Brief Summary of Minutes of Annual Meeting

Accomplishments: Outcomes, Outputs, Activities Objective 1: Develop effective and economically-viable cultural management tactics for plant-parasitic nematodes based on host resistance, nematode antagonistic rotation or cover crops, soil amendments and biological agents. Biological Control Samples from potentially Meloidogyne hapla suppressive soils in CT associated with Pasteuria were provided to Dr. Don Dickson in Florida for experimental purposes. We had previously observed a decline of populations of the northern root-knot nematode Meloidogyne hapla over time in CT field plots infested with the nematode since 1995 and repeatedly inoculated with the pathogen. Pasteuria endospores were observed on the cuticles of M. hapla juveniles exposed to soil and endospore-filled females were observed. A culture of this CT Pasteuria penetrans specific to Meloidogyne hapla is being developed by Dr. Don Dickson. Two isolates of M. hapla were collected from strawberry plantings in Florida. The Connecticut isolate of Pp attaches to both M. hapla isolates, but it has not yet been determined whether the Pp will infect and develop inside the females of M. hapla. A new species of Pasteuria, Candidatus Pasteuria aldrichii, isolated from and propagated in the bacterivore Bursilla RGD244, was characterized by gene sequencing (FL). Based upon sequences of genes encoding 16S rRNA, this Pasteuria sp. is closely related to Pasteuria penetrans and other species associated with plant parasitic nematodes. The cultivation of P. aldrichii in Bursilla RGD244 on a single bacterial species with a sequenced genome should allow the application of established high-throughput sequencing systems to provide the first genome sequence to be obtained for a species of Pasteuria. This sequence may in turn provide a template with which to decipher sequences from Pasteuria species with biocontrol potentials that are often contaminated with genes from other soil-borne bacteria. The genome of Paenibacillus sp. JDR-2, a xylanolytic spore-forming soil-borne bacterium, has been fully sequenced and annotated. Based upon 16S rRNA sequence, the isolate is most closely related with the entomopathogenic species P. popilliae and P. lentimorbus, and Paenibacillus sp. SdTc and Paenibacillus nematophilus, respectively ectoparasites of the entomoparasitic nematodes, Steinermema diaprepesi and Heterorhabditis spp. Based upon 16S rRNA sequence, these Paenibacillus species are in clades adjacent to Pasteuria species, suggesting derivation of both clades from a common progenitor during the evolution of spore-forming bacteria. The role of these bacteria in soil health and agricultural productivity deserves further study. The transcriptome of sp. JDR-2 has been determined following growth on native substrates, including methylglucuronoxylans from dicots (sweetgum wood), monocots (sorghum and sugarcane bagasse), barley glucan as well as monosaccharides. The different substrates show selective up-regulation of enzymes involved in the processing of the different plant polysaccharides coordinate with ABC transporters and transcriptional regulators required for the expression of enzymes and transporter proteins. This approach may be used to define the expression of genes in relation to nematode associations to identify environmental relationships influencing soil health. It will be used to define mechanisms of host-specific virulence in Pasteuria spp. associated with propagation and spore maturation in nematode hosts. VT - Identified plant-parasitic nematodes for a project led by Terrance Bradshaw and funded by Northeast SARE ONE14-199. The aim was to evaluate the effectiveness of two biopesticides on populations of plant-parasitic nematodes associated with apple replant disease. MeloCon and Actinovate reduced relative abundance of Meloidogyne but not Pratylenchus, compared to non-treated controls, at two sites in Vermont. Host-Plant Resistance The use of resistant varieties for nematode control related to small acreage specialty crops is underutilized, making the NE-1040 breeding work exceptionally important. Commercial cucurbit rootstocks, cucurbit Plant Introductions, and African horned cucumber (Cucumis metulifer) germplasm lines were evaluated as rootstocks for grafted cantaloupe in a root-knot nematode infested field. Melon and squash hybrid rootstocks were highly susceptible to root-knot. The C. metulifer lines were resistant and were highly compatible rootstocks for grafted cantaloupe. Advanced sweetpotato breeding lines and seedlings from the USDA program were screened for resistance to root-knot nematode. Advanced pepper germplasm lines were screened for resistance to Meloidogyne incognita, and resistant plants were selected for use in a backcross breeding program to develop resistant lines of sweet banana, sweet cherry, and Cubanelle peppers. Pepper breeding lines under development by the USDA Vegetable Lab in Charleston, SC were provided by Dr. Judy Thies for evaluation against Meloidogyne hapla in Connecticut. Two S4 lines had been screened against M. hapla in SC. We evaluated 20 replicate pots of each of these lines and a susceptible control pepper variety ‘California Wonder’ in the greenhouse in pots and in the field. All transplants were inoculated with 6,000 eggs and juveniles of Meloidogyne hapla. USDA 1348 had significantly fewer galls than USDA 1390 and California Wonder pepper, which were not different from each other. Evaluation of egg production is underway. A tomato cultivar with the Mi-1 gene for root-knot nematode (RKN) resistance and a RKN susceptible tomato cultivar were evaluated in the field in FL with and without soil fumigation. Although less than 10% of the resistant tomatoes were galled compared with nearly 100% galling on the susceptible tomatoes, neither yielded well compared with those grown in fumigated soil. Although soil temperatures were often recorded above those shown in laboratory studies to break the resistance there was no indication this occurred in these field trials. In laboratory studies fewer second-stage juveniles enter resistant cultivar roots than susceptible cultivar roots. The reason for the MI gene tomato yielding less without soil fumigation is under investigation. Through surveys we are identifying species of root-knot nematodes on root-knot nematode resistant peach rootstock. Meloidogyne arenaria and M. javanica are being found much more frequently than M. floridensis, the peach root-knot nematode. Soft-tissue X-ray imaging of the entire root volume of a plant can be captured in a single high-resolution image, allowing sophisticated comparisons of nematode numbers or egg masses with root volume and length. Experiments conducted with reniform nematode (Rotylenchulus reniformis) and resistant or susceptible cotton selections showed that nematode egg masses characteristically appeared on X-rays as hemispheres with a dark basal ring next to the root and a lighter dome, distinct from artifacts such as soil particles and root stubs. Each cotton line had a unique distribution of roots and a unique distribution of nematode feeding sites. The use of X-ray technology and digital image analysis can provide a much more accurate approach to nematode relationships with plant host roots. Host Plant Resistance and Seed Treatments In three 2013 MI field studies, soybean cyst nematode (SCN) resistant cultivars derived from a PI 548402 source of resistance yields significantly higher than susceptible cultivars. At these locations, final population densities of SCN associated with resistant varieties derived from a PI 88788 source of resistance were not significantly different from the susceptible cultivars. Seeds treated with N’Hibit (plant health regulator) or VOTiVO (Bacillus firmus) resulted in yielded greater than those associated with the non-treated seed. The seed treatments did not, however, result in lower final SCN densities compared to the non-treated controls. Eighteen sugar beet cultivars were approved by Michigan Sugar Inc. for planting in MI in 2014. Four of these are resistant to the beet cyst nematode (BCN), including the three most economically viable cultivars. In an on-farm research trial, beet yields and sugar content associated with the resistant cultivar were significantly greater than those associated with the susceptible cultivar. During the past five years, Golden Nematode (GN) resistant potato varieties have been evaluated in the Central Asia IPM CRSP funded by USAID. Dr. Murat Amitov and Dr. Nurali Saidov served as program coordinators for the Kyrgyzstan and Tajikistan projects, respectively. Potato cyst nematodes (Globodera sp?) are key pests in both countries. A number of GN resistant varieties have been developed by the Michigan State University potato breeding program and confirmed through PCR analysis. It is not possible to evaluate them under field conditions in the U.S. They have, however been evaluated in potato production systems in Central Asia. In one 2012 potato cyst nematode (PCN) site in the Kemin Region of Kyrgyzstan, tuber yields of cultivars Boulder and Missaukee were significantly greater than those associated with Picasso, an important local, but PCN susceptible cultivar. While the results in the 2012 trial with Boulder and Missaukee in Tajikistan were even better, the cultivars Dakota Diamond and Kalkaska, both, considered as PCN susceptible were the top two cultivars when grown under a PCN infestation. In the Kemin Region 2012 trial, no new PCN cysts were produced in the presence of cultivars Boulder or Missaukee, however, there were no tuber yield increases associated with cultivars Boulder and Missaukee, indicating that the nematode species might be G. pallida or some other Globodera sp. When Dakota Diamond was planted in 2014 in a PCN infested site, it was an excellent performer; whereas, the GN resistant cultivars did poorly. This is an indication that Dakota Diamond may be resistant to G. pallida or possibly multiple species of Globodera. This hypothesis will be tested molecularly in 2015. Tomato is a major vegetable crop in Uzbekistan, where Macrosiphum euphorbiae and Trialeurodes vaporariorum are devastating pests of this plant. It has been reported that the Mi gene (gene for resistance to Meloidogyne incognita) can confer cross-resistance to both M. euphorbiae and T. vaporariorum. For this to become a practical procedure for use in integrated pest management programs in Uzbekistan, the trait must be available in acceptable local tomato cultivars. The objective of this research is to determine if the resistance trait from the rootstock cv. Anahu is transferred to the scion when it is grafted to cv. Rutgers, both in the presence and absence of M. incognita. The research also involves quantification of the changes in phyllosphere volatiles in response to both grafting and the impacts of M. euphorbiae, T. vaporariorum and M. incognita. In initial studies, less than 30 volatiles were detected in the phyllosphere of cv. Rutgers tomato plants infected with M. incognita; whereas, 44 were associated with plants not infected with M. incognita. The rates of volatiles emitted into the phyllosphere ranged from 88.6 to 43,595 ng/g tissue per hour for non-infested plants and 66.6 to 66,868 ng/g tissue per hour for plants infested with M. incognia. In both cases, the greatest emission rate was for phellandrene , with the rate for the infected plants being significantly (P = 0.05) greater than that recovered from non-infected plants. It is anticipated that grafting highly desirable tomato scion cultivars to rootstocks with pest resistance properties can have a major impact on tomato IPM practices in Uzbekistan. Cover Crops In 2013, the lesion nematode (Pratylenchus sp., most likely P. penetrans) was recovered from the roots of all 11 pea varieties been evaluated for tolerance to root rot pathogens (primarily Fusarium solani f. sp. pisi) in a commercial field near LeRoy, NY with a long history of severe root rot. Interestingly, the lesion nematode was recovered from root of all the varieties at populations ranging from 15.3 to 55.3 nematodes/1 g roots. In 2014, pea plants were randomly collected from 17 pea fields in western New York in order to assess the role of the lesion nematode, if any in the increasing occurrence and damage of root rot diseases in recent years. The roots of collected plants were washed and evaluated for root rot severity on a scale of 1 (healthy) to 9 (most severe disease, >75% of root tissues affected and at a late stage of decay). Five grams of root where then selected and processed for nematode extraction for 3 days. The lesion nematode was recovered from roots of plants from 11 out of the 17 field samples. However, the population recovered from roots was rather low, except in field #11 where the number of the lesion nematode recovered was 1031/1 g root. Also, no correlation was evident between root rot severity and the number of lesion nematode recovered from roots. Trap crops are being developed for nonchemical control of cyst nematodes. A solanaceous weed, sticky nightshade (Solanum sisymbriifolium)is being evaluated to control potato cyst nematodes Globodera pallida. Because of the difficulties in working with this regulated pathogen, we are conducting experiments with the closely related tobacco cyst nematode G. tabacum as a model system. In initial experiments, we evaluated S. sisymbriifolium for ability to stimulate hatch of G. tabacum. Twenty cysts per cell were exposed to root full strength or 1 to 10 dilutions of diffusates of tobacco, sticky nightshade or distilled water. Hatch of nematodes was significantly higher for S. sisymbriifolium than from tobacco. We are currently evaluating the effects of S. sisymbriifolium as a trap crop in field microplots compared to resistant or susceptible crops. Cyst-nematode resistant tobacco lines are also being evaluated for host status and hatch stimulation in the Idaho. Mustard seed meals are byproducts of biodiesel fuel production from brassicaceous plants, and these meals can be amended into soil as fertilizers and for management of pests, pathogens and weeds. Mustard seed meal amendments (Brassica juncea and Sinapis alba) were therefore tested in the field, alone and in combinations, for management of the root-knot nematode Meloidogyne incognita on tomato, and for weed suppression. At the tested application rates, mustard seed meal from Sinapis alba showed some activity against the nematode but results were not consistent between years. No treatment affected weed population numbers. Field plots (12 ft by 50 ft) at the CAES Griswold Research Farm naturally infested with Meloidogyne hapla were planted to Pacific Gold brassica, rye or tomato in June of 2012. In September, Pacific Gold plots were either tilled in and rolled to seal, or allowed to senesce without tillage. The four resulting treatments were replicated six times. All plots were planted to San Marzano tomato transplants on 6 June 2013 and four plants per plot were dug on 25 July 2013. Roots were washed, weighed and numbers of galls counted. Gall counts from plots planted to tomato or Pacific Gold without tillage were similar, as were those planted to rye or Pacific Gold that had been tilled. In the fall of 2013, plots were either planted to Pacific Gold or winter rye as a winter cover, and tomatoes were again planted in field plots on 20 June 2014. Gall counts from end of September samples were not significantly different but additional samples are being evaluated. Pacific Gold grown over the summer may be a host to M. hapla and increase populations similar to tomato. However, tillage of shoots of this biofumigant crop results in nematode reduction similar to growing a non-host small grain rye. Growth of Pacific Gold as a winter cover crop disease not appear to increase M. hapla densities and may have been more effective than rye as a winter cover. In current mid-west agriculture, cover crops, soil health and a possible conversion to bio-agriculture are currently the hottest three topics. In a 2012-2013 trial with oil seed radish, both corn and soybean yields associated with the use of oil seed radish as a cover crop were greater than those associated with winter fallow. The opposite was true with the use of annual rye grass. The MSU program has conducted numerous cover crop trials with BCN, SCN and northern root-knot nematode. George Bird and Fred Warner intend to publish an Extension bulletin on cover crops and nematodes before the end of 2014. This is currently in very high demand. One individual has traveled from the west coast to look at the data and another is being scheduled to come from the Netherlands later this fall. Soil Amendments Soil Amendments on Heterodera glycines (MN): In a repeated greenhouse experiment, organic soil amendments were screened for effects on population density of soybean cyst nematode (SCN), Heterodera glycines, and soybean growth. Ten amendments at various rates were tested: fresh plant materials of field pennycress, marigold, spring camelina, and Cuphea; condensed distiller’s solubles (CDS), ash of combusted CDS, ash of combusted turkey manure (TMA), marigold powder, canola meal, and pennycress seed powder. Some amendments such as pennycress seed powder, and canola meal were effective for SCN management, but phytoxicity was a concern. Objective 2: Evaluate cultural management procedures for plant-parasitic nematodes in relation to their impacts on the sustainability of soil health: with special research to the utility of nematode community structure as an indicator of overall soil quality and their roles in plant nutrient cycling. Soil Health Management Practices Vermont was involved in three projects that related to this objective. First, in collaboration with University of Ghent (Belgium), nematode community indicators were validated as tools to quantify the successional maturity of compost. Second, high throughput genetic sequencing was applied to compare soils that are naturally suppressive or conducive to soybean cyst nematodes in the Midwest. Differences in community composition were not statistically different among treatments. Third, a suite of biological indicators were compared for their ability to predict suppression of Rhizoctonia damping-off disease on radish in a bioassay. The best predictors of disease suppression were respiration (immature compost) and an in vitro plate assay to quantify suppression of R. solani growth (mature compost). The sequenced genome of Paenibacillus sp. JDR-2, a xylanolytic spore-forming bacterium isolated from soil has allowed RNA-seq transcriptomic analysis to identify genes expressed in response to environmental signals. Growth with different carbohydrate substrates demonstrated expression of clusters of genes involved in the depolymerization hemicelluloses and the assimilation of the products of depolymerization. As a relative of Paenibacilllus sp. SdTc and Paenibacillus nematophilus, respectively ectoparasites of the entomoparasitic nematodes, Steinermema diaprepesi and Heterorhabditis spp., this approach may be applied to define the basis for phoretic Paenibacillus species and their nematode hosts. In 2014, we continued the monitoring of plant-parasitic and free-living nematodes in the plots of the long-term Soil Health Site on the Gates Farm near Geneva, NY. This long-term experiment was initiated in 2003, about14 acres in size, and divided into 72 plots (18 treatments replicated 4 times). The treatments are represented by three tillage systems (no-till, zone-till and plow/conventional-till systems), three cover crops (no cover, rye grain, and vetch), and two crop rotations. One rotation includes primarily high value vegetable crops (bean, beet, sweet corn, cabbage, bean, bean, peas, cucumber, bean, bean from 2003 – 2012), whereas the second rotation includes season long soil-building crops (bean, field corn, oats/sweet clover, sweet corn, bean, field corn, barley/red clover, sudangrass, bean, bean from 2003 through 2012). To-date, the no-till tillage is converted to ridge-till system whenever the plots are planted to a direct seeded vegetable crop. All the plots were planted to dry beans, as the indexing crop for assessing the impact of the various soil management practices on soil quality and productivity. Interesting results were obtained on the number of lesion nematode in roots of dry beans as impacted by cover crops (highest in the rye plots), crop rotation (higher in the diversified rotation 2), and tillage systems (highest in the reduced till-systems). However, populations of plant-parasitic and free-living nematodes were rather low to moderate in the soil of all plots and generally did not differ significantly. This site will be continued and data collection might reveal differences and significant interaction over time. Nematode Community Structure Analysis In Michigan, plant- (PC) and animal-based (AC) compost was tested for ability to suppress PPN, increase beneficial nematodes, soil conditions, and increase yield and quality of fresh market (Sugarsnax) and processing (Cupar) carrot cultivars. PC and AC, at 1, 1.5 and 2 times the standard recommendation for nitrogen source, and urea and non-amended (as controls) were tested. Preliminary analyses indicate significantly higher Maturity index (MI) in compost-treated (2.1) plots than in the controls (1.6), and fertility index (FI) in compost-treated plots was 1.0 compared to 1.3 in the controls, suggesting improved soil conditions. Abundance of omnivore-predatory nematodes was positively correlated (r = 0.53) with root to shoot dry matter ratio in the processing carrot variety plots, implying improved soil condition and more carrot root than shoot growth. Average total abundance was about 50 nematodes/100 cc of soil, with herbivores and fungivores being most abundant and omnivore and predators least abundant. Multifactor analysis of the relationship among nematode guilds, trophic groups and soil physiochemical properties shows significant correlation (r = 0.68) between c-p 1 bacteriovores and NO3-N. Soil food web structure appears to mature with time. Overall, the results support the hypothesis that compost treatment may increase beneficial nematodes and improving soil conditions. Cultural Management Long-term corn-soybean rotation effect on nematode community and the crop yields: The effects of long-term corn-soybean crop sequences on nematode community and corn soybean yield were studied in a field site that was established in 1982 in Minnesota. The crop sequences were: (i) five-year rotation of each crop such that both crops are in years 1, 2, 3, 4, and 5 of monoculture every year; (ii) annual rotation of each crop with both crops planted each year; (iii) continuous monoculture of each crop; (vi) annual rotation of two different cultivars of each crop. Beginning in 2010, half of each plot was treated with nematicide. Plant-parasitic nematodes were determined at planting, midseason, and harvest each year. In 2013 and 2014, nematode community including free-living nematodes in each plot was assessed. Nematicide (aldicarb) was effective against plant-parasitic nematodes, but also had negative effects on fungivores, shifting the nematode community to a lower ecological succession. Crop sequences strongly affected plant-parasitic nematodes, but also affected fungivores, bacterivores, nematode community diversity, maturity, and enrichment. Heterodera glycines, Pratylenchus spp., Helicotylenchus spp., and Xiphinema spp. are major plant-parasitic nematodes in the field. Crop sequence affected soybean and corn yields, which decreased with increasing number of years of monoculture. The nematicide aldicarb effectively reduced nematode population densities and increased both soybean and corn yields. However, increase of soybean and corn yield by nematicide was not related to initial densities of the major plant-parasitic nematodes, suggesting the yield penalty of inadequate crop rotation could not be fully explained by the damage caused by nematodes. This study shows the agronomic and ecological impact of crop rotation. Long-term corn-soybean rotation effect on microbial communities associated with the soybean cyst nematode: The research project was been initiated to study crop sequence effects on fungal and bacterial communities in the cysts of the SCN with cultural methods as well as metagenomic analysis. Fungi and bacteria have been isolated from the cysts in 2014, and they will be identified with morphological characteristics and DNA sequences. Objective 3: Provide educational materials and programs on cultural management of plant-parasitic nematodes based on host resistance, nematode antagonistic rotation or cover crops, soil amendments and biological agents. In 2014, two training workshops were again provided on the stem and bulb nematode (Ditylenchus dipsaci) on garlic and were attended by >100 growers, extension educators and industry personnel. The first workshop was held on the campus of the NYSAES in Geneva, NY on March 28th and the second one was held on April 9th in the Albany area. The training workshops and the subsidized diagnostic services provided under this project were funded by a grant from the Specialty Crop Program through the NYS Dept. of Agriculture and Markets. T0-date in 2014, a total of 107 garlic samples were submitted and analyzed for possible infestation by the garlic bloat nematode. These samples came from garlic growers and vendors in NY and several other states. Total of 18 samples tested positive for the garlic bloat nematode, 16.8% infection. Also, the project leader and cooperators have had extensive correspondence and visits with garlic growers throughout New York and elsewhere, discussing the biology and available management options against the bloat nematode and other diseases and pests of garlic. A pdf file of George Bird’s most recent Soil Health presentation (October 4, 2014 to the MI UP Master Gardeners) has been provided to the members of Technical Committee of NE-1040 and is available on the MSU ENT website. In addition, recommended soil health readings are made available to all meeting participants. A few of the additional outreach activities associated with NE 1040 in 2014 included: George Abawi of Cornell-Geneva held two Garlic training workshops in Geneva, NY on March 28, 2014 and Albany area, NY on April 9, 2014; The bloat Nematode on garlic and management options; >100 growers, extension educators and industry personnel were in attendance. LaMondia spoke about ‘Strawberry black root rot, nematodes and soil health’ to the New England Vegetable and Berry Growers Association in Hudson, MA (February 1, 115 persons). LaMondia was interviewed about nematode diseases of fruit and vegetable crops and management options for growers by Dorothy Noble for Growing Magazine (June 13); Neher, D.A. Nematodes as Biomonitors short course, University of Ghent, Ghent, Belgium, 1-14 May 2014. Neher, D.A. Keynote Speaker, Compost symposium, California Polytechnic State University, San Luis Obispo, 28 March 2014 Milestones Accomplished • Soil health meeting held in New York/Michigan focusing on report cards for soil health. Planning is currently underway to hold meetings immediately prior to the annual Society of Nematologists meeting in Michigan in 2015, combining both 2014 and 2015 milestones. • Molecular characterization of Melodoigyne graminis and Subanguina radicicola populations. Research is in progress.

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