The genus Morus comprises many species (Suttie 2012). The species, Morus alba is one of the most popular Morus worldwide. This plant is widely cultivated as an ornamental plant in the parks in Iran. Meloidogyne species were recovered from root samples collected from stunted and cholorotic trees growing in “Azalia” park in Teheran.

During March 2015, root samples were collected from white mulberry in a park in Tehran (N: 35°37′51″; E: 51°28′25″). Roots were washed, cut in pieces and mature female specimens removed using a scalpel under a Nikon CH-2 stereo microscope. These specimens were fixed with a hot 4 % formaldehyde solution and transferred to anhydrous glycerine using the method of De Grisse (1969). Characteristics of the perineal patterns as described by Hirschmann (1986) were used in identifying the Meloidogyne species (Fig. 1).

Fig. 1
figure 1

Meloidogyne hispanica Hirschmann 1986. a-c: Entire female. d: Anterior end of a typical female (arrow indicates lip region). e: Cuticle of a typical female. f: Vulva of a typical female. g, h: Perineal patterns of two typical females

Full size image

The molecular characterisation was based on the methodology used by Rashidifard et al. (2015). The original partial 28S (D2-D3 expansion) sequence of Meloidogyne hispanica is deposited in GenBank under accession number KT359553. Sequencing and Bayesian inference (BI) analysis of the ribosomal DNA region D2-D3 of 28S (Fig. 2) has been obtained for this species.

Fig. 2
figure 2

The Bayesian tree inferred from known and newly sequenced Meloidogyne spp. from Iran based on the 28S rDNA region

Full size image

The sequence lengths flanked by the forward primer D2A (5″-ACAAGTACCGTGAGGGAAAGTTG–3″) and the reverse primer D3B (5″-TCGGAAGGAACCAGCTACTA–3″) (according to Subbotin et al. 2006) of the 28S region of M. hispanica isolate are 694 base pairs long. The Blast test revealed that this population is seven base pairs different from the closest populations from Spain, Brazil and Portugal (EU443606, EU443607, EU443608; 98 % identity respectively). Compared with the populations from Greece (KF501128) and Spain (GQ375158), differences of eight and seven (99 % identity) base pairs, respectively, were evident.

Our phylogenetic analysis using 28S rDNA, placed the Iranian M. hispanica population in a clade together with other M. hispanica populations (Fig. 2). Molecularly characterised species of M. hispanica thus form a monophyletic group. The phylogeny of Meloidogyne spp. has been studied (De Ley et al. 2002; Tigano et al. 2005) using 18S rDNA and 18S rDNA and mtDNA, respectively. However, M. hispanica was not included in these studies. Meloidogyne hispanica and M. ethiopica are place together in a group, however, they differ in terms of the stylet of the female (small vs large), phasmids (not prominent vs prominent) and lateral line (conspicuous vs not conspicuous). Other genes e.g. mtDNA may separate these two species clearly. Two permanent microscope slides, containing the perennial patterns and females of M. hispanica, respectively, were deposited in the National Collection of Nematodes (NCN) at the Nematology Unit, Biosystematics Division, Agricultural Research Council (ARC) - Plant Protection Research Institute (PPRI) (Pretoria, South Africa) with slide numbers 50192 and 50193. According to the literature, this is the first record of M. hispanica in Iran. Morus alba seems to be affected by M. hispanica and management practices need to be put in place to control the nematode and to prevent its dispersal to other parks in Tehran.