Showing papers in "Journal of Nematology in 2005"

Incorporating Molecular Identification of Meloidogyne spp. into a Large-scale Regional Nematode Survey.

Abstract: A regional nematode survey of potato fields was conducted in the central United States during 2002 and 2003. The survey encompassed seven states and included a morphological and molecular examination of nematodes of regulatory concern from 1,929 soil samples. No regulated pest species were recovered during this survey. Meloidogyne juveniles extracted from soil were identified by mitochondrial and 18S ribosomal molecular markers. Eighty-two DNA sequences representing the two marker regions for Meloidogyne species were submitted to GenBank to facilitate evaluation of marker variability. Sufficient 18S variation was observed among some Meloidogyne species to aid in identification; however, nucleotide sequence from this highly conserved region of 18S did not discriminate among M. arenaria, M. incognita, and M. javanica. The mitochondrial gene region provided greater species discrimination and revealed intraspecific variation among many isolates. One nucleotide substitution found in a subset of M. hapla isolates from west Texas and New Mexico affected a DraI restriction site used in the PCR/RFLP diagnostic protocol. None of the mitochondrial sequence variants observed in this study compromised the PCR/RFLP identification protocol for M. chitwoodi. Additional sequence analysis is recommended for validation and evaluation of genetic markers used in diagnostic decisions.

Use of Cucumis metuliferus as a Rootstock for Melon to Manage Meloidogyne incognita

Abstract: Root-knot nematode-susceptible melons (Cantaloupe) were grown in pots with varying levels of Meloidogyne incognita and were compared to susceptible melons that were grafted onto Cucumis metuliferus or Cucurbita moschata rootstocks. In addition, the effect of using melons as transplants in nematode-infested soil was compared to direct seeding of melons in nematode-infested soil. There were no differences in shoot or root weight, or severity of root galling between transplanted and direct-seeded non-grafted susceptible melon in nematode-infested soil. Susceptible melon grafted on C. moschata rootstocks had lower root gall ratings and, at high nematode densities, higher shoot weights than non-grafted susceptible melons. However, final nematode levels were not lower on the grafted than on the non-grafted plants, and it was therefore concluded that grafting susceptible melon on to C. moschata rootstock made the plants tolerant, but not resistant, to the nematodes. Grafting susceptible melons on C. metuliferus rootstocks also reduced levels of root galling, prevented shoot weight losses, and resulted in significantly lower nematode levels at harvest. Thus, C. metuliferus may be used as a rootstock for melon to prevent both growth reduction and a strong nematode buildup in M. incognita-infested soil.

Plant-Parasitic Nematodes Associated with Reduced Wheat Yield in Oregon: Heterodera avenae.

Abstract: Heterodera avenae is widely distributed in the western United States, where most wheat is grown in non-irrigated winter wheat/summer fallow rotations in low rainfall regions. Economic and social pressures have motivated growers to pursue a transition from winter wheat/summer fallow rotation to no-till annual spring cereals. Annual cereals are also planted in some irrigated fields. The impact of H. avenae on spring wheat yield in the Pacific Northwest had been observed but not quantified. Spring wheat was planted with or without aldicarb to examine relationships between H. avenae and yield under dryland and irrigated conditions in moderately infested fields. Spring wheat yields were negatively correlated (P < 0.05) with initial populations of H. avenae. Aldicarb application improved spring wheat yield as much as 24%. The infective juvenile stage of H. avenae reached a peak density during mid-spring. Yield of irrigated annual winter wheat was also negatively correlated with initial density of H. avenae. Research priorities necessary to develop control strategies include a description of the pathotype, identification of sources for genetic resistance, and integrated practices designed to manage multiple yield-reducing pests.

Pratylenchus thornei Associated with Reduced Wheat Yield in Oregon.

Abstract: Pratylenchus thornei reaches high population densities in non-irrigated annual cropping systems in low-rainfall regions of the Pacific Northwest. Two spring wheat varieties with different levels of tolerance and susceptibility to P. thornei were treated or not treated with aldicarb in three experiments. Grain yield was inversely correlated (P < 0.05) with pre-plant populations of P. thornei in soil and with P. thornei density in mature roots. As population of P. thornei increased, yield of the moderately tolerant/moderately susceptible variety Krichauff was generally more stable than for the intolerant/susceptible variety Machete. The reproductive factor (Pf/Pi) was generally lower (P < 0.05) for Krichauff than Machete. Aldicarb improved wheat yield (P < 0.05) in highly infested fields by an average of 67% for Krichauff and 113% for Machete. Aldicarb increased (P < 0.05) numbers of headed tillers, plant height, and grain test weight and kernel weight, and reduced (P < 0.05) the density of P. thornei in mature wheat roots, variability in height of heads, and leaf canopy temperature. Aldicarb did not improve yield in a soil with a low population of P. thornei. This is the first report that P. thornei causes economic damage to wheat in the Pacific Northwest.

Detection and description of soils with specific nematode suppressiveness.

Abstract: Soils with specific suppressiveness to plant-parasitic nematodes are of interest to define the mechanisms that regulate population density. Suppressive soils prevent nematodes from establishing and from causing disease, and they diminish disease severity after initial nematode damage in continuous culturing of a host. A range of non-specific and specific soil treatments, followed by infestation with a target nematode, have been employed to identify nematode-suppressive soils. Biocidal treatments, soil transfer tests, and baiting approaches together with observations of the plant-parasitic nematode in the root zone of susceptible host plants have improved the understanding of nematode-suppressive soils. Techniques to demonstrate specific soil suppressiveness against plant-parasitic nematodes are compared in this review. The overlap of studies on soil suppressiveness with recent advances in soil health and quality is briefly discussed. The emphasis is on methods (or criteria) used to detect and identify soils that maintain specific soil suppressiveness to plant-parasitic nematodes. While biocidal treatments can detect general and specific soil suppressiveness, soil transfer studies, by definition, apply only to specific soil suppressiveness. Finally, potential strategies to exploit suppressive soils are presented.

Effect of Three Plant Residues and Chicken Manure used as Biofumigants at Three Temperatures on Meloidogyne incognita Infestation of Tomato in Greenhouse Experiments.

Abstract: Plant residues of broccoli, melon, and tomato with or without addition of chicken manure were used as biofumigants in two pot experiments with Meloidogyne incognita-infested soils. The efficacy of these biofumigants in controlling M. incognita infestation in susceptible tomato bio-assay plants was studied at soil temperatures of 20 masculine, 25 masculine, and 30 masculineC. None of the plant residues was effective at 20 masculineC, and broccoli was more effective than tomato or melon at 25 masculineC. At 30 masculineC all three plant residues reduced M. incognita infestation of tomato to very low levels. Chicken manure was effective in one of two experiments at 20 masculineC, and at 25 masculineC enhanced the efficacy of tomato and melon residue in one of two experiments. At 30 masculineC chicken manure was equally effective as the three plant residues but did not further decrease infestation levels in plant residue amended soils. It is concluded that biofumigation to control M. incognita is unlikely to be effective under cool conditions, that at soil temperatures around 25 masculineC broccoli is more effective than melon and tomato, and that the addition of chicken manure at this soil temperature may enhance the efficacy. At high soil temperatures, of approximately 30 masculineC, the biofumigant source seems of minor importance as strong reductions in tomato infestation by M. incognita were achieved by addition of each of the three plant residues as well as by addition of chicken manure.

Bionomics of a Phoretic Association Between Paenibacillus sp. and the Entomopathogenic Nematode Steinernema diaprepesi.

Abstract: Spores of an unidentified bacterium were discovered adhering to cuticles of third-stage infective juvenile (IJ) Steinernema diaprepesi endemic in a central Florida citrus orchard. The spores were cup-shaped, 5 to 6 mm in length, and contained a central endospore. Based on 16S rDNA gene sequencing, the bacterium is closely related to the insect pathogens Paenibacillus popilliae and P. lentimorbus. However, unlike the latter bacteria, the Paenibacillus sp. is non-fastidious and grew readily on several standard media. The bacterium did not attach to cuticles of several entomopathogenic or plant-parasitic nematodes tested, suggesting host specificity to S. diaprepesi. Attachment of Paenibacillus sp. to the third-stage cuticle of S. diaprepesi differed from Paenibacillus spp. associated with heterorhabditid entomopathogenic nematodes, which attach to the IJ sheath (second-stage cuticle). The inability to detect endospores within the body of S. diaprepesi indicates that the bacterial association with the nematode is phoretic. The Paenibacillus sp. showed limited virulence to Diaprepes abbreviatus, requiring inoculation of larvae with 108 spores to achieve death of the insect and reproduction of the bacterium. The effect of the bacterium on the nematode population biology was studied in 25-cm-long vertical sand columns. A single D. abbreviatus larva was confined below 15-cm depth, and the soil surface was inoculated with either spore-free or spore-encumbered IJ nematodes. After 7 days, the proportion of IJ below 5-cm depth was seven-fold greater for spore-free IJ than for spore-encumbered nematodes. Mortality of D. abbreviatus larvae was 72% greater (P = 0.01) for spore-free compared to spore-encumbered S. diaprepesi. More than 5 times as many progeny IJs (P = 0.01) were produced by spore-free compared to spore-encumbered nematodes. These data suggest that the bacterium is a component of the D. abbreviatus food web with some potential to regulate a natural enemy of the insect.

Intraspecific Variability of Rotylenchulus reniformis from Cotton-growing Regions in the United States.

Abstract: Reniform nematode (Rotylenchulus reniformis) is a major pest of cotton in the southeastern United States. The objective of this study was to examine the variation of reniform nematode populations from cotton-growing locations in the United States where it is prevalent. Multivariate analysis of variance and discriminant analysis were used to determine the variability of morphology in males and immature females. Reproduction indices of populations were measured on selected soybean and cotton genotypes in the greenhouse. High variability in morphometrics and reproduction was observed within all the populations, and several differences were found among populations. DNA sequences of the nuclear ribosomal first internal transcribed spacer region (ITS1) were compared among populations from the United States and to sequences of populations from Brazil, Colombia, Honduras, and Japan. No polymorphic nucleotide sites were observed among the amphimictic populations. Only a parthenogenic population from Japan was distinct. The phenotypic polymorphism of the species in the United States could impact the effectiveness of management strategies based on host plant resistance.

Vertical Distribution of Bacterivorous Nematodes under Different Land Uses.

Abstract: The vertical distribution of dominant genera of bacterivorous nematodes to 150-cm depth in an aquic brown soil was compared after 14 years of four contrasting land uses, i.e., cropland-rice (CR), cropland-maize (CM), abandoned cropland (AC), and woodland (WL). The study was conducted at the Shenyang Experimental Station of Ecology, a Chinese Ecosystem Research Network (CERN) site in Northeast China. Data were analyzed using two-way analysis of variance with land use and depth as independent variables. More than 70% of Chiloplacus, Eucephalobus, and Monhystera spp. were present in the uppermost soil layer (0 to 5 cm) in the CR treatment. In contrast, Chiloplacus and Prismatolaimus spp. were distributed down to 100-cm depth in the AC and CM treatments, respectively. Differences in numbers of Acrobeles, Acrobeloides, Cephalobus, Chiloplacus, Eucephalobus, Monhystera, Plectus, and Prismatolaimus were found among land uses and at various depths. Soil C and N were correlated positively with numbers of Monhystera and Plectus in the CR treatment, Acrobeloides in the CM treatment, and Acrobeles and Acrobeloides in the AC treatment. Soil pH was correlated negatively with Monhystera, Plectus (CR), and Acrobeloides (CM, AC). The relationship of pH with Acrobeles depended on land use: positive in the WL treatment and negative in the AC treatment. Our results suggested that Cephalobus and Prismatolaimus in the CR treatment, and Chiloplacus and Prismatolaimus in the WL treatment, were insensitive to soil properties measured. Differences in vertical distribution should be considered when studying dominant bacterivorous nematode genera among land uses.

Effect of Forage and Grain Pearl Millet on Pratylenchus penetrans and Potato Yields in Quebec.

Abstract: Rotation crop experiments were conducted from 1998 to 2000 to assess the impact of forage and grain pearl millet (Pennisetum glaucum) on Pratylenchus penetrans populations in three potato (Solanum tuberosum cv. Superior) fields in Quebec. These crops were compared to oats and(or) barley. Forage millet had a suppressive effect on P. penetrans populations after a 1 year rotation. The following year, marketable potato yields were negatively correlated with initial P. penetrans densities on two experimental sites (r = -0.454, P = 0.044; r = -0.426, P = 0.017). Average marketable and total yields were increased by 10% in plots previously grown in forage millet hybrid CFPM 101 when compared to oats (P = 0.017). Damage functions between preplant nematode density (Pi) and marketable yield (y = 42.0 -4.091 log₁₀ [Pi + 1]) and total yield (y = 43.9 -4.039 log₁₀ [Pi + 1]) of potato were established on pooled yield data. Forage pearl millet is an efficient and economically viable alternative for managing root-lesion nematodes and improving potato yields in Quebec.

Diversity of Meloidogyne spp. on Musa in Martinique, Guadeloupe, and French Guiana.

Abstract: Ninety-six isolates of Meloidogyne species collected from banana fields from Martinique, Guadeloupe, and French Guiana, were examined using esterase (Est) and malate dehydrogenase (Mdh) phenotypes. Adult females identified as M. arenaria, M. incognita, M. javanica, M. cruciani, M. hispanica, and Meloidogyne sp. showed species-specific phenotypes only for the esterase enzymes. Intraspecific variability among isolates of M. arenaria, M. incognita, and M. javanica was detected using Est and Mdh. Perineal patterns were used as a complementary tool together with enzyme characterization and were essential for checking the morphological consistency of the identification. The major species of M. arenaria and M. incognita were detected at 61.9% and 34.3% of the total number of isolates, respectively, and the other minor species at 3.8%. The mixed Meloidogyne species were detected in 45.1% of the samples. Genetic analysis was conducted using RAPD markers, which alone or in combination provided reliable polymorphisms both between and within species. RAPD analysis of the data resulted in clustering of species and isolates congruent with esterase phenotype characterization. The intraspecific variability in M. incognita and in M. arenaria represented 14.9% and 61.6% of the amplified polymorphic fragments, respectively. This high level of variation in M. arenaria isolates may indicate multiple origins for populations classified as M. arenaria or more than one species inside the same group, but more detailed morphological and DNA studies will be necessary to test this hypothesis.

Morphological and Molecular Characterization of a New Root-Knot Nematode, Meloidogyne thailandica n. sp. (Nematoda: Meloidogynidae), Parasitizing Ginger (Zingiber sp.).

Abstract: A root-knot nematode Meloidogyne thailandica n. sp. was discovered on roots of ginger (Zingiber spp.) intercepted from Thailand in October 2002 by the U.S. Department of Agriculture Animal and Plant Health Inspection Service at the port of San Francisco. Comparison by light microscopy (LM) and scanning electron microscopy (SEM) to five other morphologically related species (M. incognita, M. arenaria, M. microcephala, M. megatyla, and M. enterolobii) revealed that the new species differs from these by one or more of the following: body, tail and hyaline tail length, shape of head, tail and tail terminus of second-stage juveniles; stylet length and shape of spicules in males; perineal pattern, stylet length and shape of knobs in females. The distinctive perineal pattern is oval to rectangular, with smooth to moderately wavy and coarse striae, and with characteristic radial structures present underneath the pattern area; the dorsal arch is high, sometimes round to rectangular, and striae in and around the anal area form a thick network-like pattern interrupted by lateral lines and large phasmids. Second-stage juveniles have a long, slender tail and long, gradually tapering hyaline tail region ending in a rounded terminus. Male spicules commonly have an acutely angled shaft with a bidentate terminus. Molecular data from the ribosomal large subunit D3 expansion segment revealed four haplotypes, two of which were unique and distinguish M. thailandica n. sp. from M. arenaria, M. incognita, and M. javanica.

Vertical Distribution of Rotylenchulus reniformis in Cotton Fields.

Abstract: The possible impact of Rotylenchulus reniformis below plow depth was evaluated by measuring the vertical distribution of R. reniformis and soil texture in 20 symptomatic fields on 17 farms across six states. The mean nematode population density per field, 0 to 122 cm deep, ranged from 0.4 to 63 nematodes/g soil, and in 15 fields more than half of the R. reniformis present were below 30.5 cm, which is the greatest depth usually plowed by farmers or sampled by consultants. In 11 fields measured, root density was greatest in the top 15 cm of soil; however, roots consistently penetrated 92 to 122 cm deep by midseason, and in five fields in Texas and Louisiana the ratio of nematodes to root-length density within soil increased with depth. Repeated sampling during the year in Texas indicated that up to 20% of the nematodes in soil below 60 cm in the fall survived the winter. Differences between Baermann funnel and sugar flotation extraction methods were not important when compared with field-to-field differences in nematode populations and field-specific vertical distribution patterns. The results support the interpretation that R. reniformis below plow depth can significantly impact diagnosis and treatment of cotton fields infested with R. reniformis.

Bursaphelenchus anatolius n. sp. (Nematoda: Parasitaphelenchidae), an Associate of Bees in the Genus Halictus.

Abstract: Bursaphelenchus anatolius n. sp., a phoretic associate of Halictus bees from Ankara, Turkey, is described and illustrated. Bursaphelenchus anatolius n. sp. is closest to B. kevini, which is phoretically associated with Halictus bees from the Pacific Northwest. Bursaphelenchus anatolius n. sp. and B. kevini appear to be sister taxa based upon several shared morphological features, similar life histories involving phoresy with soil-dwelling Halictus bees, and molecular analysis of the near-full-length small subunit rDNA, D2D3 expansion segments of the large subunit rDNA, and partial mitochondrial DNA COI. Bursaphelenchus anatolius n. sp. can be differentiated from all other species of Bursaphelenchus based upon spicule morphology. The paired spicules are uniquely shaped and ventrally recurved, and both B. anatolius n. sp. and B. kevini possess extending flaps that open when the spicules are protracted beyond the cloaca. Population growth of B. anatolius n. sp. was measured at 23 °C in the laboratory on cultures of the fungus Monilinia fructicola grown on lactic acid-treated, 5% glycerol-supplemented potato dextrose agar. Nematode population densities rapidly increased from 110 to about 110,000/9-cm-diam. dish within 21 days.

Histological Observations of Rotylenchulus reniformis on Gossypium longicalyx and Interspecific Cotton Hybrids.

Abstract: Observations on the development of reniform nematode (Rotylenchulus reniformis) on roots of Gossypium longicalyx, G. hirsutum, and two interspecific hybrids derived from them were made by light microscopy. Gossypium longicalyx is reported to be immune to reniform nematode, but the mechanism(s) for resistance are unknown. Penetration of G. longicalyx roots by female nematodes was confirmed, and incipient swelling of the females, indicating initiation of maturation of the reproductive system, was observed. Female maturation occurred up to the formation of a single embryo inside the female body but not beyond this point. In both hybrids, development was inhibited but progressed further than in the immune parent. Reactions ranged from highly compatible, with the formation of active syncytia and full development of females, to incompatible with little or no development of the female. Compatible plants showed characteristic hypertrophied cells, enlarged nuclei, dense cytoplasm, and partial dissolution of cell walls, whereas incompatible plant reactions included lignification of the cells adjacent to the nematode head, or the complete collapse and necrosis of the cells involved. The need to characterize reactions and to carefully select among the plants descended from the hybrids during the introgression process, as well as the importance of combining the results of reproduction tests with histological observation of the plant-nematode interactions, is discussed.

Morphological and Molecular Evaluation of a Meloidogyne hapla Population Damaging Coffee (Coffea arabica) in Maui, Hawaii.

Abstract: An unusual population of Meloidogyne hapla, earlier thought to be an undescribed species, was found causing large galls, without adventitious roots, and substantial damage to coffee in Maui, Hawaii. Only in Brazil had similar damage to coffee been reported by this species. Unlike M. exigua from South and Central America, this population reproduced well on coffee cv. Mokka and M. incognita-susceptible tomato but poorly on tomato with the Mi resistance gene. Characterization included SEM images, esterase isozymes, and five DNA sequences: i) the D3 segment of the large subunit (LSU-D3 or 28S) rDNA, ii) internal transcribed spacer (ITS-1) rDNA, iii) intergenic spacer (IGS) rDNA, iv) the mitochondrial interval from cytochrome oxidase (CO II) to 16S mtDNA, and v) the nuclear gene Hsp90. Sequences for ITS-1, IGS, and COII were similar to other M. hapla populations, but within species ITS-1 variability was not less than among species. One LSU-D3 haplotype was similar to a previously analyzed population with two minor haplotypes. Hsp90 exhibited some variation between Maryland and Hawaiian populations distinct from other species. Females were narrow with wide vulval slits, large interphasmidial distances, and more posterior excretory pores; 20% of perineal patterns had atypical perivulval lines. Males had a low b ratio (<12 µm). Juveniles had a short distance between stylet and dorsal gland orifice. Juvenile body length was short (<355 µm) and was different between summer and winter populations.

Nematode gene sequences: update for december 2005.

Abstract: Molecular characterization of parasitic nematodes now relies heavily on information gained from genomic approaches. The avai l abi l i t y of t housands of new nucleotide sequences in searchable public databases ac- celerates efforts to identify and characterize genes of interest, including those encoding pathogenicity factors, diagnostic markers, targets (for vaccines, drugs, or nemat i ci des ), and i mmunomodul at ory molecules . Since 2000, we have provided periodic updates on the status of nematode sequencing projects of interest to the communities of researchers studying parasitic and free-living nematode biology (McCarter et al., 2000, 2002, 2003). Here we report on the availability of sequences from nematodes as of late 2005 and primarily cite literature from 2003–2005. Please see our 2003 update (McCarter et al., 2003) for a thorough listing of the prior literature. 2003 to 2005, EST sequencing from mammalian para- sites has slowed whereas EST generation from plant parasites has continued as the National Science Foundation (NSF)-funded collaboration between the GSC and North Carolina State University has been completed (Fig. 1). Sizable EST collections now exist for six Meloidogyne (root-knot nematode) species (Scholl and Bird, 2005), which is more than for any other nematode genus. ESTs have also been generated from three Tylenchida migratory endoparasitic species, the lesion nematodes Pratylenchus penetrans (Mitreva et al., 2004a) and Pratylenchus vulnus, and the burrowing nematode Radopholus similis. A collection of more than 9,000 ESTs also is available from the dagger nematode Xiphinema index. Being a clade I Dorylaimia nematode, this repre- sents the first EST collection from a non-Tylenchida

Reproduction of Belonolaimus longicaudatus, Meloidogyne javanica, Paratrichodorus minor, and Pratylenchus brachyurus on Pearl Millet (Pennisetum glaucum).

Abstract: Pearl millet (Pennisetum glaucum) has potential as a grain crop for dryland crop production in the southeastern United States. Whether or not pearl millet will be compatible in rotation with cotton (Gossypium hirsutum), corn (Zea mays), and peanut (Arachis hypogaea) will depend, in part, on its host status for important plant-parasitic nematodes of these crops. The pearl millet hybrid 'TifGrain 102' is resistant to both Meloidogyne incognita race 3 and M. arenaria race 1; however, its host status for other plant-parasitic nematodes was unknown. In this study, the reproduction of Belonolaimus longicaudatus, Paratrichodorus minor, Pratylenchus brachyurus, and Meloidogyne javanica race 3 on pearl millet ('HGM-100' and TifGrain 102) was compared relative to cotton, corn, and peanut. Separate greenhouse experiments were conducted for each nematode species. Reproduction of B. longicaudatus was lower on peanut and the two millet hybrids than on cotton and corn. Reproduction of P. minor was lower on peanut and TifGrain 102 than on cotton, corn, and HGM-100. Reproduction of P. brachyurus was lower on both millet hybrids than on cotton, corn, and peanut. Reproduction of M. javanica race 3 was greater on peanut than on the two millet hybrids and corn. Cotton was a nonhost. TifGrain 102 was more resistant than HGM-100 to reproduction of B. longicaudatus, P. minor, and M. javanica. Our results demonstrated that TifGrain 102 was a poor host for B. longicaudatus and P. brachyurus (Rf 1) and, relative to other crops tested, was less likely to increase densities of P. minor and M. javanica.

Alternatives to fenamiphos for management of plant-parasitic nematodes on bermudagrass.

Abstract: Plant-parasitic nematodes can be very damaging to turfgrasses. The projected cancellation of the registration for fenamiphos in the near future has generated a great deal of interest in identifying acceptable alternative nematode management tactics for use on turfgrasses. Two field experiments were conducted to evaluate the effectiveness of repeated applications of several commercially available nematicides and root biostimulants for reducing population densities of plant-parasitic nematodes and (or) promoting health of bermudagrass in nematode-infested soil. One experimental site was infested with Hoplolaimus galeatus and Trichodorus obtusus, the second with Belonolaimus longicaudatus. In both trials, none of the experimental treatments reduced population densities (P

Impacts of Sustained Use of Dairy Manure Slurry and Fertilizers on Populations of Pratylenchus penetrans under Tall Fescue

Abstract: Various manures and composts have been reported to reduce population densities of plant-parasitic nematodes. Dairy manure slurry is often used as a primary source of nitrogen for forage crops. This study was conducted to determine the effects of dairy manure on population densities of Pratylenchus penetrans parasitizing tall fescue. Beginning in 1994, dairy manure and inorganic fertilizer were applied after each harvest (2 to 4 times/year) at rates of 50 and 100 kg NH₄-N/ha; control plots were not treated. Nematode populations in soil and roots were determined at 19 sample dates during the fourth (1997), fifth (1998), and sixth (1999) years of manure and fertilizer applications. The sustained use of dairy manure and fertilizer increased population densities of P. penetrans. Our results contrast with many previous studies demonstrating that application of manures decreases population densities of plant-parasitic nematodes. Frequent applications of moderate amounts of manure to a perennial grass crop may have prevented the development of nematode-toxic levels of ammonia or other toxic substances such as nitrous acid or volatile fatty acids. Two years with no additional manure applications were required for P. penetrans population densities to return to levels similar to fertilized or untreated soil.

Molecular Polymorphism and Morphometrics of Species of the Heterodera avenae Group in Syria and Turkey

Abstract: Molecular characterization of the three most common cereal cyst nematode species of the Heterodera avenae group (H. avenae, H. filipjevi, and H. latipons), originating from various locations in major cereal-cultivating areas in Syria and Turkey, showed distinct restriction fragment patterns of the ITS-rDNA following PCR amplification and RFLP digestion with four endonucleases (Hae III, Hinf I, Ita I, and Pst I). Genetic dissimilarity within H. avenae group populations increased in comparison with H. avenae and other species; it was 0.164 with H. filipjevi and 0.354 with H. latipons populations. No intraspecific polymorphism was observed within H. latipons or H. filipjevi populations. Principal component analysis revealed contrasted correlations among 12 morphological parameters of cysts and juveniles of the three Heterodera species that separated them and distinguished differences within populations of H. latipons. Our results showed a clear separation of the three cyst nematode species on cereal using a conventional method for classification and molecular tests, and confirmed the congruence between genetics and morphological traits.

Effect of short-chain fatty acids and soil atmosphere on Tylenchorhynchus spp

Abstract: Short-chain fatty acids can be produced under anaerobic conditions by fermentative soil microbes and have nematicidal properties. We evaluated the effects of butyric and propionic acids on death and recovery of stunt nematodes (Tylenchorhynchus spp.), a common parasite of turfgrass. Nematodes in a sand-soil mix (80:20) were treated with butyric or propionic acid and incubated under air or N[sub2] for 7 days at 25 oC. Amendment of soil with 0.1 and 1.0 µmol (8.8 and 88 µg) butyric acid/g soil or 1.0 µmol (74 µg) propionic acid/g soil resulted in the death of all nematodes. The composition of the soil atmosphere had no effect on the nematicidal activity of the acids. Addition of hydrochloric acid to adjust soil pH to 4.4 and 3.5 resulted in nematode mortality relative to controls (41% to 86%) but to a lesser degree than short-chain fatty acids at the same pH. Nematodes did not recover after a 28-day period following addition of 10 µmol butyric acid/g soil under air or N[sub2]. Carbon mineralization decreased during this period, whereas levels of inorganic N and microbial biomass-N remained constant. Short-chain fatty acids appear to be effective in killing Tylenchorhynchus spp. independent of atmospheric composition. Nematode mortality appears to be a function of the type and concentration of fatty acid and soil pH.

Factors affecting the suppression of Heterodera glycines by N-viro soil

Abstract: Previous laboratory research demonstrated that N-Viro Soil (NVS), an alkaline-stabilized municipal biosolid, suppressed plant-parasitic nematodes. This study continued to explore the use of NVS as a nematode management tool specifically addressing factors that could influence its use. N-Viro Soil from different locations, the components of NVS (de-watered biosolids and fly ash admixtures), and sterilized NVS were applied to sand microcosms to determine effects on nematode survival sand solution pH and ammonia concentrations. This study confirmed the previous finding that an important mechanism of Heterodera glycines suppression by NVS was the generation of alkaline soil conditions. Only the fly ash admixture that resulted in an increase in pH to 10.0 suppressed H. glycines to the same level as NVS. Alkaline-stabilization of biosolids was necessary to achieve nematode suppression. Biosolids applied at rates <3% dry w/w did not suppress H. glycines to the same level as equivalent amounts of NVS. Sand solution pH levels after biosolid application, regardless of rate, were approximately 8.5 whereas 1% and 4% w/w NVS amendment resulted in pH levels of 10.3 and 11.6, respectively. NVS from different processing facilities were all effective in suppressing H. glycines. The NVS source that produced the highest concentration of ammonia did not reduce H. glycines survival to the same level as those sources generating pH levels above 10.1. Microbes associated with NVS appeared not to be responsible for the nematode suppressiveness of the amendment; there was no difference in nematode suppression between autoclaved and nonautoclaved NVS. The role that ammonia plays in the suppression of H. glycines by NVS is still unclear.

Rolling Circle Amplification of Complete Nematode Mitochondrial Genomes

Abstract: To enable investigation of nematode mitochondrial DNA evolution, methodology has been developed to amplify intact nematode mitochondrial genomes in preparative yields using a rolling circle replication strategy. Successful reactions were gener- ated from whole cell template DNA prepared by alkaline lysis of the rhabditid nematode Caenorhabditis elegans and a mermithid nematode, Thaumamermis cosgrovei. These taxa, representing the two major nematode classes Chromodorea and Enoplea, maintain mitochondrial genomes of 13.8 kb and 20.0 kb, respectively. Efficient amplifications were conducted on template DNA isolated from individual or pooled nematodes that were alive or stored at 80°C. Unexpectedly, these experiments revealed that multiple T. cosgrovei mitochondrial DNA haplotypes are maintained in our local population. Rolling circle amplification products can be used as templates for standard PCR reactions with specific primers that target mitochondrial genes or for direct DNA sequencing.

Root-knot Nematode Problem of Some Winter Ornamental Plants and Its Biomanagement.

Abstract: A microplot study under field conditions was carried out during 2 consecutive years to assess the effect of root-knot nematode infection (2,000 Meloidogyne incognita eggs/kg soil) on three winter ornamental plants: hollyhock (Althea rosea), petunia (Petunia hybrida), and poppy (Papaver rhoeas). Effects of root-dip treatment with the biocontrol agents Pochonia chlamydosporia, Bacillus subtilis, and Pseudomonas fluorescens and the nematicide fenamiphos were tested. The three ornamental species were highly susceptible to M. incognita, developing 397 and 285 (hollyhock), 191 and 149 (petunia), and 155 and 131 (poppy) galls and egg masses per root system, respectively, and exhibited 37% (petunia), 29% (poppy), and 23% (hollyhock) (P = 0.05) decrease in the flower production. Application of fenamiphos, P. chlamydosporia, P. fluorescens, and B. subtilis suppressed nematode pathogenesis (galls + egg masses) by 64%, 37%, 27%, and 24%, respectively, leading to 14% to 29%, 7% to 15%, 14% to 36%, and 7% to 33% increase in the flower production of the ornamental plants, respectively. Treatment with P. fluorescens also increased the flowering of uninfected plants by 11% to 19%. Soil population of M. incognita was decreased (P = 0.05) due to various treatments from 2 months onward, being greatest with fenamiphos, followed by P. chlamydosporia, B. subtilis, and P. fluorescens. Frequency of colonization of eggs, egg masses, and females by the bioagents was greatest by P. chlamydosporia, i.e., 25% to 29%, 47% to 60%, and 36% to 41%, respectively. Colonization of egg masses by B. subtilis and P. fluorescens was 28% to 31% and 11% to 13%, respectively, but the frequency was 0.3% to 1.3% in eggs. Rhizosphere population of the bioagents was increased (P = 0.05) over time, being usually greater in the presence of nematode.

Rotylenchulus reniformis below Plow Depth Suppresses Cotton Yield and Root Growth.

Abstract: Damage to cotton by Rotylenchulus reniformis below plow depth was evaluated in a sandy clay loam soil at Weslaco, Texas. In December 1999, 14 holes on 51-cm centers were dug 91 cm deep along the planting bed and adjacent furrow and 2 ml of 1,3-dichloropropene was placed 91, 61, and 30 cm deep as each hole was refilled and packed. This technique eliminated 96%, 81%, and 74% of R. reniformis down to 107 cm at distances 0, 25, and 51 cm laterally from the point of application (P = 0.05), whereas chisel fumigation at 168 liters/ha 43 cm deep reduced nematode numbers only in the top 61 cm (P = 0.001). Manual placement of fumigant increased yield 92%; chisel fumigation increased yield 88% (P = 0.005). A second experiment in February 2001 placed fumigant 43 or 81 cm deep, or at both 43 and 81 cm. Holes alone had no significant effect on nematode density at planting, midseason or harvest, on root length density at midseason, or on cotton lint yield. Fumigant at 43 cm reduced nematode numbers above fumigant application depth at planting 94% (P = 0.02), at midseason 37% (P = 0.09), and at harvest 0%, increasing yield 57% (P = 0.002). Fumigant at 81 cm reduced nematode numbers above fumigant application depth at planting 86% (P = 0.02), at midseason 74% (P = 0.02), and at harvest 48% (P = 0.01), increasing yield 53% (P = 0.002). Fumigating at both 43 and 81 cm reduced nematode numbers above 90 cm 94% at planting and 79% at midseason, increased midseason root-length density 14-fold below 76 cm, and doubled yield (P = 0.02 in all cases).

Histological Changes in Gossypium hirsutum Associated with Reduced Reproduction of Rotylenchulus reniformis.

Abstract: The reniform nematode (Rotylenchulus reniformis) is an important parasite of upland cotton (Gossypium hirsutum). Parasitism involves the formation of syncytia to provide nutrition for the female. Events that occur at the feeding site may determine the degree of susceptibility of cotton plants to reniform nematode. The objective of this work was to describe histological modifications associated with reduced reproduction of Rotylenchulus reniformis in upland cotton roots. 'Deltapine 50' cotton and a selection from this line with a moderate level of resistance were inoculated with reniform nematode in the greenhouse, and observations on roots were made 3, 6, 9, 12, and 15 days after inoculation. No differences in penetration behavior or in the formation and characteristics of syncytia were observed. Reduced reproduction was correlated with an earlier degeneration and collapse of the syncytial cells, and occasionally, with lack of hypertrophy of the pericycle cells involved. These two mechanisms accounted for 40% to 60% reduction of reproduction of reniform nematode in the plants examined.

Variability of Meloidogyne exigua on Coffee in the Zona da Mata of Minas Gerais State, Brazil.

Abstract: Minas Gerais is the major coffee-producing state of Brazil, with 28% of its production coming from the region of Zona da Mata. Four major species of root-knot nematode attacking coffee (Meloidogyne incognita, M. paranaensis, M. coffeicola, and M. exigua) have been reported from Brazil. To determine the variability in Meloidogyne spp. occurring in that region, 57 populations from 20 localities were evaluated for morphological, enzymatic, and physiological characteristics. According to the perineal pattern, all the populations were identified as M. exigua; however populations from the municipality of Sao Joao do Manhuacu exhibited patterns very similar to M. arenaria. The identity of all the populations was confirmed by the phenotypes of esterase, malate dehydrogenase, superoxide dismutase, and glutamate-oxaloacetate transaminase. Thirteen populations (22.8%) showed the typical one-band (E1) esterase phenotype, whereas the others (77.2%) had a novel two-band phenotype (E2). No intraspecies variability was found in any population. All populations were able to reproduce on tomato, pepper, beans, cacao, and soybean. Reproduction was greater on tomato and pepper than on coffee seedlings, the susceptible standard.

Recycling Potential and Fitness of Steinernematid Nematodes Cultured in Curculio caryae and Galleria mellonella.

Abstract: The recycling potential of entomopathogenic nematodes in the pecan weevil, Curculio caryae, following inundative applications is an important factor in considering whether nematodes could be incorporated into a C. caryae management strategy. Our objective was to determine the recycling potential and fitness of Steinernema carpocapsae and S. riobrave cultured in C. caryae. To estimate fitness and quality, we reared nematodes in larvae of C. caryae and in the commonly used standard host, the greater wax moth, Galleria mellonella. Nematode lipid content, infectivity (power to invade), virulence (power to kill), and reproductive capacity (yield per insect) in C. caryae larvae were compared with G. mellonella data. Lipid content was higher in S. carpocapsae cultured in C. caryae than in G. mellonella, but S. riobrave lipid content was not affected by host source. Host source did not affect subsequent infectivity or virulence to C. caryae (P > 0.05) but did affect reproductive capacity (P < 0.0001). Both nematode species produced more progeny in C. caryae when they were first cultured in G. mellonella than when they were first passed through C. caryae. In terms of potential to recycle under field conditions, we predict that nematodes resulting from one round of recycling in C. caryae larvae would be equally capable of infecting and killing more weevils, but the potential to continue recycling in C. caryae would diminish over time due to reduced reproduction in that host.

Generalized linear dynamics of a plant-parasitic nematode population and the economic evaluation of crop rotations.

Abstract: In 1-year experiments, the final population density of nematodes is usually modeled as a function of initial density. Often, estimation of the parameters is precarious because nematode measurements, although laborious and expensive, are imprecise and the range in initial densities may be small. The estimation procedure can be improved by using orthogonal regression with a parameter for initial density on each experimental unit. In multi-year experiments parameters of a dynamic model can be estimated with optimization techniques like simulated annealing or Bayesian methods such as Markov chain Monte Carlo (MCMC). With these algorithms information from different experiments can be combined. In multi-year dynamic models, the stability of the steady states is an important issue. With chaotic dynamics, prediction of densities and associated economic loss will be possible only on a short timescale. In this study, a generic model was developed that describes population dynamics in crop rotations. Mathematical analysis showed stable steady states do exist for this dynamic model. Using the Metropolis algorithm, the model was fitted to data from a multi-year experiment on Pratylenchus penetrans dynamics with treatments that varied between years. For three crops, parameters for a yield loss assessment model were available and gross margin of the six possible rotations comprising these three crops and a fallow year were compared at the steady state of nematode density. Sensitivity of mean gross margin to changes in the parameter estimates was investigated. We discuss the general applicability of the dynamic rotation model and the opportunities arising from combination of the model with Bayesian calibration techniques for more efficient utilization and collection of data relevant for economic evaluation of crop rotations.