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.     

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.     

Taxonomic and Molecular Identification of Bakernema,Criconema, Hemicriconemoides,Ogma and Xenocriconemella Species (Nematoda: Criconematidae)

Populations of Bakernema inaequale, C. petasum, C. sphagni, C. mutabile, Ogma octangulare, Xenocriconemella macrodora and Hemicriconemoides chitwoodi were identified and re-described from different geographical areas in the continental United States and molecularly characterized. Two new species of spine nematodes Criconema arkaense n. sp. from Washington County and Lee County, Arkansas and Criconema warrenense n. sp from Warren, Bradley County, Arkansas are also described and named. Criconema arkaense is characterize by having a conspicuous lip region offset from the body with two annuli, short rounded tail with a thin cuticular sheath and subterminal anus. Criconema warrenense n. sp. has two lip region annuli about the same width, first annulus directed posteriorly, separated by a narrow neck annulus and a short conoid tail, unilobed non-folded annulus. The molecular characterization of Criconema arkaense and Criconema warrenense using ITS1 rDNA gene sequence and the molecular phylogenetic relationships of these new species along with the known spines nematodes are provided.     

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.     

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.     

Morphological and Molecular Characterization of Longidorus americanum n. sp. (Nematoda: Longidoridae), a Needle Nematode Parasitizing Pine in Georgia

We describe and illustrate a new needle nematode, Longidorus americanum n. sp., associated with patches of severely stunted and chlorotic loblolly pine, (Pinus taeda L.) seedlings in seedbeds at the Flint River Nursery (Byromville, GA). It is characterized by having females with a body length of 5.4-9.0 mm; lip region slightly swollen, anteriorly flattened, giving the anterior end a truncate appearance; long odontostyle (124-165 µm); vulva at 44%-52% of body length; and tail conoid, bluntly rounded to almost hemispherical. Males are rare but present, and in general shorter than females. The new species is morphologically similar to L. biformis, L. paravineacola, L. saginus, and L. tarjani but differs from these species either by the body, odontostyle and total stylet length, or by head and tail shape. Sequence data from the D2-D3 region of the 28S rDNA distinguishes this new species from other Longidorus species. Phylogenetic relationships of Longidorus americanum n. sp. with other longidorids based on analysis of this DNA fragment are presented. Additional information regarding the distribution of this species within the region is required.     

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.     

Description of Dolichodorus cobbi n. sp. (Nematoda: Dolichodoridae) with Description of Dolichodorus cobbi n. sp. (Nematoda: Dolichodoridae) with Morphometrics and Lectotype Designation of D. heterocephalus Cobb, 1914

Dolichodorus cobbi n. sp. is described and illustrated from soil around roots of "woods" on the Stanley Farm, Scott County, Arkansas. This species is distinguishable from others of the genus by its short tail projection. It is most closely related to D. marylandicus but differs in having a short tail projection, longer stylet, greater body length, larger c value, and also in position of excretory pore. A limited number of Cobb''s original specimens and drawings of D. heterocephalus were examined. A lectotype and paralectotype were designated to establish the taxonomic base for the genus. Cobb''s original modified labeled drawing of D. heterocephalus is also included. Morphometric data on D. heterocephalus from the type locality and several other populations are given. Some variations in female tail shape, body length, and stylet length were noted.     

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.     

Descriptions of Two New Species of Tylenchorhynchus Cobb, 1913 (Nematoda: Tylenchida), with Details on Morphology and Variation of T. claytoni

Two new species of plant parasitic nematodes (Tylenchorhynchus quaidi n. sp. and T. tritici n. sp.) from Pakistan are described and illustrated. Tylenchorhynchus quaidi n. sp., from soil around roots of potato (Solanum tuberosum) from an experimental field of NNRC, Karachi, Pakistan, is distinguishable from other species by its peculiar sunken dome-shaped head. Although similar to T. goffarti, it differs by head shape, areolation of lateral field, ratios a (23-28 vs. 29-37) and c (11-14 vs. 13-20), and a vagina that is half sclerotized and half unsclerotized. Tylenchorhynchus tritici n. sp., from soil around roots of wheat (Triticum aestivum) from Campbellpur, Pakistan, is similar to T. ventrosignatus and T. nordiensis. It differs from T. ventrosignatus by a continuous lip region, number of head annules (2-3 vs. 4), coarse body annulation, absence of a wave-like structure near the vulva, and by tail shape and number of tail annules (15-23 vs. 28-32). It differs from T. nordiensis by stylet length (12.4-14.6 vs. 11-13 μm), shape of stylet knobs, number of head annules (2-3 vs. 4), non-areolated lateral field in region of phasmids, and not fusing in posterior third of tail. Morphometrics of Tylenchorhynchus claytoni from soil around stunted maize (Zea mays L.), in Muscatine County, Iowa, and several other populations are given. Detailed morphometric data on T. claytoni based on topotypes collected from type locality and several other populations revealed that this species shows variations in the shape of tail in females, number of tail annules (and sometimes annules extending further back on the terminus, almost being an annulated terminus), position of phasmid, and shape of lip region. The subgenus Bitylenchus is proposed as a new synonym of Tylenchorhynchus and its species referred to the latter genus.     

Margollus bokanicus n. sp. from Iran,the Fourth Species of a Rare Nematode Genus (Dorylaimida,Tylencholaimellidae)

Margollus bokanicus n. sp., collected from natural habitats in Khorasaneh district, Bokan, West Azarbaijan province, Iran, is described. Morphological and morphometric data are provided as well as drawings and light microscopy illustrations. The new species is characterized by a medium size body length (0.60 to 0.73 mm), labial and postlabial sclerotizations, lip region 7-μm wide, offset by constriction and long neck (167 to 207 μm), long pharyngeal basal bulb (27 to 36 μm) or 16% to 17% of total neck length, female genital system monodelphic–opisthodelphic, anterior branch reduced to a uterine sac (26–29 μm) or 1.1 to 1.3 times the body diameter, long posterior uterus (25–28 μm) or 1.1 to 1.3 times the body diameter, V = 40 to 47, cylindroid female tail (17 to 24 μm, c = 31 to 38, c’ = 1.1 to 1.4), and males unknown. This taxon is easily distinguishable from other Margollus species by its smaller general size and more posterior vulva. A compendium of Margollus species is also presented.     

Xiphinema llanosum and Trophurus vultus, Two New Plant Nematode Species from Pasture Soils in Colombia

Xiphinema llanosum n. sp. and Trophurus vultus n. sp. are described and illustrated from grass soils in Llanos Orientales, Colombia. Xiphinema llanosum is a bisexual species. The female body length is 2.3-2.7 mm, odontostyle 86-96 μm, and odontophore 58-65 μm long; vulva at 42-47%; anterior ovary is absent; the anterior uterus and oviduct are similar to the posterior branch but slightly reduced; and the tail is dorsally convex-conoid with a blunt hemispherical terminus. Male body length is 2.06-2.96 mm; spicules are 40-44 μm long; and four (rarely three or five) anterior ventromedian supplementary papillae are present. Trophurus vultus females are 0.52-0.67 mm long; vulva at 56-60%; stylet is 10.5-13.5 μm long; isthmus is as long as the basal esophageal bulb; the tail is subclavate, 1.6-2.2 times anal body width long; and the terminal cuticle thickness is about one-sixth of the tail length.     

Diversity of Xiphinema americanum-group Species and Hierarchical Cluster Analysis of Morphometrics

Of the 39 species composing the Xiphinema americanum group, 14 were described originally from North America and two others have been reported from this region. Many species are very similar morphologically and can be distinguished only by a difficult comparison of various combinations of some morphometric characters. Study of morphometrics of 49 populations, including the type populations of the 39 species attributed to this group, by principal component analysis and hierarchical cluster analysis placed the populations into five subgroups, proposed here as the X. brevicolle subgroup (seven species), the X. americanum subgroup (17 species), the X. taylori subgroup (two species), the X. pachtaicum subgroup (eight species), and the X. lambertii subgroup (five species).     

Morphological and Morphometrical Characterization of Meloidogyne incognita from Different Host Plants in Four Districts of Punjab,India

The population of M. incognita, the root knot nematode (RKN) was found infesting five different host plants (okra, banana, sunflower, bottle gourd, and brinjal) out of 24 examined from four districts of Punjab, India (Gurdaspur, Ludhiana, Patiala, and Hoshiarpur). Morphological and morphometrical characterization indicated that in the case of mature female, the characters of body length and width, neck length, ratio ‘a’, anus to tail terminus (ATT), interphasmid distance (IPD), and perineal pattern were recorded as stable characters. These taxonomic characters can be reliable for identification. All characters of second-stage juvenile (J2) such as body length, stylet length, head to median bulb length (H-MB), distance from median bulb to excretory pore (MB-EP), tail length, anal body width (ABW), and ratios C and C’ were highly variable. Analysis of interpopulation morphometric characters of mature female of M. incognita, namely, body length, width, and ratio ‘a’ were moderately variable characters (CV 0.26% to 20%) and stylet length, neck length, length of median bulb (LMB), and width of median bulb (WMB) were highly variable (CV 1.0% to 36.1%). In the perineal pattern, the two characters ATT and IPD were moderately variable (CV 8.8% to 17.6%) and two characters, anus to vulval slit (AVS) and length of vulval slit (LVS), were highly variable (CV 2.1% to 40.5%). In J2, body length, stylet length, H-MB, MB-EP, ABW, tail length, ratios C, and C’ were highly variable characters (CV > 12%).     

Morphometric Variation and Biogeography of Ogwa menzeli and Criconema sphagni

Morphometrics of Ogma menzeli from woodlands in the Adirondack Mountains of New York State and in Iowa were compared. Specimens from the Adirondacks were significantly greater in mean total body length, stylet length, the b, R, and RV values, body width, and esophagus length than specimens from Iowa. The V value was significantly greater in the Iowa than in the Adirondack specimens. The two populations are considered ecotypes of O. menzeli. Criconema sphagni morphometric measurements differed significantly for the RV value (negative) and V value (positive) relative to elevation in the Adirondacks. There was a positive regression correlation for the RV value of O. menzeli and elevation in the Adirondack Mountains.     

Xiphinema krugi,Species Complex or Complex of Cryptic Species?

Fourteen morphologically putative populations of X. krugi were clearly separated into four different profiles by RFLP analysis (Alu I and Hinf I), sequencing of the ITS-1 region, and subsequent Maximum Likelihood phylogenetic analyses. These four profiles were further supported by a principal component analysis of morphometric characters that yielded four taxonomic clusters matching those produced by the molecular data. Sequence homology was greater amongst populations that represented the same RFLP profile than between profiles and similar both between representative populations of the RFLP profiles and putative closely related Xiphinema species. This study suggests that X. krugi is a potential species complex comprised of at least four distinct genotypes.     

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.