NE171: Biologically Based IPM Systems for Management of Plant-Parasitic Nematodes

(Multistate Research Project)

Status: Inactive/Terminating

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SAES-422 Reports

Annual/Termination Reports:

[12/18/2000] [12/10/2001] [12/04/2002] [11/25/2003] [12/09/2004]

Date of Annual Report: 12/18/2000

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Annual Meeting Dates: 10/16/2000 - 10/18/2000
Period the Report Covers: 01/01/2000 - 12/01/2000

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Accomplishments

Intended outcomes and potential impacts (based on project milestones): Objective 1: Evaluate the effects of rotational crops, organic amendments and host crop genetics on nematode community structure. Complementary or collaborative studies to assess the impact of rotation and cover crops on plant-parasitic nematode populations and nematode community structure: <ul> <li>Milestone 2000: Establish a low soil volume bioassay to evaluate nematode antagonistic crops. </li> </ul> <dir> <li>(PA) A low volume soil bioassay technique has been developed (manuscript in preparation, PA) and will be used in further studies to evaluate the toxicity of green or freeze-dried plant extracts on a variety of plant parasitic nematodes using soils and nematodes supplied by cooperators in CT, MA, NY, and WV.</li> </dir> <ul> <li>Milestone 2000: Identify and evaluate nematode resistant or tolerant varieties for inclusion in an integrated management program.</li> </ul> <dir> <li>(CT) The influence of rotation and green manure crops on early dying of potato caused by Verticillium dahliae and P. penetrans was investigated in field microplots infested with both pathogens. Polynema marigold rotation crops significantly reduced the AUDPC for early dying symptoms and increased total, and marketable tuber yields.(MD) Maryland is participating in an interdisciplinary three-year grower-generated project entitled "Development and Evaluation of Management Alternatives for Root Knot Nematodes and Volunteer Potatoes." Cooperators on the project include specialists in Field Crops, Entomology, Nematology and Vegetable Pathology along with the County Extension Agent, IPM Scout and County Growers. Field and microplot evaluations initiated in 2000 include the use of cover crops, soil amendment, alternative economic crops, and tillage practices to manage root knot nematode populations. Also examined are currently used rotations, herbicide use, and tillage practices for impact on reducing volunteer potatoes. (MI) The impacts of sub-surface irrigation and tillage on nematode community structure were investigated in a corn-soybean rotation system. The water table was maintained at 50 and 75 cm below soil surface in two treatments and compared to a drainage tile system, under both conventional and reduced tillage. The sub-surface irrigated treatments had a higher percent of total population of bacterial-feeding nematodes and a lower percent of plant-parasitic and plant-associated nematode genera compared to the drainage tile treatment. The 50-cm water table treatment had a significantly higher percent of the population as bacterial feeders than the 75-cm water table system. Conventional tillage resulted in more genera of nematodes, and a higher percentage of both bacterial-feeding and plant-associate genera than reduced tillage. The percent of carnivorous nematodes was higher in the tilled than the reduced tillage system. In a 22-acre soybean variety trial to evaluate soybean varieties for resistance to Heterodera glycines, both Pratylenchus penetrans and Meloidogyne hapla were commonly associated with various varieties. It appears that all three species impact bean productivity, and could be a partial explanation to the issue of why various soybean cyst nematode resistant varieties perform well in some locations and not as well in other locations. A ten-year potato farming system trial indicated that crop rotation, variety and soil amendments influenced nematode community structure, soil quality and crop productivity. (NY, Geneva) Using perineal pattern analysis, SEM observations and host range assays, it was determined that Meloidogyne hapla was the only species of root-knot nematode currently present in the vegetable producing areas of New York State. A search was initiated for resistance to the northern root-knot (Meloidogyne hapla in onion breeding lines available at Cornell and Wisconsin as well as large number of Plant Introduction accessions of onions. To-date, 31 breeding lines and 54 Plant Introduction accessions have been tested in the greenhouse in artificially infested soil. A number of the Plant Introduction accessions evaluated appeared to be tolerant to this nematode including PI 272255, PI 273212, PI 355047, PI 546115, PI 546174. Tissue culture systems were developed using onion, dandelion and tomato root explants for the maintenance and propagation of M. hapla. (PA) Four different sources of compost were evaluated for ability to suppress plant-parasitic nematodes and weeds in a young apple orchard. Three composts were from commercial sources and one was a spent mushroom compost. The treatments consisted of four different compost sources each applied at two rates. Checks consisted of plots without compost and either left untreated or treated once with herbicide. Composts were applied at depths of 2 inches and 5 inches in late spring. Compost analysis showed minor variations in pH, mineral content and other character. Nematode assays showed that the only plant-parasitic nematode present to any significant level was Xiphinema americanum. Data showed that one application of compost was beneficial in controlling weeds throughout the growing season and performed better than a single spring herbicide application. The abundance of weeds was correlated with increases in the dagger nematode population. Neither compost source nor rate of application had any significant effect on apple size or increase in tree growth although the trend suggested that higher rates of compost were beneficial. (RI) The efficacy of direct application of butyric acid was evaluated as a preplant treatment for nematode management. An assortment of nematode species and pathogenic fungi were screened for sensitivity to butyric acid, both contact with the acid and exposure to its volatiles. To date, 2 days exposure to 8.8 or 88 mg butyric acid/10 g sand reduced densities of Hoplolaimus, Tylenchorhynchus, Helicotylenchus, Aphelenchoides (fungivore), and Heterorhabditis (bacterivore) nematodes. Volatiles of butyric acid also proved lethal to Tylenchorhynchus nematodes. The same concentrations (8.8 and 88 mg/10 g sand) effectively killed sclerotia of Rhizoctonia solani, a damping-off fungus. (USDA-SC) Developed and evaluated a core of the USDA Capsicum germplasm collection for reaction to M. hapla in greenhouse tests. The core included 440 PI accessions that comprised approximately 10 percent of each of the available Capsicum species or species/varieties selected from all available geographic origins. None of the 430 Capsicum spp. accessions evaluated exhibited high levels of resistance to M. hapla. . Eleven of the least susceptible (referred to hereafter as "best") accessions and three of the most susceptible (referred to as "worst") accessions were selected for re-evaluation in a replicated greenhouse test. The 11 best accessions exhibited intermediate to susceptible reactions to M. hapla and the three worst accessions were susceptible. Although five accessions of the best accessions had root gall severity scores <5.00 (moderately resistant reactions), the reproductive indices for these accessions ranged from 8.3 to 31.5, which indicates that the accessions were susceptible. The next step in searching for resistance to M. hapla would be to evaluate the entire Capsicum spp. collection. (WVA) A large farming systems trial was initiated to evaluate transition methods for conversion from conventional to organic farming practices. A vegetable crop trial is underway evaluating a high input, compost intensive management system versus a low-input system that relies on green manures and cover crops. In addition, earthworm densities were monitored. Earthworm species identified included Aporectodea caligenosa and Lumbricus terrestris and population densities of both species tended to be greater in compost amended plots than unamended plots. </li> </dir> Objective 2: Characterization of biological control agents for suppression of plant-parasitic nematodes. <dir> <li>(MD) Collaborative efforts of the Meyer (USDA) and Sardanelli (UMCP) laboratories continue to refine previously developed soil moisture control system materials and methods (Sardanelli and Kenworthy 1997) for ongoing evaluations of various fungi (Verticillium, Fusarium, Cladosporium) and their potential use as biocontrol agents.(NY, Geneva) A replicated field test was conducted to determine the efficacy of a drench application of abamectin (Novartis Agri-Mek, 0.15 EC; fermentation products of Streptomyces avermitilis) against M. hapla on tomato at rates of 8 and 16 oz/A (100 ml water suspension/plant). Severe incidence and damage by M. hapla was observed on tomato in this test. Agri-Mek exhibited only limited activity against M. hapla in this test. (USDA, MD) 1) Rhizosphere bacteria were tested in vitro for production of compounds inhibitory to nematodes. A number of the isolates suppressed nematode egg hatch or juvenile mobility; 2) isolated and identified active compounds from a nematode-associated fungus; 3) the gelatinous matrix of root-knot nematode was studied as a structure for inhibiting growth of bacteria and fungi on nematode eggs. </li> </dir> Obj. 2.1) Survey plant-pathogenic nematodes for occurrence of Pasteuria spp. <ul> <li>Milestone 2000: Conduct a workshop for all group members to present protocols for Pasteuria penetrans recognition, research and surveys.</li> </ul> <dir> <li>(FL) A successful workshop was conducted during the first regional meeting of the new project to teach members how to recognize Pasteuria spp. on different species of plant-parasitic nematodes. A pamphlet will be prepared and distributed to each member of the technical committee for use as a guide for proper handling of Pasteuria spp. The goal of the workshop and pamphlet was to ensure that all members follow prescribed methods in documenting the occurrence of Pasteuria on plant-parasitic nematodes in northern regions of the United States.</li> </dir> Survey plant-pathogenic nematodes for occurrence of Pasteuria spp. <dir> <li>(CT) A Pasteuria-like organism was discovered attached to males and juveniles of the turf cyst nematode Heterodera iri in samples from a commercial golf course. The course superintendent indicated that the green had previously been damaged by high nematode densities, but had recently recovered. Clean juveniles added to soil and recovered by pie pan extraction had a low incidence of infection. Infested soil was inoculated with H. iri cysts and planted with Penncross bentgrass to increase the parasite population. (FL) Pasteuria penetrans cannot be cultivated in vitro and enumerated by traditional bacteriological methods; therefore, an accurate and efficient immunoassay to quantify spore populations was sought to enhance field research and development of Pasteuria as a biocontrol agent. A monoclonal antibody (MAb) produced against whole spores was shown to bind a putative glycan epitope present as a component of the spore envelope. Following SDS-PAGE of nematode cuticle extracts and immunoblotting with spore extracts and the MAb as probes, these glycopeptides were shown to bind to discrete polypeptides derived from the nematode cuticle. Thus the MAb detects an epitope of adhesins that are involved in the attachment of Pasteuria endospores to the nematode cuticle as a prerequisite to the infection process. An indirect amplification ELISA was developed to quantify spore adhesins in a soil matrix. Linear regression analysis of spore concentration versus signal produced line fits of 0.98 and indicated a minimum detection limit of # 3000 spores/g of soil. Western blot profiles of soil-extracted antigen were the same as those provided by extracts of purified endospores, confirming that the ELISA detects Pasteuria antigens (adhesins) in soil extracts containing native endospores. The MAb did not detect antigens on cells or endospores derived from a number of endospore-forming bacteria cultured from test soils. This establishes the MAb as a definitive probe to detect adhesins as virulence determinants on Pasteuria endospores in soil. This sensitive quantitative immunoassay can be applied as a tool for evaluating chemical and biological measures required for control of root-knot nematodes. (MI) Twenty-two LTER (Long-Term Ecological Research) plots at the Kellogg Biological Research Station were surveyed for Pasteuria penetrans during the summer of 2000. The sites surveyed included old-field succession (>30 years since the last disturbance), natural succession (13 years since last disturbance), moderately disturbed (mowed annually) and highly disturbed (annual tillage) plots. P. penetrans was detected in 77% of the sites. The incidence of occurrence was 1.07% (98 of 9,183 specimens). Although it was found primarily associated with tylenchoid parasites of plants, it was also found attached to both fungivores and bacterivores. The incidence of occurrence of P. penetrans appeared to be related to the extent of ecosystem disturbance. It was most common in the highly disturbed ecosystems and least common in old fields that had not been disturbed for more than 30 years. Significant progress is being made on producing 16S sequence data for the KBS LTER strains of P. penetrans. The KBS LTER site will also be used for evaluation of molecular probes specific for isolates of Pasteuria. Soils from 15 Michigan soybean fields were analyzed for occurrence of additional parasites of Heterodera glycines second-stage juveniles. All sites were infested with H. glycines. The procedure consisted of microscopic observation followed by confirmation on potato dextrose agar. The observations were made by Dr. Senyu Chen at the University of Minnesota. Hirsutella spp. were recovered from 47% of the sites. Pasteuria was not detected in the soils from these sites. (NY, Ithaca) A survey for nematodes infected with Pasteuria-species in New York detected infected stylet-bearing nematodes from three different genera (Tylenchorhynchus spp., Tylenchus spp., Meloidogyne sp.). Also detected in this survey were infected free-living bacteriophagous nematodes from two different genera: (Cephalobus sp. and Eucephalobus sp.). The species have not been confirmed. (MA) Twenty-one soil samples received from putting greens for routine nematode assays were examined for Pasteuria-infected Tylenchorhynchus nematodes. All of the Tylenchorhynchus in each soil assay were examined. Pasteuria on other nematodes were noted but not quantified. All 21 samples had Pasteuria-infected Tylenchorhynchus. On average, 26% of Tylenchorhynchus juveniles were infected (0-82%); 40% of males (0-83%) and 40% of females (0-89%). A severity index (SI) was also calculated (total Pasteuria cells visible on cuticle divided by the total number of Tylenchorhynchus in the sample). Average SI for juveniles was 1 (0-2.9), females, 4 (0-10.6), and males, 3.6 (0-14.4). An additional putting green was comprehensively sampled (20 soil cores assayed separately, 2934 Tylenchorhynchus). Juveniles had statistically lower incidence of infection (28.8%) than males (64.45%) or females (56.66%). SI for juveniles, males and females was 1.0,1.94 and 1.25 respectively. Both Tylenchorhynchus and Pasteuria populations had a clumped distribution in the putting greens. Other nematodes with Pasteuria included Helicotylenchus, Hoplolaimus and Meloidogyne. A survey of a golf course in Rochester, New York was also conducted. The property had two 18-hole courses. Approximately 5 years ago, 18 greens (east course) were fumigated with methyl bromide; the 18 greens of the west course were not treated. Newly established turf on the fumigated greens responded well and developed a deep root system. In May 1999, assays were carried out at the University of Massachusetts on 10 greens from the east course and 3 greens from the west Course. The May 12 assay was carried out mostly on east course greens because they were in bad condition and the superintendent requested the assay. Three healthy greens from the west course were included for comparison. Green 4, which had a high population of root-knot was selected for a fenamiphos-timing study which was carried out in 1999 and 2000. During this time, 3 greens from the east course and 3 greens from the west course were monitored for root-knot activity. Root-knot nematode populations appeared to be higher in the east course versus the west course except in January and March. A possible explanation for the differences in nematode populations between the two courses is that natural enemies including some of the Pasteuria population were destroyed by the fumigation. Obj. 2.2) Determine the survivability and host preference of isolates of Pasteuria spp. from different geographic regions. (NY, Ithaca) Preliminary setup of an in vitro model system to study Pasteuria-like parasites of nematodes: A search within the Rhabditida (especially Family Cephalobidae) resulted in the discovery of a Pasteuria-like bacterium infecting Cephalobus and Eucephalobus spp. (subfamily Cephalobinae). Endospores from 10 nematodes were saved. In vitro cultures of Cephalobus and Eucephalobus have been initiated on modified media. Mutants that do not allow attachment and mutants that allow attachment but not infection will be generated and selected. Cultures of two species from subfamily Acrobelinae (viz. Zeldia punctata and Acrobeloides amurensis) have also been initiated. We recently inoculated Cephalobus nematodes in vitro with the first batch of endospores collected and are currently building up Pasteuria-nematode cultures. Observation of both infected males and females (Cephalobus and Eucephalobus) suggests factors required for endospore development may not be sex-specific. Requirements for endospore production may be confined to baseline components or conditions in the pseudocoelomic fluid that are common to all nematode species. (MD) Once plant evaluations in Objective 2.3 indicate the development of efficient methodology, populations of Meloidogyne spp. will be established and maintained in greenhouse and/or laboratory culture in cooperative studies of multistate researchers involving Pasteuria penetrans. Space allocations have been established and materials and methods are in preparation to conduct survivability and host preference studies with regional cooperation. Obj. 2.3) Evaluate different crops and methods for growing Pasteuria penetrans. (MD) Various plants will be evaluated for production of P. penetrans endospores using soil moisture control. Plants currently in evaluation for root growth and overall cultural manageability include Rutgers tomato, and Ascanthus and Tradescantia (herbaceous perennials). Additional hosts will be tested as acquired. </li> </dir> Objective 3: Comparison and evaluation of IPM system management of plant-parasitic nematodes based on crop rotation, organic amendments, host crop resistance and biological control agents. <ul> <li>Milestone 2000: Initiate collaborative rotation and cover crop system research in field plots and microplots in multiple states.</li> </ul> <dir> <li>(CT) Black root rot is a complex disease of perennial strawberry caused by Rhizoctonia fragariae and the lesion nematode Pratylenchus penetrans. Rhizoctonia infection of structural and feeder roots increases during secondary growth when the cortex is cut off by suberized tissues, during harvest, and when lesion nematodes are present in roots. The effects of inundative releases of entomopathogenic Steinernema carpocapsae and S. feltiae infective juveniles on lesion nematode (Pratylenchus penetrans) populations in strawberry roots were determined in field microplots and small plots. Application of entomopathogenic Steinernema carpocapsae and S. feltiae nematodes did not affect populations of migratory endoparasitic lesion nematodes in strawberry. We were able to demonstrate reduced mobility associated with dead insect nematodes, and this effect was greater in water than in soil. Bacteria associated with the insect parasites may cause nematode suppression, but the suppression appears to be temporary.(CT) Meloidogyne hapla can be spread in bare-root herbaceous perennial propagation material and may be difficult to control once established in new fields or in the landscape. Root pruning of bare-root plants was previously demonstrated to reduce spread and establishment of M. hapla. Planting M. hapla-resistant plants such as Rudbeckia and Aster into pots infested with M. hapla populations virtually eliminated the nematodes after 2-6 months of growth, demonstrating the potential of integrating sanitation and resistance for management of M. hapla in perennials. (MD) Survey sampling for nematode evaluations of fields using potato rotations is currently underway. One approach to reduce the survival of volunteer potato tubers is the installment of a hydraulic or chain-driven chopper situated at rear end of the harvester to cut the refuse and tubers into finer pieces. The rationale is that the smaller pieces of tuber will be more prone to desiccation and rots than whole tubers. Subsequent reductions in volunteer growth should reduce potential reservoirs for disease pathogens (i.e late blight) and CPB recruitment, as well as hosts for nematodes. (MI) The technologies of precision agriculture are being evaluated in relation to crop productivity, economics, nematode management, soil microbial populations, and soil quality parameters in both potato and soybean systems. Pratylenchus penetrans, Verticillum dahliae, potato early-die risk and soil quality characteristics for a 62-acre potato field was geopositioned on a one-acre grid basis. A replicated nematode management trial was designed using three early-die risk levels and three nematode management levels. The Michigan State University, Diagnostic Services Potato Early-Die Risk Matrix was used as the basis for the experimental design. (NY, Geneva) Limited information is available on the pathology and damage of the lesion nematode (Pratylenchus spp.) to onions. A replicated test in field microplots was established at the Geneva Station to assess the damage threshold density of this nematode to onion. The initial densities established were 0, 1, 2, 4, 8, and 16 nematodes/cc soil. The effectiveness of Vydate application against this nematode is also been assessed in microplots and in heavily infested fields. Data collection and analysis are still in-progress. A collaborative project titled "Farm ecosystem and management factors contributing to pest suppression on organic and conventional vegetable farms" was funded by NE-SARE program (2000-2002) to multidisciplinary research and extension colleagues at Cornell. Potatoes and winter squash are the primary vegetables on the selected farms. Nematode number and density are been determined at planting and harvest time on the 10 selected farms (5 organic and 5 conventional). In addition, soil suppressiveness to root pathogens is also being determined at harvest. Data collection and analysis is on-going (NY, Geneva). A multidisciplinary project titled "A partnership among eastern US carrot stakeholders to develop and implement IPM" was funded to Michigan State University in collaboration with Wisconsin, New Jersey and New York. Management options against the northern root-knot nematode is among the contributions to be researched in New York. Initial demonstration trials to be conducted in collaboration with commercial growers will focus on the use of rotational and cover crops, identified damage threshold levels, and selected nematicides. Identification of resistant carrot germplasm, evaluation of promising biological control organisms, and assessing the efficacy of natural products will be initially conducted in greenhouse, field microplots and/or experimental fields. (PA) Evaluation of green manure as a preplant treatment for replanted orchards: First year activities: An old cherry orchard with a history of tomato ringspot virus was removed and subdivided into nine plots (three treatments at three replicates each). The treatments included: soil fumigation (Telone II), green manure (Rapeseed cv. Dwarf Essex) and fallow. Nematode assays were taken prior to initiating the project. Treatment of orchard replant sites with Telone II soil fumigant reduced all plant-parasitic nematode populations to nearly undetectable levels. Rapeseed green manure treatments reduced dagger and lesion nematode populations but not as effectively as soil fumigation. Rapeseed green manure had no effect on the spiral nematode population although this nematode is not known to cause problems on fruit trees. Practices used to prepare orchard replant sites may affect soil characteristics such as compaction, organic matter content and others in subtle ways. The combined effect may affect tree establishment, vigor and early yield. In subsequent years, data will be collected on tree growth and yield along with nematode population development. (WVA) One unique finding of this research was that mulch (plastic or straw) does not prevent disease, rather, it was associated with increased disease. Mulch is promoted as a cultural practice for prevention of vegetable diseases by reducing soil splashing onto foliage thereby inhibiting the spread of inoculum. In the case of tomato diseases, particularly early blight, the use of mulch was associated with increased disease. </li> </dir> Deliverables (stakeholder items): <dir> <li>(CT) The effect of initial Pratylenchus penetrans densities in soils infested with a history of black root rot were incorporated into a strawberry economics model developed by Alison DeMarree & Regina Rieckenberg at Cornell University. Initial lesion nematode densities as low as 12 per g root resulted in an 11% loss over 4 years and densities as high as 125 per g root resulted in 100% loss in profit over the same time.(CT) Studies on the interaction of the fungal and nematode pathogens in strawberry black root rot will help to maximize sampling to best determine if nematodes are present and involved in black root rot, the contribution of the nematode to disease, and may help to manage the disease complex. The identification and use of rotation crops that reduce both pathogen populations in soil will assist in the development of nonchemical controls of black root rot of strawberry. (CT) Early dying of potato, also caused by a pathogen complex including Verticillium and Pratylenchus, is a major limiting factor of potato production and is extremely difficult to control. The potential of disease management by rotation, green manuring or organic matter amendments would be important both in potato production and the understanding of how soil ecology influences pathogens and disease development. (FL) The MAb has been established as a definitive probe to detect adhesins as virulence determinants on Pasteuria endospores in soil. This sensitive quantitative immunoassay can be applied as a tool for evaluating chemical and biological measures required for control of root-knot nematodes. (MD) The evaluation of rotational crops, organic amendments and host crop genetics on nematode community structure, and demonstration of their economic benefits, will identify alternative tactics for reducing pesticide use, lowering control costs and crop losses, and provide a sustainable strategy for managing nematodes in vegetable and field crops. (MI) The technology of precision agriculture appears to have excellent potential for use in management of plant-parasitic nematodes and the potato early-die disease complex .The data indicate that the techniques of precision agriculture had the potential to increase 2000 net profit at this site by $304 per acre. (MA) Demonstration that natural enemies for biological control, such as Pasteuria, may be destroyed by fumigation. (NY, Geneva) Assessing the genetic and pathological characteristics of nematode populations, such as those reported here for Meloidogyne hapla, is critical for identifying resistant crop germplasm, designing effective crop rotations, and developing diagnostic tools. Determining threshold damage level, identifying resistant cultivars, and searching for effective natural products against nematodes are important for the development of IPM programs against plant-parasitic nematodes. Documenting the impact of management options on nematodes and the validation of developed IPM programs in commercial fields and in collaboration with growers are much needed for the promotion of sustainable nematode management programs. (NY, Ithaca) Discovery of Pasteuria on as yet unidentified root knot and free-living nematodes. Pasteuria was found not to digest organ systems of the nematode host. Though Pasteuria penetrans is a very promising biocontrol agent for certain plant parasitic nematodes, the nature of host preference is not understood. Current findings about the nature of the nematode and bacterial surfaces will contribute to our understanding of Pasteuria attachment, infection, and requirements for endospore development and will facilitate strain enhancement and development of efficient mass culture systems for Pasteuria. (PA) Natural products, green manures and rotation crops can be useful tools for nematode management in fruit crops. These practices can be used to develop IPM and organic production programs with reduced use of synthetic pesticides. (RI) Short chain fatty acids, formed from the fermentation of organic compounds in the soil, are readily oxidized to carbon dioxide and water by soil microorganisms. Published experiments have demonstrated the nematicidal properties of organic acids, with butyric acid being the most effective. We have confirmed this effect in golf course soil. Butyric acid holds promise as a biorational alternative to synthetic fumigants. (USDA , MD) Rhizosphere bacteria possessing an ability to colonize crop plant roots have the potential to be useful biocontrol agents for plant-parasitic nematodes. In vitro assays followed by growth chamber and greenhouse tests aid in determining which microbes demonstrate activity against nematodes. Application of active natural compounds produced by microbes eliminates the need to be concerned about packaging, shipping, and shelf-life of live organisms. Studies of the gelatinous matrix and egg masses contribute knowledge about interactions between nematodes and microbes. (USDA , SC) The northern root-knot nematode causes serious damage to pepper (Capsicum spp.) in the U.S. and worldwide. The development of cultivars with genetic resistance to root knot nematodes is the most efficient and environmentally benign way of controlling this production problem. Host resistance also provides the most viable and easily implemented alternative to the use of methyl bromide soil fumigation to control root-knot nematodes. Because there are no sweet or hot pepper cultivars with resistance to M. hapla available to pepper growers, it is important to search for resistance in available pepper germplasm collections, such as the USDA pepper germplasm collection. </li> </dir> Plans for the coming year: <dir> <li>(CT) Potential biocontrol organisms isolated from Connecticut will be tested for ability to parasitize cyst, root knot or lesion nematodes in greenhouse and laboratory assays.(CT) The effects of rotation crops on black root rot pathogens in soil will continue to be evaluated. The effects of plant residues on pathogens will be distinguished from the effects of plant growth. Saia oats, sorgho-sudangrass, canola and Polynema marigold and plants in the Asteraceae will be examined. The effects of resistant perennial ornamental plants and incorporating plant residues on root-knot nematodes will be determined. (CT) The effects of rotation and antagonistic crops on potato early dying caused by Verticillium dahliae and P. penetrans will continue to be investigated in microplots. The effects of a single season of rotation crops in 1999 on early dying symptom development will be determined in 2000. (FL) Studies for the next year will seek to develop bivalent F(ab)2 and monovalent F(ab) forms of the IgM MAb to allow a better definition of the epitope recognized by the MAb. Recombinant antibodies bearing the Fv region of the MAb will be engineered and expressed in E. coli to develop alternative reagents for field tests to detect P. penetrans. (MD) Continuation of field and microplot evaluations, and expansion of host selection for maintenance and increase of Meloidogyne and Pasteuria cultures. (MA) Stunt nematodes: Six greens have been selected that have a range of Pasteuria-infected stunt nematodes. Two greens have low (3.4 and 8.3%), two have medium (14 and 22%), and two have high levels of infection (70% each). On one each of the low, medium and high sites, Nemacur-treated and non-treated plots (six replications of each) will be established. On each green, composite samples will be taken every 5 weeks from May until November to assess the population dynamics of Pasteuria and stunt nematodes. Root-knot nematodes: Six greens have been selected that represent a range of root-knot populations and Pasteuria infections. These greens will be sampled when root-knot juveniles are most prevalent in the soil, September through June. Sampling frequency will depend on weather (frozen ground) conditions. Nemacur will be applied to two of the root knot infested greens. These greens will have six treated and six untreated plots. (NY, Geneva) Onion Plant Introduction accessions tested and exhibiting tolerance to the northern root-knot nematode (Meloidogyne hapla) will be re-evaluated in the greenhouse and in field microplots. Tests will identify and will characterize suppressive soils against major plant-parasitic nematodes to vegetables in New York. Assess the effect of the inclusion of appropriate crops (rotational or antagonistic cover crops) in suppressing nematode populations and their damage to vegetables. Research efforts will be continued to identify resistance factor(s) against the northern root-knot nematode in onions and other vegetables. Efficacy of natural products and promising biological control organisms will be determined against root knot and lesion nematodes on vegetable crops. IPM programs will be designed and will be evaluated against the root knot and lesion nematodes on carrots, onions and other vegetables grown in New York. (NY, Ithaca) In vitro method for maintaining free-living nematodes infected with Pasteuria will be refined to facilitate studies of host-parasite interactions and requirements for endospore development. Studies will determine the role (if any) of sialylated residues in the epicuticle of "stripped" Meloidogyne incognita second-stage juveniles (J2). "Stripped" J2 refers to J2 with surface coat removed. Anti-P20 IgM will be tested on "stripped" Mi J2. The use of differential mass spectrometry will be explored to identify specific cuticular epitopes involved in the attachment of P. penetrans endospores to the nematode surface. (PA) Field testing green manure treatments of rapeseed and other cover crops for ability to control Xiphinema will be continued as well as evaluation of different preplant treatments on orchard establishment and early crop yields. Research on the biology and phylogenetic relationships of Xiphinema americanum - group nematodes, including virus associations, will continue. (RI) Screening nematode species and pathogenic fungi for sensitivity to butyric acid will continue. Based on those results, a greenhouse trials will be conducted to evaluate the effectiveness of butyric acid as a preplant treatment for strawberries. Parasites and pathogens of interest on strawberries include Pratylenchus penetrans, Meloidogyne hapla, Phytophthora fragariae, Pythium, Rhizoctonia solani, Fusarium oxysporum, and Verticillium dahliae. The persistence of butyric acid in soil will be quantified. (USDA, MD) Studies will continue on fungi and on rhizosphere bacteria to determine whether selected microbes produce nematode-antagonistic compounds. Promising natural compounds and/or live agents will be investigated to determine whether they are useful for management of plant-parasitic nematodes. (WVA) Population density and activity of nematode trapping fungi will be monitored in apple blocks with and without ground cover of endophyte-infected and endophyte-free fescue. Research will also continue in an effort to determine the mechanism of suppressiveness of endophyte infected fescues and to determine how best to integrate this rotation crop with orchard management systems. Farming systems research to compare compost amendments and rotations in organic vegetable and field crop production systems will be continued. Soil suppressiveness to nematodes and population dynamics of indigenous biocontrol agents will be compared in sites receiving compost amendments versus grass-legume rotations during a three-year transition phase from conventional to organic farming systems. (USDA, SC) Evaluate the next 25% of USDA pepper (Capsicum spp.) collection for resistance to northern root-knot nematode (M. hapla) and determine heat stability of N gene (for root-knot nematode resistance) in heterozygous pepper genotypes. </li> </dir>

Publications

Full Manuscripts: Abawi, G. S., T. L. Widmer, J. L. Ludwig, and N. A. Mitkowski. 1999. Biology and management of Meloidogyne hapla on carrots, onions, and lettuce in New York. J. of Nematology 31: 521. Chen, J., G. S. Abawi, and B. M. Zuckerman. 2000. Efficacy of Bacillus thuringiensis, Paecilomyces marquandii, and Streptomyces costaricanus with and without organic amendment against Meloidogyne hapla infecting lettuce. J. of Nematology 32: 70-77. Chen, J., G. S. Abawi, and B. M. Zuckerman. 1999. Suppression of Meloidogyne hapla and its damage to lettuce grown in a mineral soil amended with chitin and selected biocontrol organisms. J. of Nematology 31 (4S): 719-725. Fery, R. L. and J. A. Thies. 2000. Inheritance of resistance to the peanut root-knot nematode in hot pepper (Capsicum chinense). J. Amer. Soc. Hort. Sci. 125:615-618. Halbrendt J.M., N.O. Halbrendt and J. Brown. 1999. Screening rapeseed tissue for nematicidal activity. Journal of Nematology 31:538 (Abstr.) LaMondia, J. A. 1999. The effects of Pratylenchus penetrans on strawberry vigor and yield. Journal of Nematology 31:418-423. LaMondia, J. A. 1999. The effects of rotation crops on the strawberry pathogens Pratylenchus penetrans, Meloidogyne hapla, and Rhizoctonia fragariae. Journal of Nematology 31:600-655. LaMondia, J. A. 1999. Efficacy of insecticides for control of the foliar nematodes Aphelenchoides fragariae and Ditylenchus dipsaci in flowering perennial ornamentals. Journal of Nematology 31: 644-649. LaMondia, J. A. 1999. Seasonal population dynamics of lesion and root-knot nematodes in strawberries. Journal of Nematology 31:550. LaMondia, J. A. 2000. The effect of rotation crops on lesion nematodes, Verticillium dahliae and potato early dying severity. Phytopathology 90(6S):44. Masler, E.P., E.S. Kovaleva, and S. Sardanelli. 1999. FMRFamide-like Immunoactivity in Heterodera glycines (Nemata: Tylenchida). Journal of Nematology. 31 (2): 224-231. Masler, E.P., E.S. Kovaleva, and S. Sardanelli. 1999. Comparison of FaRP Immunoactivity in Free-Living Nematodes and in the Plant-parasitic Nematode Heterodera glycines. In Neuropeptides: Structure and Function in Biology and Behavior, volume 897 of the Annals of the New York Academy of Sciences. 253-263. Masler, E. P., E. S. Kovaleva, and S. Sardanelli. 2000. Aminopeptidase-like Activities in Caenorhabditis elegans and the soybean cyst nematode, Heterodera glycines. Manuscript submitted, Journal of Helminthology. Meyer, S., Roberts, D., Chitwood, D., Carta, L., Lumsden, R., Mao, W. Biocontrol of root-knot nematode on pepper. Phytopathology. June (Supplement) 2000. v. 90 (6): p. S53. Nitao, J. K., Meyer, S. L. F., Schmidt, W. F., Fettinger, J. C., Chitwood, D. J. Nematode-antagonistic trichothecene compounds from Fusarium equiseti. Phytopathology. June (Supplement) 2000. v. 90 (6): p. S56. Thies, J. A. and R. L. Fery. 2000. Characterization of resistance conferred by the N gene to Meloidogyne arenaria races 1 and 2, M. hapla, and M. javanica in two sets of isogenic lines of Capsicum annuum L. J. Amer. Soc. Hort. Sci. 125:71-75. Abstracts: Abawi, G. S., and J. W. Ludwig. 2000. Plant-parasitic nematodes associated with onion roots and soils in New York State. J. of Nematology 32: (Abstr., in-press). LaMondia, J. A. 1999. Rhizoctonia / Pratylenchus interactions in strawberry black root rot. APS Publication no. P-1999-0009-ssa. Mitkowski, N. A., and G. S. Abawi. 2000. Pathogenicity and virulence of Meloidogyne hapla populations from vegetables in New York state. J. of Nematology 32: (Abstr., in-press). Widmer, T. L., and G. S. Abawi. 2000. Impact of organic matter management on plant-parasitic nematodes, their damage to host crops, and soil health. J. of Nematology 32 (Abstr., in-press). Manuals, Bulletins and Reports: Abawi, G. S., J. W. Ludwig, and T. L. Widmer. 2000. Root-knot nematode and control options for carrots. NYS Veg. Conf., pp 53-56, Cornell Cooperative Extension, Ithaca, NY. Abawi, G. S., and J. W. Ludwig. 2000. Onion nematode research results, 1999: Statewide survey, varieties and Vydate. NYS Veg. Conf., pp. 151-154, Cornell Cooperative Extension, Ithaca, NY. Fery, R. L., P. D. Dukes, Sr., and J. A. Thies. 2000. Carolina Wonder and Charleston Belle: New, southern root-knot nematode resistant bell pepper cultivars for South Carolina. 1997-1998 Clemson University Vegetable Report 8:64-69. Fery, R. L. and J. A. Thies. 2000. Inheritance of resistance to southern root-knot nematode in hot pepper (Capsicum chinense). 1997-1998 Clemson University Vegetable Report 8:54-59. Kenworthy, W. J. 2000. 1999 Cyst Nematode-Resistant Soybean Variety Test. CES Agronomy Mimeo No.43. 7pp. Kenworthy, W. J. 2000. Evaluation and development of plant resistance to the soybean cyst nematode. In University of Maryland 2000 IPM Progress Report. In Press. LaMondia, J. A. 2000. Editor, Techniques in Nematode Ecology, an on-line Manual. The Society of Nematologists Ecology Committee, Lawrence, Kansas. URL: <a href="ehttp://ianrwww.unl.edu/son/Ecology_Manual_TOC.htm"e>http://ianwww.unl.edu/son/Ecology_Manual_TOC.htm</a> Meyer, S. L .F. The future of biocontrol fungi for nematode management. Mycological Society of America Program book. August 2000. p. 28. Nitao, J. K., Meyer, S. L. F., Schmidt, W .F., Fettinger, J. C., Chitwood, D. J. Nematicidal compounds from fungi: inhibitors of root-knot nematode egg hatch and juvenile mobility. BARC Poster Day Program Book. April 2000. p. 23. Sardanelli, S., Everts, K., Kratochvil, R. and G. Dively. 2000. Grower Initiated IPM- SARE Project Explores Alternative Nematode Management Tactics. In University of Maryland 2000 IPM Progress Report. In Press Thies, J.A. 1999. Southernpea Cooperative Trial Report. 1999. Nematode ratings. Pp. 26-27. In: T. E. Morelock (ed.). Composite Report of the Regional Southernpea Cooperative Trials. Dept. Horticulture, Univ. Arkansas. Fayetteville, AR. Thies, J. A. and R. L. Fery. 2000. Resistance to southern root-knot nematode in Charleston Belle and Carolina Wonder bell peppers is compromised at high soil temperatures. 1997-1998 Clemson University Vegetable Report 8:74-78. Thies, J. A., J. D. Mueller, and R. L. Fery. 2000. Use of resistant Carolina Cayenne pepper as a rotational crop to manage southern root-knot nematode. 1997-1998 Clemson University Vegetable Report 8:70-73.

Impact Statements

Date of Annual Report: 12/10/2001

Report Information

Annual Meeting Dates: 10/10/2001 - 10/12/2001
Period the Report Covers: 10/01/2000 - 09/01/2001

Participants

url

Participants Attachment:

Attachment

Brief Summary of Minutes

Minutes Attachment:

Attachment

Accomplishments

<a href="http://www.lgu.umd.edu/misc/ne171saes2001.html">NE171 Accomplishments and Impacts</a>

Publications

Full Manuscripts: Chen. S.Y., J.H. Charnecki, J.F. Preston, D.W. Dickson. 2000. Extraction and purification of Pasteuria spp. endospores. J. Nematol. 32:78-84. Elmer, W. H. and LaMondia, J. A. 2001. Plant parts and their diseases. Plant Health Instructor in press. Esnard, J. 2001. Bona fide pathogens of nematodes may hold greatest biocontrol potential. J. Nematol. Submitted; in review. Kays, S. J., W. J. McLaurin, Y. Wang, P. D. Dukes, J. Thies, J. Bohac, and D. M. Jackson. 2001. GA90-16: A nonsweet, staple-type sweetpotato breeding line. HortScience 36:175-177. Kokalis-Burelle, N., C. S. Vavrina, E. N. Rosskopf, and R. A. Shelby. 2001. Field Evaluation of PGPR Amended Transplant Mixes and Soil Solarization for Tomato and Pepper Production in Florida. Plant and Soil: In Press. Kokalis-Burelle, N., W. F. Mahaffee, R. Rodrmguez-Kabana, J. W. Kloepper, and K. L. Bowen. 2001. Effects of Switchgrass Rotations with peanut on Nematode populations and soil microflora. J. Nematology 33: In Press. Kokalis-Burelle, N., C.S. Vavrina, M. S. Reddy, and J. W. Kloepper. 2001. Effects of Plant Growth-promoting Rhizobacteria as Amendments to Muskmelon and Watermelon Transplants on Disease Resistance and Yield. HortScience: Submitted. LaMondia, J. A. 2001. Genetics of burley and flue-cured tobacco resistance to Globodera tabacum tabacum. Journal of Nematology in press. LaMondia, J. A. 2001. Broadleaf tobacco yield loss in relation to initial Globodera tabacum tabacum population density. Journal of Nematology in press. LaMondia, J. A. 2001. Registration of Scantic broadleaf tobacco. Crop Science in press. Lewis, E.E., P.S. Grewal, and S. Sardanelli.2001. Long-term interactions between the Steinernema feltiae/Xenorhabdus bovienii insect pathogen complex and the root-knot nematode, Meloidogyne incognita. In press: Biological Control Martmnez-Ochoa, N., N. Kokalis-Burelle, J. W. Kloepper, and R. Rodrmguez-Kabana. 2001. Rhizobacteria, organic amendments, and botanical aromatics for the management of Meloidogyne spp. in tomato transplants. Plant Disease: In Press. Masler, E. P., E. S. Kovaleva, and S. Sardanelli. 2001. Aminopeptidase-like Activities in Caenorhabditis elegans and the soybean cyst nematode, Heterodera glycines. In press: Journal of Helminthology. Meyer, S. L. F., Massoud, S. I., Chitwood, D. J. and Roberts, D. P. Evaluation of Trichoderma virens and Burkholderia cepacia for antagonistic activity against root-knot nematode, Meloidogyne incognita. Nematology: 871-879. 2000. Meyer, S. L. F., Roberts, D. P., Chitwood, D. J., Carta, L. K., Lumsden, R. D. and Mao, W. Application of Burkholderia cepacia and Trichoderma virens, alone and in combinations, against Meloidogyne incognita on bell pepper. Nematropica 31: 75-86. 2001. Meyer, S. L. F. Will any two biological agents manage any one nematode? Journal of Nematology 32: 445-446. 2000. Nitao, J. K., Meyer, S. L. F., Schmidt, W. F., Fettinger, J. C. and Chitwood, D. J. Nematode-antagonistic trichothecenes from Fusarium equiseti. Journal of Chemical Ecology 27: 859-869. 2001. Nitao, J. K., Meyer, S. L. F., Schmidt, W. F., Fettinger, J. C. and Chitwood, D. J. Fusarium equiseti secretes compounds antagonistic to root-knot nematode, Meloidogyne incognita. Journal of Nematology 32: 451-452. 2000. Meyer, S. L. F., Nitao, J. K. and Chitwood, D. J. Identification of nematode-antagonistic compounds from fungi. Phytopathology 91 (Supplement): S139. 2001. Preston, J.F.. D.W. Dickson, J.E. Maruniak, J.A. Brito, L.M. Schmidt, and J.M. Anderson. 2001. Pasteuria spp.: systematics-phylogeny of these unusual bacterial parasites of phytopathogenic nematodes. Phytopathology Suppl. 6, 91:S173. Thies, J. A. and R. L. Fery. 2000. Characterization and specificity of resistance of root-knot nematode (Meloidogyne spp.) resistance in Scotch Bonnet peppers (Capsicum chinense Jacq.). Proc. of the National Pepper Conference, October 1-5, 2000, Lafayette, Louisiana. P. 19. Thies, J. A. and R. L. Fery. 2000. Heat stability of resistance to Meloidogyne incognita in Scotch Bonnet peppers (Capsicum chinense Jacq.). J. Nematology 32:356-361. Thies, J. A. and R. L. Fery. 2001. Reactions of regional cooperative southernpea (cowpea) breeding lines and standard check cultivars to southern root-knot nematode race 3, 2000. B & C Tests (published by the American Phytopathological Society). On-line publication. Report # 2001:V108. Thies, J. A. and R. L. Fery. 2001. Characterization of Capsicum chinense cultigens for resistance to Meloidogyne arenaria, M. hapla, and M. javanica. Plant Dis. 85:267-270. Abstracts: Abawi, G. S., and J. W. Ludwig. 2000. Plant-parasitic nematodes associated with onion roots and soils in New York State. J. of Nematology 32: 414 (Abstr.). Fery, R. L. and J. A. Thies. 2000. Development of southernpea [Vigna unguiculata (L.) Walp.] cultivars with an enhanced persistent green seed phenotype. HortScience 35:399. Kokalis-Burelle, N. 2001. Chitin amendments for suppression of plant parasitic nematodes and fungal pathogens. Phytopathology (Abst.): In Press. LaMondia, J. A. 2000. The effect of rotation crops on lesion nematodes, Verticillium dahliae and potato early dying severity. Phytopathology 90(6S):44. (Abstract) URL:  <a href="http://www.apsnet.org/meetings/2000/search/p00ma316.htm">http://www.apsnet.org/meetings/2000/search/p00ma316.htm</a>   LaMondia, J. A. and R. S. Cowles. 2000. The effects of insect parasitic nematodes on Pratylenchus penetrans populations in strawberries. Journal of Nematology (Abstract) 32:441. LaMondia, J. A. 2001. The interaction of Rhizoctonia fragariae and Pratylenchus penetrans in strawberry black root rot. Phytopathology (Abstract) 91(6S): S195. Online Publication no. P-2001-0020-NEA URL:  <a href="http://www.apsnet.org/meetings/div/ne00abs.asp">http://www.apsnet.org/meetings/div/ne00abs.asp</a>  LaMondia, J. A. 2001. The relation between tobacco cyst nematode density and damage to broadleaf tobacco. Phytopathology (Abstract) 91:S52. Meyer, S.L.F., Nitao, J.K. and Chitwood, D.J. Isolation of nematode-antagonistic compounds from fungi. British Mycological Society International Symposium, "Bioactive Fungal Metabolites - Impact and Exploitation" Abstract Book: 18. 2001. Mitkowski, N. A., and G. S. Abawi. 2000. Pathogenicity and virulence of Meloidogyne hapla populations from vegetables in New York state. J. of Nematology 32: 446 (Abstr.). Nitao, J. K., Meyer, S. L. F., Schmidt, W. F., Fettinger, J. C. and Chitwood, D. J. Nematode-antagonistic trichothecenes from the fungus Fusarium equiseti. European Society of Nematologists 25th International Symposium on Nematology Meeting Book: 33. 2000. Schmidt, L.M., D.M. Dickson, and J.F. Preston. 2001. A monoclonal antibody for detection and quantitation of Pasteuria penetrans spores in field soil. Abstracts of the 101st General Meeting of the American Society for Micribiology. Thies, J. A. and R. L. Fery. 2000. Evaluation of a core of the USDA Capsicum germplasm collection for resistance to northern root-knot nematode, Meloidogyne hapla. Phytopathology 90:S76. Thies, J. A. 2001. Red food coloring stain: A new, safer procedure for staining nematodes in roots and egg masses on root surfaces. Phytopathology 91:S144. Thies, J. A. and R. L. Fery. 2001. Root-knot nematodes: need for host plant resistance in cowpea. HortScience 36:850. Widmer, T. L., and G. S. Abawi. 2000. Impact of organic matter management on plant-parasitic nematodes, their damage to host crops, and soil health. J. of Nematology 32: 471 (Abstr.). Manuals, Bulletins and Reports: Abawi, G. S., and J. W. Ludwig. 2001. Results of nematode research on onions and and cover crops use review. NYS Vegetable Conference, Cornell Cooperative Extension, Ithaca, NY. pp 156-160. Halbrendt J.M. 2001. Nematodes. In Concise Encyclopedia of Temperate Zone Tree Fruits. A.Basra, editor. Haworth Publishing Company (submitted). Halbrendt, J.M., L. Roth and W. Kleiner. 2000. Compost for weed and nematode management. Pennsylvania Fruit News 79:42-44. Halbrendt, J.M. 2000. Replant site preparation. Pennsylvania Fruit News 79:44-45. Kenworthy, W. J. 2001. Cyst Nematode-Resistant Soybean Variety Test. CES Agronomy Mimeo No.43. 7pp. Kenworthy, W. J. 2001. Evaluation and development of plant resistance to the soybean cyst nematode. In University of Maryland 2000 IPM Progress Report. Kokalis-Burelle, N, P. Fuentes-Bsrquez, and P. Adams. 2000. Effects of Reduced Risk Alternatives on Nematode Populations and Crop Yield. Annual International Research Conference on Methyl Bromide Alternatives and Emissions Reduction. Pages 66.1-66.2. LaMondia, J. A. 2001. Challenges in ornamental nematology. Nematology Newsletter in press. Sardanelli, S., Everts, K., Kratochvil, R. and G. Dively. 2001. Grower Initiated IPM-SARE Project Explores Alternative Nematode Management Tactics. In University of Maryland 2000 IPM Progress Report.

Impact Statements

Date of Annual Report: 12/04/2002

Report Information

Annual Meeting Dates: 10/09/2002 - 10/11/2002
Period the Report Covers: 10/01/2001 - 09/01/2002

Participants

Louis Magnarelli, Administrative Advisor
Connecticut Agricultural Experiment Stn.;
George S. Abawi * Cornell University, Geneva, NY;
George W. Bird * Michigan State University;
Don W. Dickson * University of Florida;
John M. Halbrendt * Penn State University;
Jim Kotcon * West Virginia University;
Jim LaMondia * Connecticut Agricultural Experiment Stn;
Susan L. Meyer USDA, ARS, Beltsville, MD;
James F. Preston University of Florida;
Nathaniel A. Mitkowski * University of Rhode Island;
Sandra Sardanelli * University of Maryland;
Judy A. Thies *, Secretary USDA, ARS, Charleston, SC;
Robert L. Wick * University of Massachusetts;
Janet Chen Cornell University;
Bess Dicklow University of Massachusetts;
J. Hunter Cornell University;
J. W. Ludwig Cornell University;
Karen Snover Cornell University;
Fred Warner Michigan State University;

Brief Summary of Minutes

Action Items/Assigned Responsibility/Deadlines/Target Dates

The participants introduced themselves. The minutes of the 2001 Annual Research Technical Committee meeting were approved for posting on the NE-171 website. George Abawi, Local Arrangements, welcomed everyone to New York.

Lou Magnarelli distributed a timetable for rewriting the NE-171 Project and discussed procedures for rewriting. A brief discussion ensued about the potential rewrite or revision of the project.

Jim LaMondia will be the Chair of the Committee for the next two years and Judy Thies will be the Chair-Elect. Nathan Mitkowski volunteered to be the next Secretary (replacing Judy Thies) and he was unanimously approved for the position by the Committee.

Judy Thies, Jim LaMondia, and Don Dickson were nominated and unanimously approved to serve on the writing committee. Judy Thies will chair the committee.

George Bird presented an excellent workshop titled The Identification of Free-Living Nematodes. The topic of the workshop was highly relevant to the NE-171 Committee because the first objective of the NE-171 project is to evaluate the effects of rotational crops, organic amendments and host crop genetics on nematode community structure, which includes free-living nematodes. George Abawi had arranged for a local microscope company to provide microscopes and video equipment for the workshop.

The 2003 meeting will be held in Rhode Island and Nathan Mitkowski will be the Local Arrangements Chair.

Summary of Discussions (Append distributed information):

Project members and collaborators from each of the states and USDA, ARS reported on their research during the previous year. Progress reports distributed during the meeting can be obtained from Lou Magnarelli, Sandra Sardanelli, or the respective project members.

The project milestones were reviewed and a consensus was reached that the current milestones have been met.

The theme and objectives for the new Project were discussed. Topics discussed included the role of nematode communities in soil health, sustainable soil management, and alternative strategies for managing nematodes.

Between Meeting Information and Exchange/Development Information:

The meeting report and minutes will be prepared by Jim LaMondia and Judy Thies, respectively, and sent electronically to Lou Magnarelli, Administrative Advisor, for approval. Jim LaMondia will post the approved report and minutes on the project web site.

The writing committee (Jim LaMondia, Don Dickson, and Judy Thies) will contact potential reviewers for the Project proposal in November and December 2002. Names, addresses, fax, and telephone numbers of at least 6 reviewers will be provided to Lou Magnarelli. The writing committee will also prepare and submit a request to write a proposal to Lou Magnarelli (for approval), who will then submit the approved request to the Directors Multistate Activities Committee for review in February or March 2003. After approval to write the project proposal is received, the writing committee will develop the proposal using input from NE-171 members, a CRIS search will be done, and the proposal will be distributed to NE-171 members for review (April August 2003). Lou Magnarelli will electronically submit the project proposal to National Information Management and Support System (NIMSS) and reviewers and notify all Directors to enter scientists (Appendix E). NE-171 members will ask their Deans or Directors to complete Appendix E electronically. NE-171 members will discuss final changes for new project at the annual meeting in October 2003.

Accomplishments

Objective 1: Evaluate the effects of rotational crops, organic amendments and host crop genetics on nematode community structure. Complementary or collaborative studies to assess the impact of rotation and cover crops on plant-parasitic nematode populations and nematode community structure: Dr. George Bird, Nematologist at Michigan State Univ., conducted a successful half-day workshop on the Characteristics of the Orders and Selected Taxa of the Nematoda at the 2002 NE-171 Technical Committee Meeting in Geneva, NY. The identification of free-living nematodes will assist researchers evaluating the impact of nematode management tactics on nematode community structure in soil. Dr. Bird has prepared chapters on ‘nematode biology and management‘ & ‘nematodes and soil quality‘ in two interdisciplinary MSUE Bulletins: Michigan Field Crop Ecology E-2646, and Michigan Field Crop Pest Ecology and Management E-2704. Milestone 2002: Initiate research to determine the mechanism of nematode antagonism in rotation and cover crops. (MD) An interdisciplinary IPMSARE team is now into the third year of a MD grower-generated project entitled Development and Evaluation of Management Alternatives for Root Knot Nematodes and Volunteer Potatoes. Fields in Maryland (MD) cropped repeatedly to vegetables have experienced significant losses due to root knot nematodes (RKN), Meloidogyne spp. Maryland growers have expressed concern over sporadic but widespread surges of lesion nematode populations (Pratylenchus spp.), often associated with RKN from symptomatic fields. Both RKN and lesion nematodes have broad host ranges. Cooperators on the project include specialists in Field Crops, Entomology, Nematology and Vegetable Pathology along with the Dorchester County Extension Agent, IPM Scout and Dorchester County Growers. In this project, using a combination of on-farm (natural infestations) and micro-plot (artificially infested) experiments, sustainable management alternatives and their effect(s) on the plant-parasitic nematodes are being evaluated. Of all seasonal samplings and RKN life-stage evaluations (including root gall indices and root/egg harvest), Fall soil J2 sampling is showing the greatest significance as an indicator for detection and population evaluation purposes in MD. 7Milestone 2002: Evaluate the impact of rotation and cover crops and soil amendments on nematode populations. (PA) Four different sources of compost were evaluated for ability to suppress plant-parasitic nematodes and weeds in a young apple orchard. Three composts were from commercial sources and one was a spent mushroom compost. Dagger nematode population levels were extremely variable and only very large differences were shown to be statistically different. Nevertheless, plots treated with five inches of mushroom compost consistently had the lowest nematode numbers and mushroom compost applied at two inches also showed relatively low nematode numbers throughout the study. Although no conclusions can be drawn, these data hint at the possibility that mushroom compost is suppressive to dagger nematodes. Likely candidates would be either the chemical composition or antagonistic microflora. (PA) Treatment of orchard replant sites with Telone II soil fumigant reduced all plant-parasitic nematode populations to nearly undetectable levels. Rapeseed green manure treatments also reduced dagger and lesion nematodes but not as effectively as fumigation. However, in addition to nematode suppression, the rotation also provided an opportunity to bring broadleaf weeds under control, a major concern for the prevention of Peach Stem Pitting Disease. Tree growth data indicate that tree performance is nearly the same under all preplant treatments. Another year of observation is needed to determine if stem-pitting is confirmed in untreated fallow plots. (NY) Onion germplasm consisting of >50 accessions of A. fistulosum, two sources of A. roylei, and several A. cepa were evaluated against M. hapla. All the materials evaluated were susceptible, but several accessions were found to be higher in tolerance than the commercial varieties in the tests. A cross of A. roylei and A. cepa (provided by Dr. Martha Mutchler at Cornell) was among the best materials tested. (NY) Eight selected soybean cultivars were found to be good hosts to Pratylenchus penetrans. Incorporation of these cultivars as green manures 4 weeks before planting did not reduce the number of lesion nematodes in snap bean roots. Objective 2: Characterization of biological control agents for suppression of plant-parasitic nematodes. (USDA-ARS) Strains of rhizosphere-inhabiting bacteria, known to act against certain soilborne plant-pathogenic fungi, were tested in the greenhouse for ability to suppress root-knot nematode populations on cucumber roots. The greenhouse assay system identified a potentially useful bacterium. This bacterium was tested in the lab to determine whether natural products in culture filtrates suppressed egg hatch and juvenile mobility. A strain of Trichoderma virens was also tested for activity against Meloidogyne incognita on cucumber. Collaborators: Daniel Roberts (USDA ARS Sustainable Agriculture Systems Laboratory, Beltsville). Obj. 2.1) Survey plant-pathogenic nematodes for occurrence of Pasteuria spp. (FL) Environmental detection of endospores of Pasteuria spp. Conditions have been defined for the quantification of endospores of Pasteuria spp. using a monoclonal antibody directed against an epitope associated with the parasporal fibers that cover mature endospores. An amplification ELISA assay using biotinylated secondary antibody allows quantification to a minimum of 300 endospores per gram of soil. The assay is not affected by organic matter and is applicable to soils conducive to infestations with plant-parasitic nematodes. The assay has also been used to quantify the level of endospores in root tissues, and should be useful in assessing the potential levels of soil-borne endospores in subsequent plantings. The assay should be useful in assessing the extent to which soils are or may become suppressive for nematode infestations and thereby limit the need for other control measures. Obj. 2.2) Determine the survivability and host preference of isolates of Pasteuria spp. from different geographic regions. (MD) Populations of Meloidogyne incognita have been established and are maintained in laboratory, greenhouse and field sites for in preparation for cooperative studies with multistate researchers involving Pasteuria penetrans. Once sufficient inoculum is obtained, the bacterial nematode parasite will be introduced into root-knot nematode-infested microplots in the field. Both the nematode and its parasite will be monitored over time to determine whether the parasite will increase to suppressive levels and whether it will survive the winter season. Cooperating states are FL, MD, and MI. Obj. 2.3) Evaluate different crops and methods for growing Pasteuria penetrans. Rutgers tomato and PA136 Bell pepper have been selected as hosts for in house culture of Meloidogyne incognita on the basis of root vigor and overall cultural manageability, and are being grown in a soil moisture control system (Sardanelli and Kenworthy, 1997) and in a standard greenhouse environment. Meloidogyne incognita cultures are being maintained and increased in preparation for Trichoderma evaluations and also for microplot overwintering evaluations and/or production of P. penetrans endospores planned for Florida isolates of Pasteuria penetrans. Objective 3: Comparison and evaluation of IPM system management of plant-parasitic nematodes based on crop rotation, organic amendments, host crop resistance and biological control agents. (NY) Assessing the incidence and severity of root-galling on carrots as well as conducting a soil bioassay with lettuce (indicator host) at harvest were highly effective in determining the need for root-knot nematode control on the following crop cycle of carrots or other susceptible hosts. (NY) A combined in-furrow and foliar spray application of Vydate C-LV increased potato tuber number and weight as well as slightly reduced the number of root-knot nematode in soil and roots. No root-galling symptoms were observed on infected potato roots.

Publications

Full Manuscripts: Abawi, G. S. 2002. Nematode and disease interactions. In: Encyclopedia of Plant & Crop Science. R. M. Goodman, ed. Marcial Dekker, New York (In press). Abawi, G. S., J. W. Ludwig, J. E. Carroll, and T. L. Widmer. 2001. Management of leaf blight diseases and root-knot nematode on carrots in New York. Pp. 66 67. Great Lakes Expo, MSU Extension, E. Lansig, MI. Elmer, W. H. and LaMondia, J. A. 2001. Plant parts and their diseases. The Plant Health Instructor. DOI:10.1094/PHI-K-2001-1129-02. Fery, R. L., and Thies, J. A. 2002. ‘Charleston Nemagreen‘, a root﷓knot nematode resistant, cream﷓type southernpea with a green cotyledon phenotype. HortScience 37:(Accepted 25 March 2002). LaMondia, J. A. 2002. Seasonal populations of lesion and root-knot nematodes in strawberry roots. Journal of Nematology in press. LaMondia, J. A. 2002. The interaction of Pratylenchus penetrans and Rhizoctonia fragariae in strawberry black root rot. Journal of Nematology in press. LaMondia, J. A., and R. S. Cowles. 2002. Effect of entomopathogenic nematodes and Trichoderma harzianum on the strawberry black root rot pathogens Pratylenchus penetrans and Rhizoctonia fragariae. Journal of Nematology in press. LaMondia, J. A., W. H. Elmer, T. L. Mervosh, and R. S. Cowles. 2002. Integrated management of strawberry pests by intercropping. Crop Protection accepted, in press. Li, W., Roberts, D. P., Dery, P. D., Meyer, S. L. F., Lohrke, S., Lumsden, R. D. and Hebbar, K. P. Broad spectrum anti-biotic activity and disease suppression by the potential biocontrol agent Burkholderia ambifaria Bc-F. Crop Protection Journal 21: 129-135. 2002. Mitkowski, N.A. and G.S. Abawi. 2003. Genetic diversity of New York State Meloidogyne hapla populations determined by RAPD‘s and mitochondrial DNA. Journal of Nematode Morphology and Systematics: (submitted). Mitkowski, N.A. and G.S. Abawi. 2002. Reproductive fitness on lettuce of populations of Meloidogyne hapla from New York State vegetable fields. Nematology: (accepted). Mitkowski, N.A. and G.S. Abawi. 2002. Monoxenic maintenance and reproduction of root-knot nematode (Meloidogyne hapla) on multiple-species in vitro root culture systems. Plant Cell Reports 21 (1):14-23. DOI 10.1007/s00299-002-0468-6. Mitkowski, N. A., J. G. Van der Beek, and G. S. Abawi. 2002. Characterization of root-knot nematode populations associated with vegetables in New York State. Plant Dis. 86: 840 847. Meyer, S. L. F. and Roberts, D. P. Combinations of biocontrol agents for management of plant-parasitic nematodes and soilborne plant-pathogenic fungi. Journal of Nematology: 34: 1-8. 2002. Nitao, J. K., Meyer, S. L. F., Oliver, J. E., Schmidt, W. F. and Chitwood, D. J. Isolation of flavipin, a fungus compound antagonistic to plant-parasitic nematodes. Nematology 4: 55-63. 2002. Orion, D., Kritzman, G., Meyer, S. L. F., Erbe, E. F. and Chitwood, D. J. A role of the gelatinous matrix in the resistance of root-knot nematode (Meloidogyne spp.) eggs to microorganisms. Journal of Nematology 33: 203-207. 2001. Preston, J.F., D.W. Dickson, J.E. Maruniak, G. Nong, J.A. Brito, L.M. Schmidt, and R.M. Giblin-Davis. 2003. Pasteuria spp.: Systematics and phylogeny of these bacterial parasites of phytopathogenic nematodes. Journal of Nematology Accepted. Schmidt, L.M., J.F. Preston, D.W. Dickson, J.D. Rice, and T.E. Hewlett. 2002. Environmental quantification of Pasteuria penetrans endospores using antigen extraction and immunodetection with a monoclonal antibody. FEMS Microbiology Ecology. In Press. Thies, J. A., and Fery, R. L. 2002. Evaluation of a core of the U.S. Capsicum germplasm collection for reaction to the northern root﷓knot nematode. HortScience 37: (Accepted for publication 17 October 2001). Thies, J. A., and Fery, R. L. 2002. Heat stability of resistance to southern root﷓knot nematode in bell pepper genotypes homozygous and heterozygous for the N gene. J. Amer. Soc. Hort. Sci. 127:371﷓375. 2002 Ude, G.N., J.M. Costa, W.J. Kenworthy, S. Sardanelli, and P.B. Cregan. 2002. AFLP Markers Associated with a QTL for Resistance to Soybean Cyst Nematode (Heterodera glycines) Race3. Journal of Genetics and Breeding. Accepted. Widmer, T.L., N.A. Mitkowski and G.S. Abawi. 2002. Impact of organic matter management on plant-parasitic nematodes, their damage to host crops and soil health. Journal of Nematology: (in press). Abstracts: Halbrendt, J. M. and J. A. LaMondia 2002. Nematicidal activity of selected plant residues to Xiphinema americanum. Proceedings of the Fourth International Congress of Nematology: in press. LaMondia, J. A. 2002. Nematode management on herbaceous perennial ornamentals. Phytopathology 92 (6S):107. Meyer, S. L. F., Roberts, D. P., Nitao, J. K., and Chitwood, D. J. Examination of rhizosphere-associated microbes for production of compounds active against plant-parasitic nematodes. Nematology 4: 288. 2002. Meyer, S. L. F., Roberts, D. P., Nitao, J. K., and Chitwood, D. J. Detection of root-associated microbes that produce compounds active against plant-parasitic nematodes. Phytopathology 92 (Supplement): S55. 2002. Roberts, D. P., Lohrke, S. M., Meyer, S. L. F., and Buyer, J. S. Biocontrol of seedling pathogens and root-knot on vegetables. Annual International Conference on Methyl Bromide Alternatives and Emissions Reductions, San Diego, CA, Conference Proceedings: 30. 2001. Manuals, Bulletins and Reports: Halbrendt, J.M., L. Roth and W. Kleiner. 2001. Compost for weed and nematode management. Pennsylvania Fruit News 81:48-51. Halbrendt, J.M. 2001. Replant site preparation. Pennsylvania Fruit News 81:34-35. Kenworthy, W. J. 2002 Cyst Nematode-Resistant Soybean Variety Test. CES Agronomy Mimeo No.43. 7pp. Kratochvil, R. 2001. Field Crops. In University of Maryland 2001 IPM Progress Report. p. 9. LaMondia, J. A. 2001. Nematode diseases of perennials. Yankee Grower 3(6):6-7. LaMondia, J. A. 2001. Host status of herbaceous perennial ornamentals to the northern root-knot nematode. Yankee Grower 3(6):7-10. LaMondia, J. A. 2001. Nematodes: Common and important problems in the ornamental and landscape industry. Nematology Newsletter 47(3):1-4. Mallozzi, T., Kratochvil, R., Sardanelli, S., Meyer, S., and Everts, K. 2001. Evaluating Alternatives to Methyl Bromide for Management of Root-Knot Nematode. In University of Maryland 2001 IPM Progress Report. p. 34. Mallozzi, T., Sardanelli, S., and Kratochvil, R 2002. Rearing Root-Knot Nematode Populations for Research: Utilization of an Innovative Approach. Society of Agronomy. Mitkowski, N.A. 2001. Root-knot nematodes on turf in the Northeastern United States. Turfgrass Trends 10(12):1-4. Thies, J. A., and Fery, R. L. 2002. Reactions of regional cooperative southernpea (cowpea) breeding lines and standard check cultivars to southern root﷓knot nematode race 3, 2001. B & C Tests (published by the American Phytopathological Society). On﷓line publication Thies, J. A., and Fery, R. L. Reactions of regional cooperative southernpea (cowpea) breeding lines and checks to southern root﷓knot nematode race 3, 2001. Biological and Cultural Tests for Control of Plant Diseases (Accepted 24 January 2002). Theses Agripina Gonzalez Grijalba, 2002. Nematodes associated with soils and roots of bean fields under different cultural practices. MPS Thesis, CALS, Cornell University, Ithaca, NY. 57 pp.

Impact Statements

Drs. J. A. LaMondia (CT) and J.M. Halbrendt (PA) and co-investigators were awarded a competitive grant to find alternative methods of nematode control as a part of the Methyl Bromide Transition Program. Collaborative studies from NE-171 enabled these scientists to secure additional funds for further research to help solve immediate stakeholder problems.
Drs. G.S. Abawi (NY), G.W. Bird (MI), and N.A. Mitkowski (RI) were awarded a competitive grant for research on alternative methods of nematode control as a part of the Methyl Bromide Transition Program. Their collaborative work in NE-171 helped them secure these funds for additional studies to assist stakeholders.
A workshop conducted during the NE-171 meeting provided specific training for participants to improve their skills on identifying nematodes. This knowledge will assist scientists in the evaluation of impact of nematode management tactics in growers' fields.
Field studies on mechanisms of nematode antagonism in rotation and cover crops revealed that lesion nematodes from strawberry roots were highest for Dwarf Essex canola and oats. However, the incorporation or removal of shoots had a significant effect on nematode numbers, indicating that plant breakdown products may be nematicidal. Results were shared with vegetable and fruit growers.
A bioassay technique, using freeze-dried plant material and sand, successfully detected potential nematicidal compounds, which may be useful in future control efforts.
Application of Telone II as a soil fumigant reduced all plant-parasitic nematode populations to nearly undetectable levels in orchard replant sites. Growers now have a new option for control.
The identification of root-knot resistant bell pepper cultivars provides growers with economically and environmentally compatible alternatives to the use of methyl bromide and other nematicides to control Meloidogyne incognita.
Enzyme-linked immunosorbent assays and DNA probes have been developed to detect endospores and vegetative stages of Pasteuria species, an important biocontrol agent of nematodes. These tests improve detection of Pasteuria in field plots and enable more accurate assessments of which soils are or may become suppressive for nematode infestations. Incorporation of these methods into IPM programs should help growers reduce amounts of pesticides used where Pasteuria is active.

Date of Annual Report: 11/25/2003

Report Information

Annual Meeting Dates: 10/01/2003 - 10/03/2003
Period the Report Covers: 10/01/2002 - 11/01/2003

Participants

Participants Attachment:

Attachment

Brief Summary of Minutes

Minutes Attachment:

Attachment

Accomplishments

Objective 1: Evaluate the effects of rotational crops, organic amendments and host crop genetics on nematode community structure. Milestone 2003:Integrate plant resistance with rotation crops to predict nematode population decline under integrated management systems. An interdisciplinary IPMSARE team completed a three-year MD grower-generated project entitled Development and Evaluation of Management Alternatives for Root Knot Nematodes and Volunteer Potatoes. This project evaluated on-farm and micro-plot experiments, sustainable management alternatives and their effect(s) on plant-parasitic nematodes. Of all seasonal samplings and Meloidogyne spp life-stages (gall indices and eggs/root), Fall soil J2 sampling was the most accurate indicator for population evaluation.(PA) Dagger nematodes (Xiphinema americanum) are the most important nematode problem in orchard sites in PA. Nearly 85% of samples contain dagger nematodes, which are usually below 25 per 100 cm3 soil but problematic as virus vectors. Preplant rotations with rapeseed reduce dagger nematodes, but but only after being turned in as a green manure. A bioassay technique using freeze-dried plant material was used to detect the presence of nematicidal compounds in a number of plants. (CT) In cooperation with PA, the influence of rotation and green manure crops on early dying of potato caused by Verticillium dahliae and P. penetrans was investigated. Pipersudangrass, Rudbeckia hirta, Saia oat, Dwarf Essex canola, Polynema marigold, Haychow sorgho-sudangrass, Pearl Millet 101, Trudan 8 sudangrass, Standard oat, and Kennebec potato were grown. All rotation crops but oat reduced lesion nematodes; Rudbeckia hirta, and Polynema marigold had few or undetectable levels of nematodes. (NY) Reaction of twelve commercial varieties of potatoes to M. hapla: Damage by the northern root-knot nematode to onion, carrots, lettuce and other susceptible crops is most severe when grown after potatoes, especially on organic soils. Potato is a host for M. hapla, but root-galling symptoms and yield losses have not been observed in New York. Thus, 12 varieties of potatoes were rated for root-galling severity on a scale of 1 (no galling) to 9 (>80% roots with galls). Eggs were extracted. All the potatoes exhibited extensive root-galling on the fine fibrous roots, but not coarse roots. (NY) Evaluation of twenty-one commercial carrot varieties for resistance to M. hapla: The northern root-knot nematode is a major pathogen of carrots grown in New York. Twenty one carrot varieties were evaluated for M. hapla. Six weeks after inoculation, carrots were rated for root-galling severity, and eggs were extracted from roots. All varieties were susceptible hosts to M. hapla. (RI) The effect of nematicidal applications of the organic butyric and propionic acids against Meloidogyne hapla and Pratylenchus penetrans was investigated. Concentrations of butyric acid and propionic acid were employed as a preplant nematicidal treatment, in conjunction with an untreated and a Vydate control. Butyric acid acted as a biocide, and nematode control was equivalent to Vydate. (USDA-ARS SC) Root-knot nematodes (Meloidogyne arenaria, M. incognita, M. javanica and M. hapla) reduce yields of vegetables worldwide. Resistant cultivars of many crops to different root-knot species are not available. There was significant genetic variability within the U.S. PI Citrullus germplasm collection for resistance to M. arenaria race 1. The C. lanatus var. citroides accessions are potential sources for resistance to M. arenaria race 1 in improved watermelon cultivars. Peppers with the N gene for resistance to southern root-knot nematode have been effective in nematode management. A pepper PI with moderate resistance to the northern root-knot nematode M. hapla has been identified and may be a useful management alternative in the Northeast. Objective 2: Characterization of biological control agents for suppression of plant-parasitic nematodes. (CT) A possible northern root-knot nematode-suppressive soil was identified. Soils with M. hapla present at high densities for over a decade exhibited a population decrease after being planted to hosts. M. hapla juveniles were exposed to soil for 24 hours and examined microscopically. Pasteuria endospores were present on the cuticles of 60% of the J2, perhaps explaining the reduction in nematodes. (USDA-ARS) Strains of rhizosphere-inhabiting bacteria, known to act against certain soilborne plant-pathogenic fungi, were tested for ability to suppress root-knot nematodes on cucumber roots. A greenhouse assay system identified a potentially useful bacterium, which was tested in the lab to determine whether natural products in culture filtrates suppressed egg hatch and juvenile mobility. A strain of Trichoderma virens was also tested for activity against Meloidogyne incognita on cucumber. Collaborators: Daniel Roberts (USDA ARS Sustainable Agriculture Systems Laboratory, Beltsville). (USDA-ARS, FL) Field trials were conducted in Florida to evaluate the effects of tomato and pepper transplants treated with biological control agents and planted into solarized soil on root-knot nematodes, plant growth and yield. Formulations of plant growth-promoting rhizobacteria (PGPR) increased tomato and pepper transplant growth. Transplant vigor and survival in the field were improved by PGPR in both tomato and pepper and on pepper, one PGPR formulation reduced root-knot galling. Yield of pepper was increased with two PGPR. Soil solarization did not control the root-knot nematode on tomato. However, solarization combined with PGPR on pepper produced yields comparable to untreated plants grown in methyl bromide fumigated soil. (FL) Long-term effects of peanut, Bahiagrass, and weed fallow on the levels of Meloidogyne arenaria race 1 and Pasteuria penetrans were investigated to determine the persistence of P. penetrans after 9 years. A soil test was developed to determine the suppressiveness of P. penetrans to M. arenaria. Over 4 years, the mean number of endospores per J2 and percentage of J2 with endospores increased greatly. Weed fallow plots had the highest percentage of J2 with endospores attached (75%). Bahiagrass and rhizomal peanut had lower percentages (7 and 2%, respectively). When water percolated through soils, most endospores stayed on the top layer of soil. The incidence of the bacterium is related to the density of the peanut root knot nematode. (WV) Perennial ryegrass lines were inoculated with endophytic Neotyphodium lolii isolates genetically modified to suppress production of specific toxins for nematode suppression. P. scribneri increased on endophyte-free plants, but declined on all endophyte infected plants. Populations on plants where the endophyte was modified by a Lysergyl Peptide Synthetase gene knockout (LPS) were as low as on wild-type endophyte-infected plants. Plants where the endophyte was modified with either a failed LPS gene insert, or with a Di-Methyl Allyll-tryptophan synthetase gene knockout had intermediate populations, suggesting that nematode suppression is not related specifically to ergovaline production. Obj. 2.1) Survey plant-pathogenic nematodes for occurrence of Pasteuria spp. (FL) Development of DNA probes for the detection of vegetative stages of Pasteuria spp.is ongoing. Based upon 80% of sequence determined for the sigE gene in P. penetrans, primers were designed for the PCR amplification of this gene in M. arenaria derived from tomato plants. Using Real-Time PCR, the level of infection of M. arenaria by vegetative cells of P. penetrans was determined. An assay is available to complement the immunoassay that detects mature endospores through the recognition of an epitope present only on the mature spores. Obj. 2.2) Determine the survivability and host preference of isolates of Pasteuria spp. from different geographic regions. Milestone 2003: Evaluate the winter survival of Pasteuria penetrans in northern states. (MA) Pasteuria penetrans parasitic on Meloidogyne graminis and Tylenchorhynchus has been found overwintering on turf in golf greens in MA. A correlation between the percent of Pasteuria-encumbered J2 and the density of Meloidogyne graminis J2 was present at high but not low densities. A correlation between Pasteuria- encumbered Tylenchorhynchus spp. and density was evident. In addition, Pasteuria spores appeared to be present on the cuticles of the M. hapla J2 in field microplots that had been exposed to winter conditions in CT. Pasteuria endospores appear to readily survive the winter in CT and MA. Objective 3: Comparison and evaluation of IPM system management of plant-parasitic nematodes based on crop rotation, organic amendments, host crop resistance and biological control agents. Efficacy of control products against M. hapla or Pratylenchus penetrans in field microplots. Researchers in MI, NY and RI, funded by the methyl bromide alternative program, was initiated in 2003. In spring 2003, plots were Methyl bromide-fumigated (350 lbs/A, under plastic), treated with Basamid (granular, 350 lbs./A), Vydate L (2,25 gal./A), Fosthiazate (11.5 pts./A), Agri-Mek (16 ozs/A, 2 apps) or an Untreated check. Tomatoes grown in the methyl bromide-treated plots did not exhibit root-galling symptoms and had the lowest number of M. hapla eggs. Root-galling ratings and egg recovery were reduced by Basamid, Fosthiazate, and Vydate. Only methyl bromide increased number and weight of fruit. (MI) The persistence of Heterodera glycines under various crop rotation and H. glycines sources of resistance were studied at the Greg Mahoney Farm in St. Charles, Michigan in 1995. Soybean yields were greater after three years of corn, three years of clover, or one year of corn followed by two years of a resistant soybean cultivar, compared to yields after five years of continuous susceptible soybeans. There were no differences among the at-harvest nematode densities among any of the systems following a single year of a resistant cultivar. The at-harvest density of H. glycines associated with the mixture of PI 88788, Peking and Kenwood 94 was not different from any of the other systems in 2001. Assessing the occurrence, damage and need for controlling M. hapla on carrots: The northern root-knot nematode has a wide distribution and causes yield losses to carrots in New York. A collaborative project was started to train carrot and onion growers to index their fields for root-knot. The latter involve collecting appropriate soil samples from fields and conducting a soil bioassay with lettuce for 28-42 days. The need to treat is based on the root-galling severity observed on lettuce roots {for carrots is only an average of >1 gall/root system; a rating of >2 (1-3% of roots galled)} on the 1 to 9 scale. The latter scheme is being compared to the extraction of J2 of M. hapla from soil sample to assess the need to treat. (WV) The economic returns of low versus high input farming systems for transition to organic practices were comapared for a Market Garden system. Yields of various crops in organic farming systems were generally greater with the high input (with compost) system than in low input (cover crop only) farming systems. Yields were not correlated with any nematode pest, but were related to soil factors such as organic matter content and pH. Although the with compost treatment was highly significant in predicting yields, soil organic matter was not increased, nor was pest density or biocontrol agent activity affected. Leaf petiole nitrate in potato was lower in plots with compost, indicating that this nutrient does not explain crop yields.

Publications

Brito, J. A, J. F. Preston, D. W.Dickson, R. M. Giblin-Davis, D. S. Williams, H. C. Aldrich, and J. D. Rice. 2003. Temporal Production and Immunolocalization of an Epitope During Pasteuria penetrans sporogenesis. Journal of Nematology 35(3):in press Edson, C., S. S. Swinton, J. Nugent, G. W. Bird, A. Coombs and D. Epstein. 2003. Cherry orchard floor management: Opportunities to Improve Profit and Stewardship. Michigan State University Extension, Bulletin E?2890. 6 pp. Giblin-Davis, R.M., D.S. Williams, S. Bekal, D.W. Dickson, J.A. Brito, J.O. Becker, and J.F. Preston. 2003. Candidatus Pasteuria usgae, sp.nov., an obligate endoparasite of the phytopathogenic nematode Belanolaimus longicaudatus. Int. J. Syst. Evol. Microbiol. 53:197-200. Kokalis-Burelle, N. and D. W. Dickson. 2003. Effects of soil fumigants and plant growth-promoting rhizobacteria on disease and yield of tomato. Proceedings of the Annual International Research Conference on Methyl Bromide Alternatives and Emissions Reductions: In Press. Kokalis-Burelle. 2003. Effects of transplant type and soil fumigant on growth and yield of strawberry in Florida. Plant and Soil: In Press. Kokalis-Burelle, N., C. S. Vavrina, M. S. Reddy, and J. W. Kloepper. 2003. Amendment of muskmelon and watermelon transplant media with plant growth-promoting rhizobacteria: Effects on disease and nematode resistance. HortTechnology 13:476-482. Kokalis-Burelle, N., N. Martinez-Ochoa, R. Rodrmguez-Kabana, and J. W. Kloepper. 2002. Development of multi-component transplant mixes for suppression of Meloidogyne incognita on tomato (Lycopersicon esculentum). Journal of Nematology 34: 362-369. Kokalis-Burelle, N., C. S. Vavrina, D. S. Kenney, E. N. Rosskopf, and R. A. Shelby. 2002. Field Evaluation of Plant Growth-Promoting Rhizobacteria Amended Transplant Mixes and Soil Solarization for Tomato and Pepper Production in Florida. Plant and Soil 238: 257-266. Kokalis-Burelle, N., W. F. Mahaffee, R. Rodrmguez-Kabana, J. W. Kloepper, and K. L. Bowen. 2002. Effects of Switchgrass (Panicum virgatum) Rotations with peanut (Arachis hypogaea L.) on Nematode populations and soil microflora. J. Nematology 34 (2): 98-105. Kloepper, J. W., M. S. Reddy, D. S. Kenney, N. Kokalis-Burelle, N. Martinez-Ochoa, and C. S. Vavrina. 2002. Theory and application for rhizobacteria in transplant production and yield enhancement. 26th International Horticulture Congress, International Soc. of Hort. Soc., Aug. 13-19, Toronto, Canada, Acta Horticulturae. Landis, J. N., J. E. Sanchez, G. W. Bird, C. E. Edson, I, Issacs, R. H. Lehnert, A. M. C. Schilder and S. M. Swinton (eds). 2002. Fruit Crop Ecology and Management. MI State University Ext. Bull. E?2759. E. Lansing, MI. 101 pp. LaMondia, J. A. 2003. Interaction of Pratylenchus penetrans and Rhizoctonia fragariae in strawberry black root rot. Journal of Nematology 35:17-22. LaMondia, J. A. 2002. Seasonal populations of lesion and root-knot nematodes in strawberry roots. Journal of Nematology 34(4):409-413. LaMondia, J. A., and R. S. Cowles. 2002. Effect of entomopathogenic nematodes and Trichoderma harzianum on the strawberry black root rot pathogens Pratylenchus penetrans and Rhizoctonia fragariae. Journal of Nematology 34(4):351-357. LaMondia, J. A., W. H. Elmer, T. L. Mervosh, and R. S. Cowles. 2002. Integrated management of strawberry pests by intercropping. Crop Protection 21:837-846. Mitkowski, N.A. and K. Jordan. 2003. Organic acids as preplant nematicides for use against Meloidogyne hapla and Pratylenchus penetrans. Phytopathology 93:S62. Mitkowski, N. A. and N. Jackson. 2003. Subanguina radicicola, the root-gall nematode, infecting Poa annua in New Brunswick, Canada. Plant Disease 87 (10): 1263. Mitkowski, N.A. and G.S. Abawi. 2003. Genetic diversity of New York State Meloidogyne hapla populations determined by RAPD‘s and mitochondrial DNA. Journal of Nematode Morphology and Systematics 5(2):191-202. Mitkowski, N.A. and G.S. Abawi. 2003. Reproductive fitness on lettuce of populations of Meloidogyne hapla from New York State vegetable fields. Nematology 5(1):77-83. Mitkowski, N.A., H. Van der Beek and G.S. Abawi. 2002. Characterization of root-knot nematode populations associated with vegetables in New York State. Plant Disease 86 (8):840-847. Mitkowski, N.A. and G.S. Abawi. 2002. Monoxenic maintenance and reproduction of root-knot nematode (Meloidogyne hapla) on multiple-species in vitro root culture systems. Plant Cell Reports 21 (1):14-23. DOI 10.1007/s00299-002-0468-6. Mitkowski, N.A. 2001. Root-knot nematodes on turf in the Northeastern United States. Turfgrass Trends 10(12):1-4. Nelson, S. D., S. J. Locascio, L. H. Allen, Jr., D. W. Dickson, D. J. Mitchell. 2002. Soil flooding and fumigant alternatives to methyl bromide in tomato and eggplant production. HortScience 37:1057-1060. Padgham, J. L., G. S. Abawi, J. M. Duxbury, and M. A. Mazid. 2002. Impact of Meloidogyne graminicola on yield of lowland rice in Bangladesh. Suppl. Phytopathology 92:S62. Preston, J. F., D. W. Dickson, J. E. Maruniak, G. Nong, J. A. Brito, L. M. Schmidt, and R. M. Giblin-Davis. 2003. Pasteuria spp. : Systematics and phylogeny of these bacterial parasites of phytopathogenic nematodes. Journal of Nematology. 35: 198-207. Sanchez, J.E., C.E. Edson, G.W. Bird, M.E. Whalon, T.C. Willson, R.R. Harwood, K. Kizilkaya, J.E. Nugent, W. Klein, A. Middleton, T.L. Loudon, D.R. Mutch, and J. Scfimger. 2003. Orchard floor and nitrogen management influences soil and water quality and tart cherry yields. Journal American Society Horticultural Science. 128:277?284. Sanchez, J. E., T. C. Willson, R. J. Zoppolo, D. Stefanelli and G. W. Bird. 2002. The Soil. pp. 21?28. (in) Fruit Crop Ecology and Management, Landis, J. N., J. E. Sanchez, G. W. Bird, C. E. Edson, I, Issacs, R. H. Lehnert, A. M. C. Schilder and S. M. Swinton (eds). MI State Univ. Ext. Bull. E?2759. E. Lan., MI. 101 pp. Schmidt, L. M., J. F. Preston, D. W. Dickson, J. D. Rice, T. W. Hewlett. 2003. Environmental quantification of Pasteuria penetrans endospores using in situ antigen extraction and immunodetection with a monoclonal antibody. FEMS Microbiol. Ecol. 44:17-26. Ude, G.N., J.M. Costa, W.J. Kenworthy, S. Sardanelli, and P.B. Cregan. 2002. AFLP Markers Associated with a QTL for Resistance to Soybean Cyst Nematode (Heterodera glycines) Race3. Journal of Genetics and Breeding 56: 213-220. Vavrina, C.S., P.D. Roberts and N. Kokalis-Burelle. 2002. Use of Commercial Systemic Acquired Resistance (SAR) Inducers in the Stand Establishment of Tomato; Impact on Plant Growth, Disease and Nematode Suppression. 26th International Horticulture Congress, International Soc. of Hort. Soc., Aug. 13-19, Toronto, Canada, Acta Horticulturae. Widmer, T.L., N.A. Mitkowski and G.S. Abawi. 2002. Soil organic matter and management of plant-parasitic nematodes. Journal of Nematology 34(4):289-295. Abstracts : Abawi, G. S., and J. W. Ludwig. 2003. Research results on the management of nematodes on onions. Pp. 167-172, NYS Veg. Conf. Proc., Cornell Coop. Ext., Ithaca, NY. Abawi, G. S., and J. W. Ludwig. 2003. Host efficiency of selected soybeans to Meloidogyne hapla and Pratylenchus penetrans. J. of Nematology 35: 321 (Abstr.). Abawi, G. S., and J. W. Ludwig. 2003. Evaluation of onion germplasm for resistance to Meloidogyne hapla. J. of Nematology 35: 321 (Abstr.). Abawi, G. S., J. W. Ludwig, C. R. MacNeil, and J. J. Vanderheide. 2003. Damage and management of the northern root-knot nematode on onions in New York State. Pp. 251, Proc. Of the ICPP, Christchurch, NZ, Feb. 2-7, 2003. Halbrendt, J. M. and J. A. LaMondia 2002. Nematicidal activity of selected plant residues to Xiphinema americanum. Proceedings of the Fourth International Congress of Nematology; Nematology 4:285. Kenworthy, W. J. 2002. Cyst Nematode-Resistant Soybean Variety Test. Maryland CES Agronomy Mimeo No.43. 7pp. Kokalis-Burelle, N. and P. D. Adams. 2002. Effects of transplant type and soil treatment on strawberry rhizosphere fungal populations. Proceedings of the Annual International Research Conference on Methyl Bromide Alternatives and Emissions Reductions Pages 86.1-86.4. Kovaleva,E. s., E.P. Masler, S. Sardanelli, and D. J. Chitwood. 2003 Heat Shock Proteins 70 in Heterodera Glycines. Society of Nematologists 42nd Annual Meeting, Cornell University, Ithaca, NY. July, 2003. LaMondia, J. A. and J. M. Halbrendt. 2002. Rotation crops for control of nematodes pathogenic to tree and small fruits (A progress report). Proceedings of the 78th Cumberland-Shenandoah Fruit Workers Conference. 180-183. LaMondia, J. A. and J. M. Halbrendt 2003. Differential host status of rotation crops to dagger, lesion and root-knot nematodes. Journal of Nematology 35:349. LaMondia, J. A. and T. L. Mervosh. 2003. Strawberry black root rot - lack of association with terbacil application. Phytopathology 93 (6):S48. Mallozzi, T., Sardanelli, S., Kratochvil, R., and Everts, K. 2002. Evaluating Biocontrol Alternatives to Methyl Bromide for Management of Root-Knot Nematode. In press Maryland IPM 2002 Annual Report. p. 20. Mitkowski, N. A. and G. S. Abawi. 2003. Root-knot nematodes. The Plant Health Instructor. DOI:10.1094/PHI-I-2003-0917-01. Rungrassamee, W. and Wick, R. L. 2003. Relationship of the nematode hyperparasite Pasteuria spp. To Meloidogyne graminis and Tylenchorhynchus spp. in golf greens. Journal of Nematology 35:361. Sardanelli, S., Mallozzi, T., Everts, K. and Kratochvil, R. 2002. Sustainable Management Alternatives for Root Knot Nematodes In Maryland IPM 2002 Annual Report. p. 19. Troccoli, A., G. S. Abawi, J. W. Ludwig, and F. Lamberti. 2003. Morpho-anatomical notes on populations of Pratylenchus penetrans and P. crenatus from New York state (USA). J. of Nematology 35: 367-368 (Abstr.). LaMondia, J. A. 2003. Influence of rotation crops on lesion nematode infection of strawberry, 2002. Biological and Cultural Tests for Control of Plant Diseases Vol. 18:N002. LaMondia, J. A. 2003. The association of lesion nematodes with strawberry black root rot. Nourse Commercial Newsletter. Workshops: Mallozzi, T., and S. Sardanelli. 2003. The Moisture Replacement System- a versatile tool facilitating the evaluation of a potential biocontrol agent in the search for root-knot nematode management. Biocontrol Workshop, Society of Nematologists 42nd Annual Meeting, Cornell University, Ithaca, NY. July, 2003. J. M. Halbrendt and J. A. LaMondia presented a workshop entitled "A bioassay test for nematicidal activity in plant tissue" at the Biocontrol Workshop at the Society of Nematologists 42nd Annual Meeting, Cornell University, Ithaca, NY. July, 2003.

Impact Statements

Validation of alternative nematode management tactics and demonstration of their economic benefits provides growers with additional tools for reducing pesticide use, lowering control costs and crop losses, reducing human health risk, increasing food safety and providing a more sustainable strategy for managing nematodes in vegetable and field crop rotation.
Effective nonchemical controls will help to maintain production in the absence of methyl bromide, and enhance economic opportunity in rural communities. For example, the current application of Vydate (up to 4 gal/acre rate) on carrot for nematode control costs growers approximately $250.00 per acre per year. There is a great opportunity to develop a safer alternative to Vydate at a lower cost.
Training of onion, carrot, and other vegetable growers to conduct their own bioassays for root-knot nematodes will allow targeted nematode management only in fields with damaging nematodes, reducing human health risk, environmental exposure, pesticide residues in food, and reduced production costs. The use of Vydate (up to 4 gal/acre) on carrot costs growers about $250.00 per acre per year.
Cooperative research on PGPR formulations with Gustafson LLC has resulted in a new commercial product for use on transplanted crops, BioYieldTM, consisting of a formulation of plant growth and yield enhancing bacteria, reducing human health risk, environmental exposure, pesticide residues in food.
Root-knot nematodes (Meloidogyne spp.) are a major constraint to production of vegetables. Host resistance is the most economical, environmentally benign alternative to methyl bromide. Root-knot nematode resistance conferred by the N gene is effective in field?planted bell pepper and should provide economical and environmentally compatible alternatives to methyl bromide and other nematicides for managing root?knot nematodes.
The demonstration that rotation crops can control lesion nematodes and potato early dying disease may lead to effective non-chemical management, increasing yield and reducing pesticide use. The identification of strawberry cultivars resistant or tolerant to black root rot would greatly assist disease management.
Validation of alternative nematode management tactics and demonstration of their economic benefits will provide growers with additional tools for reducing pesticide use, lowering control costs and crop losses, and providing a more sustainable strategy for managing nematodes in vegetable and field crop rotation.
The immunoassay using a monoclonal antibody against an adhesin epitope on the surface of the endospores of P. penetrans provides a quantitative measure of the densities of endospores in soil and in infected nematodes. The assay should be useful to other scientists assessing the extent to which soils are or may become suppressive for nematode infestations and thereby limit the need for other control measures.
The development of oligonucleotides complementary to DNA sequences of the sporulation genes of P. penetrans has provided probes and primers for quantifying P. penetrans infections in planta. A rapid method using real-time PCR has been developed. This provided an approach that is complementary to the immunoassay, allowing detection and quantification of P. penetrans at all stages of development.

Date of Annual Report: 12/09/2004

Report Information

Annual Meeting Dates: 10/14/2004 - 10/15/2004
Period the Report Covers: 10/01/1999 - 09/01/2004

Participants

Magnarelli, Louis (Louis.Magnarelli@po.state.ct.us) - Connecticut Agric. Exp. Station;
Abawi, George (Gsa1@nysaes.cornell.edu) - Cornell University, Geneva, NY;
Bernard, Ernest - University of Tennessee;
Bird, George (birdg@msu.edu) - Michigan State University;
Desaeger, Johan - University of Georgia;
Dickson, Don (Dwd@ufl.edu) - University of Florida;
Green, James , CSREES Advisor - USDA-CSREES, Washington, DC;
Halbrendt, John (Jmh23@psu.edu) - Penn State University, Fruit Res & Ed Ctr;
Huettel, Robin - Auburn University;
Kotcon, Jim (Jkotcon@wvu.edu) - West Virginia University;
LaMondia, Jim, Chair (James.LaMondia@po.state.ct.us) - Connecticut Agric. Exp. Station;
Meyer, Susan (Smeyer@asrr.arsusda.gov) - USDA-ARS, Beltsville, MD;
Preston, James (Jpreston@ufl.edu) - University of Florida;
Thies, Judy (Jthies@saa.ars.usda.gov) - USDA-ARS, Charleston, SC;
Zasada, Inga , Secretary - USDA-ARS, Beltsville, MD;

Brief Summary of Minutes

Minutes Attachment:

Attachment

Accomplishments

2000: " Establish a low soil volume bioassay to evaluate nematode antagonistic crops. A low volume soil bioassay developed in PA was used to evaluate toxicity of green or freeze-dried plant extracts on plant parasitic nematodes in CT, MA, NY, and WV. " Conduct a workshop for all group members to present protocols for Pasteuria penetrans recognition, research and surveys. A workshop conducted in FL to taught members to recognize Pasteuria spp. on different species of nematodes with the goal that all members follow prescribed methods in working with Pasteuria. " Initiate collaborative rotation and cover crop system research in field plots and microplots in multiple states. Collaborative multi-state interdisciplinary competitive projects were awarded to NE171 scientists. Maryland participated in a grower-generated project on the use of cover crops, soil amendment, and tillage to manage root knot. A multidisciplinary project was funded to Michigan, Wisconsin, New Jersey and New York. Grower trials were conducted on rotation and cover crops, resistance, biological control, and natural products. The host status of nematode-suppressive rotation crops or green manures was investigated on nematodes in CT and PA. " Identify and evaluate nematode resistant or tolerant varieties for inclusion in an integrated management program. A number of crops were evaluated for resistance or tolerance to nematodes, including the USDA Capsicum germplasm collection for reaction to Meloidogyne (USDA-SC); strawberry cvs were evaluated against black root rot pathogens in CT; Onion germplasm (>50 accessions of Allium spp.) were evaluated against M. hapla (NY); carrot cvs were evaluated against M. hapla (NY); and selected soybean cvs were found to be hosts to P. penetrans. Bell pepper (Capsicum annuum) cvs near isogenic for the N gene for resistance to root knot were evaluated (USDA-ARS-SC); and C. colocynthis (21) and C. lanatus var. citroides (88) accessions, and C. lanatus var. lanatus (155) accessions from the U.S. Citrullus collection were evaluated against M. arenaria race I. Results demonstrated resistance, tolerance, or genetic variability within the cultivars or PI's tested. 2001: " Survey northern states for the presence of Pasteuria penetrans using protocols identified in 2000. Pasteuria penetrans or Pasteuria-like organisms were discovered attached to Heterodera iri from turf (CT). Pasteuria were detected in 77% of Long-Term Ecological Research plots in MI. Pasteuria spp. were associated with plant nematodes, fungivores and bacterivores. A New York survey detected Pasteuria from Tylenchorhynchus spp., Tylenchus spp., and Meloidogyne sp. Free-living Cephalobus sp. and Eucephalobus sp. were also found with endospores. Samples from putting greens in MA all had Pasteuria-infected Tylenchorhynchus. Other nematodes with Pasteuria included Helicotylenchus, Hoplolaimus and Meloidogyne. Conditions were defined for detection of Pasteuria spp endospores using a monoclonal antibody against an epitope of the parasporal fibers that cover mature endospores. An amplification ELISA using biotinylated secondary antibody quantified a minimum of 300 endospores per g soil, and may also quantify endospores in roots. FL developed DNA probes for detection of vegetative stages of Pasteuria using published genome sequences for Bacillus subtilis and B. halodurans, free-living endospore-forming bacteria, as a basis for designing primers for PCR amplification of orthologous genes from Pasteuria. Degenerate primers and anealing conditions were used to amplify PCR products using DNA templates obtained from P. penetrans P20 endospores. Using Real-Time PCR, M. arenaria infection level by vegetative cells of P. penetrans was determined. The assay selectivity was confirmed as primers did not detect DNA in uninfected M. arenaria. This work identified gene sequences unique to P. penetrans. Using these primers, the level of infection by P. penetrans may be determined before sporulation. This will be complementary to the immunoassay recognizing an epitope present on mature spores. " Evaluate crops and greenhouse or growth chamber systems for the production of large numbers of Pasteuria penetrans endospores. Rutgers tomato and PA136 Bell pepper were selected as hosts for culture of Meloidogyne incognita based on root vigor and cultural manageability, and grown in a greenhouse soil moisture control system (Sardanelli and Kenworthy, 1997). Root-knot density and Nemacur treatment were evaluated in MA for impact on the incidence of infection by Pasteuria. There was an inverse relationship between nematode numbers and Pasteuria. Nematicide did not affect the incidence of Pasteuria infection. 2002: " Initiate research to determine the mechanism of nematode antagonism in rotation and cover crops. A bioassay developed in PA using freeze-dried plant material was used to detect the presence of potential nematicidal compounds. Xiphinema americanum were exposed to freeze-dried leaf and root tissue. Nematodes were not killed in sterile sand or water, and mortality increased with amounts of tissue. The LC50 ranged from 0.6 mg (Nasturtium) to 13.2 mg (Sunn hemp) per cc of sand. Nasturtium leaves showed potent nematicidal activity with an LC50 of 0.6 mg. (CT) In cooperation with PA, the host status of several rotation crops was evaluated against Pratylenchus penetrans and Meloidogyne hapla. Shoot incorporation indicated that plant breakdown products may be nematicidal. " Compare isolates of Pasteuria penetrans for host preference and biocontrol potential. The long-term persistence and suppression of Pasteuria penetrans against Meloidogyne arenaria were investigated over 9 years of crop cultivation and weed fallow in FL. FL developed a monoclonal to quantify Pasteuria endospores in soil. We also developed assays for DNA sequences unique to different species of Pasteuria to quantify host specificities. The incidence of the bacterium was related to the density of the peanut root knot nematode, but not M. javanica. (MA) The relationship of Pasteuria penetrans to Meloidogyne graminis and Tylenchorhynchus spp was investigated on golf greens. A correlation between percent Pasteuria- encumbered juveniles and the density of M. graminis and Tylenchorhynchus was found. " Evaluate the impact of rotation and cover crops and soil amendments on nematodes. Experiments were conducted combining an alkaline-stabilized biosolid (ASB) amendment with Streptomyces sp. for M. incognita control. Only ASB at the highest rate (3% wt/g soil) reduced M. incognita. No suppression was achieved by applying Streptomyces sp. to soil (USDA ARS Beltsville). (PA) Compost was evaluated for ability to suppress dagger nematodes in an apple orchard. Plots treated with five inches of mushroom compost consistently had the lowest nematodes, suggesting that mushroom compost is suppressive to dagger nematodes. (CT) Rotation and green manure crops resulted in lower lesion nematode densities. Incorporation of crop shoots further reduced nematodes. Rudbeckia and marigold controlled nematodes without incorporation. A single season of marigold controlled P. penetrans and increased tuber yields in potato plots. The effects of the organic butyric and propionic acids against Meloidogyne hapla on tomatoes and Pratylenchus penetrans on strawberries in RI indicated that Vydate and butyric acid treatments were similar. 2003: " Evaluate the winter survival of Pasteuria penetrans in northern states. (MA) Pasteuria penetrans on Meloidogyne graminis and Tylenchorhynchus has overwintered on turf in golf greens in Massachusetts. Pasteuria spores were present on the cuticles of M. hapla juveniles after exposure to winter conditions in CT. Pasteuria endospores appear to readily survive the winter in CT, MA, NY and MI. " Determine the sequence of events required for formation of endospore-associated proteins and adhesion to nematodes. (FL) Pasteuria penetrans cannot be grown in vitro or enumerated by bacteriological methods; therefore, an immunoassay was developed to quantify spores. A monoclonal antibody (MAb) produced against whole spores was shown to bind a putative glycan epitope present on the spore envelope. Following SDS-PAGE of cuticle extracts and immunoblotting with spore extracts and the MAb as probes, the glycopeptides were shown to bind to discrete polypeptides derived from the nematode cuticle. The MAb detects an epitope of adhesins that are involved in the attachment of Pasteuria endospores to the nematode cuticle. An indirect amplification ELISA was developed to quantify spore adhesins. Regression of spore concentration versus signal produced line fits of 0.98 and indicated a detection limit of 3000 spores/g soil. Western blot profiles of soil-extracted antigen were the same as those provided by extracts of purified endospores, confirming that the ELISA detects Pasteuria adhesins in soil. The MAb did not detect antigens on cells or endospores derived from other endospore-forming bacteria. This establishes the MAb as a definitive probe to detect adhesins as virulence determinants on Pasteuria endospores in soil. " Integrate plant resistance with rotation crops to predict nematode population decline under integrated management systems. Potato cvs were rated for root-galling severity in NY. All potatoes tested had galls on fine fibrous, but not coarse roots. Large numbers of eggs were recovered from all cvs, which were hosts of M. hapla. Carrot cvs were evaluated against M. hapla. All cultivars tested were hosts of M. hapla. Black root rot is a strawberry disease caused by Rhizoctonia fragariae and Pratylenchus penetrans. Cultivars that performed well in yield and vigor after 3 years in infested soils in CT were Latestar, Primetime, Annapolis, Lester, Idea, Winona and Earliglow. Results from USDA-ARS SC demonstrated genetic variability within the U.S. PI Citrullus germplasm collection for resistance to M. arenaria race 1. The C. lanatus var. citroides accessions are sources of resistance to M. arenaria race 1 in watermelon. Peppers with the N gene for resistance to southern root-knot nematode have been useful in management. A pepper PI with moderate resistance to M. hapla has been identified for use in the Northeast. 2004: " Evaluate rotation and cover crop effects on nematode community structure in soil. A workshop on identification of free-living nematodes at the 2002 Technical Committee Meeting was held to assist researchers evaluating the impact of nematode management on nematode community structure. Dr. Bird has prepared chapters on 'nematode biology and management' & 'nematodes and soil quality' in two interdisciplinary MSUE Bulletins. The effects of seasonal changes in nematode community structure were determined in long-term ecological plots in MI. " Determine the morphological and phylogenetic relations among Pasteuria penetrans isolates. Phylogenetic analysis of the sigE gene from gram-positive endospore-forming bacteria finds Pasteuria penetrans most closely related to Paenbacillus polymyxa. SigE is a sporulation gene with sequence diversity for species and biotype differentiation of Pasteuria. The development of a DNA library from Pasteuria represents a special case. The GenomiPhi process was evaluated from Bacillus subtilus cells and indicates that a complete Pasteuria library may be prepared from as little as 1000 bacterial genomes from single nematodes. " Develop an economic analysis of rotation and soil amendment tactics for nematode control. The analysis of nematode community structure in long-term ecological sites in MI has led to 5 proposed categories of soil health and quality based on disturbance, soil nutrient/amendment inputs, and system degradation. Within a category, rotation with nematode-resistant soybeans has shown an economic advantage of 9 to 26%.

Publications

Bird, G. W. 2003. Role of integrated pest management (in) Integrated pest management in the global arena, pp. 73-85 (in) Maredia, K. S., D. Dakouo and D. Mota-Sanchez (eds). CABI Publishing, Cambridge. 512.pp. Brito, J. A., J. F. Preston, D. W. Dickson, R. M. Giblin-Davis, D. S. Williams, H. C. Aldrich, and J. D. Rice. 2003. Temporal production and immunolocalization of an epitope during Pasteuria penetrans sporogenesis. J. Nematol. 35:278-288. Fery, R. L., J. A. Thies and A. G. Gillaspie. 2004. 'KnuckleHull-VNR', a crowder-type southernpea resistant to blackeye cowpea mosaic virus and root-knot nematode. HortScience 39:183-184. Halbrendt, J. M., and J. A. LaMondia. 2004. Crop rotation and other cultural practices. Pp.273-294 in, Nematology, Advances and Perspectives. Z. X. Chen, S. Y. Chen, and D. W. Dickson, eds. Kokalis-Burelle, N., and D. W. Dickson. 2004. Evaluation of Plantpro 45 and Plantpro 20EC as alternatives to methyl bromide soil fumigation for tomato production in Florida. Nematropica 33:171-178. Kratochvil, R. J., S. Sardanelli, K. E. Everts, and L. E. Gallagher. 2004. Evaluation of Crop Rotation and other Control Practices for Management of Root-knot and Lesion Nematodes. Agron. J. 2004; 96:1419-1428. LaMondia, J. A. 2004. Field performance of twenty-one strawberry cultivars in a black root rot-infested site. Journal of the American Pomological Society 58(4):226-232. Levi, A., C. E. Thomas, J. A. Thies, A. M. Simmons, Y. Xu, X. Zhang, O. U. K. Reddy, A. R. Davis, and T. Wehner. 2004. Developing a genetic linkage map for watermelon: polymorphism, segregation, and distribution of markers. Progress in Cucurbit Genetics and Breeding Research. A. Lebeda and H. S. Paris (Eds.) p. 515-523. Mervosh, T. L., and J. A. LaMondia. 2004. Strawberry black root rot and berry yield are not affected by use of terbacil herbicide. HortScience 39(6):1339-1342. Meyer, S.L.F., R.N. Huettel, X.-Z. Liu, R.A. Humber, J. Juba and J. Nitao. 2004. Activity of fungal culture filtrates against soybean cyst nematode and root-knot nematode egg hatch and juvenile motility. Nematology 6: 23-32. Padgham, J. L., J. M. Duxbury, A. M. Mazid, G. S. Abawi, and M. Hossain. 2004. Yield loss caused by Meloidogyne graminicola on lowland rainfed rice in Bangladesh. J. of Nematology 36: 42-48. Padgham, J. L., G. S. Abawi, and J. M. Duxbury. 2003. Survival and infectivity of Meloidogyne graminicola in flooded and non-flooded soils. Nematol. Medit. 31:225-230. Schmidt, L. M., J. F. Preston, G. Nong, D. W. Dickson, and H. C. Aldrich. 2004. Detection of Pasteuria penetrans infection in Meloidogyne aerenaria race 1 in planta by polymerase chain reaction. FEMS Microbiol. Ecol. 48:457-464. Smith. J. and G. W. Bird. 2003. Impact of physical and chemical and biological disturbances on nematode community structure. Journal of Nematology 35:364-365. Thies, J. A., R. L. Fery, J. D. Mueller, G. Miller, and J. Varne. 2003. Response of bell pepper cultivars near-isogenic for the N gene to Meloidogyne incognita in field trials. HortScience 38:1394-1396. Thies, J. A., R. F. Davis, J. D. Mueller, R. L. Fery, D. B. Langston, and G. Miller. 2004. Double cropping cucumbers and squash after resistant bell pepper for root-knot nematode management. Plant Disease 88:589-593. Thies, J. A. and A. Levi. 2003. Resistance of watermelon germplasm to the peanut root-knot nematode (Meloidogyne arenaria race 1). HortScience 38:1417-1421. Walia, R. K., T. E. Hewlett, and D. W. Dickson. 2004. Microwave treatment of Pasteuria penetrans parasite preparation for selective elimination of undesired microorganism. Nematologia Mediterranea 32:15-17.

Impact Statements

(CT) The effect of initial Pratylenchus penetrans densities in soils infested with a history of black root rot were incorporated into a strawberry economics model developed by Alison DeMarree & Regina Rieckenberg at Cornell University. Initial lesion nematode densities as low as 12 per g root resulted in an 11% loss over 4 years and densities as high as 125 per g root resulted in 100% loss in profit over the same time.
(CT) Studies on the interaction of the fungal and nematode pathogens in strawberry black root rot will help to maximize sampling to determine if nematodes are involved in black root rot, the contribution of the nematode to disease, and may help to manage the disease complex.
(CT) The identification and use of rotation crops that reduce both strawberry black root rot pathogen populations in soil will reduce the use of soil fumigants. The identification of strawberry cultivars resistant or tolerant to black root rot would further reduce pesticide use and economic losses to disease.
(MI) The technology of precision agriculture appears to have excellent potential for use in management of plant-parasitic nematodes and the potato early-die disease complex. Precision agriculture techniques had the potential to increase net profit at the study site by $304 per acre.
(MA) The demonstration that natural enemies for biological control, such as Pasteuria, may be destroyed by fumigation may encourage growers to use fewer broad spectrum pesticides and adopt biological or cultural controls.
(NY, Geneva) Assessing the genetic and pathological characteristics of nematode populations, such as Meloidogyne hapla, is critical for identifying resistant crop germplasm, designing effective crop rotations, and developing diagnostic tools.
(NY, Geneva) The development of a simple bioassay to predict damaging nematode population levels by growers has reduced crop losses, environmental exposure to pesticides and human health risks.
(PA) Natural products, green manures and rotation crops can be useful tools for nematode management in fruit crops. These practices can be used to develop IPM and organic production programs with reduced use of synthetic pesticides.
(USDA, SC) The northern root-knot nematode causes serious damage to pepper in the U.S. and worldwide. The development of cultivars with genetic resistance to root knot is the most efficient and environmentally benign way of controlling this problem without to the use of soil fumigation. As there are no pepper cultivars with resistance to M. hapla, the identification of a PI with moderate resistance to M. hapla is a significant and important finding.
(FL) The monoclonal antibody has been established as a definitive probe to detect adhesins as virulence determinants on Pasteuria endospores in soil. This sensitive quantitative immunoassay can be applied as a tool by researchers for evaluating chemical and biological measures required for control of root-knot nematodes.
(FL) The immunoassay using a monoclonal antibody against an adhesin epitope on the surface of the endospores of P. penetrans provides a quantitative measure of the densities of endospores in soil and in infected nematodes. The assay should be useful in assessing the extent to which soils are or may become suppressive for nematode infestations and thereby limit the need for other control measures.
(FL) The development of oligonucleotides complementary to DNA sequences of the sporulation genes of P. penetrans has provided probes and primers for quantifying P. penetrans infections in planta. A rapid method using real-time PCR has been developed. This provided an approach that is complementary to the immunoassay, allowing detection and quantification of P. penetrans at all stages of development.
(FL) The determination of the sequences of genes from other species/strains of Pasteuria should allow the development of probes with which to distinguish and identify host specific strains in different ecological settings.
(MD) The refinement of nematode sampling recommendations, validation of alternative nematode management tactics and demonstration of their economic benefits provide growers with additional tools for reducing pesticide use, lowering control costs and crop losses, and providing a more sustainable strategy for managing nematodes in vegetable and field crops.
(MI) Data on the influence of management systems on nematode community structure confirm that impacts on soil biology can be detected and predicted through the use of nematode community structure analysis.
(USDA-ARS, FLA) Cooperative research on PGPR formulations with Gustafson LLC has resulted in a new commercial product for use on transplanted crops, BioYieldTM, consisting of a formulation of plant growth and yield enhancing bacteria. This product will be a low toxicity alternative to pesticides, reducing environmental and human exposure.
(USDA-ARS, SC) None of the bell pepper cultivars currently available have sufficient resistance to Meloidogyne incognita. Results of controlled environment studies indicate that root-knot resistant cultivars such as Charleston Belle that are homozygous (NN) for the N gene value as parental lines for developing resistant F1 hybrid (Nn) bell pepper cultivars. These hybrids should exhibit the same level of resistance as the Charleston Belle parent.