their constructive suggestions and helpful comments. This study is a part of the Biodiversity and Conservation project under a joint program by the Ministry of Science, Research and Technology of Iran and the Leibniz Association, Germany. Financial support for this study was provided by the Iran University of Science and Technology, International Affairs, Shahrekord University, Iran and the Leibniz Association, Germany.