Garra Roseae, a New Species from the Makran Region in Southern Iran (Teleostei: Cyprinidae)
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Zootaxa 4671 (2): 223–239 ISSN 1175-5326 (print edition) https://www.mapress.com/j/zt/ Article ZOOTAXA Copyright © 2019 Magnolia Press ISSN 1175-5334 (online edition) https://doi.org/10.11646/zootaxa.4671.2.3 http://zoobank.org/urn:lsid:zoobank.org:pub:B98ADFCA-4211-496A-B047-4F72CC4CAB0D Garra roseae, a new species from the Makran region in southern Iran (Teleostei: Cyprinidae) HAMED MOUSAVI-SABET1,2, MARYAM SAEMI-KOMSARI1, IGNACIO DOADRIO3 & JÖRG FREYHOF4 1Department of Fisheries Sciences, Faculty of Natural Resources, University of Guilan, Sowmeh Sara, Guilan, Iran. E-mails: [email protected], [email protected] 2The Caspian Sea Basin Research Center, University of Guilan, Rasht, Guilan, Iran. 3Biodiversity and Evolutionary Group, Museo Nacional de Ciencias Naturales, CSIC, Madrid, Spain. E-mail: [email protected] 4Museum für Naturkunde, Leibniz Institute for Evolution and Biodiversity Science, 10115 Berlin, Germany. E-mail: [email protected] Abstract Garra roseae, new species, is described from the stream Tang-e-Sarhe in the Iranian Makran region. It is distinguished from its congeners in the Middle East by lacking barbels, having a small mental disc, 42–58 total scales along the lateral line, 24–30 scales along the predorsal midline, and 20–24 circumpeduncular scales. It is further characterised by having five diagnostic nucleotide substitutions and a minimum K2P distance of 5.39% to G. rossica and 5.49% to G. nudiventris in the mtDNA COI barcode region. Garra phryne from eastern Iran is considered to be a synonym of G. nudiventris. Key words: Freshwater fish, Taxonomy, Cytochrome oxidase I, Middle East Introduction Labeonine cyprinids of the genus Garra are widespread in the Middle East, South- and East Asia and tropical Africa (Menon 1964). In the Middle East, the genus Garra is well separated in two species groups (Hamidan et al. 2014): the Garra rufa group and the G. variabilis group. In the G. variabilis group, Hamidan et al. (2014) recognised four species, G. variabilis, G. rossica, G. kemali and G. klatti. Esmaeili et al. (2016) also recognised G. nudiventris as a valid species in this group. Interestingly, the G. variabilis species group has been only known from the Middle East and Central Asia until now and further research is needed to reveal its relationships with the Oriental Garra species. The Makran region is geographically at the border between the Palearctic and Oriental biogeographic realms. The Oriental starts here and reaches then through Pakistan and India to the east. The Makran region, which is shared by Iran and Pakistan, is located along the coastal region in the north of the Oman Gulf between the Cape Dschask in the Iranian Hormozgan province and the Sonmiani bay about 25 km north of Karachi. The Minab River in the west and the Sarbaz River in the east are the two most important rivers in the Iranian Makran. There are also some small streams between the two river systems, which often do not have perennial or continuous flow. The Makran has bio- geographical affinity to the South Asian fauna by containing oriental species, which are not much further distributed in the Middle East such as Cabdio morar, Tariqilabeo diplochilus, Bangana dero, and Glossogobius giuris. The Garra populations of the Iranian Makran region was identified as Garra persica and G. rossica by Sayyadzadeh et al. (2015), Esmaeili et al. (2016) and those from the Pakistani Makran was identified as G. rossica and G. gotyla by Mirza (1972). Nebeshwar & Vishwanath (2013) re-described G. gotyla, which is a species with four barbels and an elevated proboscis and restrict its distribution to Sikkim in north-eastern India. These authors also pointed out that many Garra species had been misidentified as G. gotyla and this was for sure also the case by Mirza (1972). We were able to examine Garra populations from the Makran region including one from the stream Tang-e- Sarhe, in southern Iran (Fig. 1). A detailed morphological study in combination with DNA barcoding routines sup- port the hypothesis that the population represents an undescribed species, which is diagnosed below. Furthermore, Accepted by R. Pethiyagoda: 19 Aug. 2019; published: 17 Sept. 2019 223 we test, (a) if G. rossica and G. nudiventris represent two species as already suggested by Esmaeili et al. (2016), despite their low genetic distance and (b) if G. phryne is a synonym of G. rossica as considered by Coad (1981) FIGURE 1. Records of the Garra variabilis species group in Iran. See below for the sources of records the map is based on. Material and methods After anaesthesia, fishes were fixed in 5% formaldehyde and stored in 70% ethanol or directly fixed in 99% ethanol. Measurements were made with a dial calliper and recorded to 0.1 mm. All measurements were made point to point, never by projections. Methods for counts and measurements follow Kottelat & Freyhof (2007). The terminology of the snout morphology and the oromandibular structures follow Stiassny & Getahun (2007) and Nebeshwar & Vish- wanath (2013). Standard length (SL) was measured from the tip of the snout to the end of the hypural complex. The length of the caudal peduncle is measured from behind the base of the last anal-fin ray to the end of the hypural com- plex, at mid-height of the caudal-fin base. The last two branched rays articulating on a single pterygiophore in the dorsal and anal fins are counted as “1½”. The holotype is included in the mean and SD calculation of morphometric characters. Scales along the lateral line are counted from the first one (the first one to touch the shoulder girdle) to the last one on the caudal-fin base (total lateral line scales). Gill rakers are counted on the outer margin of the first gill arch. Garra kemali and G. klatti are redescribed by Küçük et al. (2015) and Yoğurtçuoğlu et. al. (2018) and we base our comparison on these re-descriptions and own materials examined. Except those published by Annandale (1919), Esmaeili et al. (2016), and our own data points, all records of Garra species shown in Figure 1 were identi- fied as G. rossica in the publications or databases these were taken from. As we were not able to identify these from the materials, we re-identify them based on their geographic range derived from properly identified materials. Abbreviations used. SD, standard deviation, SL, standard length; K2P, Kimura 2-parameter. Collection codes: GUIC, Guilan University Ichthyological Collection, Sowmeh-Sara; VMFC, Vatandoust & Mousavi-Sabet Fish Col- lection, Tehran; FSJF, Fischsammlung J. Freyhof, Berlin. DNA extraction and PCR: Total genomic DNA was extracted from fin tissue of the specimens, using the 224 · Zootaxa 4671 (2) © 2019 Magnolia Press SABET ET AL. commercial kit Biosprint15 for tissue and blood (Qiagen). The COI gene was amplified using primers FishF1-(5’- TCAACCAACCACAAAGACATTGGCAC-3’) and FishR1-(5’-TAGACTTCTGGGTGGCCAA AGAATCA-3’) (Ward et al. 2005). PCR mixtures were prepared in 25 μ with a final concentration of 0.5 mM each primer, 0.2 mM dNTP, 1.5 mM MgCl2, and 1-unit Taq DNA polymerase (Invitrogen). Amplification cycles followed as denaturation for 2 min at 95°C; 30 cycles at 94°C for 30 seconds, 54°C for 30 seconds, 72°C for 1 min and a final extension for 10 min at 72°C. Afterward, PCR products has been checked on 1% agarose gels, then purified by ExoSAPITTM (USB, Cleveland, USA) then sent to MACROGEN Inc (Madrid, Spain; https://dna.macrogen.com) for sequencing. FIGURE 2. Phylogenetic tree of mitochondrial COI barcode region rendered by Bayesian inference (BI). Numbers on branches indicate posterior probability values for BI and Maximum Likelihood (ML) bootstrap. Model of rate heterogeneity: Gamma with 4 categories Gamma shape alpha: 0.1905. Black solid bars right to the specimen labels indicate species delimitation results from mPTP followed by the results of the PTP approach as gray solid bars. The tree is drawn to scale with branch lengths depict- ing number of substitutions per site. GARRA ROSEAE Zootaxa 4671 (2) © 2019 Magnolia Press · 225 TABLE 1. List of COI-sequences downloaded from NCBI Genbank with information on drainage and country of ori- gin. Species Drainage Country Genbank Reference Garra amirhosseini Tigris Iran KX570873 Esmaeili et al. 2016 Garra amirhosseini Tigris Iran KX570874 Esmaeili et al. 2016 Garra barreimiae Shawkah UAE KM214783 Hamidan et al. 2014 Garra barreimiae Wurayah UAE KM214767 Hamidan et al. 2014 Garra barreimiae Shawkah UAE KM214779 Hamidan et al. 2014 Garra elegans Tigris Iraq KM214702 Hamidan et al. 2014 Garra elegans Tigris Iraq KM214721 Behrens-Chapuis et al. 2015 Garra ghorensis Ghor Jordan KJ553520 Geiger et al. 2014 Garra ghorensis Wadi Alhass Jordan KM214788 Hamidan et al. 2014 Garra gymnothorax Tigris Iran KM214803 Behrens-Chapuis et al. 2015 Garra gymnothorax Tigris Iran KM214735 Behrens-Chapuis et al. 2015 Garra gymnothorax Tigris Iran KM214719 Hamidan et al. 2014 Garra jordanica Mujib Jordan KM214756 Hamidan et al. 2014 Garra jordanica Wadi Jallain Syria KM214788 Hamidan et al. 2014 Garra kemali Tuz Turkey KJ553337 Geiger et al. 2014 Garra klatti Isikli Turkey KJ553545 Geiger et al. 2014 Garra klatti Isikli Turkey KJ553625 Geiger et al. 2014 Garra klatti Isikli Turkey KJ553344 Geiger et al. 2014 Garra klatti Köpru Turkey KJ553379 Geiger et al. 2014 Garra longipinnis Jabal Al Akhdar Oman KM214752 Hamidan et al. 2014 Garra longipinnis Jabal Al Akhdar Oman KM214756 Hamidan et al. 2014 Garra lorestanensis Tigris Iran MG852049 Hashemzadeh Segherloo et al. 2018 Garra lorestanensis Tigris Iran MG852033 Hashemzadeh Segherloo et al. 2018 Garra lorestanensis Tigris Iran MG852038 Hashemzadeh Segherloo et al. 2018 Garra mondica Mond Iran MG852031 Hashemzadeh Segherloo et al. 2018 Garra mondica Mond Iran MG852030 Hashemzadeh Segherloo et al.