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2.  Detection and Investigation of Soil Biological Activity against Meloidogyne incognita 
Journal of nematology  2008;40(2):109-118.
Greenhouse experiments with two susceptible hosts of Meloidogyne incognita, a dwarf tomato and wheat, led to the identification of a soil in which the root-knot nematode population was reduced 5- to 16-fold compared to identical but pasteurized soil two months after infestation with 280 M. incognita J2/100 cm3 soil. This suppressive soil was subjected to various temperature, fumigation and dilution treatments, planted with tomato, and infested with 1,000 eggs of M. incognita/100 cm3 soil. Eight weeks after nematode infestation, distinct differences in nematode population densities were observed among the soil treatments, suggesting the suppressiveness had a biological nature. A fungal rRNA gene analysis (OFRG) performed on M. incognita egg masses collected at the end of the greenhouse experiments identified 11 fungal phylotypes, several of which exhibited associations with one or more of the nematode population density measurements (egg masses, eggs or J2). The phylotype containing rRNA genes with high sequence identity to Pochonia chlamydosporia exhibited the strongest negative associations. The negative correlation between the densities of the P. chlamydosporia genes and the nematodes was corroborated by an analysis using a P. chlamydosporia-selective qPCR assay.
PMCID: PMC2586535  PMID: 19259527
biological control; dwarf tomato; Meloidogyne incognita; Pochonia chlamydosporia; root-knot nematode; Solanum lycopersicon; suppressive soil; Triticum aestivum; wheat
3.  Host-Finding and Invasion by Entomopathogenic and Plant-Parasitic Nematodes: Evaluating the Ability of Laboratory Bioassays to Predict Field Results 
Journal of nematology  2008;40(2):93-98.
Directly viewing soil-dwelling entomopathogenic and plant-parasitic nematodes in situ is difficult, if not impossible. As a result, researchers have developed a diverse array of bioassays which assess nematode behavioral traits within arenas designed to simulate various aspects of the natural habitat. However, reliably rendering what we can see in the laboratory into accurate predictions of how nematodes achieve their objectives in the field is challenging. In the current review, we systemically assessed the goals and attributes of several of the assays most commonly used to investigate nematode host finding and host invasion behavior. By illuminating the relative strengths and limitations of each assay, we hope to improve our ability to develop meaningful predictions for the field.
PMCID: PMC2586539  PMID: 19259525
behavior; chemotaxis; ecology; electrophysiology; olfactometer; soil column
4.  From Augmentation to Conservation of Entomopathogenic Nematodes: Trophic Cascades, Habitat Manipulation and Enhanced Biological Control of Diaprepes abbreviatus Root Weevils in Florida Citrus Groves 
Journal of nematology  2008;40(2):73-84.
The use of entomopathogenic nematodes (EPN) for management of the root weevil, Diaprepes abbreviatus, in Florida citrus groves is considered a biological control success story and typically involves augmentation in which EPN are applied inundatively as biopesticides to quickly kill the pest. However, recent evidence indicates that efficacy of EPN applications in Florida citrus depends on soil type. They are very effective in the well drained coarse sands of the Central Ridge but often less so in poorly drained fine-textured soils of the Flatwoods. Moreover, groves on the Central Ridge can harbor rich communities of endemic EPN that might often suppress weevil populations below economic thresholds, whereas Flatwoods groves tend to have few endemic EPN and frequent weevil problems. Current research is examining the ecological dynamics of EPN in Florida citrus groves, the potential impact of EPN augmentation on soil food webs, especially endemic EPN, and whether habitat manipulation and inoculation strategies might be effective for conserving and enhancing EPN communities to achieve long-term control in problem areas. Conservation biological control could extend the usefulness of EPN in Florida citrus and be especially appropriate for groves with persistent weevil problems.
PMCID: PMC2586536  PMID: 19259523
biological control; citrus; conservation; Diaprepes abbreviatus; ecology; entomopathogenic nematode; Florida; food web; Heterorhabditis; management; manure; mulch; root weevil; soil; Steinernema; suppression; trophic cascades
5.  Potential for Entomopathogenic Nematodes in Biological Control: A Meta-Analytical Synthesis and Insights from Trophic Cascade Theory 
Journal of nematology  2008;40(2):61-72.
Entomopathogenic nematodes (EPN) are ubiquitous and generalized consumers of insects in soil food webs, occurring widely in natural and agricultural ecosystems on six continents. Augmentative releases of EPN have been used to enhance biological control of pests in agroecosystems. Pest managers strive to achieve a trophic cascade whereby natural-enemy effects permeate down through the food web to suppress host herbivores and increase crop production. Although trophic cascades have been studied in diverse aboveground arthropod-based systems, they are infrequently investigated in soil systems. Moreover, no overall quantitative assessment of the effectiveness of EPN in suppressing hosts with cascading benefits to plants has been made. Toward synthesizing the available but limited information on EPN and their ability to suppress prey and affect plant yield, we surveyed the literature and performed a meta-analysis of 35 published studies. Our analysis found that effect sizes for arthropod hosts as a result of EPN addition were consistently negative and indirect effects on plants were consistently positive. Results held across several different host metrics (abundance, fecundity and survival) and across measures of plant performance (biomass, growth, yield and survival). Moreover, the relationship between plant and host effect sizes was strikingly and significantly negative. That is, the positive impact on plant responses generally increased as the negative effect of EPN on hosts intensified, providing strong support for the mechanism of trophic cascades. We also review the ways in which EPN might interact antagonistically with each other and other predators and pathogens to adversely affect host suppression and dampen trophic cascades. We conclude that the food web implications of multiple-enemy interactions involving EPN are little studied, but, as management techniques that promote the long-term persistence of EPN are improved, antagonistic interactions are more likely to arise. We hope that the likely occurrence of antagonistic interactions in soil food webs should stimulate researchers to conduct field experiments explicitly designed to examine multiple-enemy interactions involving EPN and their cascading effects to hosts and plants.
PMCID: PMC2586540  PMID: 19259522
biological control; crop yield; EPN; food-web dynamics; intraguild predation; interspecific competition; meta-analysis; multiple-enemy interactions; pest suppression; trophic cascade
6.  Effect of a Terminated Cover Crop and Aldicarb on Cotton Yield and Meloidogyne incognita Population Density 
Journal of nematology  2008;40(2):147-151.
Terminated small grain cover crops are valuable in light textured soils to reduce wind and rain erosion and for protection of young cotton seedlings. A three-year study was conducted to determine the impact of terminated small grain winter cover crops, which are hosts for Meloidogyne incognita, on cotton yield, root galling and nematode midseason population density. The small plot test consisted of the cover treatment as the main plots (winter fallow, oats, rye and wheat) and rate of aldicarb applied in-furrow at-plant (0, 0.59 and 0.84 kg a.i./ha) as subplots in a split-plot design with eight replications, arranged in a randomized complete block design. Roots of 10 cotton plants per plot were examined at approximately 35 days after planting. Root galling was affected by aldicarb rate (9.1, 3.8 and 3.4 galls/root system for 0, 0.59 and 0.84 kg aldicarb/ha), but not by cover crop. Soil samples were collected in mid-July and assayed for nematodes. The winter fallow plots had a lower density of M. incognita second-stage juveniles (J2) (transformed to Log10 (J2 + 1)/500 cm3 soil) than any of the cover crops (0.88, 1.58, 1.67 and 1.75 Log10(J2 + 1)/500 cm3 soil for winter fallow, oats, rye and wheat, respectively). There were also fewer M. incognita eggs at midseason in the winter fallow (3,512, 7,953, 8,262 and 11,392 eggs/500 cm3 soil for winter fallow, oats, rye and wheat, respectively). Yield (kg lint per ha) was increased by application of aldicarb (1,544, 1,710 and 1,697 for 0, 0.59 and 0.84 kg aldicarb/ha), but not by any cover crop treatments. These results were consistent over three years. The soil temperature at 15 cm depth, from when soils reached 18°C to termination of the grass cover crop, averaged 9,588, 7,274 and 1,639 centigrade hours (with a minimum threshold of 10°C), in 2005, 2006 and 2007, respectively. Under these conditions, potential reproduction of M. incognita on the cover crop did not result in a yield penalty.
PMCID: PMC2586531  PMID: 19259531
Aldicarb; conservation tillage; cotton; Gossypium hirsutum; Meloidogyne incognita; root-knot nematode
7.  Nutsedge Counts Predict Meloidogyne incognita Juvenile Counts in an Integrated Management System 
Journal of nematology  2008;40(2):99-108.
The southern root-knot nematode (Meloidogyne incognita), yellow nutsedge (Cyperus esculentus) and purple nutsedge (Cyperus rotundus) are important pests in crops grown in the southern US. Management of the individual pests rather than the pest complex is often unsuccessful due to mutually beneficial pest interactions. In an integrated pest management scheme using alfalfa to suppress nutsedges and M. incognita, we evaluated quadratic polynomial regression models for prediction of the number of M. incognita J2 in soil samples as a function of yellow and purple nutsedge plant counts, squares of nutsedge counts and the cross-product between nutsedge counts . In May 2005, purple nutsedge plant count was a significant predictor of M. incognita count. In July and September 2005, counts of both nutsedges and the cross-product were significant predictors. In 2006, the second year of the alfalfa rotation, counts of all three species were reduced. As a likely consequence, the predictive relationship between nutsedges and M. incognita was not significant for May and July. In September 2006, purple nutsedge was a significant predictor of M. incognita. These results lead us to conclude that nutsedge plant counts in a field infested with the M. incognita-nutsedge pest complex can be used as a visual predictor of M. incognita J2 populations, unless the numbers of nutsedge plants and M. incognita are all very low.
PMCID: PMC2586538  PMID: 19259526
alfalfa; crop rotation; Cyperus esculentus; Cyperus rotundus; interaction; Medicago sativa; Meloidogyne incognita; method; perennial weed; Poisson regression; predictive modeling; purple nutsedge; southern root-knot nematode; yellow nutsedge
8.  Mortality of Pratylenchus penetrans by Volatile Fatty Acids from Liquid Hog Manure 
Journal of nematology  2008;40(2):119-126.
As part of our research program assessing the use of liquid hog manure (LHM) to control root-lesion nematodes, Pratylenchus penetrans, a series of acute toxicity tests was conducted to: (i) examine if non-ionized forms of volatile fatty acids (VFA) are responsible for the mortality of P. penetrans exposed to LHM under acidic conditions, (ii) determine if Caenorhabditis elegans can be a surrogate for P. penetrans in screening tests by comparing their sensitivities to VFA, (iii) characterize the nematicidal effect of individual VFA in LHM to P. penetrans, and (iv) determine whether individual VFA in LHM interact in their toxicity to P. penetrans. LHM was significantly (P ≤ 0.05) more toxic to P. penetrans than a mixture of its main VFA components at concentrations of 5% and 10% (vol. VFA or LHM /vol. in buffer). Pratylenchus penetrans was more sensitive to acetic acid than C. elegans, whereas the sensitivity of both nematode species to n-caproic acid was similar. Individual VFA vary in their lethality to P. penetrans. n-valeric acid was the most toxic (LC95= 6.8 mM), while isobutyric acid was the least toxic (LC95 = 45.7 mM). Individual VFA did not interact in their toxicity to P. penetrans, and their effects were considered additive. VFA account for the majority of the lethal effect of LHM to P. penetrans under acidic conditions. Caenorhabditis elegans cannot be used as a surrogate to P. penetrans in toxicity studies using VFA. The efficacy of LHM to control P. penetrans can be evaluated by assessing its VFA content prior to application, and this evaluation is facilitated by the fact that the interaction of individual VFA appears to be simply additive.
PMCID: PMC2586537  PMID: 19259528
bioassay; Caenorhabditis elegans; LHM; liquid hog manure; LSM; liquid swine manure; management; nematode; potato; Pratylenchus penetrans; root-lesion nematode; VFA; volatile fatty acids
9.  Effect of Liquid Swine Manure on Hatch and Viability of Heterodera glycines 
Journal of nematology  2008;40(2):152-160.
Experiments were conducted in the laboratory and greenhouse to determine the effect of raw and anaerobically digested liquid swine manures on the hatch and viability of Heterodera glycines, the soybean cyst nematode. Anaerobic digestion was performed for 15 and 35 days to enrich volatile fatty acids (VFA) and ammonium (NH4 +), respectively. All filtrates of the raw, VFA-enriched, and NH4 +-enriched manures at 10−1 to 250−1 dilutions inhibited H. glycines hatch, and the reduction of hatch was increased with increasing concentration of the manure. Cumulative hatch at day 21 was only 2.1% to 3.7% in the 10−1 dilution manures, while the hatch in water was 21% to 27.3%. The high concentrations appeared to be lethal to some eggs. Most second-stage juveniles (J2) of H. glycines were killed when incubated for 8 hours in the manure filtrate at the original concentration (>90% mortality) or for 48 hours at the 64−1 dilution (> 82% mortality). When J2 were treated with the manures at 10−1 to 250−1 dilutions for 4 hours, only the 10−1 dilution of VFA-enriched and raw manures resulted in a lower number of J2 that penetrated soybean roots as compared with lower concentrations. The VFA-enriched manure was the best, raw manure intermediate, and NH4 +-enriched manure the least effective in inhibiting H. glycines hatch and killing eggs and J2.
PMCID: PMC2586534  PMID: 19259532
ammonia; fatty acid; Glycine max; hatch; Heterodera glycines; mortality; swine manure; soybean; soybean cyst nematode
10.  Dynamics of a Subterranean Trophic Cascade in Space and Time 
Journal of nematology  2008;40(2):85-92.
Trophic cascades, whereby predators indirectly benefit plant biomass by reducing herbivore pressure, form the mechanistic basis for classical biological control of pest insects. Entomopathogenic nematodes (EPN) are lethal to a variety of insect hosts with soil-dwelling stages, making them promising biocontrol agents. EPN biological control programs, however, typically fail because nematodes do not establish, persist and/or recycle over multiple host generations in the field. A variety of factors such as local abiotic conditions, host quantity and quality, and rates of movement affect the probability of persistence. Here, we review results from 13 years of study on the biology and ecology of an endemic population of Heterorhabditis marelatus (Rhabditida: Heterorhabditidae) in a California coastal prairie. In a highly seasonal abiotic environment with intrinsic variation in soils, vegetation structure, and host availability, natural populations of H. marelatus persisted at high incidence at some but not all sites within our study area. Through a set of field and lab experiments, we describe mechanisms and hypotheses to understand the persistence of H. marelatus. We suggest that further ecological study of naturally occurring EPN populations can yield significant insight to improve the practice and management of biological control of soil-dwelling insect pests.
PMCID: PMC2586530  PMID: 19259524
entomopathogenic nematode; Hepialus californicus; Heterorhabditis marelatus; long-term persistence; Lupinus arboreus; metapopulation dynamics; biocontrol; trophic cascade
11.  Molecular Phylogeny of Geographical Isolates of Bursaphelenchus xylophilus: Implications on the Origin and Spread of this Species in China and Worldwide 
Journal of nematology  2008;40(2):127-137.
The genetic diversity and phylogeny of 26 isolates of Bursaphelenchus xylophilus from China, Japan, Portugal and North America were investigated based on the D2/3 domain of 28S rDNA, nuclear ribosomal Internal Transcribed Spacer (ITS) sequences, and random amplified polymorphic DNA (RAPD) analysis. The genetic diversity analysis showed that the D2/3 domain of 28S rDNA of isolates of B. xylophilus from China, Portugal, Japan and the US were identical and differed at one to three nucleotides compared to those from Canada. ITS sequences of isolates from China and Portugal were the same; they differed at one or two nucleotides compared to those of Japanese isolates and at four and 23 nucleotides compared to those from the US and Canada, respectively. The phylogenetic analysis indicated that Chinese isolates share a common ancestor with one of the two Japanese clades and that the Canadian isolates form a sister group of the clade comprised of isolates from China, Portugal, Japan, and the US. The relationship between Japanese isolates and those from China was closer than with the American isolates. The Canadian isolates were the basal group of B. xylophilus. This suggests that B. xylophilus originated in North America and that the B. xylophilus that occurs in China could have been first introduced from Japan. Further analysis based on RAPD analysis revealed that the relationship among isolates from Guangdong, Zhejiang, Shandong, Anhui provinces and Nanjing was the closest, which suggests that pine wilt disease in these Chinese locales was probably dispersed from Nanjing, where this disease first occurred in China.
PMCID: PMC2586533  PMID: 19259529
Bursaphelenchus xylophilus; ITS; phylogeny; pinewood nematode; RAPD; systematics; D2/3 regions of 28S rDNA
12.  Protease Inhibitor Expression in Soybean Roots Exhibiting Susceptible and Resistant Interactions with Soybean Cyst Nematode 
Journal of nematology  2008;40(2):138-146.
Protease inhibitors play a role in regulating proteases during cellular development and in plant defense. We cloned and sequenced cDNA encoding six protease inhibitors expressed in soybean roots infected with soybean cyst nematode (SCN) and determined their expression patterns. Four of these protease inhibitors are novel and have not been reported previously. Using RT-PCR, we measured the relative transcript levels of each protease inhibitor in roots of the soybean cv. Peking inoculated with either SCN TN8 to examine the expression of protease inhibitors during the susceptible interaction or with SCN NL1-RHg representing the resistant interaction. Within 12 to 24 hours, mRNA transcripts encoding five of the six protease inhibitors were more highly elevated in soybean roots exhibiting the susceptible interaction than the resistant interaction. Transcripts encoding two protease inhibitors possessing Kunitz trypsin inhibitor domains were induced 37- and 27-fold in the susceptible interaction within 1 dpi, but were induced only 5- to 7-fold in roots displaying the resistant interaction. Our results indicate that soybean roots recognize differences between these two SCN populations before the nematodes initiate a feeding site, and accordingly the roots express transcripts encoding soybean protease inhibitors differentially. These transcripts were generally less abundant in roots exhibiting the resistant interaction.
PMCID: PMC2586541  PMID: 19259530
host-parasitic relationship; molecular biology; protease inhibitors; resistance; RT-PCR; soybean; soybean cyst nematode
13.  The Ecological Complexities of Biological Control: Trophic Cascades, Spatial Heterogeneity, and Behavioral Ecology 
Journal of nematology  2008;40(2):59-60.
Biological control can be considered an intentional induction of a trophic cascade, whereby the addition of herbivores’ natural enemies or other habitat manipulations effectively enhance natural enemy populations, lead to reduced herbivore populations or feeding damage, and indirectly improve or protect plant health, agricultural yield, or the condition of some other biotic population or community of interest to man. The following set of papers (Denno et al., 2008; Ram et al., 2008; Stuart and Duncan, 2008; Spence et al. 2008) offer insights into the broad- and fine-scale factors that ultimately contribute to the success of biological control efforts. Many of the ideas herein were presented and discussed during a special session at the 2007 Annual Meeting of the Society of Nematologists. The goal of this session was to examine explicitly the ramifications of spatial and temporal heterogeneity in the context of effective biological control. The biological focus was primarily on interactions involving entomopathogenic nematodes (EPN), although many of the authors’ conclusions are applicable to other types of nematodes, soil fauna and natural enemies in general.
PMCID: PMC2586532  PMID: 19259521
Bioassay; biological control; bush lupine; citrus; entomopathogenic nematodes; meta-analysis; plant parasitic nematodes; trophic cascade
14.  Early Crop Root Destruction for Management of Tobacco Cyst Nematodes 
Journal of nematology  2008;40(1):26-29.
Prompt tillage after crop harvest was investigated as a cultural control for the tobacco cyst nematode, Globodera tabacum tabacum, on stalk-cut broadleaf cigar wrapper tobacco. Stalk stumps and roots remaining after harvest were destroyed by tilling immediately or from 2 to 6 wk after harvest in field experiments over 4 yr. Cyst nematode Pf/Pi ratios ranged from 0.65 to 1.62 when plants were tilled immediately after harvest and 1.13 to 5.88 when tillage was delayed. Nematode population development was monitored by inoculating plants in pots placed in fields with J2 in eggs and sampling over time (8 to 18 wk). Three generations per year were observed, and G. t. tabacum generation time was as short as 6 wk for each generation. Destroying stalks and root systems remaining after harvesting stalk-cut broadleaf cigar wrapper tobacco removes the host to preclude development of nematodes at the end of the second and entire third generation. Early tillage resulted in consistently lower tobacco cyst nematode populations than allowing viable roots to remain in fields for an additional 8 to 18 wk. This management tactic reduces the need for nematicide application to slow nematode population increases over time and can reduce losses due to infection by G. t. tabacum.
PMCID: PMC2586525  PMID: 19259515
Cultural control; Globodera tabacum tabacum; management; Nicotiana tabacum; Nemacur; root destruction; tillage; tobacco; tobacco cyst nematode
15.  Exposure Time to Lethal Temperatures for Meloidogyne incognita Suppression and Its Implication for Soil Solarization 
Journal of nematology  2008;40(1):7-12.
Meloidogyne incognita eggs or J2 were incubated in test tubes containing sand:peat mix and immersed in a water bath heated to 38, 39, 40, 41, 42, 43, 44 and 45°C for a series of time intervals. Controls were maintained at 22°C. Nematodes surviving or hatching were collected from Baermann trays after three weeks of incubation. Regression analyses between percent survival or egg hatch and hours of heat treatment were performed for each temperature. Complete suppression of egg hatch required 389.8, 164.5, 32.9, 19.7 and 13.1 hours at 38, 39, 40, 41 and 42°C, respectively. Complete killing of J2 required 47.9, 46.2, 17.5 and 13.8 hours at 39, 40, 41 and 42°C, respectively. J2 were not completely killed at 38°C within 40 hours of treatment, but were killed within one hour at 44 and 45°C. Effect of temperature on nematode killing is not determined by heat units. Oscillating temperature between cool and warm did not interfere with the nematode suppressive effect by the heat treatment. Six-week solarization in the field during the summers of 2003 and 2004 in Florida accumulated heat exposure times in the top 15 cm of soil that surpassed levels required to kill M. incognita as determined in the water bath experiments. Although near zero M. incognita were detected right after solarization, the nematode population densities increased after a cycle of a susceptible pepper crop. Therefore, future research should address failure of solarization to kill nematodes in the deeper soil layers.
PMCID: PMC2586528  PMID: 19259512
Capsicum annuum; bell pepper; soil temperature; heat units; Meloidogyne incognita, solarization; root-knot nematodes
16.  Correlation of Female Indices From Virulence Assays on Inbred Lines and Field Populations of Heterodera glycines  
Journal of nematology  2008;40(1):39-45.
A lack of diversity and durability of resistant soybean varieties complicates management of the soybean cyst nematode (SCN), Heterodera glycines, exemplified by the current overdependence on the PI 88788 source of resistance. Of interest is the effect of adaptation of a SCN population to a source of resistance on its subsequent ability to develop on others. Female indices (FI) from virulence assays (race, HG Type and SCN Type tests) for SCN field populations and inbred lines were analyzed. Female indices on PI 88788, PI 209332 and PI 548316 were highly correlated, as were those of PI 548402, PI 90763, PI 89772 and PI 438489B. Previous studies on resistant SCN-infected soybean roots indicated that the cellular resistance response was similar within these two groups of soybean genotypes. In field populations, highly significant correlations were also found between FI on PI 88788 and PI 548402 and those on PI 89772 and PI 437654. In inbred lines, FI on PI 437654 were correlated with PI 90763 and PI 438489B. To avoid further adaptation, rotation of cultivars with resistance from these groups should be carefully monitored, including those from the most promising source of resistance, PI 437654, such as CystX. In a separate test, 10 soybean varieties developed from CystX were tested against HG Type 0, HG Type 2.5.7 and HG Type 1–7. Female development occurred in all tests but one. Although identification and deployment of unique resistance is needed, management strategies to prevent and detect adaptation should be emphasized.
PMCID: PMC2586527  PMID: 19259518
Adaptation; genetic diversity; Heterodera glycines; management; resistance; soybean cyst nematode
17.  Detection of Suppressiveness against Rotylenchulus reniformis in Soil from Cotton (Gossypium hirsutum) Fields in Texas and Louisiana 
Journal of nematology  2008;40(1):35-38.
Rotylenchulus reniformis is a major problem confronting cotton production in the central part of the cotton belt of the United States of America. In this study, the hypothesis that natural antagonists in some cases are responsible for unusually low densities of the nematode in certain fields was tested by assaying soils from 22 selected fields for the presence of transferable agents in pots containing cotton plants. In one field, soil from four different depth ranges was tested. In the first of two types of assays, 1 part nematode infested soil was added to 9 parts test soil that was left untreated or autoclaved before mixing; this mixture was used to fill pots. In the second type of assay, 1 part test soil was added to 9 or 19 parts pasteurized fine sand, and nematodes were introduced in aqueous suspension. In three experiments representing both types of assay, transferable or autoclavable agent(s) from four fields in South Texas suppressed nematode populations by 48, 78, 90 and 95%. In one experiment, transferable agents in five fields in Louisiana suppressed populations from 37 to 66%. Identification and evaluation of these agents for biological control of R. reniformis merits further study.
PMCID: PMC2586520  PMID: 19259517
biological control; cotton; Gossypium hirsutum; Rotylenchulus reniformis; reniform nematode; soil suppressiveness
18.  Effects of host nutrition on virulence and fitness of entomopathogenic nematodes: Lipid- and protein-based supplements in Tenebrio molitor diets 
Journal of nematology  2008;40(1):13-19.
Entomopathogenic nematodes, Heterorhabditis indica and Steinernema riobrave, were tested for virulence and reproductive yield in Tenebrio molitor that were fed wheat bran diets with varying lipid- and protein-based supplements. Lipid supplements were based on 20% canola oil, peanut, pork or salmon, or a low lipid control (5% canola). Protein treatments consisted of basic supplement ingredients plus 0, 10, or 20% egg white; a bran-only control was also included. Some diet supplements had positive effects on nematode quality, whereas others had negative or neutral effects. All supplements with 20% lipids except canola oil caused increased T. molitor susceptibility to H. indica, whereas susceptibility to S. riobrave was not affected. Protein supplements did not affect host susceptibility, and neither lipid nor protein diet supplements affected reproductive capacity of either nematode species. Subsequently, we determined the pest control efficacy of progeny of nematodes that had been reared through T. molitor from different diets against Diaprepes abbreviatus and Otiorhynchus sulcatus. All nematode treatments reduced insect survival relative to the control (water only). Nematodes originating from T. molitor diets with the 0% or 20% protein exhibited lower efficacy versus D. abbreviatus than the intermediate level of protein (10%) or bran-only treatments. Nematodes originating from T. molitor lipid or control diets did not differ in virulence. Our research indicates that nutritional content of an insect host diet can affect host susceptibility to entomopathogenic nematodes and nematode fitness; therefore, host media could conceivably be optimized to increase in vivo nematode production efficiency.
PMCID: PMC2586524  PMID: 19259513
diet; entomopathogenic nematode; Heterorhabditis; in vivo, mass production; Steinernema
19.  Comparative Efficiency of the Fenwick Can and Schuiling Centrifuge in Extracting Nematode Cysts from Different Soil Types 
Journal of nematology  2008;40(1):30-34.
The Fenwick can and Schuiling centrifuge are widely used to extract nematode cysts from soil samples. The comparative efficiencies of these two methods during cyst extraction have not been determined for different soil types under different cyst densities. Such information is vital for statutory laboratories that must choose a method for routine, high-throughput soil monitoring. In this study, samples of different soil types seeded with varying densities of potato cyst nematode (Globodera rostochiensis) cysts were processed using both methods. In one experiment, with 200 ml samples, recovery was similar between methods. In a second experiment with 500 ml samples, cyst recovery was higher using the Schuiling centrifuge. For each method and soil type, cyst extraction efficiency was similar across all densities tested. Extraction was efficient from pure sand (Fenwick 72%, Schuiling 84%) and naturally sandy soils (Fenwick 62%, Schuiling 73%), but was significantly less efficient from clay-soil (Fenwick 42%, Schuiling 44%) and peat-soil with high organic matter content (Fenwick 35%, Schuiling 33%). Residual moisture (<10% w/w) in samples prior to analyses reduced extraction efficiency, particularly for sand and sandy soils. For each soil type and method, there were significant linear relationships between the number of cysts extracted and the numbers of cysts in the samples. We discuss the advantages and disadvantages of each extraction method for cyst extraction in statutory soil laboratories.
PMCID: PMC2586521  PMID: 19259516
Fenwick can; potato cyst nematode; Globodera rostochiensis; Schuiling centrifuge; soil monitoring; method; statutory testing
20.  Assessment of Parasitic Activity of Fusarium Strains Obtained from a Heterodera schachtii-Suppressive Soil 
Journal of nematology  2008;40(1):1-6.
This study assessed the potential impact of various Fusarium strains on the population development of sugarbeet cyst nematodes. Fungi were isolated from cysts or eggs of Heterodera schachtii Schmidt that were obtained from a field suppressive to that nematode. Twenty-six strains of Fusarium spp. were subjected to a phylogenic analysis of their rRNA-ITS nucleotide sequences. Seven genetically distinct Fusarium strains were evaluated for their ability to influence population development of H. schachtii and crop performance in greenhouse trials. Swiss chard (Beta vulgaris) seedlings were transplanted into fumigated field soil amended with a single fungal strain at 1,000 propagules/g soil. One week later, the soil was infested with 250 H. schachtii J2/100 cm3 soil. Parasitized eggs were present in all seven Fusarium treatments at 1,180 degree-days after fungal infestation. The percentage of parasitism ranged from 17 to 34%. Although the most efficacious F. oxysporum strain 471 produced as many parasitized eggs as occurred in the original suppressive soil, none of the Fusarium strains reduced the population density of H. schachtii compared to the conducive check. This supports prior results that Fusarium spp. were not the primary cause of the population suppression of sugarbeet cyst nematodes at this location.
PMCID: PMC2586523  PMID: 19259511
biological control; Beta vulgaris; egg parasitism; Fusarium spp; Heterodera schachtii; nematophagous fungi; population suppression; sugarbeet cyst nematode; Swiss chard
21.  Characterization of Root-Knot Nematode Resistance in Medicago truncatula  
Journal of nematology  2008;40(1):46-54.
Root knot (Meloidogyne spp.) and cyst (Heterodera and Globodera spp.) nematodes infect all important crop species, and the annual economic loss due to these pathogens exceeds $90 billion. We screened the worldwide accession collection with the root-knot nematodes Meloidogyne incognita, M. arenaria and M. hapla, soybean cyst nematode (SCN-Heterodera glycines), sugar beet cyst nematode (SBCN-Heterodera schachtii) and clover cyst nematode (CLCN-Heterodera trifolii), revealing resistant and susceptible accessions. In the over 100 accessions evaluated, we observed a range of responses to the root-knot nematode species, and a non-host response was observed for SCN and SBCN infection. However, variation was observed with respect to infection by CLCN. While many cultivars including Jemalong A17 were resistant to H. trifolii, cultivar Paraggio was highly susceptible. Identification of M. truncatula as a host for root-knot nematodes and H. trifolii and the differential host response to both RKN and CLCN provide the opportunity to genetically and molecularly characterize genes involved in plant-nematode interaction. Accession DZA045, obtained from an Algerian population, was resistant to all three root-knot nematode species and was used for further studies. The mechanism of resistance in DZA045 appears different from Mi-mediated root-knot nematode resistance in tomato. Temporal analysis of nematode infection showed that there is no difference in nematode penetration between the resistant and susceptible accessions, and no hypersensitive response was observed in the resistant accession even several days after infection. However, less than 5% of the nematode population completed the life cycle as females in the resistant accession. The remainder emigrated from the roots, developed as males, or died inside the roots as undeveloped larvae. Genetic analyses carried out by crossing DZA045 with a susceptible French accession, F83005, suggest that one gene controls resistance in DZA045.
PMCID: PMC2586522  PMID: 19259519
root-knot nematode; cyst nematode; resistance; Medicago truncatula; host range; genetics
22.  Description of Tylenchorhynchus qasimii sp. n. with a New Report of T. kegasawai Minagawa, 1995 from Pakistan 
Journal of nematology  2008;40(1):20-25.
A new stunt nematode, from soil around the roots of coconut (Cocos nucifera L.) and rice (Oryza sativa L.) from Karachi, Pakistan, is described and illustrated as Tylenchorhynchus qasimii n. sp. This new species is characterized by having females with 3–4 head annules, anteriorly directed stylet knobs, absence of post anal extension, presence of rounded sperm filled spermatheca and conoid to bluntly rounded hemispherical tail terminus. Males are common. Also included is the record of T. kegasawai from around the roots of rice (O. sativa L.), a new report from Sindh, Pakistan.
PMCID: PMC2586526  PMID: 19259514
Cocos nucifera L.; Oryza sativa L.; taxonomy; Tylenchorhynchus qasimii n. sp.; Tylenchorhynchus kegasawai
24.  An Aphelenchoides sp. nematode Parasitic of Polianthes tuberosa in the Mekong Delta 
Journal of nematology  2007;39(3):248-257.
Polianthes tuberosa is a commercially valuable flower crop in the Mekong Delta of Vietnam that is propagated by the harvesting and planting of bulbs. The cultivation of P. tuberosa is complicated by an endemic nematode infection that damages a high proportion of the plants. Based on morphological criteria and ribosomal RNA gene sequencing, we have determined that the infection is caused by an Aphelenchoides sp. nematode and is most likely Aphelenchoides besseyi. By scoring various parts of harvested plants with flowers for the presence of viable nematodes over a period of six months, we determined that the nematode is an ectoparasite that can survive the intercrop periods, especially in the bulbs that are used to plant new crops. A comparison of farming practices in the Mekong Delta failed to identify useful control methods, but rather indicated that fields that have cultivated P. tuberosa for the longest periods suffer the worst damage from the nematode infection. Finally, we demonstrated that the nematode is capable of infecting 30 rice cultivars but does not cause the white tip disease usually associated with A. besseyi infection.
PMCID: PMC2586507  PMID: 19259495
Aphelenchoides besseyi; biological control; host-parasitic relationship; Mekong Delta; plant disease loss; Polianthes tuberosa; rRNA sequence; survival; Vietnam
25.  Two Cyclic Dipeptides from Pseudomonas fluorescens GcM5-1A Carried by the Pine Wood Nematode and Their Toxicities to Japanese Black Pine Suspension Cells and Seedlings in vitro 
Journal of nematology  2007;39(3):243-247.
Pseudomonas fluorescens GcM5-1A, isolated from the pine wood nematode (PWN), Bursaphelenchus xylophilus, was cultured in Luria Broth medium (LB). The clarified culture was extracted with ethyl acetate, and two dipeptides were purified from the extract. The chemical structures of 1 and 2 were identified as cyclo(-Pro-Val-)and cyclo(-Pro-Tyr-), respectively, by MS, 1H NMR, 13C NMR,1H-1H COSY, 1H -13C COSY spectra. Bioassay results showed that the two compounds were toxic to both suspension cells and seedlings of Pinus thunbergii, which may offer some clues to research the mechanism of pine wilt disease caused by PWN.
PMCID: PMC2586505  PMID: 19259494
Bursaphelenchus xylophilus; cyclo(-Pro-Tyr-); cyclo(-Pro-Val-); interaction; Pinus thunbergii; Pseudomonas fluorescens GcM5-1A; toxicity

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