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Nematology 21 (2019) 333-335 brill.

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The first molecular phylogenetic study of Xiphinema robbinsi


Pedram, Niknam & Decraemer, 2008 (Nematoda: Longidoridae)
based on specimens from the type locality
Farahnaz JAHANSHAHI AFSHAR ∗

Six species of the dagger nematode genus Xiphinema dition, the vulva was located more anteriorly (44-46 vs
Cobb, 1913 have been described from Iran, viz., X. rob- 48-56% in the type) (Guesmi-Mzoughi et al., 2017). Dur-
binsi Pedram, Niknam & Decraemer, 2008, X. iran- ing the present study, a live population of the species
icum Pedram, Niknam, Robbins, Ye & Karegar, 2009, X. was kindly provided from the type locality by Dr M. Pe-
mazandaranense Pedram, Pourjam, Robbins, Ye, Atighi dram, one of the original proposers of the species. The
& Decraemer, 2012, X. granatum Pedram, Pourjam, morphological and morphometric characters of a recov-
Palomares-Rius, Ghaemi, Cantalapiedra-Navarrete & ered female (together with a juvenile specimen, mounted
Castillo, 2012, X. zagrosense Ghaemi, Pourjam, Pedram, on permanent slides using standard methods (De Grisse,
Robbins, Ye & Decraemer, 2012, and X. castilloi Roshan- 1969)) were in accordance with the type population de-
Bakhsh, Pourjam, Pedram, Robbins & Decraemer, 2014, scribed by Pedram et al. (2008) (major morphometrics
all of which, except for X. robbinsi, were established as follows: L = 3.5 mm, a = 73.8, b = 7.7, c = 93,
based upon both morphological and molecular phylo-
c = 1.3, V = 48, lip region diam. = 13 μm, lip re-
genetic criteria. Xiphinema robbinsi is an amphimictic
gion height = 7 μm, odontostyle = 128 μm, odon-
species belonging to morphospecies group 6, charac-
tophore = 74 μm, neck (anterior end to base of pharyn-
terised by equally developed genital branches, absence of
geal bulb) = 457 μm, vulval body diam. = 48 μm, anal
Z- or pseudo-Z-organ but presence of spines (Loof & Luc,
body diam. = 30 μm, and tail = 38 μm; Fig. 1). Two
1990), and mainly characterised by 3.0-3.6 mm long fe-
males having an anteriorly broad rounded lip region with additional live individuals were used for DNA extraction.
a depression at the junction with the body, odontostyle Recovery of the live material enabled molecular phylo-
107.5-127.0 μm long, tail mainly with a wide, slightly genetic analyses based upon two genomic markers: 28S
bulging terminus with a blind canal, four juvenile devel- rDNA D2-D3 and internal transcribed spacer 1 (ITS1) for
opmental stages, and males common with 52.5-56.2 μm the first time. DNA extraction from live individuals, the
long spicules (Pedram et al., 2008). The species is cur- polymerase chain reaction (PCR) and molecular phyloge-
rently known only from the type locality and recently netic analyses were performed according to Gharibzadeh
from Tunisia, although the Tunisian female specimens et al. (2018). A total number of 82 species/isolates of
were longer (3.7-4.5 mm), the single male specimen pos- Xiphinema non-americanum species group (including two
sessing shorter spicules 49 μm long, differences that were dorylaim outgroup taxa) were selected for 28S phylogeny,
attributed to geographical intraspecific variability. In ad- and a total number of 37 species/isolates of Xiphinema

Iranian Research Institute of Plant Protection, Agricultural Research, Education and Extension Organization (AREEO),
Tehran, Iran

E-mail: Afshar_fj@yahoo.com
Received: 19 August 2018; revised: 1 October 2018
Accepted for publication: 1 October 2018; available online: 30 October 2018
Keywords: 28S, dagger nematode, Dorylaimida, Iran, ITS1, morphometrics, taxonomy, Tunisia.

© Koninklijke Brill NV, Leiden, 2019 DOI 10.1163/15685411-00003219

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Short communication

Fig. 1. Xiphinema robbinsi Pedram, Niknam & Decraemer, 2008 from the type locality, Female. A: Typical morphology of lip region;
B: Tubular part of uterus with small crystalloids (arrowheads); C: Tail; D: Vagina and ovejector with sperm. (Scale bar = 10 μm.)

non-americanum species group (including three america-


num-group species as outgroup taxa) were selected for
ITS1 phylogeny. The inferred trees using the aforemen-
tioned large datasets are not shown herein and are avail-
able from the author on request. In the 28S tree (inferred
using the large dataset), the presently sequenced popula-
tion of X. robbinsi (MH744579) formed a clade with X.
granatum, and these two species fell into a clade including
X. vuittenezi Luc, Lima, Weischer & Flegg, 1964, X. her-
akliense Tzortzakakis, Archidona-Yuste, Cantalapiedra-
Navarrete, Nasiou, Palomares-Rius & Castillo, 2015, X.
zagrosense, and X. israeliae Luc, Brown & Cohn, 1982,
whereas the Tunisian population (KX062685) occupied a
distant placement from this clade; the relationships of the
other species are in agreement with Guesmi-Mzoughi et
al. (2017). In the ITS1 tree the placement of these two
populations (the presently sequenced population and the
Tunisian population) was also distant. In Figure 2, the
pruned 28S and ITS1 trees are represented. In the pruned
28S tree (Fig. 2A), in accordance with the original tree
inferred using the large dataset, the presently sequenced
Iranian population of X. robbinsi from the type local-
ity (MH744579) occupied a distant placement from the Fig. 2. Pruned Bayesian phylogenetic trees inferred using 28S
Tunisian population (KX062685), and in the pruned ITS1 rDNA D2-D3 (A) and ITS1 rDNA (B) of Xiphinema robbinsi
Pedram, Niknam & Decraemer, 2008 from the type locality
tree (Fig. 2B), in accordance with the original tree inferred
under the GTR + I + G model. The newly generated sequences
using the large dataset, it also occupied a distant place- are in bold font.
ment from the Tunisian population (KX062701). The two
28S D2-D3 sequences of the Iranian population from type matches and seven gaps). Remarkable differences were
locality and the Tunisian population had remarkable dif- also observed between the ITS1 sequences of both popu-
ferences in their overlapping region (more than 50 mis- lations (more than 50 mismatches and nine gaps). Tak-

334 Nematology
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Short communication

ing into account the morphological differences between Gharibzadeh, F., Pourjam, E. & Pedram, M. (2018). Description
the Tunisian population and the original description of X. of Longidorus azarbaijanensis n. sp. (Dorylaimida: Longi-
robbinsi discussed by Guesmi-Mzoughi et al. (2017), the doridae) from Iran. Journal of Nematology 50, 207-218. DOI:
exact identity of the former clearly needs to be rechecked. 10.21307/jofnem-2018-009
Guesmi-Mzoughi, I., Archidona-Yuste, A., Cantalapiedra-
Navarrete, C., Palomares-Rius, J.E., Regaieg, H., Horrigue-
Acknowledgement Raouani, N. & Castillo, P. (2017). Integrative identification
and molecular phylogeny of dagger and needle nematodes as-
I appreciate the kind help of Dr Pedram for providing sociated with cultivated olive in Tunisia. European Journal
the live material for this molecular study and for checking of Plant Pathology 147, 389-414. DOI: 10.1007/s10658-016-
the species identity. 1011-x
Loof, P.A.A. & Luc, M. (1990). A revised polytomous key
for the identification of species of the genus Xiphinema
References Cobb, 1913 (Nematoda: Longidoridae) with exclusion of the
Xiphinema americanum-group. Systematic Parasitology 16,
De Grisse, A.T. (1969). Redescription ou modifications de 35-66. DOI: 10.1007/BF00009600
quelques techniques utilisées dans l’étude des nématodes Pedram, M., Niknam, G. & Decraemer, W. (2008). Xiphinema
phytoparasitaires. Mededelingen Faculteit Landbouwweten- robbinsi sp. n. (Nematoda, Dorylaimida), an amphimictic
schappen Rijksuniversiteit Gent 34, 351-369. species from Iran. Russian Journal of Nematology 16, 49-57.

Vol. 21(3), 2019 335


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