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An intracellular bacterium, strain IAS T , was observed to infect several species of the plant-parasitic nematode genus Xiphinema ( Xiphinema astaregiense , Xiphinema incertum , Xiphinema madeirense , Xiphinema pachtaicum , Xiphinema parapachydermum and Xiphinema vallense ). The bacterium could not be recovered on axenic medium. The 16S rRNA gene sequence of IAS T was found to be new, being related to the family Burkholderiaceae, class Betaproteobacteria. Fungal endosymbionts Mycoavidus cysteinexigens B1-EB T (92.9 % sequence identity) and ' Candidatus Glomeribacter gigasporarum' BEG34 (89.8 % identity) are the closest taxa and form a separate phylogenetic clade inside Burkholderiaceae. Other genes ( atpD , lepA and recA ) also separated this species from its closest relatives using a multilocus sequence analysis approach. These genes were obtained using a partial genome of this bacterium. The localization of the bacterium (via light and fluorescence in situ hybridization microscopy) is in the X. pachtaicum females clustered around the developing oocytes, primarily found embedded inside the epithelial wall cells of the ovaries, from where they are dispersed in the intestine. Transmission electron microscopy (TEM) observations supported the presence of bacteria inside the nematode body, where they occupy ovaries and occur inside the intestinal epithelium. Ultrastructural analysis of the bacterium showed cells that appear as mostly irregular, slightly curved rods with rounded ends, 0.8-1.2 µm wide and 2.5-6.0 µm long, possessing a typical Gram-negative cell wall. The peptidoglycan layer is, however, evident only occasionally and not detectable by TEM in most cells. Another irregularly occurring shell surrounding the endosymbiont cells or the cell clusters was also revealed, probably originating from the host cell membrane. Flagella or spore-like cells do not occur and the nucleoid is diffusely distributed throughout the cell. This endosymbiont is transmitted vertically through nematode generations. These results support the proposal of IAS T as a new species, although its obligate intracellular and obligate endosymbiont nature prevented isolation of a definitive type strain. Strain IAS T is therefore proposed as representing ' Candidatus Xiphinematincola pachtaicus' gen. nov., sp. nov.

Sheath nematodes of the genus Hemicycliophora are migratory root-ectoparasites of many crops, but the damage is documented for only a few species. Hemicycliophora subbotini n. sp., isolated in Hangzhou, Zhejiang, China, from the rhizosphere of camphor tree (Cinnamomum camphora (L.) Presl), is described and characterized morphologically and molecularly with important morphological details elucidated by SEM studies. The new species can be characterized by the conoid lip region, with bulging oral disc protruding beyond the first annulus in some specimens. Oral disc narrow ovate, amphidial apertures covered by lateral plates, lying in the same plane as oral disc when observed en face. Stylet 118 (108-125) μm long and flexible, stylet knobs concave shaped having a slight cavity. The excretory pore is located 1-2 annuli posterior to the base of pharyngeal bulb. Vulval lips slightly modified with small vulval sleeve, spermatheca rounded to oblong. Anus located 18 (15-22) annuli posterior to vulva. Tail elongated, with conoid or slightly offset spike having a narrowly rounded terminus. The study provides the first description of a new Hemicycliophora species from China. Phylogenetic analyses based on D2-D3 expansion segments of the 28S and ITS fragments revealed the new species forming a separate clade from other Hemicycliophora species which supported the status as a new species of this genus.

Bacterial endosymbionts have been detected in some groups of plant-parasitic nematodes, but few cases have been reported compared to other groups in the phylum Nematoda, such as animal-parasitic or free-living nematodes. This study was performed on a wide variety of plant-parasitic nematode families and species from different host plants and nematode populations. A total of 124 nematode populations (previously identified morphologically and molecularly) were screened for the presence of potential bacterial endosymbionts using the partial 16S rRNA gene and fluorescence in situ hybridization (FISH) and confocal microscopy. Potential bacterial endosymbionts were only detected in nematode species belonging to the genus Xiphinema and specifically in the X. americanum group. Fifty-seven partial 16S rRNA sequences were obtained from bacterial endosymbionts in this study. One group of sequences was closely related to the genus 'Candidatus Xiphinematobacter' (19 bacterial endosymbiont sequences were associated with seven nematode host species, including two that have already been described and three unknown bacterial endosymbionts). The second bacterial endosymbiont group (38 bacterial endosymbiont sequences associated with six nematode species) was related to the family Burkholderiaceae, which includes fungal and soil-plant bacterial endosymbionts. These endosymbionts were reported for the first time in the phylum Nematoda. Our findings suggest that there is a highly specific symbiotic relationship between nematode host and bacterial endosymbionts. Overall, these results were corroborated by a phylogeny of nematode host and bacterial endosymbionts that suggested that there was a high degree of phylogenetic congruence and long-term evolutionary persistence between hosts and endosymbionts., (© 2016 John Wiley & Sons Ltd.)

The genus Xiphinema includes a remarkable group of invertebrates of the phylum Nematoda comprising ectoparasitic animals of many wild and cultivated plants. Damage is caused by direct feeding on root cells and by vectoring nepoviruses that cause diseases on several crops. Precise identification of Xiphinema species is critical for launching appropriate control measures. We make available the first detailed information on the diversity and distribution of Xiphinema species infesting wild and cultivated olive in a wide-region in southern Spain that included 211 locations from which 453 sampling sites were analyzed. The present study identified thirty-two Xiphinema spp. in the rhizosphere of olive trees, ten species belonging to Xiphinema americanum-group, whereas twenty-two were attributed to Xiphinema non-americanum-group. These results increase our current knowledge on the biodiversity of Xiphinema species identified in olives and include the description of four new species (Xiphinema andalusiense sp. nov., Xiphinema celtiense sp. nov., Xiphinema iznajarense sp. nov., and Xiphinema mengibarense sp. nov.), and two new records for cultivate olives (X. cadavalense and X. conurum). We also found evidence of remarkable prevalence of Xiphinema spp. in olive trees, viz. 85.0% (385 out of 453 sampling sites), and they were widely distributed in both wild and cultivated olives, with 26 and 17 Xiphinema spp., respectively. Diversity indexes (Richness, Hill´s diversity, Hill´s reciprocal of D and Hill´s evenness) were significantly affected by olive type. We also developed a comparative morphological and morphometrical study together with molecular data from three nuclear ribosomal RNA genes (D2-D3 expansion segments of 28S, ITS1, and partial 18S). Molecular characterization and phylogenetic analyses allowed the delimitation and discrimination of four new species of the genus described herein and three known species. Phylogenetic analyses of Xiphinema spp. resulted in a general consensus of these species groups. This study is the most complete phylogenetic analysis for Xiphinema non-americanum-group species to date., Competing Interests: The authors have declared that no competing interests exist.

The genus Longidorus includes a remarkable group of invertebrate animals of the phylum Nematoda comprising polyphagous root-ectoparasites of numerous plants including several agricultural crops and trees. Damage is caused by direct feeding on root cells as well as by transmitting nepoviruses that cause disease on those crops. Thus, correct identification of Longidorus species is essential to establish appropriate control measures. We provide the first detailed information on the diversity and distribution of Longidorus species infesting wild and cultivated olive soils in a wide-region in southern Spain that included 159 locations from which 449 sampling sites were analyzed. The present study doubles the known biodiversity of Longidorus species identified in olives by including six new species (Longidorus indalus sp. nov., Longidorus macrodorus sp. nov., Longidorus onubensis sp. nov., Longidorus silvestris sp. nov., Longidorus vallensis sp. nov., and Longidorus wicuolea sp. nov.), two new records for wild and cultivate olives (L. alvegus and L. vineacola), and two additional new records for wild olive (L. intermedius and L. lusitanicus). We also found evidence of some geographic species associations to western (viz. L. alvegus, L. intermedius, L. lusitanicus, L. onubensis sp. nov., L. vineacola, L. vinearum, L. wicuolea sp. nov.) and eastern distributions (viz. L. indalus sp. nov.), while only L. magnus was detected in both areas. We developed a comparative study by considering morphological and morphometrical features together with molecular data from nuclear ribosomal RNA genes (D2-D3 expansion segments of 28S, ITS1, and partial 18S). Results of molecular and phylogenetic analyses confirmed the morphological hypotheses and allowed the delimitation and discrimination of six new species of the genus described herein and four known species. Phylogenetic analyses of Longidorus spp. based on three molecular markers resulted in a general consensus of these species groups, since lineages were maintained for the majority of species. This study represents the most complete phylogenetic analysis for Longidorus species to date.

Knowledge of rRNA structure is increasingly important to assist phylogenetic analysis through reconstructing optimal alignment, utilizing molecule features as an additional source of data and refining appropriate models of evolution of the molecule. We describe a procedure of optimization for alignment and a new coding method for nucleotide sequence data using secondary structure models of the D2 and D3 expansion fragments of the LSU-rRNA gene reconstructed for fifteen nematode species of the agriculturally important and diverse family Hoplolaimidae, order Tylenchida. Using secondary structure information we converted the original sequence data into twenty-eight symbol codes and submitted the transformed data to maximum parsimony analysis. We also applied the original sequence data set for Bayesian inference. This used the doublet model with sixteen states of nucleotide doublets for the stem region and the standard model of DNA substitution with four nucleotide states for loops and bulges. By this approach, we demonstrate that using structural information for phylogenetic analyses led to trees with lower resolved relationships between clades and likely eliminated some artefactual support for misinterpreted relationships, such as paraphyly of Helicotylenchus or Rotylenchus. This study as well as future phylogenetic analyses is herein supported by the development of an on-line database, NEMrRNA, for rRNA molecules in a structural format for nematodes. We also have developed a new computer program, RNAstat, for calculation of nucleotide statistics designed and proposed for phylogenetic studies.

A new species from the family Tylenchidae was recovered in the grasslands of Isfahan province, Iran, and is herein described based on morphological and molecular data. Ottolenchus isfahanicus n. sp. is mainly characterized by having a faintly annulated cuticle; elongated, slightly sigmoid amphidial apertures developed metacorpus with distinct valve under LM; vulva located at 69.472.3% of body length; large spermatheca about 2.75 times of corresponding body width; and elongated conoid tail with broadly rounded tip. SEM observations showed that the lip region is smooth; the amphidial apertures are elongated, slightly sigmoid slits; and the lateral field is a simple band. It is further characterized by 477-to-515-μm-long females with delicate 5.7-to-6.9-μm-long stylets with small, slightly posteriorly sloping knobs, as well as functional males, in the population. The new species closely resembles O. facultativus, but is separated from it based on morphological and molecular data. It was further morphologically compared with O. discrepans, O. fungivorus, and O. sinipersici. The phylogenetic relationships of the new species with other relevant genera and species were reconstructed using near-full-length sequences of small subunit and D2-D3 expansion segments of large subunit (SSU and LSU D2-D3). In the inferred SSU phylogeny, the newly generated sequence of Ottolenchus isfahanicus n. sp. formed a clade with two sequences of O. sinipersici and sequences assigned to O. facultativus and O. fungivorus. In the inferred LSU phylogeny, the three newly generated sequences of the new species and LSU sequences of O. sinipersici and O. discrepans formed a clade.

During a recent inventory survey of the nematofauna of tea plantation at Zhejiang Province, China, Discocriconemella parasinensis n. sp. was detected in the rhizosphere of Camellia sinensis. The new species can be characterized by having the uninterrupted rounded labial disc, en face view showing rectangular-rounded labial plate without submedian lobes, R = 82.6 (80–86), Rex = 22 (21–23), stylet length of 68.3 (59–76) μm, excretory pore located 1–2 annuli posterior to the esophageal bulb, vulva open, postvulval body elongated conoid, and tail conoid with bilobed terminus. Morphologically, the species shares the same lip-type with D. discolabia, D. mauritiensis, D. mineira, D. perseae, and D. sinensis. Phylogenetic relationships of the new species based on D2–D3 expansion segments of 28S, ITS, and 18S rRNA genes revealed that D. parasinensis n. sp. formed a separated clade from other criconematid species, thereby supporting its status as a new species of the genus. The new species showed close phylogenetic relationships with Criconemoides geraerti.

Summary: A population of Xiphinema artemisiae was recovered from Iran, representing the first report after its original description from a meadow pasture in northern Caucasus, Russia. The Iranian population was recovered from the rhizosphere of grasses in the Damavand region, Tehran province. The females of the recovered populations are characterised by a lip region separated from the rest of body by a depression, a 5.1-5.6 mm long body, a 129-138 μ m long odontostyle, 76-85 μ m long odontophore, a uterus having a pseudo-Z-organ and crystalloid bodies observed in some specimens, and a short, rounded, dorsally more convex tail with a subcentral mucron, four juvenile developmental stages (the first stage was not recovered) and functional males. The Iranian population is morphologically and morphometrically similar to the type population and minor morphological differences can be attributed to geographical interpopulation differences. Molecular phylogenetic relationships of the Iranian population of X. artemisiae with other species were reconstructed using sequences of three genomic ribosomal markers, viz. small, large subunit D2-D3 and internal transcribed spacer (SSU, LSU D2-D3 and ITS) rDNA. [ABSTRACT FROM AUTHOR]

Summary: Cyst-forming nematodes of the genus Heterodera are highly derived and economically important plant parasites. The Schachtii group of this genus is one of the largest ones with a total of 18 species parasitising dicotyledons. In this study, we provided comprehensive phylogenetic analyses of several hundred COI and ITS rRNA gene sequences of selected species from the Schachtii group, including H. betae , H. cajani , H. ciceri , H. galeopsidis , H. glycines , H. medicaginis , H. mediterranea , H. schachtii , H. sonchophila and H. trifolii , using Bayesian inference, maximum likelihood, and statistical parsimony. One hundred and twenty four new COI , 57 ITS rRNA and eight hsp90 gene sequences from 81 nematode populations collected in 19 countries were obtained in this study. Our study showed that the ITS rRNA gene has limited discrimination power compared to the COI gene. However, our analysis also revealed that partial COI gene sequences were identical for H. trifolii , H. betae and H. galeopsidis. Based on the results of phylogeographical analysis and age estimation of clades with a molecular clock approach, it was hypothesised that the majority of the Schachtii group species originated and diversified in the Mediterranean Basin biodiversity hotspot during the Pleistocene and then dispersed from this region across the world. The Sino-Japanese Floristic Region is likely one of the centres of diversification for the soybean cyst nematode, which showed distinct population structure. The possible role of hybridisation and polyploidisation in the evolution of species of the Schachtii group is discussed. [ABSTRACT FROM AUTHOR]

Three populations of Xiphinema non-americanum group species were detected in agricultural and natural ecosystems, during routine surveys for plant-parasitic nematodes in Spain. Based on morphological and molecular analyses, the species were identified as Xiphinema histriae and Xiphinema lapidosum, being this the first record and molecular characterization of both species in Spain. The morphometrics and morphology of the Spanish populations agree with those of the original description and paratype specimens and the present study provided a first description of the second to fourth juvenile stages of both species. A detailed study on the morphology in the Spanish populations of X. histriae, as well as in paratypes, showed a pseudo-Z-organ with weakly muscularized wall and containing numerous small dense granular bodies, which was different to the original description by Lamberti et al. (1993). This new finding suggests that X. histriae must be considered a member of the morphospecies Group 5 of X. non-americanum. Phylogenetic analysis based on D2 to D3 expansion segments of 28S gene, ITS1 and partial CoxI gene indicated that X. histriae and X. lapidosum are phylogenetically related with other Xiphinema non-americanum group spp. reported from Spain. Considering the pathological and economic importance of this group of nematodes, the combination of morphological characters, measurements, and molecular analysis is crucial for accurate identification of these species.

Summary: The study aims to provide the morphological and molecular characterisation of two newly discovered Cryptaphelenchus species, namely: C. curvatum n. sp. and C. tumidus n. sp. Morphologically both species are minute nematodes, possess medium-sized stylets with basal knobs laterally directed, and short post-uterine sacs (PUS). Cryptaphelenchus curvatum n. sp. was detected from the local Pinus branch samples and is characterised by having an equal proportion of adult males and females, medium-sized body lengths of females 353 (320-388) μ m and males 277 (256-293) μ m, lateral fields with four lateral lines, 8-10 μ m long stylet, PUS about one vulva body width long and conical posterior body end with a blunt pointed tip. Cryptaphelenchus tumidus n. sp. was detected in a packaging wood sample from Japan and is characterised by having rare male, short body lengths of females 236 (217-270) μ m and males 180 μ m, obscure lateral fields, stylet 6.8 (6.0-7.5) μ m, PUS about half vulva body width long and a slim conical posterior body end with a finely rounded tip. The phylogenetic trees were also constructed based on rRNA gene sequences (D2-D3 segments of the 28S, ITS and 18S) and discussed in detail. Since Cryptaphelenchus species offer multitrophic relationships among their insect vectors and associated microbiota, species recognition and molecular studies of recovered populations will facilitate researchers to determine whether these species might play any ecological roles in insect bionomics or the residing host tree. [ABSTRACT FROM AUTHOR]

Bacteria of the genus "Candidatus Cardinium" and related organisms composing the Cardinium clade are intracellular endosymbionts frequently occurring in several arthropod groups, freshwater mussels and plant-parasitic nematodes. Phylogenetic analyses based on two gene sequences (16S rRNA and gyrB) showed that the Cardinium clade comprised at least five groups: A, B, C, D and E. In this study, a screening of 142 samples of plant-parasitic nematodes belonging to 93 species from 12 families and two orders using PCR with specific primers and sequencing, revealed bacteria of Cardinium clade in 14 nematode samples belonging to 12 species of cyst nematodes of the family Heteroderidae. Furthermore, in this study, the genome of the Cardinium cHhum from the hop cyst nematode, Heterodera humuli, was also amplified, sequenced and analyzed. The comparisons of the average nucleotide identity (ANI) and digital DNA–DNA hybridization (dDDH) values for the strain Cardinium cHhum with regard to related organisms with available genomes, combined with the data on 16S rRNA and gyrB gene sequence identities, showed that this strain represents a new candidate species within the genus "Candidatus Paenicardinium". The phylogenetic position of endosymbionts of the Cardinium clade detected in nematode hosts was also compared to known representatives of this clade from other metazoans. Phylogenetic reconstructions based on analysis of 16S rRNA, gyrB, sufB, gloEL, fusA, infB genes and genomes and estimates of genetic distances both indicate that the endosymbiont of the root-lesion nematode Pratylenchus penetrans represented a separate lineage and is designated herein as a new group F. The phylogenetic analysis also confirmed that endosymbionts of ostracods represent the novel group G. Evolutionary relationships of bacterial endosymbionts of the Cardinium clade within invertebrates are presented and discussed. [ABSTRACT FROM AUTHOR]

Summary: Hemicycliophora species are migratory root ectoparasites of higher plants, commonly known as sheath nematodes. The present study describes a new species, Hemicycliophora geraerti n. sp., recovered from the rhizosphere of peach tree (Prunus persica) in Zhejiang Province, China. This species is characterised by a cuticular sheath loosely fitting to the body, a lip region slightly set-off with body contour bearing 3-4 annuli, oral plate narrow, ovate, and non-protruding, amphidial apertures plugged, dorsoventrally flanked in the first lip annulus, stylet 110 (103-116) μ m long and flexible, excretory pore located 4-7 annuli posterior to the base of the pharyngeal bulb; vulval lips modified with a small vulval sleeve, anus located 16 (13-21) annuli posterior to vulva, tail elongated, having a narrowly rounded slightly offset spike. The molecular characterisation and phylogenetic analyses based on D2-D3 expansion segments of the 28S rRNA, ITS rRNA, and COI mtDNA gene indicated the unique status and support the novelty of this population of Hemicycliophora , therefore herein described as H. geraerti n. sp. [ABSTRACT FROM AUTHOR]

Summary: Specimens of a thin longidorid species collected in Peñalba (Huesca), north-west Spain, were previously described as Paralongidorus iberis. However, we conclude, through scanning electron microscopy and molecular studies on a population from about 15 km from the type locality and on paratype specimens, that this species was originally placed in the wrong genus. Both populations have pore-like amphidial apertures, not slit-like as in Paralongidorus , and the species is therefore transferred to Longidorus. Longidorus iberis n. comb. is regarded as a valid species and is clearly different from closely related species such as L. tabernensis , L. iliturgiensis , L. alvegus and L. indalus in morphometrics and molecular markers. Molecular data are reported for the first time, including the D2-D3 expansion segments of 28S rRNA, ITS1 rRNA, partial 18S rRNA and partial mitochondrial coxI regions. These molecular markers were used for inferring the phylogenetic relationships with other species within Longidorus and Paralongidorus , all clearly separating L. iberis n. comb. from other related taxa and placing the species in the Longidorus clade, rather than with Paralongidorus. [ABSTRACT FROM AUTHOR]

Summary: Two new and one known species of Atetylenchus recovered from Dezful, Khuzestan Province, south-western Iran, are illustrated and described using morphological, morphometric and molecular data. Atetylenchus dezfuliensis n. sp. is characterised by medium-sized body 700-998 μ m long, stylet 10.0-11.5 μ m long, characteristic tail shape, first tapering regularly but then cylindrical to slightly clavate and 90-136 μ m long, phasmids located at one-third anterior part of tail and short spicules 18.7 μ m long. Atetylenchus longilabiatus n. sp. is characterised by small-sized body 593-720 μ m long, prominent lip region, stylet 9.5-12.5 μ m long, conical tail with hook-shaped terminus and 42-63 μ m long, phasmids located near mid-tail and short spicules 17.5 μ m long. Atetylenchus cf. graminus is characterised by small-sized body 643-787 μ m long, stylet 10.7-11.7 μ m long, vulva located at 51.7-55.1% of the body length, and conical tail with finely rounded end and 63-69 μ m long. Molecular data are provided for the characterisation of this genus from these two new and one known species using ribosomal genes (18S and 28S rRNA). This genus is molecularly separated from Psilenchus , but closely related in both genes. These molecular markers showed different phylogenies for both genera; partial 18S showed their relationship with Tylenchidae, while 28S rRNA showed their relationship with Merliniidae. A key to the genus is provided for species identification. [ABSTRACT FROM AUTHOR]

Summary: Sigmolenchus n. gen., is proposed as a new member of the Tylenchidae. The new genus is proposed based upon light and scanning electron microscopic (LM and SEM) observations and phylogenetic analyses based upon small, large and internal transcribed spacer sequences of ribosomal DNA (SSU, LSU D2-D3, ITS rDNA) and mitochondrial cytochrome c oxidase I gene (COI mtDNA) sequences. It is characterised partly by its slender body, but mainly by having a dorso-ventrally flattened, smooth and elevated cephalic region, unique elongate sinuous amphidial openings, each appearing as a wavy slit with three peaks, originating from the vicinity of the labial plate, extending posteriorly and reaching and passing the first body annulus, and a spacious amphidial fovea. It has an anteriorly rounded outline under LM, knobbed stylet, its conus comprising about one-third of the total length, lateral field with a single band, metacorpus with a distinct valve, vulva with small lateral flaps, and males common with tylenchoidiform spicules. The new genus is morphologically compared with seven tylenchid genera, viz., Chilenchus , Ecphyadophoroides , Filenchus , Labrys , Lelenchus , Sakia , and Tenunemellus , mainly by having small slender body and dorso-ventrally flat cephalic region, and is placed under the subfamily Ecphyadophorinae because of its slender body, elevated smooth cephalic region, long amphidial openings and spacious amphidial fovea. It was recovered from marine muddy sediments taken 2 m deep in mangrove forests off southern Iran. Currently S. sinuosus n. gen., n. sp. is monotypic. In SSU phylogeny, the new genus formed an unsupported sister relationship with Filenchus discrepans , and in LSU phylogeny it occupied a placement inside a major clade including several Tylenchinae genera. [ABSTRACT FROM AUTHOR]

The tea plant is native to China; the country has the greatest tea production areas in the world. In an attempt to investigate the nematode biodiversity associated with the tea plantations of Hangzhou, Zhejiang Province, a population of stunt nematode was detected. This group of nematodes is comprised of migratory ecto-parasites of roots and can subsist on a variety of host plants. Therefore, the detected population was studied carefully using the integrative taxonomy approach and identified as a new species of genus Geocenamus. Geocenamus chengi n. sp. can be characterized by females having six incisures in the lateral field; labial region is dome shaped and slightly offset from the rest of the body having four to five annuli; head framework is weakly developed; deirids are absent; excretory pore is located at the anterior region of basal pharyngeal bulb. Under SEM, the vulva is a transverse slit, vulval lips are elongated and ellipsoidal with epiptygma. The tail is annulated, elongated, and conical having bluntly pointed tip and a terminal hyaline region that forms 21 to 33% of the tail length. Spicule is 22 to 25 μm long, gubernaculum is saucer shaped; bursa is crenated covering the tail until the hyaline tail region. Morphologically, the species is close to G. circellus, G. joctus, G. loofi, G. ordinarius, G. processus, G. tetyllus, and G. tortilis. Phylogenetic relationships of the new species based on D2-D3 expansion domains of 28 S, ITS, and 18 S rRNA genes indicated that G. chengi n. sp. clustered in a separate clade with G. vietnamensis. [ABSTRACT FROM AUTHOR]

The Family Criconematidae is commonly referred as ring nematodes that include some members with economic importance as plant parasites. During a recent nematode inventory survey at Zhejiang Province, China, a new species of genus Nothocriconemoides was detected in the rhizosphere of elm tree. Nothocriconemoides hangzhouensis n. sp. can be characterized by the female body having annuli with fine longitudinal striations and 2 to 3 anastomoses at the posterior half of the body. The first cephalic annulus is rounded and expanded enclosing the lip region, and the second annulus is narrow, offset, collar like. En face view shows a central elevated labial disk bearing four distinct equal-sized submedian lobes and “I” shaped oral aperture. Excretory pore is located 3–4 annuli posterior to esophageal bulb. Vagina is straight and vulva closed. The ventral side of postvulval annuli is inverted, in majority of individuals. Anus is indistinct and located on the next annuli posterior to vulva. Tail is short, conoid, with forked or branched terminus. Juveniles are devoid of collar-shaped annuli in the lip region. The cephalic region has two rounded annuli where the first annulus shows slight depression in the middle. Body annuli are finely crenated. Anus is indistinct and located 3 to 4 annuli from tail terminus. Tail is short ending in a single lobed terminus. Phylogenetic studies based on analysis of the D2–D3 expansion segments of the 28 S rRNA, ITS rRNA, partial 18 S rRNA, and coxI gene revealed that the new species formed a separate clade from other criconematid species, thereby supporting its status as a new species of the genus. The new species showed close relationships with Discocriconemella sinensis. Additionally, this is the first record of genus Nothocriconemoides from China. [ABSTRACT FROM AUTHOR]

Summary: Populations of Hemicriconemoides chitwoodi , H. fujianensis , H. parasinensis , and Criconemoides myungsugae isolated in China from the rhizosphere soil of woody perennials were characterised molecularly, important morphological details being elucidated by SEM observations. The morphometric data of the Chinese populations were compared with all previously reported populations. The SEM observations of en face views indicated that H. chitwoodi and H. parasinensis have lip patterns belonging to type 1 and phylogenetically both species clustered with other Hemicriconemoides species that showed the same kind of lip pattern. On the other hand, H. fujianensis showed a lip pattern belonging to type 2 and phylogenetically appears basal to the above-mentioned clade. SEM observation of C. myungsugae showed that the first lip annulus forms a non-projecting uninterrupted disc and the labial annulus is a rectangular shape with slight dorsal and ventral indentations. Phylogenetic relationships among Criconemoides spp. are apparently not well resolved. The present study provides updated morphological descriptions, molecular diagnostics and phylogenetic relationships of H. chitwoodi , H. fujianensis , H. parasinensis , and C. myungsugae , the last species being the first report from China. [ABSTRACT FROM AUTHOR]

Three populations of Xiphinema non- americanum group species were detected in agricultural and natural ecosystems, during routine surveys for plant-parasitic nematodes in Spain. Based on morphological and molecular analyses, the species were identified as Xiphinema histriae and Xiphinema lapidosum, being this the first record and molecular characterization of both species in Spain. The morphometrics and morphology of the Spanish populations agree with those of the original description and paratype specimens and the present study provided a first description of the second to fourth juvenile stages of both species. A detailed study on the morphology in the Spanish populations of X. histriae, as well as in paratypes, showed a pseudo- Z-organ with weakly muscularized wall and containing numerous small dense granular bodies, which was different to the original description by Lamberti et al. (1993). This new finding suggests that X. histriae must be considered a member of the morphospecies Group 5 of X. non-americanum. Phylogenetic analysis based on D2 to D3 expansion segments of 28S gene, ITS1 and partial CoxI gene indicated that X. histriae and X. lapidosum are phylogenetically related with other Xiphinema non-americanum group spp. reported from Spain. Considering the pathological and economic importance of this group of nematodes, the combination of morphological characters, measurements, and molecular analysis is crucial for accurate identification of these species. [ABSTRACT FROM AUTHOR]

Females and juveniles from a population of Xiphinema ifacolum from Sri Lanka are described based on morphology, morphometrics and molecular analyses. Morphologically, females and juveniles from Sri Lanka are similar to original descriptions and other reports from Brazil, Cameroon, Liberia, and São Tomé and Príncipe. The identity of the species was also confirmed by 18S rRNA gene sequences deposited in NCBI from Brazil (AY297826). Integrative diagnosis was completed with molecular data using D2-D3 expansion segments of 28S rRNA, ITS1 region, partial 18S-rRNA and the partial mitochondrial gene cytochrome c oxidase subunit 1 (coxI). This is the third molecular characterisation for a species of the X. non- americanum Group 4, after X. oleae and X. tica. The use of different ribosomal and mitochondrial markers in this study, particularly, D2-D3, ITS1 and partial coxI , provided a precise and unequivocal tool for the identification of X. ifacolum and contributes to a better knowledge of the diversity within Xiphinema. Morphospecies Group 4 appears to be a paraphyletic group within the X. non- americanum assemblage. [ABSTRACT FROM AUTHOR]

Longidorids are economically important plant-parasitic nematodes because several species are virus vectors. Populations of Paralongidorus sali and Longidorus jonesi, isolated from woody perennials of Hangzhou, Zhejiang, China, were characterized molecularly and morphologically. The morphometric data of the Chinese populations of both species were compared with other populations. The present study provided a first record of the occurrence of Paralongidorus in China coupled with description of the first-stage juvenile and male of L. jonesi. Phylogenetic analysis based on 18S and D2-D3 expansion segments of 28S gene indicated that L. jonesi clustered with L. jonesi reported from Japan and P. sali grouped with P. bikanerensis from Iran. Considering the pathological and economic importance of this group of nematodes, the study emphasized the need of updated descriptions from accurately identified specimens, isolation of sufficient material for examination, and molecular and phylogenetic analysis for a better understanding and diagnostics of Longidorid nematodes. [ABSTRACT FROM AUTHOR]

Sheathoid nematodes of the genus Hemicriconemoides are migratory root-ectoparasites of many crops but damage is documented for only a few species. Hemicriconemoides paracamelliae sp. n., isolated in Hangzhou, Zhejiang, China, from the rhizosphere of Cinninghamia lanceolata, is described together with H. kanayaensis and H. parataiwanensis. These three species are characterised morphologically and molecularly with important morphological details elucidated by SEM photographs. The new species can be characterised by the en face view having a dorsoventrally orientated oral disc with slit-like opening, and the labial plate being composed of two lateral semi-globular shaped projections on the lateral sides of the oral disc. This new species has the first lip annulus expanded and slightly wider than the second, stylet 83 (80-85) µm long, excretory pore located 5-6 annuli posterior to the base of the pharyngeal bulb, vulva slit-like, vulval flaps absent, spermatheca rounded to oblong-shaped, and anus located 5-8 annuli posterior to vulva. The tail is elongated, conoid, with the terminal annuli curving dorsally or ventrally to form a finely rounded tip. The study provides the first record of H. kanayaensis from mainland China, a new host association of H. parataiwanensis, and molecular sequencing data of the 18S, 28S D2-D3 and ITS sequences. [ABSTRACT FROM AUTHOR]

A new and a known longidorid nematode, Paralongidorus lusitanicus n. sp. and Paralongidorus plesioepimikis, are described and illustrated from populations extracted from soil associated with grapevine (Vitis vinifera L.) from Escaroupim and Pó (central-Western Portugal), respectively. The new needle nematode P. lusitanicus n. sp. is characterised by a very large body size (8072-12,022 μm), an expanded and rounded lip region, ca 30 μm wide, with a clear constriction followed by a depression posterior to the amphidial aperture, amphidial fovea very large (11.0-19.0 μm), stirrup-shaped, with conspicuous slit-like aperture as shown in scanning electron microscopy studies, a very long and flexible odontostyle (180.0-223.0 μm), guiding ring located at 28.0-41.5 μm from anterior end, vulva anterior to the mid-body (34-41%), a dorsally convex-conoid tail with rounded terminus (29-42 μm long), bearing two or three pairs of caudal pores and males common (ratio 1:1.6 females) with spicules ca 80 μm long. Morphological and morphometric traits for P. plesioepimikis fit well with the original description, and is reported for the first time in Portugal. Integrative diagnosis of both species was completed with molecular data obtained using D2-D3 expansion segments of 28S rDNA, ITS1-rDNA and partial 18S-rDNA. The phylogenetic relationships of these species with other Paralongidorus spp. using these three molecular markers indicated that P. lusitanicus n. sp. clustered together with other Paralongidorus spp. forming a sister clade with P. plesioepimikis, both of them sharing a large body, long odontostyle, an anteriorly located vulva and an expanded and rounded lip region with a clear constriction followed by a depression posterior to the amphidial aperture. [ABSTRACT FROM AUTHOR]

Hemicycliophora presently contains 132 valid species of sheath nematodes. Within several samples obtained from surveys in Canada, South Africa, Spain and the USA, we distinguished six valid and six putative unidentified species by integrating the results of morphological and molecular analyses. Valid species included: H. californica, H. gracilis, H. parvana, H. poranga, H. raskii, and H. signata. The putative unidentified species were indicated as Hemicycliophora sp. 10, sp. 12, sp. 15, sp. 16, sp. 17, and sp. 18. Two new species of sheath nematodes from Spain and the USA were described and named as H. onubensis sp. n. and H. robbinsi sp. n., respectively. Hemicycliophora wyei is proposed as a junior synonym of H. parvana and H. ripa is proposed as a junior synonym of H. poranga. Eighteen valid and 13 unidentified species of sheath nematodes were characterised using the partial COI mtDNA gene. A total of 94 new sequences of which 77 were for the COI mtDNA gene were obtained in this study. Phylogenetic relationships within Hemicycliophora, using the D2-D3 expansion segments of 28S rDNA, ITS rRNA and COI gene sequences, are presented as inferred from Bayesian analysis. [ABSTRACT FROM AUTHOR]

A new dagger nematode, Xiphinema poasense n. sp., is described and illustrated from three populations extracted from soil associated with a combined plantation of Eucalyptus sp., Cupressus sp. and Pennisetum sp. and wild plants from a tropical premontane forest in Costa Rica. The new dagger nematode is characterised by a moderate body size 2612 (2416-3042) µm long, a rounded lip region 15.0 (13.5-16.5) µm broad, separated from the body contour by a shallow depression, amphidial fovea large, stirrupshaped, a very long odontostyle (175 (164-188) µm), stylet guiding ring located 167 (136-181) µm from anterior end, vulva situated anterior to mid-body (36-40%), anterior genital branch complete but strongly reduced, without uterine differentiation, female tail short, hemispherical to convex-conoid with a c' ratio = 0.7 (0.6-0.8) and bearing two pairs of caudal pores, and male absent. Integrative diagnosis was completed with molecular data using D2-D3 expansion segments of 28S rRNA, ITS1 region, partial 18S-rRNA and the partial mitochondrial gene cytochrome c oxidase subunit 1 (coxI). The phylogenetic relationships based on D2-D3 segments of this species with other Xiphinema spp. of the X. non-americanum group indicated that X. poasense n. sp. clustered with other species with a reduced anterior genital branch from the morphospecies Group 2, viz., X. costaricense and X. krugi. However, the phylogeny of coxI and partial 18S rRNA gene revealed that the new species did not cluster with Xiphinema species having the anterior genital branch absent or reduced (i.e., morphospecies Groups 1 and 2, respectively). [ABSTRACT FROM AUTHOR]

The reniform nematodes of the genus Rotylenchulus are semi-endoparasites of numerous herbaceous and woody plant roots and distributed in regions with Mediterranean, subtropical and tropical climates. In this study, we provide morphological and molecular characterisation of three out of 11 valid species of the genus Rotylenchulus: R. macrodoratus, R. macrosoma, and R. reniformis from Greece (Crete), Italy and Spain. The overall prevalence of reniform nematodes in wild and cultivated olives in Greece, Italy, and Spain was 11.5%, 19.0% and 0.6%, respectively. In Greece, R. macrodoratus and R. macrosoma were detected in cultivated olive with a prevalence of 8.2% and 6.2%, respectively, but none of them were found in wild olive. This is the first report of R. macrosoma in Greece. Only one reniform nematode species was detected in olive from Italy and Spain, viz. R. macrodoratus and R. macrosoma, respectively. The parasitism of R. macrosoma on hazelnut in northern Spain was also confirmed for the first time. This study demonstrates that R. macrodoratus and R. macrosoma have two distinct rRNA gene types in their genomes, specifically the two types of D2-D3 for R. macrosoma and R. macrodoratus, the two types of ITS for R. macrodoratus and the testing of the ITS variability in other R. macrosoma populations in different countries. Rotylenchulus macrosoma from Greece and Spain showed differences in nucleotide sequences in the ITS region and D2-D3 of 28S rRNA gene. [ABSTRACT FROM AUTHOR]

Radopholoides japonicus n. sp. isolated from rhizospheric soil samples associated with Podocarpus macrophyllus imported from Japan into Ningbo Port of China is described and illustrated. It is characterised by four lateral lines, lip region low, convex with three annuli. Stylet 16.5-18.5 µm long with anteriorly sloping rounded knobs. Excretory pore at the level of pharyngo-intestinal junction. Vulva posteriorly located, vulval lips not protruding, spermatheca squarish. Post-vulval uterine sac 35-44 µm long. Tail conoid with broadly rounded terminus, phasmid 6-8 annuli posterior to anus. Among five species of genus Radopholoides, the new species is close to R. antoni but can be differentiated from it by lip and tail morphology. This is the first Radopholoides species that provided detailed morphological and molecular characterisation. Maximum Likelihood analysis using the Shimodaira-Hasegawa test for the validity of Radopholoides was performed and showed the validity of the genus using the partial 18S and D2-D3 expansion segments of 28S. [ABSTRACT FROM AUTHOR]