Longidorus paravineacola n. sp. (Nematoda: Longidoridae), a New Species from Arkansas

Longidorus paravineacola n. sp., described herein, was found in a survey of longidorids of Arkansas. It is a parthenogeneticspecies characterized by its long body (6.68-9.85 mm); slightly expanded and rounded head, head width 21-27 µm; odontostyle length 95-114 µm; guide ring 28-37 µm posterior to the head end; short rounded tail, and c'' = 0.6-1.0. Longidorus paravineacola n. sp. is similar to the amphimictic species L. vineacola Sturhan &Weischer, 1964; L. balticus Brzeski, Peneva &Brown, 2000; L. kuiperi Brinkman, Loof &Barbez, 1987; and parthenogenetic species L. crassus Thorne, 1974, which also occurred in the type locality.     

Influence of Temperature on Multiplication and Egg Hatching of Longidorus africanus

Longidorus africanus multiplication on tomato was highest at 29 °C. Few nematodes were recovered after 6 weeks at soil temperatures of 35 °C or below 23 °C. The time to egg hatching was shortest and the percentage of eggs hatching was highest at 29 °C. The minimum temperature and the heat sum above this temperature required for egg development were calculated to be 14.3 °C and 94.08 degree-days, respectively. The thermal times required for egg development by L. africanus and L. elongatus were nearly identical. For both species the product of the base temperature and the heat sum was near constant, and at a temperature of 22.3 °C the rates of egg development were equal.     

Longidorus grandis n. sp. and L. paralongicaudatus n. sp. (Nematoda: Longidoridae), Two Parthenogenetic Species from Arkansas

Two new parthenogenetic species of Longidorus were found in Arkansas. Longidorus grandis n. sp. is characterized by its body (5.80-8.24 mm), slightly offset head, head width 20-27 µm, odontostyle 86-100 µm, guide ring 26-35 µm posterior to the anterior end, short conoid to mammiliform tail. Longidorus grandis n. sp. is similar to L. vineacola Sturhan &Weischer, 1964; L. lusitanicus Macara, 1985; L. edmundsi Hunt &Siddiqi, 1977; L. kuiperi Brinkman, Loof &Barbez, 1987; L. balticus Brzeski, Peneva &Brown, 2000; L. closelongatus Stoyanov, 1964; and L. seinhorsti Peneva, Loof &Brown, 1998. Longidorus paralongicaudatus n. sp. is characterized by its body length (2.60-5.00 µm), anteriorly flattened and offset head region 13-18 µm wide, odontostyle length 92-127 µm, guide ring 21-30 µm posterior to the anterior end, tail elongate-conical, and c'' = 1.2-2.6. Longidorus paralongicaudatus n. sp. most closely resembles L. longicaudatus Siddiqi, 1962; L. socialis Singh &Khan, 1996; L. juvenilis Dalmasso, 1969; and L. curvatus Khan, 1986.     

Stepwise and Canonical Discriminant Analysis of Longidorus Species (Nematoda: Longidoridae) from Arkansas

During a 1998-to-2001 survey from Arkansas, nine distinct species of Longidorus were found including five new species. Morphometrics of these nine species were used in a stepwise and canonical discrimination to select a subset of characteristics that best identified each species. Student''s t test was applied to compare Longidorus breviannulatus Norton &Hoffman, 1975; L. crassus Thorne, 1974; L. diadecturus Eveleigh &Allen, 1982; L. fragilis Thorne, 1974; L. biformis Ye &Robbins, 2004; L. glycines Ye &Robbins, 2004; L. grandis Ye &Robbins, 2003; L. paralongicaudatus Ye &Robbins, 2003; and L. paravineacola Ye &Robbins, 2003 to examine interspecies variation and test for the most useful morphometric characters in species discrimination. Most of the morphometric characters were useful to differentiate species, but species identification could not be based on a single character because the morphometric character ranges often overlap. Stepwise discriminant analysis indicated that the guide ring position, head width, tail length, body length, odontostyle length, and anal body width were the most important variables. These were used to generate canonical variables in discriminating the species. The first three canonical variables accounted for 95% of the total variance. The scatterplots by the first three canonical variables grouped and separated the Longidorus species from Arkansas. Stepwise and canonical discriminant analyses were useful for examining the groupings and morphometric relationships of the nine Longidorus species.     

Longidorus biformis n. sp. and L. glycines n. sp. (Nematoda: Longidoridae): Two Amphimictic Species from Arkansas

Two new amphimictic species of Longidorus were found in Arkansas. Longidorus biformis n. sp., found in the rhizosphere of hardwood trees along streams in sandy soil in 14 Arkansas locations, is characterized by its long body (5.42-9.50 mm), wide expanded flattened head end, head width 20.0 to 26.0 µm, odontostyle 96 to 125 µm, guide ring 29 to 38 µm posterior to the anterior end, elongate conoid tail, and c'' = 0.9-2.1. Females with 2 to 11 vetromedian supplement-like structures were found in 2 of 14 populations of this new species. Longidorus biformis n. sp. is closest to L. seinhorsti Peneva, Loof &Brown, 1998 and L. closelongatus Stoyanov, 1964. Among North American species it is closest to L. glycines n. sp. A distinguishing feature of L. biformis n. sp. is the presence of supplement-like organs in some females. Longidorus glycines n. sp., found in soybean microplots at the Main Research Station, Fayetteville, Arkansas, is characterized by its long body (6.14-8.31 mm), wide offset flattened head end, head width 20.3 to 23.3 µm, odontostyle 87.3 to 99.5 µm, guide ring 22.3 to 26.4 µm posterior to the anterior end, short conoid tail with rounded terminus, and c'' = 0.9-1.4. Longidorus glycines n. sp. is closest to L. lusitanicus Macara, 1985. Among North American species it is close to L. biformis n. sp., L. breviannulatus Norton and Hoffman, 1975, and L. crassus Thorne, 1964. Both new species are believed to have four juvenile stages; the first stage was not found for L. biformis n. sp.     

Observations on the Feeding and Symptomatology of Xiphinema and Longidorus on Selected Host Roots

In vitro feeding of Xiphinema brevicolle, X. index and Longidorus africanus on roots of host seedlings is described. Both Xiphinema spp. fed mainly along roots rather than at tips and up to several days at a single site. Feeding of L. africanus was confined to root tips and lasted up to 15 min. No visible short term reaction of roots parasitized by the Xiphinema spp. could be discerned, but both swelling and cessation of growth of root tips were observed within 20 hr after feeding by L. africanus. Long-term (12-month) symptoms on roots of several host plants caused by cultured populations of X. brevicolle, X. index, X. italiae, L. africanus and L. brevicaudatus are described. All the Xiphinema spp. caused a thinning and distinct darkening of root systems and, at some sites, a breakdown of the cortex. Both species of Longidorus caused stubby and swollen root tips. Root symptom severity was in proportion to nematode population levels.     

Descriptions of Three New Longidorus Species from Alaska (Nematoda: Longidoridae)

Three new Longidorus species, L. alaskaensis n. sp., L. paralaskaensis n. sp., and L. bernardi n. sp., are described from specimens collected near Fairbanks, Alaska. Longidorus alaskaensis differs from all species of Longidorus by the presence of a caecum-like structure situated at the reflex of the oviduct. Longidorus paralaskaensis most closely resembles L. alaskaensis n. sp., L. crassus Thorne, L. picenus Roca, Lamberti &Agostinelli, and L. silvae Roca, differing from the last three of these species by having a parallel vs. a tapered lip region, and from all four by having a more narrowly rounded tail tip. Longidorus paralaskaensis differs from L. alaskaensis by having a longer odontostyle (119-128 vs. 110-118 μm) and by lacking the caecum-like structure found at the reflex of the oviduct. Longidorus bernardi n. sp. most closely resembles L. mirus Khan, Chawla &Seshadri, from which it differs by having a longer tail with a more acutely rounded tip, a longer body length (3.5-4.6 vs. 3.0-3.6 μm), and a larger c'' value (1.6-1.8 vs. 1.3-1.6). Longidorus bernardi differs from L. sylphus Thorne, L. africanus Merny, L. auratus Jacobs &Heyns, and L. conicaudatus Khan by having a slightly expanded lip region vs. a lip region with parallel body walls and a more finely rounded tail tip.     

Ingestion,Retention, and Transmission of two Strains of Raspberry Ring-spot Virus by Longidorus macrosoma

The transmission of two strains of raspberry ringspot virus (RRV) by small numbers of nematodes was compared. A strain of RRV from Scotland (RRV-S), originally found in the field associated with Longidorus elongatus, was transmitted frequently by L. elongatus but only once by L. macrosoma. A strain from England (RRV-E) associated with L. macrosoma in the field was transmitted infrequently by each species of nematode. The reasons why L. macrosoma infected only a small proportion of bait plants with virus were investigated, and it was found that most of the nematodes tested had fed on the source plants and many had ingested virus. Most nematodes exposed to RRV-E or RRV-S had fed on the roots of the bait plants and, when thin sections were examined by electron microscope, had retained particles (thought to be those of the virus) in the region of the anterior odontostyle, Thus, most nematodes seem to have had ample opportunity to transmit virus, and the low frequency of transmission may have been due to a failure of the virus particles to be released from the site of retention or to a lack of infectivity of the virus when L. macrosoma was the vector and Petunia hybrida was the host.     

Descriptions of Three New Longidorus Species from Slovakia (Nemata: Longidoridae)

Three new Longidorus species from Slovakia are described. Longidorus carpathicus n. sp. most closely resembles Longidorus silvae but differs by having a longer odontostyle, odontophore, and total stylet; smaller a and c ratios; and longer distance to the guide ring. This new species also resembles L. picenus, L. macrosoma, and L. major but differs by having a narrower lip width. It further differs from L. picenus by having a longer odontostyle and smaller c ratio, and by lacking males; from L. macrosoma by having a longer odontostyle, smaller c ratio, by lacking males, and a more pronounced J1 tail peg; and from L. major by having a shorter body length, longer odontostyle, longer odontophore, and longer J1 tail peg. Longidorus piceicola n. sp. most closely resembles L. eridanicus, from which it differs by having a greater lip width, longer tail, smaller c ratio, larger c'' ratio, shorter hyaline tail length, and a conically rounded vs. hemispherical tail. This new species differs from L. cylindricaudatus by having a larger lip width, longer odontostyle and odontophore, and a greater distance to the guide ring; from L. nevesi by having a shorter body length, longer odontostyle, larger c'' ratio, and shorter hyaline tail length. Longidorus juglansicola n. sp. most closely resembles L. athesinus but differs by its longer body, wider lips, and larger a and c ratios. It closely resembles L. vineacola but differs by its shorter body length, smaller c ratio, and an almost parallel lip outline vs. an expanded lip outline; from L. lusitanicus by a longer odontophore and tail, and an almost parallel lip outline vs. an expanded lip outline.     

Cluster Analysis of Longidorus Species (Nematoda: Longidoridae), a New Approach in Species Identification

Hierarchical cluster analysis based on female morphometric character means including body length, distance from vulva opening to anterior end, head width, odontostyle length, esophagus length, body width, tail length, and tail width were used to examine the morphometric relationships and create dendrograms for (i) 62 populations belonging to 9 Longidorus species from Arkansas, (ii) 137 published Longidorus species, and (iii) 137 published Longidorus species plus 86 populations of 16 Longidorus species from Arkansas and various other locations by using JMP 4.02 software (SAS Institute, Cary, NC). Cluster analysis dendograms visually illustrated the grouping and morphometric relationships of the species and populations. It provided a computerized statistical approach to assist by helping to identify and distinguish species, by indicating morphometric relationships among species, and by assisting with new species diagnosis. The preliminary species identification can be accomplished by running cluster analysis for unknown species together with the data matrix of known published Longidorus species.     

Phylogenetic Relationships and Genetic Variation in Longidorus and Xiphinema Species (Nematoda: Longidoridae) Using ITS1 Sequences of Nuclear Ribosomal DNA

Genetic analyses using DNA sequences of nuclear ribosomal DNA ITS1 were conducted to determine the extent of genetic variation within and among Longidorus and Xiphinema species. DNA sequences were obtained from samples collected from Arkansas, California and Australia as well as 4 Xiphinema DNA sequences from GenBank. The sequences of the ITS1 region including the 3'' end of the 18S rDNA gene and the 5'' end of the 5.8S rDNA gene ranged from 1020 bp to 1244 bp for the 9 Longidorus species, and from 870 bp to 1354 bp for the 7 Xiphinema species. Nucleotide frequencies were: A = 25.5%, C = 21.0%, G = 26.4%, and T = 27.1%. Genetic variation between the two genera had a maximum divergence of 38.6% between X. chambersi and L. crassus. Genetic variation among Xiphinema species ranged from 3.8% between X. diversicaudatum and X. bakeri to 29.9% between X. chambersi and X. italiae. Within Longidorus, genetic variation ranged from 8.9% between L. crassus and L. grandis to 32.4% between L. fragilis and L. diadecturus. Intraspecific genetic variation in X. americanum sensu lato ranged from 0.3% to 1.9%, while genetic variation in L. diadecturus had 0.8% and L. biformis ranged from 0.6% to 10.9%. Identical sequences were obtained between the two populations of L. grandis, and between the two populations of X. bakeri. Phylogenetic analyses based on the ITS1 DNA sequence data were conducted on each genus separately using both maximum parsimony and maximum likelihood analysis. Among the Longidorus taxa, 4 subgroups are supported: L. grandis, L. crassus, and L. elongatus are in one cluster; L. biformis and L. paralongicaudatus are in a second cluster; L. fragilis and L. breviannulatus are in a third cluster; and L. diadecturus is in a fourth cluster. Among the Xiphinema taxa, 3 subgroups are supported: X. americanum with X. chambersi, X. bakeri with X. diversicaudatum, and X. italiae and X. vuittenezi forming a sister group with X. index. The relationships observed in this study correspond to previous genera and species defined by morphology.     

Abundance and Vertical Distribution of Longidorus breviannulatus Associated with Corn and Potato

Longidorus breviannulatus was detected in a field planted to corn after 13 years of potato. Nematode populations were maintained in this field in adjacent corn and potato plots for 2 years but did not increase significantly on either crop. Population levels increased until approximately 60 days after planting and then declined until the end of the growing season. Overwinter mortality was negligible. The vertical distribution of the nematode population changed during the course of the season. More nematodes were recovered from depths of 0-15 cm in early season samples and from depths of 15-30 cm in late season samples. Data indicated that this redistribution was due to nematode migration.     

Morphological Variation in Xiphinema spp. from New York Orchards

Xiphinema specimens were collected from orchards in southeastern, northeastern, and western New York. Total length, distance of vulva from anterior end, spear length (odontostyle plus odontophore), body diameter at vulva, tail length, anal body diameter, and length and diameter of hyaline tail tip were measured on fixed, glycerol-infiltrated adult females. Most specimens were identified as X. americanum or X. rivesi, but one western New York population was identified as X. californicum (a new record for New York). Multivariate analyses indicated that, with one exception, western New York populations of both X. americanum and X. rivesi were smaller and slimmer than their eastern counterparts. Regional differences were generally larger than differences attributed to host species.     

Morphological Comparisons Between Xiphinema rivesi Daimasso and X. americanum Cobb Populations from the Eastern United States

Though in the past Xiphinema americanum has been the most commonly reported dagger nematode in the eastern United States, our studies revealed the presence in Pennsvlvania of a previously unrecognized and unreported species related to X. americanum, Morphometric data and photomicrographs establish the identity of this form as X. rivesi and show expected variations in populations of this species from various locations. Similar data and illustrations are given for X. americanum populations from Pennsylvania and other areas, showing variations and relationships. Xiphinema rivesi is widely distributed in the fruit producing area of south-central Pennsylvania and is also reported herein from raspberry in Vermont and apple in Maryland and New York. This species is frequently found in fruit growing areas of Pennsylvania associated with tomato ringspot virus-induced diseases and is also found associated with corn, bluegrass sod, and alfalfa.     

Scanning Electron Microscopy of Xiphinema,Longidorus, and Californidorus Stylet Morphology

Stylet ultrastructure of five Xiphinema, four Longidorus, and three Californidorus species was compared by scanning electron microscopy. Morphological differences were seen in the odontophores and odontostyle bases between the genera and some of the species. All Xiphinema studied had well-developed odontophore flanges; the Longidorus species lacked flanges, except for weakly developed ones in L. diadecturus; and none of the Californidorus had flanges. Three sinuses were present in the odontophores of all species. The sinuses varied in length depending upon species. In Xiphinema and Californidorus the odontostyle bases had distinct overlapping collars, but in Longidorus the collars were absent except for L. diadecturus. The odontostyle-odontophore junction from a lateral view appeared as a slanted transverse line in all the species, but in a dorsal view of Xiphinema and Californidorus it was V-shaped. Dorsal longitudinal seams of the odontostyle and odontophore were observed in all the species. The dorsally located odontostyle aperture was ca. 1 μm from the anterior end in all species, except in one Longidorus sp. it was ca. 4 μm from the end.     

Characteristics and Efficacy of a Sterile Hyphomycete (ARF18), a New Biocontrol Agent for Heterodera glycines and Other Nematodes

A filamentous, nonsporulating fungus, designated Arkansas Fungus 18 (ARF18), was isolated from 9 of 95 populations of Heterodera glycines, the soybean cyst nematode, in Arkansas. In petri dishes, ARF18 parasitized 89% of H. glycines eggs in cysts. The fungus also infected eggs of Meloidogyne incognita and eggs in cysts of Cactodera betulae, H. graminophila, H. lespedezae, H. leuceilyma, H. schachtii, and H. trifolii. In pot tests, reproduction of SCN was 70% less in untreated field soil that was naturally infested by ARF18 than in autoclaved field soil. Although ARF18 grew well at 25 C on cornmeal agar over a wide pH range, it did not sporulate on 28 media and thus could not be identified to genus or species.     

Distribution of Races of Heterodera glycines in the Central United States

A total of 62 populations of Heterodera glycines were collected in 10 states along the Mississippi and Missouri rivers, and 206 populations were collected in Arkansas. Among the 62 populations, races 2, 3, 4, 5, 6, 9, and 14 were found south of 37°N latitude, and races 1 and 3 were found north of 37°N latitude. In Arkansas samples, races 2, 4, 5, 6, 9, and 14 comprised 87% of the populations. In both groups of samples, H. glycines populations with genes that enabled the population to parasitize cv. Pickett occurred the most frequently, followed by those with genes for parasitism of cv. Peking, then PI88.788, and the fewest with genes for parasitism of PI90.763. The diversity of races in this study raises questions about the effectiveness of race-specific cultivars for the management of soybean cyst nematodes. The greater diversity of races of H. glycines in the southern United States may be because of a longer history of planting resistant cultivars.     

Influence of Edaphic Factors and Previous Crop on Pratylenchus spp. Population Densities in Potato

Root and soil samples from commercial potato fields were assayed for nematodes in 1983 and 1984. Pratylenchus spp. population densities in Suffolk County, New York, were consistently, though not always statistically, higher in potato fields that had been planted to rye or wheat rather than potatoes during the previous growing season. Regardless of the previous crop, population densities in the two potato production areas in Suffolk County differed significantly: population densities on the south fork were 1.9-5.5 times higher than those on the north fork. Species prevalence differed significantly on the two forks but was not related to the previous year''s crop. P. penetrans and P. crenatus were found primarily on the north and south forks, respectively. Differences in species distribution were associated with differences in soil types. P. crenatus was usually found on loams and silt loams, but P. penetrans was found more frequently on sandy soils.