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Eryx Miliaris (PALLAS, 1773), from Sistan, East Iran 133-144 Sheikh Etal Oraie Eryx Miliaris Iran:HERPETOZOA.Qxd 12.02.2019 15:08 Seite 1 ZOBODAT - www.zobodat.at Zoologisch-Botanische Datenbank/Zoological-Botanical Database Digitale Literatur/Digital Literature Zeitschrift/Journal: Herpetozoa Jahr/Year: 2019 Band/Volume: 31_3_4 Autor(en)/Author(s): Sheikh Younes, Oraie Hamzeh, Rastegar-Pouyani Eskandar Artikel/Article: Morphometric versus genomic evidence. The systematic status of Eryx miliaris (PALLAS, 1773), from sistan, east Iran 133-144 Sheikh_etal_Oraie_Eryx_miliaris_Iran:HERPETOZOA.qxd 12.02.2019 15:08 Seite 1 HerPeTozoa 31 (3/4): 133 - 144 133 Wien, 28. februar 2019 Morphometric versus genomic evidence. The systematic status of Eryx miliaris (PaLLas , 1773 ), from sistan, east Iran (squamata: serpentes: Boidae) der taxonomische status von Eryx miliaris (PaLLas , 1773 ) von sistan, ostiran Morphometrische und genetische Hinweise im Vergleich. (squamata: serpentes: Boidae) younes sHeIkH & H aMzeH oraIe & e skandar rasTegar -P ouyanI kurzfassung exemplare von Eryx miliaris (PaLLas , 1773 ) aus sistan, ost-Iran, wurden morphologisch und genetisch untersucht. dabei zeigte sich eine hohe Variabilität in anordnung und zahl bei kopfschilden, während die dns- sequenzen des mitochondrialen Cytochrom b wenig unterschiedlich waren. der Vergleich der sequenzen mit denen von nordost-Iranischen E. tataricus (L ICHTensTeIn , 1823) aus genBank spricht für die artliche Identität von E. tataricus (L ICHTensTeIn , 1823) mit E. miliaris und in der folge die Bewertung von E. tataricus als subjektives Juniorsynonym von E. miliaris . die autoren sprechen sich dagegen aus, morphometrische Merkmale als alleini - ge Informationsquelle für systematische zuordnungen zu verwenden. aBsTraCT specimens of Eryx miliaris (PaLLas , 1773 ) from sistan, east Iran, were investigated morphologically and genetically. The results demonstrated a high variation in topology and number of head scales, while mtdna Cytochrome b sequences showed little differences. Comparison with sequences from ne-Iranian E. tataricus (L ICHTensTeIn , 1823) taken from genBank provides support for the identity at species level of E. tataricus (L ICHTensTeIn , 1823) with E. miliaris . as a consequence, E. tataricus is identified a subjective junior synonym of E. miliaris . Moreover, the authors strongly suggest not to use morphometric characters as the only source of infor - mation for systematic decisions. key Words reptilia: squamata: serpentes: Boidae; Eryx miliaris , Eryx tataricus , systematics, taxonomy, cytochrome b, molecular genetics, morphology, pholidosis, intraspecific plasticity, sistan, Iran InTroduCTIon In parallel with the scientific advances western China in the east ( Lanza & n IsTrI based on comparative genomics, morpho - 2005) . according to the literature, the genus logical data are still being widely used to Eryx is represented by six species in Iran: E. determine the systematic position of many jaculus (LInnaeus , 1758), E. miliaris (PaL- taxa. Conflicts between molecular and mor - Las , 1773), E. johnii (russeLL , 1801), E. ta - phological studies can arise from morpho - taricus (LICHTensTeIn , 1823), E. elegans logical characters which show homoplastic (gray , 1849) and E. jayakari BouLenger , evolution. In such cases, morphological data 1888 (e.g., LaTIfI 2000; r asTegar -P ouyanI may mislead phylogenetic and systematic et al . 2008; safaeI -M aHroo et al . 2015). interpretations ( Hedges & M axson 1996; However, there is disagreement on the WIens et al. 2003). correct identification of some specimens or The main distribution of the genus systematic status of some nominal taxa of Eryx daudIn , 1803, includes a wide range the genus. Morphological and molecular from southeastern europe, north and east studies of Eryx specimens in ne Iran inde - africa, over southwestern asia as far as pendently, but in confirmation of each other Sheikh_etal_Oraie_Eryx_miliaris_Iran:HERPETOZOA.qxd 12.02.2019 15:08 Seite 2 134 y . s HeIkH & H. o raIe & e. r asTegar -P ouyanI fig. 1: sampling areas of sand boas morphologically classified as Eryx miliaris (PaLLas , 1773) and E. tataricus (LICHTensTeIn , 1823) included in this study. 1 ‒ fariman, sarakhs and Torbate-Jam (razavi khorasan Province), 2 ‒ sabzevar, neyshabour, Parvand and Joghtay (razavi khorasan Province), 3 ‒ Birjand and shaskooh (south khorasan Province), 4 ‒ Various locations in sistan (sistan & Blochestan Province). details are presented in Tables 1 and 2. abb. 1: fundgebiete von sandboas, die morphologisch Eryx miliaris (PaLLas , 1773) und E. tataricus (LICHTensTeIn , 1823) zugeordnet waren und in der vorliegenden arbeit untersucht wurden. 1 ‒ fariman, sarakhs und Torbate-Jam (Provinz razavi khorasan), 2 ‒ sabzevar, neyshabour, Parvand und Joghtay (Provinz razavi khorasan), 3 ‒ Birjand und shaskooh (Provinz süd-khorasan), 4 ‒ verschiedene orte in sistan (Provinz sistan & Belutschistan). zu einzelheiten siehe Tabellen 1 und 2. (eskandarzadeH et al . 2013; rasTegar - tionable. The systematic status of E. tatari - PouyanI et al. 2014), suggested that snakes cus and E. miliaris was already debated in previously assigned to E. elegans are con - preceding molecular studies, which consid - specific with E. jaculus , and populations ered the relationship between E. tataricus earlier identified as E. tataricus cannot be and E. miliaris to be on a subspecific level differentiated from E. miliaris . These find - (Pyron et al. 2013), while the nominal sub - ings are in disagreement with several stud - species E. tataricus vittatus (ČernoV , 1959), ies (e.g., TerenT ’eV & C HernoV 1965; was found to be more distant from these sørensen 1988; L aTIfI 2000; r eynoLds et (reynoLds et al. 2014). al. 2014; zarrInTaB et al. 2017). a recent since E. miliaris was reported from review of the genus Eryx in Iran exclusive - sistan (LaTIfI 2000), the purpose of the ly based on external morphological traits present study was to (i) add information on (zarrInTaB et al . 2017 ) was controversial, the systematic status and phylogenetic at least as to the E. tataricus /miliaris com - affinities of E. miliaris from sistan using plex ( sens. Tokar 1990). according to this mtdna Cytochrome b sequences and (ii) study, several Iranian specimens were describe the degree of intraspecific variation assigned to E. tataricus , whereas the occur - at the external morphology level in these rence of E. miliaris was classified as ques - specimens. Sheikh_etal_Oraie_Eryx_miliaris_Iran:HERPETOZOA.qxd 12.02.2019 15:08 Seite 3 The systematic status of Eryx miliaris (PaLLas , 1773 ) from sistan, east Iran 135 MaTerIaLs and MeTHods Morphological analyses .- Ma - out in a total volume of 20 µl, containing 10 terial for the morphological examinations µl ready to use Prime Taq premix (am - included 22 alcohol-preserved specimens of pliqon, Cat number: a180301), 1 µl of E. miliaris from sistan, eastern Iran (fig. genomic template dna (approximately 100 1). This sample belongs to the herpetologi - ng), 1 µl of each primer (10 pm/ml) and 7 µl cal collection of shahrekord university, distilled deionized water with the following shahrekord, Iran. details of the localities, conditions: an initial cycle at 94 °C for 4 museum and genBank accession numbers minutes, followed by 36 cycles at 94 °C for of sequences are given in Tables 1 and 2. 30 seconds, 58 °C for 40 seconds and 72 °C specimens were identified morphologically for 90 seconds, and a final cycle at 72 °C for based on TzareWsky (1916), T erenT ’eV & 10 minutes. sequences were derived from CHernoV (1965) and Tokar (1991). The these products in an aBI 3700 automated following morphological characters were sequencer at Macrogen, Inc. (seoul, korea). measured using Vernier calipers with an aligning and editing the chromato - accuracy to the nearest 0.01 mm: snout-vent graphs was done manually using Bioedit v. length (sVL) from tip of snout to vent; tail 7.0.5 ( HaLL 1999). To avoid amplification length (TL) from vent to tip of tail; interor - of pseudogenes, sequences were translated bital width (IoW); head width (HW) at the into amino-acids and checked for stop- widest point; and distance between posteri - codons. The cyt- b fragments were aligned or edge of eye and corner of mouth (deM). using MaffT v. 7 ( kaToH & s TandLey In addition to the metric variables, the fol - 2013) applying default parameters (auto lowing meristic variables (pholidosis strategy, gap opening penalty: 1.53, offset counts) were determined using a stereomi - value: 0.0). uncorrected p-distances with croscope: numbers of loreal scales right and complete deletion of positions with ambigu - left (Ls, r/L), supralabial scales right and ities and alignment gaps were calculated left (sL, r/L), infralabial scales right and using Mega7 v. 7.0 ( kuMar et al. 2016). left (IL, r/L), longitudinal dorsal scale rows additional sequences representing individu - around the middle of the body (sM), ventral als of E. miliaris and E. tataricus were down - scales (Ven), subcaudal scales (sCd), pre - loaded from genBank and added to the ventral scales (PVen), scales between eye dataset (Table 2). Eryx elegans (kJ841063 and nasal (Ben), scales posterior to inter - and kJ841061) were included in the analy - nasal as far as to the eye (PIn), scales sis as an outgroup taxon, since it was between eyes counted along a straight line revealed as sister taxon of the E. miliaris/E. (Bes), scales around eye right and left tataricus complex ( rasTegar -P ouyanI et (sae, r/L), longitudinal dorsal scales rows, al. 2014). counted right behind the vent (dsV). The authors used maximum likelihood Molecular
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