Morphological Variation among 23 Xiphinema americanum Populations

Morphometrics of 23 United States populations of Xiphinema americanum sensu lato, sharing the characteristics of an offset lip region and conoid tail, were examined and analyzed statistically by canonical discriminant analysis (CDA). Specimens were collected from Arkansas, Georgia, Tennessee, Mississippi, Florida, Oklahoma, California, and North Dakota. Eleven measurements and body ratios obtained from female specimens were used in the analysis. Xiphinema americanum, X. bricolensis, X. californicum, X. citricolum, X, intermedium, X. tarjanense, and X. thornei, and one undescribed species were identified among the 23 populations. Three groups -- X. americanum-group, X. californicum-group, and X. intermedium-group (X. intermedium and X. tarjanense) -- were formed and four populations belonging to four different species were separated consistently from these groups in CDA scatterplots of the 23 populations. Composition of the groups was somewhat related to the geographical origins of the populations in the groups. A population from California had morphometrics intermediate between X. americanum and X. californicum. Separation between the X. americanum-group and X. californicum-group in the CDA scatterplots was not as distinct as that between them and the X. intermedium-group or between any of the three groups and the four single outlying populations.     

Morphometric Evidence for Three Juvenile Stages in Some Species of Xiphinema americanum sensu lato

One to two hundred nematodes from each of seven Xiphinema americanum-group populations were measured to determine the range of stylet and body lengths for juveniles and adults. First-stage juveniles were identified by the position of the replacement odontostyle (i.e., the tip of the replacement odontostyle overlapped the base of the odontophore). Nematodes were identified as second stage if the functional odontostyle was the same length as the replacement odontostyle of the first stage. Subsequent stages were similarly identified by establishing the range of corresponding replacement and functional odontostyle lengths. In all populations examined, this procedure created natural divisions that clearly grouped nematodes by stylet and body length. Presumably these groups identified all juvenile and adult stages. Populations of X. americanum, X. rivesi, and X. californicum from the United States had three juvenile stages, but a population of X. pachtaicum from Bulgaria had four juvenile stages.     

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.     

Seasonal Population Fluctuation of Xiphinema americanum and X. rivesi in New York and Pennsylvania Orchards

The population fluctuation and composition of Xiphinema americanum (sensu stricto) and X. rivesi were studied in a New York apple orchard (only X. americanum present), a Pennsylvania apple orchard (both X. americanum and X. rivesi present), and a Pennsylvania peach orchard (X. americanum, X. rivesi, and X. californicum present). Few clear trends in population fluctuation or composition were observed. The adult female was the predominant stage in most sample periods, and the reproductive period was limited to late spring and early summer. Only a few of the females at any sample period were gravid. All stages were present throughout the year, and all stages overwintered. Eggs in soil were not monitored. In the Pennsylvania apple orchard, X. americanum and X. rivesi were easily separated by morphological characteristics; however, the two species did not display differences in population structure or composition. The predominance of adults, the relatively low reproductive rates, and the association of these species with stable habitats suggest that the life strategies of X. americanum and X. rivesi are K-selected as opposed to r-selected.     

Transmission of Nepoviruses by Xiphinema americanum-group Nematodes

The transmission of North American nepoviruses by putative species belonging to the Xiphinema americanum-group is reviewed. Xiphinema americanum sensu stricto, X. californicum, and X. rivesi each transmit cherry rasp leaf (CRLV), tobacco ringspot (TobRSV), and tomato ringspot nepovirus (TomRSV), and X. bricolensis is a vector of TomRSV. The apparent lack of specificity in the transmission of North American nepoviruses by X. americanum-group species markedly contrasts with the specific associations between European nepoviruses and their vector nematode species. Two complementary projects are described examining the taxonomic identity of putative species in the X. americanum-group, their morphological and genetic relationships, their ontogeny, and their ability to transmit viruses.     

Effect of Three Plant Species on Population Densities of Xiphinema americanum and X. rivesi

A taxonomic revision of the Xiphinema americanum species complex has necessitated a reexamination of the host range of species in the complex before recommendations can be made with confidence on the likelihood that specific crops will be damaged. Toward this end, populations of X. americanum and X. rivesi collected from apple orchards in eastern and western New York state were evaluated after 3 months in pots planted with cucumber, apple, or dandelion seedlings. Eastern and western New York populations of both nematode species declined on cucumber but increased to similar final densities on apple and dandelion.     

Biotin-Avidin ELISA Detection of Grapevine Fanleaf Virus in the Vector Nematode Xiphinema index

The value of biotin-avidin (B-A) ELISA for the detection of grapevine fanleaf virus (GFLV) in Xiphinema was estimated with field populations and greenhouse subpopulations. Samples consisted of increasing numbers of adults ranging from 1 to 64 in multiples of two. Tests with virus-free X. index populations reared on grapevine and fig plants as negative controls did not reveal a noticeable effect of the host plant. ELISA absorbances of virus-free X. index samples were greater than corresponding absorbances of X. pachtaicum samples. Differences occurred between two X. index field populations from GFLV-infected grapevines in Champagne and Languedoc. In most tests, 1-, 2-, 4-, and 8-nematode samples of virus-free and virus-infected populations, respectively, could not be separated. Consequently, B-A ELISA was not a reliable method for GFLV detection in samples of less than 10 X. index adults, but comparison of the absorbances obtained with increasing numbers may allow differentiation of the viral infectious potential of several populations.     

Cryofracturing for Scanning Electron Microscope Observations of Internal Structures of Nematodes

Nematodes were prepared for scanning electron microscopy by cryofracturing in ethanol and then by critical-point drying in carbon dioxide. Cross sections of Caenorhabditis briggsae and Xiphinema americanum showed the arrangement of the intestine, ovaries, muscle cells, and some layers of the cuticle. The technique is complementary to transmission electron microscopy and facilitates the interpretation of results from thin sections.     

Distribution of Xiphinema americanum and Related Species in North America

All species of the Xiphinema americanum-group and their synonyms are listed. The North American species reported are listed by state or province. Among these species, X. rivesi has the most widely reported distribution. Six species (X. diffusum, X. floridae, X. laevistriatum, X. luci, X. shell, and X. tarjanense) have been reported from only Florida. The reports of X. pachtaicum, X. sheri, and X. luci did not include morphometrics and need to be confirmed; X. brevicolle from California was identified before Lamberti and Bleve-Zacheo described 15 new species in 1979 and similarly needs to be confirmed. Because of the proliferation of species in this group, reports of X. americanum (sensu stricto) before 1979 are questionable. Extraction techniques for longidorids are discussed.     

Factors Affecting Population Trends of Plant-Parasitic Nematodes on Rangeland Grasses

The effects of environmental conditions on population trends of plant-parasitic nematodes were studied in experimental plots of five wheatgrasses in the western Utah desert. In a 3-year (1984-86) field study, soil water and temperature affected the population trends of the ectoparasites, Tylenchorhynchus acutoides and Xiphinema americanum, and the migratory endoparasite, Pratylenchus neglectus, on Fairway crested wheatgrass, Agropyron cristatum; ''Hycrest'' crested wheatgrass, A. cristatum X A. desertorura; ''Rosana'' western wheatgrass, Pascopyrum smithii; ''Oahe'' intermediate wheatgrass, Thinopyrum intermedium; and RS-1 hybrid (Elytrigia repens X Pseudoroegneria spicata). The largest soil populations of these nematode species were collected in 1984 under good plant-growth conditions. A reduction in nematode populations occurred in 1985 and 1986, possibly because of low soil-water conditions. There was a positive relationship between high soil water and maximum population densities of T. acutoides in the spring and fall of 1984, and between low soil water and minimum population densities of the nematode in 1985 and 1986. Pratylenchus neglectus populations were affected by soil water, although to a lesser degree than the ectoparasitic nematodes. Population densities of the three nematode species were significantly lower in the drier years of 1985 and 1986 than in 1984. Nematode populations were greater at the lower soil depths in the fall than in the spring or summer.     

Plant-parasitic Nematodes in Loess Toposequences Planted with Corn

Populations of plant-parasitic nematodes in an Iowa cornfield were studied along north- and west-facing toposequences. Samples were collected monthly during the growing season. The greatest biomass for Xiphinema americanum occurred at the footslope on the north face. Paratylenchus microdorus had its greatest biomass at the summit position, generally more in the west- than in the north-facing slope. Pratylenchus spp. in the roots peaked at the toeslope in the north-facing slope, but at the footslope in the west-facing slope. Helicotylenchus pseudorobustus peaked at the backslope and the toeslope along the north- and west-facing slopes, respectively. Diversity, as computed for each plot by the Shannon-Weiner diversity index, was highest at the backslope in both toposequences.     

Age Structure and Community Diversity of Nematodes Associated with Maize in Iowa Sandy Soils

Age structure of nematode populations around maize growing in sandy soils in Iowa was studied at soil depths of 0-15and 15-30 cm for 2 years. Numbers of Longidorus breviannulatus were generally greater at 0-15 cm than at 15-30 cm deep until mid to late season. The decline in numbers of females as the season progressed indicates that fecundity slowed and is evidence of only one generation per year. Peak populations of Pratylenchus scribneri and Xiphinema americanum occurred in late August or early September. Adults of Hoplolaimus galeatus were few in the roots but common in the soil, indicating that fertilization occurred mostly in the soil. Numbers of P. scribneri were generally greater at the lower depth, especially late in the season. Community diversity (H'') was less when nematode biomass was used instead of numbers. Numbers of H. galeatus did not decline over the winter. Numbers of L. breviannulatus, P. scribneri, and X. americanum declined significantly over the winter, but not between spring cultivation and planting.     

Transmission of Tomato Ringspot Virus by Xiphinema americanum and X. rivesi from New York Apple Orchards

Populations of Xiphinema americanum and X. rivesi were collected from apple orchards in eastern and western New York and tested in the laboratory for ability to transmit tomato ringspot virus (TmRSV) to cucumber and dandelion. Populations varied in the frequency with which they transmitted TmRSV, but this variation did not correspond to variation in disease prevalence in the orchard. The lower prevalence of TmRSV-incited disease in apple trees in western New York cannot be attributed to inability of the local Xiphinema spp. to transmit TmRSV.     

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.     

Community Structure of Plant-parasitic Nematodes Related to Soil Types in Illinois and Indiana Soybean Fields

Comparisons of plant parasitic nematode populations using a resemblance equation and community ordination showed that community structure tends to be similar on dark-colored, highly productive soybean soils throughout Indiana and Illinois. On lighter-colored soils community structures differed somewhat from those of darker soils and from each other.     

First report of Xiphinema brevicolle Lordello et Costa, 1961 (Nematoda, Longidoridae) in Japan

Mixed populations of Xiphinema americanum-group species were detected from a root zone soil sample of Japanese holly, Ilex crenata, during a survey for plant-parasitic nematodes of commercial ornamental plant nurseries in Chiba Prefecture, Japan. From the result of the morphological study, the species were identified as Xiphinema brevicolle and Xiphinema sp. This is the first record of Xiphinema brevicolle in Japan. Morphometrics of Xiphinema brevicolle generally agree with those of the type specimens and the topotype specimens. Xiphinema sp. morphometrically resembles Xiphinema paramonovi except for tail length. The mitochondrial COI region, the nuclear 18S rDNA and the nuclear large subunit rDNA D2/D3 region of the species were sequenced and compared in the molecular study. For the COI region, PCR primers were newly designed to obtain longer sequences, ca. 900 bp, than previously used. Sequence identities of COI, 18S and D2/D3 regions between these two populations were 84.0-84.1%, 99.9% and 98.1-98.2%, respectively. Phylogenetic analyses of maximum likelihood trees were carried out to compare genetic relationships among the group and some suggestions were made on the Xiphinema brevicolle-subgroup.     

Antagonists of Plant-parasitic Nematodes in Florida Citrus

In a survey of antagonists of nematodes in 27 citrus groves, each with a history of Tylenchulus semipenetrans infestation, and 17 noncitrus habitats in Florida, approximately 24 species of microbial antagonists capable of attacking vermiform stages of Radopholus citrophilus were recovered. Eleven of these microbes and a species of Pasteuria also were observed attacking vermiform stages of T. semipenetrans. Verticillium chlamydosporium, Paecilomyces lilacinus, P. marquandii, Streptomyces sp., Arthrobotrys oligospora, and Dactylella ellipsospora were found infecting T. semipenetrans egg masses. Two species of nematophagous amoebae, five species of predatory nematodes, and 29 species of nematophagous arthropods also were detected. Nematode-trapping fungi and nematophagous arthropods were common inhabitants of citrus groves with a history of citrus nematode infestation; however, obligate parasites of nematodes were rare.     

Identification of Cyst Nematodes of Agronomic and Regulatory Concern with PCR-RFLP of ITS1

The first internally transcribed spacer region (ITS1) from cyst nematode species (Heteroderidae) was compared by nucleotide sequencing and PCR-RFLP. European, Asian, and North American isolates of five heterodefid species were examined to assess intraspecific variation. PCR-RFLP patterns of amplified ITS1 DNA from pea cyst nematode, Heterodera goettingiana, from Northern Ireland were identical with patterns from Washington State. Sequencing demonstrated that ITS1 heterogeneity existed within individuals and between isolates, but did not result in different restriction patterns. Three Indian and two U.S. isolates of the corn cyst nematode, Heterodera zeae, were compared. Sequencing detected variation among ITS1 clones from the same individual, between individuals, and between isolates. PCR-RFLP detected several restriction site differences between Indian and U.S. isolates. The basis for the restriction site differences between isolates from India and the U.S. appeared to be the result of additional, variant ITS1 regions amplified from the U.S. isolates, which were not found in the three India isolates. PCR-RFLP from individuals of the U.S. isolates created a composite pattern derived from several ITS1 types. A second primer set was specifically designed to permit discrimination between soybean (H. glycines) and sugar beet (H. schachtii) cyst nematodes. Fok I digestion of amplified product from soybean cyst nematode isolates displayed a uniform pattern, readily discernible from the pattern of sugar beet and clover cyst nematode (H. trifolii).