Characterization of a population of Pelodera strongyloides (Nematoda: Rhabditidae) associated with the beetle Lucanus ibericus (Coleoptera: Lucanidae) from Georgia.

During a field survey for entomopathogenic nematodes in Georgia, in the territory of Borjomi-Kharagauli, a nematode population was isolated from the body of single dead beetle of Lucanus ibericus Motschulsky 1845 (Coleoptera: Lucanidae). Based on morphological characters and molecular analyses, the nematode species is identical to Pelodera strongyloides (Schneider, 1860, 1866), being the first record of this species in Georgia. Morphometrics of the Georgian population agree with the original description, and females differ from males by very few characters. The Georgian population of P. strongyloides was molecularly characterized by sequencing the D2 to D3 expansion domains of the 28S rRNA gene and two new molecular markers as the ITS and the mitochondrial COI gene. Phylogenetic analyses revealed that all P. strongyloides sequences grouped together along with all other Pelodera species.

Selecting optimal hyperspectral bands to discriminate nitrogen status in durum wheat: a comparison of statistical approaches.

Hyperspectral data can provide prediction of physical and chemical vegetation properties, but data handling, analysis, and interpretation still limit their use. In this study, different methods for selecting variables were compared for the analysis of on-the-ground hyperspectral signatures of wheat grown under a wide range of nitrogen supplies. Spectral signatures were recorded at the end of stem elongation, booting, and heading stages in 100 georeferenced locations, using a 512-channel portable spectroradiometer operating in the 325-1075-nm range. The following procedures were compared: (i) a heuristic combined approach including lambda-lambda R(2) (LL R(2)) model, principal component analysis (PCA), and stepwise discriminant analysis (SDA); (ii) variable importance for projection (VIP) statistics derived from partial least square (PLS) regression (PLS-VIP); and (iii) multiple linear regression (MLR) analysis through maximum R-square improvement (MAXR) and stepwise algorithms. The discriminating capability of selected wavelengths was evaluated by canonical discriminant analysis. Leaf-nitrogen concentration was quantified on samples collected at the same locations and dates and used as response variable in regressive methods. The different methods resulted in differences in the number and position of the selected wavebands. Bands extracted through regressive methods were mostly related to response variable, as shown by the importance of the visible region for PLS and stepwise. Band selection techniques can be extremely useful not only to improve the power of predictive models but also for data interpretation or sensor design.

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.

Morphological and molecular characterization of Pratylenchus lentis n. sp. (Nematoda: Pratylenchidae) from Sicily.

Pratylenchus lentis n. sp. parasitizing roots of lentil in Sicily, Italy, is described and illustrated. The new species is characterized by a relatively high lip region with three annuli, mean stylet length of 16 mum, with anteriorly flattened knobs, cylindrical body with a relatively anterior vulva, large and ovoid spermatheca full of sperm, plump tail with truncate, irregularly annulated terminus, and by the presence of males. Molecular ITS-RFLP and sequencing analyses of the new species showed clear differences from other most morphologically similar species, such as P. thornei and P. mediterraneus. Preliminary host range tests revealed that chickpea, pea, faba bean and durum wheat are good hosts of P. lentis n. sp., whereas common bean, alfalfa and barley are less robust hosts and tomato, bell pepper, eggplant, melon and sunflower are poor hosts for the nematode.

Pathogenicity and Host-Parasite Relationships of Meloidogyne arenaria in Sweet Basil.

Stunted growth of sweet basil (cv. Genovese) associated with large patches and severe soil infestations by the root-knot nematode Meloidogyne arenaria race 2 were observed in open fields at the end of the crop-growing season in Albenga, province of Savona, Northern Italy by early September 2007. Observed morphological traits of nematode life stages and results of analysis of isozyme electrophoretic patterns and differential host tests were used for nematode species and race identification. Nematode-induced mature galls (either in naturally infected plants or in artificially inoculated root tissues) were spherical or ellipsoidal and confluent along the root axis, containing usually several females, males, and egg-masses with eggs. Feeding sites were characterized by the development of giant cells that contained several hypertrophied nuclei and nucleoli. Giant cell cytoplasm was aggregated along the thickened cell walls. Vascular elements within galls appeared disorganized and disrupted. Due to the heavy damage observed, the relationships between the initial population density and growth of basil plants was also tested in a greenhouse experiment in which inoculum levels varied from 0 to 512 eggs and juveniles/cm3 of soil. Height and top fresh weight data of the inoculated and control plants was fitted to Seinhorst's model. Tolerance limits with respect to plant height and fresh top weight of basil cv. Genovese plants to M. arenaria race 2 were estimated as 0.15 eggs and juveniles/cm3 of soil. The minimum relative values (m) for plant height and top fresh weight were 0.39 and 0.19 at initial nematode population density (Pi) ≥16 eggs and juveniles/cm3 of soil, respectively. The maximum nematode reproduction rate (Pf/Pi) was 448.7 times at an initial population density of 4 eggs and second-stage juveniles/cm3 of soil.

A New Root-Knot Nematode Parasitizing Sea Rocket from Spanish Mediterranean Coastal Dunes: Meloidogyne dunensis n. sp. (Nematoda: Meloidogynidae).

High infection rates of European sea rocket feeder roots by an unknown root-knot nematode were found in a coastal dune soil at Cullera (Valencia) in central eastern Spain. Morphometry, esterase and malate dehydrogenase electrophoretic phenotypes and phylogenetic trees demonstrated that this nematode species differs clearly from other previously described root-knot nematodes. Studies of host-parasite relationships showed a typical susceptible reaction in naturally infected European sea rocket plants and in artificially inoculated tomato (cv. Roma) and chickpea (cv. UC 27) plants. The species is herein described and illustrated and named as Meloidogyne dunensis n. sp. The new root-knot nematode can be distinguished from other Meloidogyne spp. by: (i) perineal pattern rounded-oval, formed of numerous fine dorsal and ventral cuticle striae and ridges, lateral fields clearly visible; (ii) female excretory pore at the level of stylet knobs, EP/ST ratio 1.6; (iii) second-stage juveniles with hemizonid located 1 to 2 annuli anteriorly to excretory pore and long, narrow, tapering tail; and (iv) males with lateral fields composed of four incisures anteriorly and posteriorly, while six distinct incisures are observed for large part at mid-body. Phylogenetic trees derived from distance and maximum parsimony analyses based on 18S, ITS1-5.8S-ITS2 and D2-D3 of 28S rDNA showed that M. dunensis n. sp. can be differentiated from all described root-knot nematode species, and it is clearly separated from other species with resemblance in morphology, such as M. duytsi, M. maritima, M. mayaguensis and M. minor.

Nematode biology and larval development of Thelazia callipaeda (Spirurida, Thelaziidae) in the drosophilid intermediate host in Europe and China.

Thelazia callipaeda, commonly known as the 'oriental eyeworm', has been recently reported in Italy and other European countries. The insect/s that act as intermediate hosts and details of larval development inside the vector remain unclear. In order to (1) demonstrate the species of fly that may act as vector/s for T. callipaeda in southern Italy (Site A) and China (Site B) and (2) describe the larval development of the nematode in the body of flies, 847 Phortica (Drosophilidae) flies were collected from the above two sites, each with a history of human and/or canine thelaziosis. Flies were identified as Phortica variegata (245 - site A) and Phortica okadai (602 - site B), experimentally infected by 1st-stage larvae (L1), kept at different temperatures and dissected daily until day 180 post-infection (p.i.). Dead flies from site A were subjected to specific polymerase chain reaction (PCR) assay to detect T. callipaeda. To demonstrate the role of Phortica as vectors of T. callipaeda, 3rd-stage larvae (L3) recovered from the proboscis of flies were deposited onto the cornea of the eyes of dogs and rabbits. Following dissection, 3 (2.9%) of P. variegata in site A were found to be infected by L3 in the proboscis on days +14, +21 and +53 p.i., compared with 26 (18.4%) of Phortica flies recorded as being positive by PCR. Sequences from positive PCR products were 99% identical to sequences of the corresponding species available in GenBank (AY207464). At site B, 106 (17.6%) of 602 dissected P. okadai were found to be infected by T. callipaeda larvae (different stages) and in total 62 L3 were recovered from the proboscis of 34 (5.6%) flies. The shortest time in which L3 were found was at day +14, +17, +19, and +50 p.i. respectively, depending on the environmental temperatures. Of 30 flies overwintered for 6 months, 6 L3 were detected at day +180 p.i. in 3 flies (10%). The biology of larval development was reconstructed on the basis of the dissection of 602 P. okadai-infected flies and the morphology of larval stages in the insect body described. The present work provides evidence that P. variegata and P. okadai act as vectors for T. callipaeda in southern Europe and in China, respectively. The phenomenon of overwintering is described here for the first time for T. callipaeda and discussed. Finally, the relationship between T. callipaeda and its fly vector is considered in light of disease prophylaxis and to model its dissemination into habitats and environments favourable to Phortica flies.

Detection of grain protein content QTLs across environments in tetraploid wheats.

Grain protein content (GPC) is an important quality factor in both durum and bread wheats. GPC is considered to be a polygenic trait influenced by environmental factors and management practice. The objectives of this study were both to compare the quantitative trait loci (QTL) for GPC in a population of 65 recombinant inbred lines of tetraploid wheats evaluated in three locations for several years (eight data sets), and to investigate the genetic relationship among GPC and grain yield. QTLs were determined based on the Messapia x dicoccoides linkage map which covers 217 linked loci on the 14 chromosomes with 42 additional loci as yet unassigned to linkage groups. The map extends to 1352 cM; the average distance between adjacent markers was 6.3 cM. Seven QTLs for GPC, located on the chromosome arms 4BS, 5AL, 6AS (two loci), 6BS, 7AS and 7BS, were detected that were significant in at least one environment at P<0.001 or in at least two environments at P<0.01. One QTL was significant in all but one environment, two were significant in four or five environments, and four were significant in two out of eight environments. Six out of seven protein content QTLs had pleiotropic effects or were associated to QTLs for grain yield and explained the negative correlation among GPC and yield components. The present results support the concept that studies conducted in a single environment are likely to underestimate the number of QTLs that can influence a trait and that the phenotypic data for a quantitative trait should be collected over a range of locations to identify putative QTLs and determine their phenotypic effects.

Schistonchus africanus n. sp. (Aphelenchida: Aphelenchoididae) Associated with Ficus thonningii (Moraceae) and its Pollinator Wasp Elisabethiella stuckenbergi (Chalcidoidea: Agaonidae).

Syconia ("figs") from Ficus thonningii and adults of its pollinator wasp Elisabethiella stuckenbergi were dissected to elucidate their association with a new species of Schistonchus (Aphelenchoididae). Schistonchus africanus n. sp. is characterized by a short stylet (13-16 mum long); position of the excretory pore opening in both sexes at a level just behind the stylet knobs; and short post-uterine branch, one body-width long. Schistonchus africanus n. sp. parasitizes F. thonningii florets and is transported by the winged females of E. stuckenbergi. Juveniles, females, and males of the nematode were found in the female and male fig florets and in the abdomen of the vector. Nematode populations extracted from female wasps or fig floret tissues did not differ in their morphology. No association was observed with the wingless males of the pollinator wasp.

Pratylenchoides hispaniensis n. sp. (Nemata: Pratylenchidae).

Pratylenchoides hispaniensis n. sp. is described and illustrated from a bisexual population found in a natural habitat at Santa Elena, Jaen, central Spain. Its main distinctive characters are very long esophageal gland lobe (81-117 mum; N' = 51-71) overlapping the intestine 3 to 5 times the body width; lateral field with six incisures; stylet knobs sloping posteriorly; labial disc encircled by the irregular sectors of the first annule; tail cylindrical, extremity annulated, and frequently with a slight dorsal indentation of the hyaline portion at the end of the lateral field. Pratylcnchoides hispaniensis n. sp. appears closely related to P. megalobatus and P. nevadensis. It differs from the former primarily by its longer body length (761-998 vs. 430-621 mum), longer stylet length (20.5-24.4 vs. 18-21 mum), six incisures in the lateral field vs. four for P. megalobatus, and posteriorly sloping stylet knobs vs. rounded or anteriorly flattened knobs in P. megalobatus. It differs from P. nevadensis mainly by the shape of the stylet knobs (sloping in P. hispaniensis vs. rounded in P. nevadensis), length of esophageal lobe (81-117 vs. 34-82 mum), and position of esophageal gland nuclei (all posterior to esophago-intestinal junction in P. hispaniensis vs. at least one nucleus anterior to junction in P. nevadensis).

Description of Rotylenchus graecus n. sp. from Greece (Nematoda: Hoplolaimidae).

Rotylenchus graecus n. sp. is described and illustrated from a bisexual population found in Filippias, Epirus, northwestern Greece, in the rhizosphere of ivy (Hedera helix L.) and giant reed (Arundo donax L.) grown on the edge of irrigation canals. Rotylenchus graecus n. sp. is characterized by a lip region with five annules, stylet length of 29-32 mum, vulva position at 53-65%, female tail pointed-conoid with 15-23 annules, and bursa with regularly annulated margins enveloping tail. The new species differs from all other Rotylenchus species with couoid tails in having a greater number of tail annules. Rotylenchus graecus n. sp. appears closely related to R. acuspicaudatus Van den Berg &Heyns, 1974, but differs from it primarily by the greater number of female tail annules (15-23 vs. 13-16), stylet length (29--32 vs. 25.7-28.3 mum), and the less ventrally protruding bursa, which has smooth posterior margins rather than the finger-like appearance of the posterior bursal annules in R. acuspicaudatus.