A nematicidal tannin from Punica granatum L. rind and its physiological effect on pine wood nematode (Bursaphelenchus xylophilus).

The ethanol extract of Punica granatum L. rind was tested to show significant nematicidal activity against pine wood nematode. Three nematicidal compounds were obtained from the ethanol extract by bioassay-guided fractionation and identified as punicalagin 1, punicalin 2, and corilagin 3 by mass and nuclear magnetic resonance spectral data analysis. Punicalagin 1 was most active against PWN among the purified compounds with the LC50 value of 307.08µM in 72h. According to the enzyme assays in vitro, punicalagin 1 could inhibit the activity of acetylcholinesterase, amylase and cellulase from PWN with IC50 value of 0.60mM, 0.96mM and 1.24mM, respectively. The morphological structures of PWNs treated by punicalagin 1 were greatly changed. These physiological effects of punicalagin 1 on PWN may helpful to elucidate its nematicidal mechanism.

A new stem nematode, Ditylenchus oncogenus n. sp. (Nematoda: Tylenchida), parasitizing sowthistle from Adriatic coast dunes in southern Italy.

Morphological and molecular analyses of a stem nematode causing a severe disease on infected sowthistle (Sonchus bulbosus) plants, involving the formation of gall-like structures on infected leaves and stems, have led to the description of a new species named Ditylenchus oncogenus n. sp. Morphologically, the new species is characterized by a medium to large body size (all adults more than 1 mm in length); a delicate stylet (9.0-11.0 µm long) with minute, rounded knobs; a long post-vulval uterine sac (c. 65% of the vulva-anus distance); six incisures at the lateral fields and characteristic D. destructor-pattern of spicules (with pronounced ventral tumulus and anteriorly pointed, less sclerotized, cuticle parts present within the lamina). The results of molecular analysis of rRNA gene sequences, including the D2-D3 expansion regions of 28S rRNA, internal transcribed spacer (ITS) rRNA, partial 18S rRNA gene, the protein-coding mitochondrial gene, cytochrome oxidase c subunit I (COI), and the heat-shock protein 90 (hsp90) gene, support the new species status. The results of a host-suitability test indicated that the new species does not parasitize potato (Solanum tuberosum) tubers and broad bean (Vicia faba) seedlings. Histopathological observations on naturally infected sowthistle tissues revealed that D. oncogenus n. sp. causes floral stem neoplasia and midrib leaf gall formation on the type, and to date only known, host. The galls were characterized by extensive hyperplasia, where several necrotic cells in the neoplasic area were directly damaged by feeding of the nematode, whereas a number of adjacent cells showed typical cytological changes, such as granulated cytoplasm with hypertrophied nuclei and nucleoli.

Comparative, three-dimensional anterior sensory reconstruction of Aphelenchus avenae (nematoda: Tylenchomorpha).

The anterior sensory anatomy (not including amphids) of the nematode Aphelenchus avenae (Tylenchomorpha) has been three-dimensionally reconstructed from serial, transmission electron microscopy thin sections. Models, showing detailed morphology and spatial relationships of cuticular sensilla and internal sensory receptors, are the first computerized reconstruction of sensory structures of a Tylenchomorpha nematode. Results are analyzed with respect to similarly detailed reconstructions of Rhabditida outgroup nematodes, Acrobeles complexus (Cephalobomorpha) and Caenorhabditis elegans (Rhabditomorpha). Homologies identified in A. avenae demonstrate the general conservation of the anterior sensory system between freeliving nematodes and the largely plant parasitic Tylenchomorpha. A higher degree of similarity is shown between A. avenae and A. complexus, with common features including: the presence of a second, internal outer labial dendrite (OL1); a second cephalic dendrite in the female (CEP2/CEM); an accessory process loop of inner labial dendrite 1; and terminus morphology and epidermal associations of internal sensory receptors BAG and URX. Unique to A. avenae is a pair of peripheral, lateral neurons of unknown homology but with axial positions and intercellular relationships nearly identical to the "posterior branches" of URX in A. complexus. Knowledge of homologies and connectivity of anterior sensory structures provides a basis for expansion of the experimental behavioral model of C. elegans to the economically important nematodes of Tylenchomorpha.

Three-dimensional reconstruction of the stomatostylet and anterior epidermis in the nematode Aphelenchus avenae (Nematoda: Aphelenchidae) with implications for the evolution of plant parasitism.

A three-dimensional model of the stomatostylet and associated structures has been reconstructed from serial thin sections of Aphelenchus avenae, a representative of Tylenchomorpha, a group including most plant parasitic nematodes. The reconstruction is compared with previous work on bacteriovorous cephalobids and rhabditids to better understand the evolution of the stylet and its associated cells. Two arcade syncytia ("guide ring") line the stylet shaft, supporting the hypothesis that the stylet shaft and cone (into which the shaft extends and which is not lined by syncytia) are homologous with the gymnostom of cephalobids, the sister taxon of tylenchids. Epidermal syncytia, HypA, HypB, HypC, and HypE, line the cephalic framework, vestibule, and vestibule extension, congruent with the hypothesis that these components are homologous with the cephalobid cheilostom. Relative to outgroups, HypC is expanded in A. avenae, enclosing sensilla that fill most of the cephalic framework. The homolog of syncytium HypD in the cephalobid Acrobeles complexus is not observed in A. avenae. Arcade syncytia are reduced compared with those of cephalobids. Stylet protractor muscles in A. avenae are homologous with the most anterior set of radial muscles of cephalobids. Observations to date test and verify our previous hypotheses of homology of the stomatostylet with respect to the stoma of bacteriovorous outgroups. Reconstruction of the stegostom and pharynx will provide further tests of homology and evolution of feeding structure adaptations for plant parasitism.

Sphaerularia vespae sp. nov. (Nematoda, Tylenchomorpha, Sphaerularioidea), an endoparasite of a common Japanese hornet, Vespa simillima Smith (Insecta, Hymenoptera, Vespidae).

Sphaerularia vespae sp. nov., an endoparasite of a common Japanese hornet, Vespa simillima is described from Hokkaido, Japan, and its molecular sequence profiles are given. This newly discovered nematode appears to belong to the genus Sphaerularia, judging from its characteristic parasitic form, the uterium, which looks like a sausage with many verrucae on its surface. Sphaerularia vespae sp. nov. is distinguished from S. bombi, the only other nominal species of the genus, by the morphology of the male bursa, female tail, and anterior end of parasitic juveniles. SSU, ITS, and D2D3 (LSU) DNA sequences were determined for S. vespae sp. nov. and compared with those from related nematodes obtained from the GenBank database. The sequences from S. vespae sp. nov. were close to those of S. bombi and several another tylenchid entomoparasitic nematodes. Sphaerularia vespae sp. nov. parasitizes overwintering gynes of the hornet and practically sterilizes them, as S. bombi does for bumblebee gynes.

Metaparasitylenchus hypothenemi n. sp. (Nematoda: Allantonematidae), a parasite of the coffee berry borer, Hypothenemus hampei (Curculionidae: Scolytinae).

Metaparasitylenchus hypothenemi n. sp. (Nematoda: Allantonematidae) is described from the coffee berry borer, Hypothenemus hampei (Ferrari) (Curculionidae: Scolytinae), in Chiapas, Mexico. This species differs from other members of the genus by its small size, annulated cuticle, lateral fields with 3 ridges, free-living stages with an excretory pore located between the pharyngeal gland orifices, a distinct stylet with basal swellings in free-living females, a postvulval uterine extension, a thin stylet lacking basal swellings in males, 2 separate spicules, a gubernaculum, and a peloderan bursa. Parasitic females are white, with a straight or slightly curved body and are ovoviviparous. Third-stage juveniles emerge from parasitized beetles and molt twice before reaching the adult stage. Because the coffee berry borer is the most important pest of coffee throughout the world and this parasite partially or completely sterilizes female beetles, it is worthy of further investigation as a potential biological control agent.

Tylenchida associated with different crops in Sennar State (Sudan).

A study was done on the taxonomy and morphology of plant parasitic nematodes (Tylenchida) found in Sennar State (Sudan). Sixty samples of different crops were collected in the sugarcane area. Thirty samples originated from soil around the roots of Saccharum officinarum (sugarcane) from different ratoons and thirty samples were collected from other crops (Mangifera indica; Citrus limon; Citrus aurantifolia; Citrus paradisi; Citrus sinensis, Phoenix dactylifera, Musa sapentium; Cassia italica, Capsicum annuum, Sorghum bicolor, Sorghum sudanensis, Gossypium barbadense, Ficus nitida, Khaya senegalensis, Eucalyptus microtheca, Acacia nilotica, Acacia seyal, Azardichta indica, Cajanus cajana, Caltropsis spp. and Liguster ovalifolium). Seven species belonging to seven different genera of Tylenchida were identified: Paratrophurus lobatus, Scutellonema clathricaudatum, Hoplolaimus aegypti and Filenchus cylindricus. Helicotylenchus plumariae, Pratylenchus thornei and Malenchus andrassyi are new records for Sudan. These seven species were compared with the descriptions given in the literature and differences and variations were discussed. Additional morphological data were described by means of SEM microscopy.