Deladenus bonabensis n. sp. (Tylenchomorpha: Neotylenchidae) from East Azarbaijan province, northwestern Iran: A morphological and molecular phylogenetic study

A population of Deladenus, representing a new species, was recovered from Bonab’s Ghara-Gheshlagh lagoon. It is mainly characterized by having a long body (1051–1185 μm), long distance of anterior end to pharyngeal glands end (270–312 μm), six lines in lateral fields, and a short mucro-like differentiation on the tail tip. Furthermore, it has a small-sized stylet (8.5–11.0 μm) with three knobs, no postvulval uterine sac, and males with 24- to 28-μm-long tylenchoid spicules and penial tube. With six lateral lines, the new species is comparable with seven species of the genus: D. apopkaetus, D. brevis, D. cocophilus, D. durus, D. obtusicaudatus, D. processus, and D. ulani. It was furthermore compared with D. oryzae with an unknown number of lateral lines and D. aridus, D. obesus, and D. parvus having a different number of lateral lines but similar morphology. In the molecular phylogenic analyses using small and large subunit ribosomal DNA (SSU and LSU D2-D3 rDNA) sequences, the relationships of the new species with other species and genera were not resolved in SSU phylogeny. However, it formed a clade with Deladenus sp. and D. brevis in LSU phylogeny

Description of Ektaphelenchoides fuchsi n. sp. (Nematoda: Ektaphelenchinae) from western Iran

Esmaeili, Mehrab, Heydari, Ramin, Pourjam, Ebrahim, Atighi, Mohammad Reza (2014): Description of Ektaphelenchoides fuchsi n. sp. (Nematoda: Ektaphelenchinae) from western Iran. Zootaxa 3846 (3): 430-438, DOI: 10.11646/zootaxa.3846.3.

Ektaphelenchoides fuchsi Esmaeili, Heydari, Pourjam & Atighi, 2014, n. sp.

Ektaphelenchoides fuchsi n. sp. Figs 1 & 2 Measurements. See Table 1. Material examined. Holotype, female, from soil samples associated with the rhizosphere of Cucurbita maxima D. in Gilangharb region, Kermanshah province, Western Iran. Collected by Mehrab Esmaili in August 2013. It has been deposited in the Nematode Collection of the Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran (Slide no. Ektaphelenchoides fuschi n. sp., Holotype 1). Paratypes, one male and one female deposited at Nematode Collection of the Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran. Three female and two male paratypes (Slides AEF001, AEF002) deposited in nematode collection of the Department of Plant Protection, College of Agriculture and Natural Resources, University of Tehran, Karaj, Iran. Three female paratypes deposited at each of the following collections: CABI Europe– UK, Egham, Surrey, UK; USDA Nematode Collection, Beltsville, MD, USA and Department of Nematology, WANECO collection, Wageningen (http://www.waneco.eu/). 1 Length of conus as percentage of total stylet length. 2 Distance between anterior end of body and centre of median pharyngeal bulb as percentage of pharyngeal length. 3 Distance from pharyngo–intestinal junction to tip of dorsal gland. Description. Female. Body cylindrical, slightly ventrally curved when heat–killed. Cuticle mostly less than 1 Μm thick, finely annulated. Lateral fields with three incisures. Lip region slightly set–off, separated from body contour with a weak constriction, 3–4 µm high, 7–8 µm wide. Stylet 14–18 Μm long, with basal knobs, conus occupying ca 34–41 % of its total length. Procorpus cylindrical, ending in an elongate rectangular metacarpus with anterior granular part, occupying 35–40 % of its total length. Pharyngeal glands well-developed, overlapping intestine dorsally, 4–7 times as long as body diameter at level of median bulb. Excretory pore 8–22 Μm posterior to the base of metacorpus, hemizonid 13–20 Μm posterior to the excretory pore and 70–84 Μm from anterior end. Reproductive tract prodelphic, gonad outstretched, occupying 40–48 % of the body length, oocytes mostly in two rows, spermatheca irregular, filled with rounded sperm, vagina not sclerotised, straight, without flap, post-uterine sac short, less than half body diameter at vulva region. Intestine apparently ending in a blind sac at about one third to half distance from vulva to posterior end, no rectum or functional anus. Posterior end (tail) filiform. Male. Rare. Body slender, arcuate to J-shaped after heat relaxation, posterior end more ventrally curved. Anterior region similar to that of female. Testis single, expanded anteriorly, not reflexed. Spicules arcuate, separate, ca 3.2 –4.0 times longer than capitulum width, lamina/calomus smoothly and symmetrically curved, rostrum sharply pointed, condylus well-developed and more developed than rostrum. Cucullus and bursa absent. Five papillae present with their arrangement as follows: a single mid-ventral pre-cloacal papilla (P 1) about 7 µm anterior to the cloacal opening, one pair of sub-ventral papillae (P 2) at level of or just posterior to the cloacal opening, and one sub-ventral pair of post-cloacal papillae (P 3) at 16–17 µm posterior to the cloacal opening. Tail conoid with sharply pointed filiform terminus. FIGURE 1. Ektaphelenchoides fuchsi n. sp. A: Female entire body; B: Male entire body; C: Female anterior region; D: Female posterior body; E: Female anterior end; F: Position of excretory pore and Hemizonid. G: Male posterior body showing caudal papillae; H: Female stylet; I: Spicule in detail; J: Cross section of female. FIGURE 2. Ektaphelenchoides fuchsi n. sp. A, B: Female anterior body; C: Female stylet; D: Cross section of female showing lateral field; E, F: Female genital tract; G: Blind end of the intestine; H–J: Male posterior body showing spicules and papillae arrangement. FIGURE 3. Bayesian 50 % majority rule consensus tree inferred from 51 sequences of D 2 /D 3 expansion segments of 28 S rRNA gene under the GTR + I + G model. Posterior probabilities and Maximum Likelihood bootstrap values are given on appropriate clades in the form Bayesian posterior probabilities /Maximum Likelihood bootstrap. The new and previously described species from Iran are in bold. Type locality and habitat. Collected from rhizosphere of C. maxima in Gilangharb region, Kermanshah province, Western Iran (GPS coordinates: N 33 ˚ 59 ', E 46 ˚ 12 ', 1248 meters above sea level). Diagnosis and relationships. Ektaphelenchoides fuchsi n. sp. is characterized by its body length (529–712 Μm long in females), stylet 14–18 Μm long with distinct basal knobs, lateral fields with three incisures, position of excretory pore at level of posterior part of the metacorpus, 70–84 Μm from anterior end, hemizonid 87–96 Μm from anterior end, post-uterine sac short, less than half body diameter at vulva region (6–9 µm) and posterior body end (tail) filiform in females, spicules with sharply pointed rostrum and rounded condylus, and conoid tail with a sharply pointed terminus in males. The male genital papillae comprise a single precloacal papilla and an adcloacal and a post-cloacal pair. The new species is close to E. attenuata (Massey, 1974) Baujard, 1984, E. musae Baujard, 1984 and E. kelardashtensis in morphology and morphometrics. It differs from the closest species, E. kelardashtensis, by its longer stylet (14–18 vs 13–16 Μm) with basal knobs vs none, longer post-uterine sac (6–9 vs 3–6 Μm), more posterior location of the excretory pore and hemizonid (70–84 vs 55–66 and 87–96 vs 67–78 µm posterior to anterior end, respectively), and longer spicules (12–13 vs 8–10 Μm). It differs from E. attenuata by its smaller body length (529–712 vs 959 Μm), lower a ratio (33–42 vs 42), shorter post-uterine sac (6–9 vs 23–28 µm), slightly vs clearly offset lip region, stylet with basal knobs vs not, shape of the tail (conical with slight transition to filiform shape vs conical with sharp transition to filiform shape). Compared with E. musae, the new species has a shorter stylet (14–18 vs 18.5 –22.0 Μm) with basal knobs vs not, more posterior location of the excretory pore and hemizonid (70–84 vs 60–74 and 87–96 vs 78–89 Μm from the anterior end, respectively), shorter post-uterine sac (6–9 vs 9–19 Μm) and more posterior location of vulva (V= 64.2–87.5 vs 64–69). Etymology. Named in honor of Gilbert Fuchs, a pioneer in the taxonomy of aphelenchid nematodes. Molecular phylogenetic relationships. For molecular analyses, the 631 bp ribosomal DNA large subunit (LSU) D 2 /D 3 (accession number: KJ 190832) of the new species was sequenced. A BlastN search of these sequences revealed the highest match was to aphelenchids, but with nothing identical. The species with the highest identity matches were included in the phylogenetic analyses. The dataset was composed of 837 total characters from which 619 characters were variable and 534 characters were parsimonious informative after manual editing. The average nucleotide composition was as follows: 22.7 % A, 19.2 % C, 30.9 % G and 27.2 % T. Figure 3 presents a phylogenetic tree based on the LSU D 2 /D 3 of various aphelenchids and one outgroup taxon. Using Poikiliolaimus piniperdae (DQ059060) as an outgroup, Ektaphelenchoides fuchsi n. sp. formed a monophyletic clade with high posterior probability (1.00) with species of Ektaphelenchoides and Devibursaphelenchus Kakuliya, 1967. The new species is unique and different from four (characterized by molecular data) other described species from Iran with its partial sequence of 28 S rDNA D 2 /D 3.Published as part of Esmaeili, Mehrab, Heydari, Ramin, Pourjam, Ebrahim & Atighi, Mohammad Reza, 2014, Description of Ektaphelenchoides fuchsi n. sp. (Nematoda: Ektaphelenchinae) from western Iran, pp. 430-438 in Zootaxa 3846 (3) on pages 431-436, DOI: 10.11646/zootaxa.3846.3.6, http://zenodo.org/record/22452

Abursanema Yaghoubi, Pourjam, Pedram, Siddiqi & Atighi, 2014, n. gen.

Genus Abursanema n. gen. Description. Body straight or slightly ventrally arcuate after fixation, less than 1mm long. Cuticle thin, outer cuticle smooth, inner cuticle finely annulated. Lateral field narrow, with two incisures. Head low, flattened, framework six-sectored, lateral sectors narrower than submedian ones. Amphidial apertures not clearly observed. Stylet short, without basal knobs, dorsal arm of the shaft slightly longer than the ventral one. Orifice of dorsal pharyngeal gland located close to stylet base, at ca 1–2 µm distance. Corpus cylindroid, metacorpus spindleshaped, lacking cuticular thickening or valve, isthmus short, terminal bulb large, pyriform, with three pharyngeal glands and a stem-like extension projecting into the lumen of intestine. Excretory pore opening from near base of isthmus to opposite basal bulb. Vulva at 66–75 % of total body length, more than two body widths distance from anus in type species. Ovary single, prodelphic, outstretched anteriorly, usually reflexed twice at proximal end, cells in single row, spermatheca rounded to ellipsoid, filled with spheroid sperm, postvulval uterine sac less than corresponding body width in length. Tail conoid with narrow terminus, similar in both sexes. Spicules simple, slightly arcuate, 18–23 Μm long in type species. Gubernaculum arcuate. Bursa absent. Entomoparasitic infective females not found. Diagnosis and relationships. In the above-mentioned morphological and biological characters, Abursanema n. gen. belongs to the superfamily Sphaerularioidea and family Paurodontidae. The new genus is distinguished from all other valid genera in the family Paurodontidae by the absence of a bursa. It can also be distinguished from Paurodontus by having a stylet without basal knobs (vs stylet with rounded and symmetrical knobs), and by having two incisures in lateral field (vs four or six). The new genus differs from Misticius by the absence of stylet knobs (vs stylet with prominent symmetrical basal knobs), having two incisures in lateral field (vs absence of lateral field), and by having the excretory pore opening near base of isthmus to opposite basal bulb (vs near the stylet base). When compared with Neomisticius, the new genus differs by the absence of stylet knobs (vs stylet with prominent symmetrical basal knobs), excretory pore opening opposite terminal bulb (vs near the base of stylet) and by having a longer tail (vs a short, subcylindrical one with a small projection at its tip). It differs from Paurodontella by the absence of stylet knobs (vs stylet with basal knobs) and the presence of short postvulval uterine sac (vs absence). Abursanema n. gen. is close to Paurodontoides, but differs by the absence of stylet knobs (vs stylet with asymmetrical basal knobs) and by having a head with six sectors (vs eight). From Luella, the new genus differs by having two incisures in the lateral field (vs absence), and structure of the base of the terminal bulb (vs bulb offset and stem-like, basal extension absent). The genus Bealius has a stylet with knobs, a stem-like basal pharyngeal bulb extension projecting into the intestine dorsally, no lateral field or postvulval uterine sac, dissimilar tail shape in female and male, and spicules with a bifurcate tip in comparison with the new genus. Based on molecular data, the new genus has sequences similar to those of Sphaerularia Dufour, 1837. However, it differs from the free-living phase by absence of a bursa in male and by having a stem-like basal extension in pharyngeal bulb projecting into the intestine (vs basal pharyngeal bulb abutting or slightly overlapping the intestine). Based on male characters (especially lacking of bursa), the new genus is similar to Gymnotylenchus Siddiqi, 1961, but is clearly different in having a stylet without basal knobs, extension of the basal pharyngeal bulb into the intestine (vs pharyngeal glands forming an elongate diverticulum extending over intestine dorsally or subdorsally), junction of pharynx and intestine posterior to nerve ring (vs anterior to nerve ring), two incisures in lateral field vs six, and structure of the spicules (tylenchoid vs prominently cephalated) and presence of a gubernaculum (vs absence). Type habitat and locality: Recovered from bark samples of dead Populus alba L. collected in village of Sorkh Abad, Hamedan province, western Iran, during April 2013. Type material: Holotype male, 7 paratype males and 17 females deposited in the Nematode Collection of the Faculty of Agriculture, Tarbiat Modares University, Tehran, Iran. Three male and three female paratypes deposited in each of the following collections: CABI Europe-UK, Egham, Surrey, UK; USDA Nematode Collection, Beltsville, MD, USA; Department of Nematology, Agricultural University, Wageningen, the Netherlands.Published as part of Yaghoubi, Ali, Pourjam, Ebrahim, Pedram, Majid, Siddiqi, Mohammad Rafiq & Atighi, Mohammad Reza, 2014, Description of Abursanema iranicum n. gen., n. sp. (Nematoda: Hexatylina, Sphaerularioidea) from Iran and its phylogenetic relationships, pp. 301-314 in Zootaxa 3826 (2) on pages 304-305, DOI: 10.11646/zootaxa.3826.2.1, http://zenodo.org/record/25457

Morphological and Molecular Characterization of Filenchus multistriatus n. sp. (Tylenchomorpha: Tylenchidae) and Data on a Known Species of the Genus from Bushehr Province, Southern Iran

During a nematological survey in southern Iran, a population belonging to the family Tylenchidae was recovered from a tomato field in Bushehr province. The recovered population belongs to the genus Filenchus, was described and illustrated herein as F. multistriatus n. sp. It is mainly characterized by having a wide and low annulated lip region continuous with adjacent body; amphidial openings confined to the labial plate; four lines in lateral fields forming three bands, with the two outer bands broken by transverse, and the inner one broken by both transverse and longitudinal lines; and median bulb oval with visible valve and elongate-conoid tail uniformly and gradually narrowing toward the distal region, ending in a widely rounded tip. Its morphological and morphometric differences with three closely similar species were discussed. The phylogenetic relationships of the new species with other relevant genera and species were reconstructed using partial sequences of small, and large subunit ribosomal DNA (SSU and LSU rDNA) sequences. Morphometric and morphological data were also provided for an Iranian population of F. sandneri recovered from Bushehr province. Both populations were characterised using SEM data

Molecular and morphological characterisations of two new species of Rotylenchus (Nematoda: Hoplolaimidae) from Iran

Two new amphimictic species, Rotylenchus iranicus n. sp. and R. conicaudatus n. sp., are described. Rotylenchus iranicus n. sp. is characterised mainly by an offset hemispherical lip region with 5-6, rarely 7, annuli, stylet 39-44 μm long, vulva located at 53-65% and rounded tail with 4-9 annuli and typical smooth tip. Morphologically this species is related to R. montanus, R. provincialis and R. aqualamus. Rotylenchus conicaudatus n. sp. is characterised mainly by a slightly offset conoid-rounded lip region with four or five, rarely six annuli, stylet 27-32 μm long, vulva located at 52-63% and conoid-rounded tail with 10-16 annuli and a typical annulated tip. Morphologically this species is related to R. pumilus, R. abnormecaudatus, R. acuspicaudatus and R. provincialis. The results of the phylogenetic analysis based on sequences of the D2-D3 expansion regions of 28S and ITS1-rRNA genes confirmed the species differentiation. Phylogenetic relationships with other species were difficult to assign using D2-D3 expansion regions of 28S. However, using ITS1-rRNA, R. conicaudatus n. sp. presented a close relationship with R. unisexus, whilst R. iranicus n. sp. was closely related to R. conicaudatus n. sp., R. unisexus, R. incultus and R. laurentinus.Peer Reviewe

Reaction of Commercial Cultivars of Kiwifruit to Infection by Root-knot Nematode and Its Biocontrol Using Endophytic Bacteria

Root-knot nematodes (RKN) cause considerable economic losses to kiwifruit production annually. Screening of resistant cultivars has been one of the long-standing methods to manage root-knot nematodes. Here, the reaction of the four most common commercial cultivars of kiwifruit, namely, Actinidia chinensis var. deliciosa cv. Hayward, A. chinensis var. deliciosa cv. Abbott, A. chinensis var. deliciosa cv. Bruno, and A. chinensis var. chinensis cv. Haegeum (commonly known as ‘Golden’ kiwifruit) to infection by the RKN, Meloidogyne incognita, was evaluated. Among examined cultivars ‘Golden’ was the most susceptible, having on average 52.8 galls, 56.1 egg masses per gram of root, and 642 J2 population per 200 gram of soil. ‘Bruno’ showed the highest resistance, with 3.3 galls, 4.1 egg masses per gram of root, and 79 J2 in 200 g of soil. Then, two potential biological control agents, namely Priestia megaterium 31.en and Agrobacterium tumefaciens 19.en were used on ‘Hayward’ seedlings against M. incognita and showed a significant reduction in the number of galls and egg masses on roots, juvenile population in the soil, and increased the growth parameters of the plants compared to non-treated seedlings. We demonstrated that integrated management using resistant cultivars and biological control can provide a safe and economic method to control RKN, and these resistant cultivars can be used in breeding programs

Molecular and morphological characterization of Veleshkinema iranicum n. gen., n. sp. (Nematoda: Hexatylina, Sphaerularioidea) from Iran

Miraeiz, Esmaeil, Heydari, Ramin, Álvarez-Ortega, Sergio, Pedram, Majid, Atighi, Mohammad Reza (2015): Molecular and morphological characterization of Veleshkinema iranicum n. gen., n. sp. (Nematoda: Hexatylina, Sphaerularioidea) from Iran. Zootaxa 4000 (5): 531-546, DOI: http://dx.doi.org/10.11646/zootaxa.4000.5.