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1.  First Report of Korean Cyst Nematode, Heterodera koreana, Parasitic on Bamboo, Phyllostachys nigra, from Iran 
Journal of Nematology  2015;47(3):167-168.
Bamboo is grown sporadically in the north of Iran and is confined to very limited areas. The history of growing bamboo was to some extent simultaneous with the entrance, commencement, and growth of the tea industry in the north about a century ago. The bamboo was used for making baskets to transfer the harvested tea foliage from farm to the factory and other linked functions. A main area allocated for bamboo growing is located in Lahidjan Agricultural Research Station (LARS) in the north of Iran, where several species of bamboo were cultivated in an area of 5 ha. The species include five species of Phyllostachys (viz., P. aurea, P. bambusoides, P. decora, P. nigra, P. vivax) and one species of Arundinaria gigantean, Pleioblastus fortune, and Semiarundinaria fastuosa; however, only P. aurea and P. nigra have been precisely identified. A survey on plant parasitic nematodes associated with bamboo mainly on P. nigra in LARS revealed second-stage juveniles of cyst forming nematode in soil samples. Further analysis of root and soil samples led to recovery of a cyst nematode belonging to the genus Heterodera and the Afenestrata group. Cysts, vulval cone, and second-stage juveniles were studied for morphological and morphometric features. The classical identification was followed by amplification of the ribosomal RNA-ITS region and the D2-D3 expansion segments of 28S large-subunit rRNA gene; the amplified fragments were sequenced, edited, and compared with those of the corresponding published gene sequences. New D2-D3 and rRNA-ITS gene sequences were deposited in the GenBank database under the accession numbers KR818910 and KR818911, respectively. Based on the morphological and molecular data, the species of the cyst-forming nematode was identified as H. koreana (Vovlas et al., 1992; Mundo-Ocampo et al., 2008). The body contour of cysts was mainly subspherical, vey often with irregular shape (Fig. 1A), yellowish to light brown, thin cuticle with fine zigzag pattern, without fenestration, lacking bulla, and underbridge. Vulval lips protruded, cuticular pattern of vulval cone with a tuberculate area (Fig. 2B), and vagina embedded into vulval lips. The second-stage juveniles cylindrical and slender, hemispherical cephalic framework, with three lines in lateral field, well-developed rounded stylet knobs, tail conoid tapring to fine rounded terminus, phasmids posterior to anus. The cyst measurements were (n = 21) length 502 ± 70 (420 to 640) µm; width = 408 ± 60 (320 to 520) µm; length/width = 1.23 ± 0.09 (1.07 to 1.5) µm. The morphometric characters of vulval cone were measured (n = 7): fenestral length = 62.4 ± 6.5 (51 to 71) µm; fenestral width = 50.7 ± 3.2 (45 to 54) µm; vulval slit = 51.9 ± 4.3 (46 to 59) µm; distance from vulva to anus = 51.3 ± 4.4 (43 to 56) µm. Second-stage juveniles showed the following morphometric characters (n = 14): L = 455 ± 11.3 (437 to 472) µm; a = 29.9 ± 0.9 (28.3 to 31.5); b΄ = 2.7 ± 0.4 (2.2 to 3.5); c = 7.4 ± 0.9 (6 to 8.9); ć = 6.1 ± 0.4 (5.1 to 6.7); lip region height = 3 µm; lip region width = 7.5 ± 0.5 (7 to 8) µm; stylet length = 18.1 ± 0.5 (17 to 19) µm; anterior end to median bulb = 72.2 ± 1.7 (70 to 75) µm; anterior end to secretory-excretory pore = 99.7 ± 2.5 (96 to 103) µm; maximum body width = 15.2 ± 0.4 (15 to 16) µm; body width at anus = 10.1 ± 1 (8 to 11) µm; tail length = 62.0 ± 6.9 (51 to 74) µm; hyaline part of tail = 44.0 ± 1.8 (40 to 47) µm. The egg measurements for 11 individuals were length = 102.5 ± 7.9 (93 to 119) µm; width = 39.3 ± 4.2 (33 to 46) µm; length/width = 2.6 ± 0.3 (2.0 to 3.1). The morphology, morphometric characters and molecular data of the population of H. koreana isolated from bamboo in Iran are in agreement with those previously reported for this species (Vovlas et al., 1992; Mundo-Ocampo et al., 2008). At present, five species of Heterodera belonging to the Cyperi and Afenestrata groups were reported from bamboo, H. bamboosi (Kaushal and Swarup, 1988; Wouts and Baldwin, 1998) on Bambusa sp. from India; H. koreana on P. pubescence, P. aurea, and P. nigra from South Korea and the United States; and H. hainanensis (Zhuo et al., 2013), H. fengi (Wang et al., 2013), and H. guangdongensis (Zhuo et al., 2014) on P. pubescence from China; thus showing host suitability of bamboo for at least five species of cyst-forming nematodes. A greenhouse test performed by planting rice seed cv. Hashemi in soil containing H. koreana showed successful multiplication of Korean cyst nematode on rice seedlings after 2 mon. The exact date of the establishment of bamboo plantation in LARS is not precisely clear, but it indicates that the Korean cyst nematode was most likely brought with the imported bamboo seedlings from unknown origin several decades ago. According to our best knowledge, this is the first report of occurrence of H. koreana from Iran. So far the Korean cyst nematode was reported from South Korea, Thailand, and the United States, Florida (from nurseries); this study includes the distribution of this cyst-forming nematode in Iran and expands the information of the occurrence of H. koreana for the world.
PMCID: PMC4612185  PMID: 26527836
bamboo; detection; Heterodera koreana; Iran
2.  Molecular Characterization of Meloidogyne christiei Golden and Kaplan, 1986 (Nematoda, Meloidogynidae) Topotype Population Infecting Turkey Oak (Quercus laevies) in Florida 
Journal of Nematology  2015;47(3):169-175.
Meloidogyne christiei isolated from turkey oak, Quercus laevies, from the type locality in Florida was characterized using isozyme profiles and ribosomal and mitochondrial gene sequences. The phenotype N1a detected from a single egg-laying female of M. christiei showed one very strong band of malate dehydrogenase (MDH) activity; however, no esterase (EST) activity was identified from macerate of one or even 20 females per well. Phylogenetic relationships within the genus Meloidogyne as inferred from Bayesian analysis of partial 18S ribosomal RNA (rRNA), D2-D3 of 28S rRNA, internal transcribed spacer (ITS) rRNA, and cytochrome oxidase subunit II (COII)-16S rRNA of mitochondrial DNA (mtDNA) gene fragments showed that M. christiei formed a separate lineage within the crown group of Meloidogyne and its relationships with any of three Meloidogyne clades were not resolved.
PMCID: PMC4612186  PMID: 26527837
18S rRNA; 28S rRNA; isozymes; ITS rRNA; Quercus laevis; Meloidogyne christiei; mtDNA; oak; phylogeny; root-knot nematodes
3.  Sex Attraction and Mating in Bursaphelenchus okinawaensis and B. xylophilus 
Journal of Nematology  2015;47(3):176-183.
The fungal feeding, hermaphroditic Bursaphelenchus okinawaensis is a laboratory model to understand the biology of Bursaphelenchus. The extent to which B. okinawaensis can be used to model Bursaphelenchus xylophilus mating was investigated. A chemotaxis assay was conducted to examine whether B. xylophilus and B. okinawaensis produce and respond to volatile sex attractants. Unmated B. xylophilus females were found to attract B. xylophilus males. Similarly, old (sperm depleted) but not young (sperm repleted) B. okinawaensis hermaphrodites attract B. okinawaensis males. Thus, in both species, sperm status corresponds to its ability to attract males. B. xylophilus males also produce a volatile pheromone that attracts both mated and unmated females. A second assay, in which the behavior of males on petri plates in the presence of different females or hermaphrodites of Bursaphelenchus was observed, revealed that B. xylophilus unmated females attract B. okinawaensis males, and B. okinawaensis old hermaphrodites attract B. xylophilus males. These observations suggested that the pheromones of Bursaphelenchus work to some extent across species. Mating behavior through spicule insertion occurs across species, suggesting that postcopulatory mechanisms prevent production of interspecific progeny. The hermaphroditic B. okinawaensis will be a useful model to conduct genetic studies for the understanding of the molecular mechanisms underlying mating behavior in Bursaphelenchus nematodes.
PMCID: PMC4612187  PMID: 26527838
behavior; mating behavior; reproductive isolation; sex pheromone; sperm status
4.  Viability and Virulence of Entomopathogenic Nematodes Exposed to Ultraviolet Radiation 
Journal of Nematology  2015;47(3):184-189.
Entomopathogenic nematodes (EPNs) can be highly effective biocontrol agents, but their efficacy can be reduced due to exposure to environmental stress such as from ultraviolet (UV) radiation. Our objectives were to 1) compare UV tolerance among a broad array of EPN species, and 2) investigate the relationship between reduced nematode viability (after exposure to UV) and virulence. Nematodes exposed to a UV radiation (254 nm) for 10 or 20 min were assessed separately for viability (survival) and virulence to Galleria mellonella. We compared 9 different EPN species and 15 strains: Heterorhabditis bacteriophora (Baine, fl11, Oswego, and Vs strains), H. floridensis (332), H. georgiana (Kesha), H. indica (HOM1), H. megidis (UK211), Steinernema carpocapsae (All, Cxrd, DD136, and Sal strains), S. feltiae (SN), S. rarum (17C&E), and S. riobrave (355). In viability assessments, steinernematids, particularly strains of S. carpocapsae, generally exhibited superior UV tolerance compared with the heterorhabditids. However, some heterorhabditids tended to be more tolerant than others, e.g., H. megidis and H. bacteriophora (Baine) were most susceptible and H. bacteriophora (Vs) was the only heterorhabditid that did not exhibit a significant effect after 10 min of exposure. All heterorhabditids experienced reduced viability after 20 min exposure though several S. carpocapsae strains did not. In total, after 10 or 20 min exposure, the viability of seven nematode strains did not differ from their non-UV exposed controls. In virulence assays, steinernematids (particularly S. carpocapsae strains) also tended to exhibit higher UV tolerance. However, in contrast to the viability measurements, all nematodes experienced a reduction in virulence relative to their controls. Correlation analysis revealed that viability among nematode strains is not necessarily related to virulence. In conclusion, our results indicate that the impact of UV varies substantially among EPNs, and viability alone is not a sufficient measure for potential impact on biocontrol efficacy as other characters such as virulence may be severely affected even when viability remains high.
PMCID: PMC4612188  PMID: 26527839
biocontrol; entomopathogenic nematode; Heterorhabditis; Steinernema; tolerance; ultraviolet
5.  A New Species of Pristionchus (Rhabditida: Diplogastridae) and Its Bacterial Symbiont from Yixing, China 
Journal of Nematology  2015;47(3):190-197.
A new nematode species, Pristionchus entomophilus n. sp., was collected during a soil sample survey in Yixing of Jiangsu province, eastern China. P. entomophilus n. sp. is distinguished by its unique characteristics. This new species is mainly hermaphroditic, with males seldom found. The new nematode has a similar body length but has much narrower body width compared with P. pacificus. Its body is covered with longitudinal ridges: 12 ridges on head, 13 or 14 ridges in the middle, 11 and 7 ridges in front and rear of the anus, respectively. The eurystomatous form mouth includes a triangular dorsal tooth, a large claw-like right subventral tooth, and a row of five ventral denticles placed opposite the dorsal tooth. Only eight pairs of genital papillae and a pair of phasmids are present in the tail of the male as the sixth pair of papillae having seemingly been degenerated and lost. Molecular phylogenetic trees based on 18S rDNA confirmed that the new species belongs to the genus Pristionchus and is most closely related to P. pacificus. Moreover, the new species was found to be occasionally associated with the entomopathogenic bacterial strain 09FLYB1 of Serratia nematodophila and be able to stably transfer the bacterial strain for several generations.
PMCID: PMC4612189  PMID: 26527840
insect phoretic; phylogenetic; Pristionchus; saprophagous; taxonomy
6.  Marine Nematode Taxonomy in Africa: Promising Prospects Against Scarcity of Information 
Journal of Nematology  2015;47(3):198-206.
From the late 19th century, Africa has faced heavy exploitation of its natural resources with increasing land/water pollution, and several described species have already become extinct or close to extinction. This could also be the case for marine nematodes, which are the most abundant and diverse benthic group in marine sediments, and play major roles in ecosystem functioning. Compared to Europe and North America, only a handful of investigations on marine nematodes have been conducted to date in Africa. This is due to the scarcity of experienced taxonomists, absence of identification guides, as well as local appropriate infrastructures. A pivotal project has started recently between nematologists from Africa (Tunisia), India, and Europe (Italy) to promote taxonomic study and biodiversity estimation of marine nematodes in the African continent. To do this, as a first step, collection of permanent slides of marine nematodes (235 nominal species and 14 new to science but not yet described) was recently established at the Faculty of Sciences of Bizerte (Tunisia). Capacity building of next generation of African taxonomists have been carried out at level of both traditional and molecular taxonomy (DNA barcoding and next-generation sequencing [NGS]), but they need to be implemented. Indeed, the integration of these two approaches appears crucial to overcome lack of information on the taxonomy, ecology, and biodiversity of marine nematodes from African coastal waters.
PMCID: PMC4612190  PMID: 26527841
Africa; barcoding; biodiversity; integrative taxonomy; molecular tools; morphology; next-generation sequencing
7.  Pasteuria penetrans for Control of Meloidogyne incognita on Tomato and Cucumber, and M. arenaria on Snapdragon 
Journal of Nematology  2015;47(3):207-213.
Meloidogyne incognita and Meloidogyne arenaria are important parasitic nematodes of vegetable and ornamental crops. Microplot and greenhouse experiments were conducted to test commercial formulations of the biocontrol agent Pasteuria penetrans for control of M. incognita on tomato and cucumber and M. arenaria on snapdragon. Three methods of application for P. penetrans were assessed including seed, transplant, and post-plant treatments. Efficacy in controlling galling and reproduction of the two root-knot nematode species was evaluated. Seed treatment application was assessed only for M. incognita on cucumber. Pasteuria treatment rates of a granular transplant formulation ranged from 1.5 × 105 endospores/cm3 to 3 × 105 endospores/cm3 of transplant mix applied at seeding. Additional applications of 1.5 × 105 endospores/cm3 of soil were applied as a liquid formulation to soil post-transplant for both greenhouse and microplot trials. In greenhouse cucumber trials, all Pasteuria treatments were equivalent to steamed soil for reducing M. incognita populations in roots and soil, and reducing nematode reproduction and galling. In cucumber microplot trials there were no differences among treatments for M. incognita populations in roots or soil, eggs/g root, or root condition ratings. Nematode reproduction on cucumber was low with Telone II and with the seed treatment plus post-plant application of Pasteuria, which had the lowest nematode reproduction. However, galling for all Pasteuria treatments was higher than galling with Telone II. Root-knot nematode control with Pasteuria in greenhouse and microplot trials varied on tomato and snapdragon. Positive results were achieved for control of M. incognita with the seed treatment application on cucumber.
PMCID: PMC4612191  PMID: 26527842
Antirrhinum majus; biological control; Cucumis sativus; cucumber; Meloidogyne; Pasteuria penetrans; Solanum lycopersicum; root-knot nematodes; snapdragon; tomato
8.  Potential of Sahelian Native Shrub Materials to Suppress the Spiral Nematode Helicotylenchus dihystera 
Journal of Nematology  2015;47(3):214-217.
Pearl millet (Pennisetum glaucum) is a drought-tolerant cereal commonly grown for grain and fodder in arid areas throughout the world. Senegalese millet fields are infested with Helicotylenchus. The native evergreen woody shrub Piliostigma reticulatum is widely distributed in sub-Saharan Africa. Its coppiced residues are used by small farmers as mulch in crop fields. The shrub’s nematicidal effect on the spiral nematode Helicotylenchus dihystera was evaluated in a pearl millet pot experiment. The abundance of nematodes decreased by 64% after application of either leaf powder or a pulverized mixing of leaves and stems, suggesting the use of aboveground materials of P. reticulatum as a potential nematicide. The results show promise for use of a local resource by subsistence farmers in the Sahel. Further research is needed on application to fully develop this approach as a biopesticide.
PMCID: PMC4612192  PMID: 26527843
Biological control; Helicotylenchus dihystera; millet intercropping; native evergreen woody shrub; nematicidal activity; Pennisetum glaucum; Piliostigma reticulatum; plant-parasitic nematode; Sahelian agrosystem management; soil organic amendment; spiral nematode; western Africa
Journal of Nematology  2015;47(3):218-281.
PMCID: PMC4612193
10.  Diplogasteroides asiaticus n. sp. is Associated with Monochamus alternatus in Japan 
Journal of Nematology  2015;47(2):105-115.
Diplogasteroides asiaticus n. sp. is described and illustrated, and its molecular profile and phylogenetic status within the family Diplogastridae are inferred. Morphologically, the new species is characterized by its stomatal structure, a tube-like stoma with three small, rod-like dorsal teeth and two subventral ridges; a spicule clearly ventrally bent at 1/3 from the anterior end; a gubernaculum with a rounded anterior end and sharply pointed distal end in lateral view; nine pairs of genital papillae with an arrangement of ; a short tail spike in males; and a well-developed receptaculum seminis, i.e., the antiparallel blind sacs of the uteri beyond the vulva region and elongated conical tail in females. This new species is morphologically similar to D. haslacheri, but it can be distinguished by the morphology of the somewhat shorter tail in females. D. asiaticus n. sp. shares high sequence conservation with D. andrassyi as there is only one base pair difference in the nearly full-length 18S rDNA and seven base pair differences in the D2-D3 expansion segments of the 28S rDNA. Despite this sequence conservation, the species status of D. asiaticus n. sp. was confirmed using the biological species concept, as D. asiaticus n. sp. and D. andrassyi failed to generate viable F2 progeny in hybridization tests.
PMCID: PMC4492285  PMID: 26170472
Diplogasteroides asiaticus n. sp.; Japan; Monochamus alternatus; morphology; phylogeny; Pinus densiflora; taxonomy
11.  Biochemistry of Anhydrobiosis in Beddingia siricidicola, a Biological Control Agent of Sirex noctilio 
Journal of Nematology  2015;47(2):116-125.
Proto-anhydrobiosis of the nematode, Beddingia siricidicola, was achieved by incubation in polyethylene glycol or various concentrations up to 4 M of glycerol. The associated changes in the levels of glycerol, unbound proline, trehalose, lipids, and glycogen were determined by alkylation strategies, followed by gas chromatography or gas chromatography/mass spectrometry. The level of glycerol reached 8.9% of dry weight, proline 2.4% of dry weight, and trehalose 8.0% of dry weight within B. siricidicola that were incubated in 1.5 M glycerol over 6 d, while glycerol reached 17.9% of dry weight after incubation for the same period in 4 M glycerol. Movement was thereby reduced but the nematodes from 1.5 M glycerol revived after a few minutes upon rehydrating and they were able to avoid osmotic damage by rapidly excreting the glycerol, much of it being expelled within the first hour. The potential for storage and transport of this nematode for the biological control of the pine-killing wasp, Sirex noctilio, was greatly improved when nematode suspensions were maintained in 1.5 M glycerol under refrigeration.
PMCID: PMC4492286  PMID: 26170473
anhydrobiosis; Beddingia siricidicola; biochemistry; derivatization; gas chromatography; glycerol; osmosis; proline; protectants; Sirex noctilio; trehalose
12.  Efficacy of Four Nematicides Against the Reproduction and Development of Pinewood Nematode, Bursaphelenchus xylophilus 
Journal of Nematology  2015;47(2):126-132.
To understand the efficacy of emamectin benzoate, avermectin, milbemectin, and thiacloprid on the reproduction and development of Bursaphelenchus xylophilus, seven parameters, namely population growth, fecundity, egg hatchability, larval lethality, percent larval development, body size, and sexual ratio, were investigated using sublethal (LC20) doses of these compounds in the laboratory. Emamectin benzoate treatment led to a significant suppression in population size, brood size, and percent larval development with 411, 3.50, and 49.63%, respectively, compared to 20850, 24.33, and 61.43% for the negative control. The embryonic and larval lethality increased obviously from 12.47% and 13.70% to 51.37% and 75.30%, respectively. In addition, the body length was also significantly reduced for both males and females in the emamectin benzoate treatment. Avermectin and milbemectin were also effective in suppressing population growth by increasing larval lethality and reducing larval development, although they did not affect either brood size or embryonic lethality. Body length for both male and female worms was increased by avermectin. Thiacloprid caused no adverse reproductive effects, although it suppressed larval development. Sexual ratio was not affected by any of these four nematicides. Our results indicate that emamectin benzoate, milbemectin, and avermectin are effective against the reproduction of B. xylophilus. We think these three nematicides can be useful for the control of pine wilt disease.
PMCID: PMC4492287  PMID: 26170474
avermectin; Bursaphelenchus xylophilus; emamectin benzoate; milbemectin; pine wilt disease; thiacloprid
13.  Mi-1-Mediated Nematode Resistance in Tomatoes is Broken by Short-Term Heat Stress but Recovers Over Time 
Journal of Nematology  2015;47(2):133-140.
Tomato (Solanum lycopersicum L.) is among the most valuable agricultural products, but Meloidogyne spp. (root-knot nematode) infestations result in serious crop losses. In tomato, resistance to root-knot nematodes is controlled by the gene Mi-1, but heat stress interferes with Mi-1-associated resistance. Inconsistent results in published field and greenhouse experiments led us to test the effect of short-term midday heat stress on tomato susceptibility to Meloidogyne incognita race 1. Under controlled day/night temperatures of 25°C/21°C, ‘Amelia’, which was verified as possessing the Mi-1 gene, was deemed resistant (4.1 ± 0.4 galls/plant) and Rutgers, which does not possess the Mi-1 gene, was susceptible (132 ± 9.9 galls/plant) to M. incognita infection. Exposure to a single 3 hr heat spike of 35°C was sufficient to increase the susceptibility of ‘Amelia’ but did not affect Rutgers. Despite this change in resistance, Mi-1 gene expression was not affected by heat treatment, or nematode infection. The heat-induced breakdown of Mi-1 resistance in ‘Amelia’ did recover with time regardless of additional heat exposures and M. incognita infection. These findings would aid in the development of management strategies to protect the tomato crop at times of heightened M. incognita susceptibility.
PMCID: PMC4492288  PMID: 26170475
heat stress; Meloidogyne incognita; Mi-1 gene; resistance; root-knot nematode; Solanum lycopersicum
14.  Impact of Grapevine (Vitis vinifera) Varieties on Reproduction of the Northern Root-Knot Nematode (Meloidogyne hapla) 
Journal of Nematology  2015;47(2):141-147.
One of the most commonly encountered plant-parasitic nematodes in eastern Washington Vitis vinifera vineyards is Meloidogyne hapla; however, limited research exists on the impact of this nematode on V. vinifera. The objectives of this research were to determine the impact of M. hapla on Chardonnay and Cabernet Sauvignon vine establishment and to determine the host status of V. vinifera varieties/clones predominantly grown in Washington to M. hapla. In a microplot experiment, Chardonnay and Cabernet Sauvignon vines were planted into soil inoculated with different densities of M. hapla; population dynamics of M. hapla and vine performance were monitored over 3 yr. In greenhouse experiments, several clones representing five V. vinifera varieties, Chardonnay, Riesling, Cabernet Sauvignon, Merlot, and Syrah, were evaluated as hosts for M. hapla. In both microplot and greenhouse experiments, white varieties were significantly better hosts than red varieties. In the greenhouse experiments, Chardonnay and Riesling had 40% higher reproduction factor values than Syrah and Merlot, however, all varieties/clones screened were good hosts for M. hapla (reproduction factors > 3). In the microplot experiment, M. hapla eggs/g root were 4.5 times greater in Chardonnay compared to Cabernet Sauvignon 3 yr after planting but there was no evident impact of M. hapla on vine establishment.
PMCID: PMC4492289  PMID: 26170476
host-parasite interaction; host status; M. hapla; microplots; own-rooted; Washington
15.  Further Studies on Soil Nematode Fauna in North Western Iran with the Description of One New Species 
Journal of Nematology  2015;47(2):148-152.
Heterodorus youbertghostai n. sp. is described and illustrated based on morphological, morphometric, and molecular data. The new species was found in two geographically distant points in northwestern Iran and is characterized by having angular lip region, separated from the rest body by a constriction, body length of 1,432.5 to 1,751.3 µm, odontostyle length of 24 to 28 µm, rod-like odontophore, 37.0 to 42.5 µm long, lacking flanges at base, double guiding ring at 14 to 16 µm distance from anterior end, pharyngeal bulb comprising 40% to 48% of pharynx, intestine usually containing green material, female reproductive system amphidelphic with less divided short uterus, specific structure of pars distalis vaginae, bluntly conical tail, dorsally convex and ventrally flat, with rounded tip and saccate bodies in ventral side. The new species comes more close to H. conicaudatus and H. irregularis by its morphology and morphometric characters. Compared to former, it has remarkable difference in vulva position and tail characters, and compared to the latter, it could be separated by shorter body, posteriorly located vulva, wider lip region, and longer tail. In phylogenetic analyses using partial sequences of 28S rDNA D2-D3, the new species formed a fully supported clade with several isolates of H. brevidentatus, prevalent in Iran. The other nordiid taxon, Enchodorus dolichurus, already reported from Iran, was also sequenced for the same genomic region and included in phylogenetic analyses.
PMCID: PMC4492290  PMID: 26170477
28S rDNA; city of Kaleibar; description; Eastern Azarbayjan; Nordiidae; Sabalan; taxonomy
16.  Description of Sclerorhabditis miniata n. sp. and First Description of Male of Diploscapter coronatus Cobb, 1913 (Nematoda: Rhabditidae) 
Journal of Nematology  2015;47(2):153-158.
A population of Sclerorhabditis miniata n. sp. is described and illustrated from Poonch district of Jammu and Kashmir State, India. The new species is characterized by small body size, with an annulated cuticle, offset labial region, crown shaped, strongly sclerotized lips, thin lateral lips, membranous, stegostom without glottoid apparatus, cheilostom rod shaped, sclerotized, spicules free, strong and thick, gubernaculum simple, bent proximally, bursa open, peloderan with seven pairs of bursal papillae in 1+1/1+1+2+1 pattern. The males of Diploscapter coronatus are described for the first time. They are usually smaller than the females and have labial region similar to females. Spicules separate, with a small dorsal velum, gubernaculum simple, almost straight, bursa open, pseudopeloderan with seven pairs of bursal papillae in 1+1/1+2+1+1 pattern.
PMCID: PMC4492291  PMID: 26170478
Diploscapter; India; morphology; new species; Sclerorhabditis; taxonomy
17.  Sequence and Spatiotemporal Expression Analysis of CLE-Motif Containing Genes from the Reniform Nematode (Rotylenchulus reniformis Linford & Oliveira) 
Journal of Nematology  2015;47(2):159-165.
The reniform nematode, Rotylenchulus reniformis, is a sedentary semi-endoparasitic species with a host range that encompasses more than 77 plant families. Nematode effector proteins containing plant-ligand motifs similar to CLAVATA3/ESR (CLE) peptides have been identified in the Heterodera, Globodera, and Meloidogyne genera of sedentary endoparasites. Here, we describe the isolation, sequence analysis, and spatiotemporal expression of three R. reniformis genes encoding putative CLE motifs named Rr-cle-1, Rr-cle-2, and Rr-cle-3. The Rr-cle cDNAs showed >98% identity with each other and the predicted peptides were identical with the exception of a short stretch of residues at the carboxy(C)-terminus of the variable domain (VD). Each RrCLE peptide possessed an amino-terminal signal peptide for secretion and a single C-terminal CLE motif that was most similar to Heterodera CLE motifs. Aligning the Rr-cle cDNAs with their corresponding genomic sequences showed three exons with an intron separating the signal peptide from the VD and a second intron separating the VD from the CLE motif. An alignment of the RrCLE1 peptide with Heterodera glycines and Heterodera schachtii CLE proteins revealed a high level of homology within the VD region associated with regulating in planta trafficking of the processed CLE peptide. Quantitative RT-PCR (qRT-PCR) showed similar expression profiles for each Rr-cle transcript across the R. reniformis life-cycle with the greatest transcript abundance being in sedentary parasitic female nematodes. In situ hybridization showed specific Rr-cle expression within the dorsal esophageal gland cell of sedentary parasitic females.
PMCID: PMC4492292  PMID: 26170479
CLE; effector; host-parasite relationship; reniform nematode; Rotylenchulus
18.  Belonolaimus longicaudatus: An Emerging Pathogen of Peanut in Florida 
Journal of Nematology  2015;47(2):87-96.
Sting nematode (Belonolaimus longicaudatus) is an economically important ectoparasitic nematode that is highly pathogenic on a wide range of agricultural crops in sandy soils of the southeastern United States. Although this species is commonly found in Florida in hardwood forests and as a soilborne pathogen on turfgrasses and numerous agronomic and horticultural crops, it has not been reported infecting peanut. In the summers of 2012 and 2013, sting nematode was found infecting three different peanut cultivars being grown on two separate peanut farms in Levy County, FL. The damage consisted of large irregular patches of stunted, chlorotic plants at both farms. The root systems were severely abbreviated and there were numerous punctate-like isolated lesions observed on pegs and pods of infected plants. Sting nematodes were extracted from soil collected around the roots of diseased peanut over the course of the peanut season at both farm sites. Peanut yield from one of these nematode-infested sites was 64% less than that observed in areas free from sting nematodes. The morphological characters of the nematode populations in these fields were congruous with those of the original and other published descriptions of B. longicaudatus. Moreover, the molecular analyses based on the sequences of D2/D3 expansion fragments of 28S rRNA and internal transcribed spacer (ITS) rRNA genes from the nematodes further collaborates the identification of the sting nematode isolates as B. longicaudatus. The sequences were deposited in GenBank (accession no. KF963097, KF963098 for ITS, and KF96399, KF963100 for D2-D3). The results of the phylogenetic analysis using the sequences of these isolates from peanut compared with those of other isolates from Florida suggests that the sting nematode from both peanut farms are genetically close to B. longicaudatus populations occurring in the state. Peanut plants inoculated with both nematode isolates showed punctate-like isolated lesions on pods and pegs, and an abbreviation of their root systems, whereas those symptoms were not observed on noninoculated peanut plants. To our knowledge, this is the first report of large-scale field damage caused by sting nematode infecting peanut grown under field conditions in Florida.
PMCID: PMC4492293  PMID: 26170470
Arachis hypogaea; Belonolaimus longicaudatus; emerging pathogen; morphology; pathogenicity; peanut; phylogenetics; sting nematode
19.  A First Report of Anguina pacificae in Ireland 
Journal of Nematology  2015;47(2):97-104.
Anguina pacificae is a significant pest of Poa annua golf course greens in northern California. This study presents the first confirmed case of an A. pacificae infestation outside of North America, where the nematode’s distribution is further restricted to a relatively limited coastal region. Species confirmation was made by morphometric and molecular methods and comparisons to closely related species including the European species, Anguina agropyri. The A. pacificae population detected on an Irish golf course was monitored over a 2-yr period and the life cycle compared with Californian population dynamics. A. pacificae was assessed for the potential risk of spreading to the local agricultural sector, in addition, the biosecurity risks from A. pacificae and plant parasitic nematodes in general were reviewed for northwest Europe.
PMCID: PMC4492294  PMID: 26170471
Agrostis; Anguina pacificae; annual bluegrass; biosecurity; detection; diagnosis; host range; morphology; morphometrics; Pacific shoot-gall nematode; Poa annua; population dynamics; stem gall
20.  Utility of Classical α-Taxonomy for Biodiversity of Aquatic Nematodes 
Journal of Nematology  2015;47(1):1-10.
“Classical α-taxonomy” has different interpretations. Therefore, within the framework of an integrated taxonomic approach it is not relevant to divide taxonomy in different components, each being allocated a different weight of importance. Preferably, taxonomy should be seen in a holistic way, including the act of delimiting and describing taxa, based on different features and available methods, and taxonomy can not be interpreted without looking at evolutionary relationships. The concept of diversity itself is quite diverse as is the measure of diversity. Taxonomic descriptions of free-living aquatic nematodes are very valuable as they provide basic phenotypic information that is necessary for the functional ecological, behavioral, and evolutionary interpretation of data gathered from molecular analyses and of the organism as a whole. In general, molecular taxonomic analyses have the advantage of being much faster and of being able to deal with a larger number of specimens but also possess the important advantage of dealing with a huge amount of features compared to the morphology-based approach. However, just as morphological studies, molecular analyses deal only with partial of an organism.
PMCID: PMC4388575  PMID: 25861112
morphology; molecular analyses; taxonomic descriptions
21.  Experimental Studies with Nematodes in Ecotoxicology: An Overview 
Journal of Nematology  2015;47(1):11-27.
With respect to their high abundances, their role as intermediaries between microorganisms and higher trophic levels, and their ubiquitous occurrence in all habitats, nematodes are of strong potential interest as environmental indicators. Ecotoxicological methods to evaluate the risk of anthropogenic pollutants on ecosystems require both in vitro and in vivo toxicity tests to investigate either mechanisms or pathways of toxicity and to set accurate toxicity thresholds. For this, the interest in nematodes as model organisms in ecotoxicology increased over the past few decades and existing appropriate experimental methods are reviewed in this manuscript. An overview of the various existing ecotoxicological tools for nematodes, ranging from molecular laboratory methods to experimental model ecosystem approaches, and their role as indicator organisms is given. The reviewed studies, approaches that range from species-based to community-based methods, reveal exciting possibilities for the future use of nematodes in ecotoxicological studies. Suitable ecotoxicological tools and ecological indices for nematodes should be integrated in weight-of-evidence approaches for assessing the ecological risk of contamination.
PMCID: PMC4388576  PMID: 25861113
chemicals; contamination; ecology; ecotoxicology; free-living; freshwater; marine; methods; microcosms; model ecosystems; molecular; NemaSPEAR[%] index; review; single species; soil
22.  Free-Living Nematodes in the Freshwater Food Web: A Review 
Journal of Nematology  2015;47(1):28-44.
Free-living nematodes are well-recognized as an abundant and ubiquitous component of benthic communities in inland waters. Compelling evidence from soil and marine ecosystems has highlighted the importance of nematodes as trophic intermediaries between microbial production and higher trophic levels. However, the paucity of empirical evidence of their role in freshwater ecosystems has hampered their inclusion in our understanding of freshwater food web functioning. This literature survey provides an overview of research efforts in the field of freshwater nematode ecology and of the complex trophic interactions between free-living nematodes and microbes, other meiofauna, macro-invertebrates, and fishes. Based on an analysis of the relevant literature and an appreciation of the potential of emerging approaches for the evaluation of nematode trophic ecology, we point out research gaps and recommend relevant directions for further research. The latter include (i) interactions of nematodes with protozoans and fungi; (ii) nonconsumptive effects of nematodes on microbial activity and the effects of nematodes on associated key ecosystem processes (decomposition, primary production); and (iii) the feeding selectivity and intraspecific feeding variability of nematodes and their potential impacts on the structure of benthic communities.
PMCID: PMC4388577  PMID: 25861114
algae; bacteria; ecology; fish; food web; free-living; freshwater; fungi; interaction; macrofauna; meiofauna; method; organic matter; predation; protozoa; selectivity
23.  Effect of Storage Environment on Hatching of the Cyst Nematode Globodera ellingtonae 
Journal of Nematology  2015;47(1):45-51.
Globodera spp. eggs go through a diapause, which remains dormant until favorable hatching conditions are reached. Because of the regulatory concerns with cyst nematodes, it is often only possible to rear eggs for research in the greenhouse. However, hatch is often lower for greenhouse-produced eggs than for eggs obtained from the field. The goal of this research was to determine storage conditions for Globodera ellingtonae eggs produced in the greenhouse that would increase percentage hatch. Over 3 yr, G. ellingtonae greenhouse-produced eggs were stored in different environments (−20°C, 4°C, room temperature, and the field) in either dry or moist soil. Percentage hatch after exposure to the different environments was determined in potato root diffusate. Across two experiments, field-produced eggs had higher hatch rates (65.2%) than greenhouse-produced eggs (10.4%). Temperature did not have an appreciable influence on hatch of eggs stored dry in two experiments (2.8% to 8.4% and 3.8% to 8.6%), but hatch of eggs stored in moist soil was significantly higher than in dry soil at all temperatures except −20°C (26.8% and 28.7%). However, the ability of G. ellingtonae greenhouse-, microplot-, and field-produced eggs to reproduce on potato in field microplots was not different. Although it may not be possible to produce G. ellingtonae eggs in the greenhouse that have the magnitude of hatch as those produced in the field, hatching can be greatly increased by storing eggs in moist soil at either 4°C or room temperature.
PMCID: PMC4388578  PMID: 25861115
diapause; dormancy; hatching; potato root diffusate; quiescence; rearing; senescence; storage
24.  Descriptions of Deladenus albizicus n. sp. and D. processus n. sp. (Nematoda: Hexatylina) from Haryana, India 
Journal of Nematology  2015;47(1):52-59.
Two different nematodes were isolated from the bark of Albizia lebbeck trees; one from insect infested and another from noninfested, healthy tree. Based on the biological, morphological, and molecular evidences, the nematodes are described as Deladenus albizicus n. sp. and D. processus n. sp. (Nematoda: Hexatylina). Deladenus albizicus n. sp., isolated from insect-infested tree, multiplied on the fungus Nigrospora oryzae. Myceliophagous females of this nematode reproduced by parthenogenesis and spermathecae were indistinct. Infective females, readily produced in the cultures, are dorsally curved. Only one type of males containing small-sized sperms in their genital tracts were produced in the culture. Myceliophagous females: L = 0.75 to 1.71 mm, a = 32.3 to 50.8, b = 9.3 to 11.2, b’ = 5.2 to 7.3, c = 27.2 to 35.6, V = 91.0 to 93.3, c’ = 2.0 to 2.9, stylet = 11 to 12 µm, excretory pore in the region of median pharyngeal bulb, 43 to 47 µm anterior to hemizonid. Deladenus processus n. sp., isolated from bark of healthy A. lebbeck tree, was cultured on Alternaria alternata. Myceliophagous females reproduced by amphimixis and their spermathecae contained rounded sperms. Infective females were never produced, even in old cultures. Myceliophagous females: L = 0.76 to 0.99 mm, a = 34 to 49, b = 13.3 to 17.7, b’ = 3.8 to 5.8, c = 19.6 to 22.8, V = 92.2 to 93.5, c’ = 2.7 to 3.5, stylet = 6 to 7 µm, excretory pore in the proximity of hemizonid, tail conoid, tapering from both sides to a long pointed central process. It is proposed to classify Deladenus species in three groups: durus, siricidicola, and laricis groups based on female and spermatogonia dimorphism, mode of reproduction, and insect parasitism.
PMCID: PMC4388579  PMID: 25861116
Albizia lebbeck; Alternaria alternata; Beddingia; biology; COI; D2D3; Deladenus processus n. sp.; D. albizicus n. sp.; entomoparasitic nematode; ITS; Nigrospora oryzae; Phaenopsitylenchus; taxonomy
25.  Pathogenicity of Two Species of Entomopathogenic Nematodes Against the Greenhouse Whitefly, Trialeurodes vaporariorum (Hemiptera: Aleyrodidae), in Laboratory and Greenhouse Experiments 
Journal of Nematology  2015;47(1):60-66.
The greenhouse whitefly Trialeurodes vaporariorum (Hemiptera: Aleyrodidae) is a polyphagous pest in greenhouse crops. The efficacy of two entomopathogenic nematodes (EPN), Steinernema feltiae and Heterorhabditis bacteriophora, as biological control agents against T. vaporariorum was evaluated using two model crops typical of vegetable greenhouse productions: cucumber and pepper. Laboratory tests evaluated adults and second nymphal instars for pest susceptibility to different EPN species at different concentrations of infective juveniles (IJ; 0, 25, 50, 100, 150, 200, and 250 IJ per cm2); subsequent greenhouse trials against second nymphal instars on cucumber and pepper plants evaluated more natural conditions. Concentrations were applied in combination with Triton X-100 (0.1% v/v), an adjuvant for increasing nematode activity. In laboratory studies, both life stages were susceptible to infection by the two nematode species, but S. feltiae recorded a lower LC50 than H. bacteriophora for both insect stages. Similarly, in greenhouse experiments, S. feltiae required lower concentrations of IJ than H. bacteriophora to reach the same mortality in nymphs. In greenhouse trials, a significant difference was observed in the triple interaction among nematode species × concentration × plant. Furthermore, the highest mortality rate of the second nymphal instars of the T. vaporariorum was obtained from the application of S. feltiae concentrated to 250 IJ/cm2 on cucumber (49 ± 1.23%). The general mortality caused by nematodes was significantly higher in cucumber than in pepper. These promising results support further investigation for the optimization of the best EPN species/concentration in combination with insecticides or adjuvants to reach a profitable control of this greenhouse pest.
PMCID: PMC4388580  PMID: 25861117
biological control; entomopathogenic nematodes; greenhouse; insect pathology; Trialeurodes vaporariorum

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