Aphelenchus yinyuensis n. sp. (Tylenchina: Aphelenchoididae) found in Terminalia sp. in China.

During a nematode biodiversity survey in Hainan Province, China, Aphelenchus yinyuensis n. sp. was detected in the rhizosphere of Terminalia sp. It is characterized by medium-sized a body of adult nematodes, i.e. 793 (639-877) µm and 756 (647-863) µm for females and males, respectively, with low, rounded, not offset lip region. The lateral field has 10 incisures. The excretory pore is located posterior to the nerve ring. The vagina is not sclerotised and the vulva has simple lips without a flap. The PUS is well developed and forms ca 45 to 83% of the vulva to anus distance. Female tail is straight, cylindrical, ca 2.7 times longer than anal body diam, tail tip broad, and bluntly rounded. Males have four pairs of caudal papillae and spicule 28.7 (25.8-32.3) µm long in the chord and well developed bursa, extending to the tail tip. 18S and 28S rRNA phylogenetic analyses were performed for the new species, and the ITS analysis was not performed due to low posterior probability support. Phylogenetically, the new species grouped with Aphelenchus avenae and this is the first new Aphelenchus species ever described from China.

Description of Laimaphelenchus sinensis n. sp. (Nematoda: Aphelenchoididae) from declining Chinese pine, Pinus tabuliformis in Beijing, China.

Laimaphelenchus sinensis n. sp. isolated from declining Chinese pine, Pinus tabuliformis Carrière, is described and characterized morphologically and molecularly. The new species has four incisures in the lateral field and the excretory pore situated posterior to the nerve ring; the female has a vulval flap and vaginal sclerotization is quite prominent in majority of specimens. The female tail is conoid, ventrally curved having a single stalk-like terminus with 8 to 10 projections. The male spicules are 14.0 (13.2-15) µm long along curved median line and tail is ventrally curved typical of the genus; however, the projections are less prominent as compared to those of female. The male has two pairs of caudal papillae and Bursa is absent. Phylogenetically, the ribosomal DNA sequences of the new species placed it within Laimaphelenchus clade and are morphologically similar to L. persicus, L. preissii, L. simlaensis and L. unituberculus.Laimaphelenchus sinensis n. sp. isolated from declining Chinese pine, Pinus tabuliformis Carrière, is described and characterized morphologically and molecularly. The new species has four incisures in the lateral field and the excretory pore situated posterior to the nerve ring; the female has a vulval flap and vaginal sclerotization is quite prominent in majority of specimens. The female tail is conoid, ventrally curved having a single stalk-like terminus with 8 to 10 projections. The male spicules are 14.0 (13.2-15) µm long along curved median line and tail is ventrally curved typical of the genus; however, the projections are less prominent as compared to those of female. The male has two pairs of caudal papillae and Bursa is absent. Phylogenetically, the ribosomal DNA sequences of the new species placed it within Laimaphelenchus clade and are morphologically similar to L. persicus, L. preissii, L. simlaensis and L. unituberculus.

Description of Seinura italiensis n. sp. (Tylenchomorpha: Aphelenchoididae) found in the medium soil imported from Italy.

Seinura italiensis n. sp. isolated from the medium soil imported from Italy is described and illustrated using morphological and molecular data. The new species is characterized by having short body (477 (407-565) µm and 522 (469-590) µm for males and females, respectively), three lateral lines, stylet lacking swellings at the base, and excretory pore at the base or slightly anterior to base of metacorpus; females have 58.8 (51.1-69.3) µm long post-uterine sac (PUS), elongate conical tail with its anterior half conoid, dorsally convex, and ventrally slightly concave and the posterior half elongated, narrower, with finely rounded to pointed tip and males having seven caudal papillae and 14.1 (12.6-15.0) µm long spicules. Morphologically, the new species is similar to S. caverna, S. chertkovi, S. christiei, S. hyrcania, S. longicaudata, S. persica, S. steineri, and S. tenuicaudata. The differences of the new species with aforementioned species are discussed. In molecular phylogenetic analyses using near full-length small and large subunit ribosomal DNA (SSU and LSU rDNA D2-D3 expansion segments) sequences, the new species fell into a clade including three previously described/sequenced species of the genus in both SSU and LSU Bayesian phylogenetic trees.Seinura italiensis n. sp. isolated from the medium soil imported from Italy is described and illustrated using morphological and molecular data. The new species is characterized by having short body (477 (407-565) µm and 522 (469-590) µm for males and females, respectively), three lateral lines, stylet lacking swellings at the base, and excretory pore at the base or slightly anterior to base of metacorpus; females have 58.8 (51.1-69.3) µm long post-uterine sac (PUS), elongate conical tail with its anterior half conoid, dorsally convex, and ventrally slightly concave and the posterior half elongated, narrower, with finely rounded to pointed tip and males having seven caudal papillae and 14.1 (12.6-15.0) µm long spicules. Morphologically, the new species is similar to S. caverna, S. chertkovi, S. christiei, S. hyrcania, S. longicaudata, S. persica, S. steineri, and S. tenuicaudata. The differences of the new species with aforementioned species are discussed. In molecular phylogenetic analyses using near full-length small and large subunit ribosomal DNA (SSU and LSU rDNA D2-D3 expansion segments) sequences, the new species fell into a clade including three previously described/sequenced species of the genus in both SSU and LSU Bayesian phylogenetic trees.

A new rare nematode Nothocriconemoides hangzhouensis n. sp. (Nematoda: Criconematidae) from Hangzhou, China.

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.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 3­4 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 D2­D3 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.

A new stunt nematode, Geocenamus chengi n. sp. (Nematoda: Merliniinae) in the rhizosphere of tea (Camellia sinensis) from Zhejiang Province, China.

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.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.

Description of Ektaphelenchus koreanus n. sp. (Nematoda: Ektaphelenchidae) with morphometrical notes on the Ektaphelenchus species.

Ektaphelenchus koreanus n. sp. is isolated from Pinus packaging wood from Korea in Ningbo customs. The new species can be characterized by having four incisures in the lateral field, the excretory pore located posterior to nerve ring, female devoid of vulval flap, and having long post-vulval intestinal sac (55-106 µm), tail with a finely rounded terminus. Six caudal papillae in male and spicule short (12.7-13.7 µm), having broad squared to rounded condylus, triangular rounded rostrum, cucullus absent. Morphologically the species is most similar with E. berbericus, E. joyceae, E. oleae, E. ibericus, and E. taiwanensis but it can be differentiated by plenty of morphometrical and morphological characters. In addition, a morphometry table of Ektaphelenchus is also presented.

Updated description of Paratylenchus lepidus Raski 1975 and P. minor Sharma, Sharma and Khan, 1986 by integrating molecular and ultra-structural observations.

Two populations of Paratylenchus lepidus and P. minor were detected in the rhizosphere of Elaeocarpus sp. and Chinese red pine from Taizhou and Hangzhou, Zhejiang Province, China. Previously, P. lepidus has been reported from China whereas P. minor was originally described from India decades ago in the rhizosphere of peach but was never reported thereafter. In this study, both species were characterized morphologically and molecularly coupled with SEM observations. Morphologically, both species have four incisures in the lateral field, vulval present (SEM observations), stylet less than 30 µm long and cephalic region without submedian lobes. Phylogenetically, both species grouped with paratylenchid species having short stylets. Both species can be differentiated from each other by the shape of lip region (rounded in P. lepidus and narrow truncated in P. minor) and tail terminus (pointed in P. lepidus and a broadly rounded in P. minor) and several morphomemtrical values.The study provided an updated description of P. lepidus and P. minor and a first record of P. minor from China. In addition, both species are the first paratylenchid species recorded from Elaeocarpus sp. and Pinus tabuliformis, respectively.

Description of Xiphinema parachambersi sp. n. (Nematoda: Longidoridae) from Imported Ornamental Plants in Japan with a Key to Xiphinema Species in Group 1.

Xiphinema parachambersi n. sp. recovered in Ningbo, China, from the rhizosphere of ornamental plants ( Gardenia jasminoides and Euonymus hamiltonianus ) imported from Japan is described. The new species is characterized by a long female body 1,830 to 2,109 µ m long, odontostyle 105 to 116 µ m long, reproductive system mono-opisthodelphic, vulva located anteriorly at 25.2 to 27.7% of total body, long ovary 119 to 292 µ m with simple uterus and Z-organ absent, female tail elongated conoid with a rounded terminus. Only two juvenile stages were available to study and no male was found. The polytomous identification codes for this new species are A1, B4, C2, D23, E1, F2, G2, H2, I2, J2, K?, L1 and it belongs to the morphospecies group 1. Phylogenetic analysis based on the 18S, ITS1 and 28S D2/D3 sequences of the new species showed close relationships with X. chambersi . Morphologically, the new species is similar to X. chambersi , X. hangzhouense , and X. winotoi but can be differentiated by morphological characters and DNA sequences. To help identify the species, a diagnostic key to the group 1 species is presented.

Morphological and Molecular Characterization of Paralongidorus sali Siddiqi, Hooper, and Khan, 1963 with a Description of the First-Stage Juvenile and Male of Longidorus jonesi Siddiqi, 1962 from China.

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.

Description of Gracilacus paralatescens n. sp. (Nematoda:Paratylenchinae) found from the rhizosphere of Bamboo in Zhejiang, China.

Gracilacus paralatescens n. sp., isolated in Anji County, Zhejiang Province, China from the rhizospheric soil of bamboo. The new species can be characterized by the female lateral field with three incisures, stylet 71.5 to 78.8 µm long, lip region truncated, excretory pore located anterior to basal knobs. Vulval lips non-protruding and without vulval flap, spermatheca large, elongated squarish shaped filled with sperms. Tail slender, relatively straight having wedge shape terminus. The spicule slender, slightly curved and 17.5 to 18.9 µm long. In the phylogenetic analysis based on 18S, D2-D3 of 28S and ITS regions of rDNA, the new species is clustered with Paratylenchid species having longer stylet length. Morphologically, the new species belongs to Group 9 of Paratylenchus sensu lato and is most similar to G. latescens.Gracilacus paralatescens n. sp., isolated in Anji County, Zhejiang Province, China from the rhizospheric soil of bamboo. The new species can be characterized by the female lateral field with three incisures, stylet 71.5 to 78.8 µm long, lip region truncated, excretory pore located anterior to basal knobs. Vulval lips non-protruding and without vulval flap, spermatheca large, elongated squarish shaped filled with sperms. Tail slender, relatively straight having wedge shape terminus. The spicule slender, slightly curved and 17.5 to 18.9 µm long. In the phylogenetic analysis based on 18S, D2-D3 of 28S and ITS regions of rDNA, the new species is clustered with Paratylenchid species having longer stylet length. Morphologically, the new species belongs to Group 9 of Paratylenchus sensu lato and is most similar to G. latescens.

Xiphinema japonicum n. sp. (Nematoda: Longidorinae) from the Rhizosphere of Japanese Podocarpus macrophyllus (Thunb.), a Cryptic Species Related to Xiphinema bakeri Williams, 1961.

Xiphinema japonicum n. sp., isolated in Ningbo, China, from the rhizosphere of Podocarpus macrophyllus (Thunb.) imported from Japan is described. The new species belongs to Xiphinema non-americanum group 7 and is characterized by medium body length (3.0-3.7 mm), total stylet length 190-201 µm, vulva located anteriorly (V = 30.5%-35.3%), two equally developed female genital branches without uterine differentiation (no Z or pseudo-Z organ and/or spines in the uteri), short tail, convex-conoid with subdigitate peg in terminus, and absence of males. The species has four juvenile developmental stages (J1 was not found). The polytomous identification codes of the new species are (codes in parentheses are exceptions) A4-B4-C4-D5(4)-E2(3)-F3(4)-G2(3)-H2-I3-J4-K?-L1. Morphologically, the new species is mainly characterized by combination of the codes C4 and E2(3), making the species unique and different from other species in the genus. It is most similar to the North American species Xiphinema bakeri, herein considered as its cryptic species by the nature of high morphological similarity, but with significant differences in DNA sequences in nearly full length 18S, ITS1, 28S D2/D3, and cytochrome c oxidase subunit 1 sequences. The second morphologically close species is Xiphinema setariae from which the new species could be separated based on morphological and molecular characters. Morphological comparisons with phylogenetically related species are also discussed.

Distribution of trichodorid species in mainland China with description of Trichodorus hangzhouensis sp. nov. (Nematoda, Triplonchida).

Seven trichodorid species including a new one (Trichodorus hangzhouensis sp. nov., T. nanjingensis, T. pakistanensis, T. cedarus, Paratrichodorus porosus, Nanidorus renifer and N. minor) were recovered from the rhizosphere of different hosts in 13 provinces of China. Each of the recovered species was characterized based on morphology and molecular data using rRNA gene sequences. Trichodorus hangzhouensis sp. nov. is characterized by its males having medium-sized onchiostyle (46-49 µm) and three ventromedian cervical papillae (CP) anterior to the secretory-excretory (S-E) pore, CP1 located opposite the anterior part of isthmus, S-E pore opposite the isthmus or anterior end of pharyngeal bulb, spicules slightly ventrally curved, relatively small, 33.2 (32.0-34.5) µm long, wider slightly marked capitulum, lamina partially striated without bristles at striation; and females having rounded triangular sclerotized vaginal pieces with tips directed towards vulva, 1.5-2.0 µm sized, at about 1 µm apart, vulva pore-like in ventral view. Phylogenetic analysis based on D2-D3 28S rRNA gene sequences differentiated the new species among Trichodorus species from Europe, Asia and USA which formed a large clade. A review of the distribution of Trichodorus, Nanidorus and Paratrichodorus species revealed that T. cedarus, T. nanjingensis, T. pakistanensis and P. porosus are the most widespread species recorded from different provinces of China. This is the first extensive study of trichodorid species occurring in China.

Meloidogyne graminicola protein disulfide isomerase may be a nematode effector and is involved in protection against oxidative damage.

The rice root-knot nematode, Meloidogyne graminicola, is a serious pest in most rice-growing countries. Usually, nematodes employ antioxidants to counteract the harm of reactive oxygen species (ROS) and facilitate their infection. Here the gene encoding M. graminicola protein disulphide isomerase (MgPDI) was identified. The deduced protein is highly conserved in the putative active-site Cys-Gly-His-Cys. In situ hybridization showed that MgPDI was specifically localized within esophageal glands of pre-parasitic second stage juveniles (J2s). MgPDI was significantly up-regulated in the late parasitic J2s. Characterization of the recombinant protein showed that the purified MgPDI exhibited similar activities to other oxidases/isomerases such as the refolding of the scrambled RNase and insulin disulfide reductase and the protection of plasmid DNA and living cells from ROS damage. In addition, silencing of MgPDI by RNA interference in the pre-parasitic J2s lowered their multiplication factor. MgPDI expression was up-regulated in the presence of exogenous H2O2, whereas MgPDI silencing resulted in an increase in mortality under H2O2 stress. MgPDI is localized in the apoplast when transient expression in Nicotiana benthamiana leaves. The results indicated that MgPDI plays important roles in the reproduction and pathogenicity of M. graminicola and it also contributes to protecting nematodes from exogenous H2O2 stress.

Morphological and molecular characterization of Hemicycliophora subbotini n. sp. (Tylenchida: Hemicycliophoridae) from China.

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.