Brief Summary of Minutes of Annual Meeting

NE-1040 2011 Annual Meeting Minutes Project Title: Plant-Parasitic Nematode Management as a Component of Sustainable Soil Health Programs in Horticultural and Field Crop Production Systems Administrative Advisor: Magnarelli, Louis (Louis.Magnarelli@po.state.ct.us)  Connecticut Agric. Exp. Station Chair: George Bird (birdg@msu.edu)  Michigan State University Recording Secretary Senyu Chen (chenx099@umn.edu)  University of Minnesota Technical Committee Members Abawi, George (Gsa1@nysaes.cornell.edu)  Cornell University, Geneva, NY Bernard, Ernest (ebernard@utk.edu)  University of Tennessee Burelle Nancy (nancy.burelle@ars.usda.gov)  USDA ARS, Fort Pierce, Florida Dickson, Donald (dwd@ufl.edu)  University of Florida Grewal, Parwinder (grewal.4@osu.edu)  Ohio State University Halbrendt, John (Jmh23@psu.edu)  Penn State University, Fruit Res & Ed Ctr Huettel, Robin (Huettro@auburn.edu)  Auburn University Kotcon, Jim (Jkotcon@wvu.edu)  West Virginia University LaMondia, Jim (James.LaMondia@po.state.ct.us)  Connecticut Agric. Exp. Station Meyer, Susan (susan.l.meyer@ars.usda.gov)  USDA ARS, Beltsville, MD Melakeberhan, Haddish (melakebe@msu.edu) - Michigan State University Mitkowski, Nathaniel (mitkowski@uri.edu)  University of Rhode Island Neher, Deborah (Deborah.neher@uvm.edu)  University of Vermont Preston, James (Jpreston@ufl.edu)  University of Florida Thies, Judy (judy.thies@ars.usda.gov)  USDA ARS, Charleston, SC Wick, Robert (rwick@pltpath.umass.edu)  University of Massachusetts Meeting Location and Date: Burlington, Vermont, October 10-11, 2011 Meeting attendants: Members: George Abawi, Ernest Bermard, George Bird, Senyu Chen, Don Dickson, Parwinder Grewal, John Halbrendt, Jim LaMondia, Louis Magnarelli, Haddish Melakeberhan, Nathaniel Mitkowski, Deborah Neher, James Preston. Other attendants: Tharshani Nishanthan, and Jie Zhao  Graduate students, Univ. VT. Monday, October 10, 2011 The meeting was started at 8:15 a.m. in the conference room in the Department of Plant and Soil Science, University of Vermont. Deb Neher, the meeting host, welcomed the group, and announced local arrangements. Tom Vogelmann, Dean of UVM College of Agriculture and Life Science, welcomed the meeting attendants, and introduced the college. The meeting was called to order by Chair George Bird. The minutes of last year meeting were reviewed and there were no changes. George requested that the state written reports be submitted to him within a week. Louis Magnarelli, administrative advisor, discussed issues related to funding in federal agencies and regional levels. He announced that the deadline for submitting the project report is Dec 11, 2011. The report should be concise including key results, and mile stones. This is a 7-year project, and mid-term will be 3.5 years. We may need to consider a mid-term report at next years meeting. It was announced that Michigan AgBioResearch (formerly Michigan Agricultural Experiment Station) is in the process of appointing Haddish Melakeberhan to NE-1040, and that Robin Huettel has retired from her position at Auburn University.1 George Bird reviewed the project objectives and milestones before the state reports began. State reports were presented by the following presenters: Jim LaMondia (CT), Tharshani Nishanthan (VT), Senyu Chen (MN), John Halbrendt (PA), George Abawi (NY), Don Dickson (FL), Jim Preston (FL), Haddish Melakeberhan (MI) and Nathaniel Mitkowski (RI). During the afternoon break, the group visited Deb Nehers new laboratory, department and greenhouse facilities. Tuesday, October 11, 2011 The final three state reports were presented by Ernest Bernard (TN), Parwinder Grewal (OH), and George Bird (MI). The business meeting was called to order by George Bird. George put forward a resolution to thank Deb Neher for the excellent local arrangements and the resolution was agreed to by all meeting members. Ernest Bernard invited NE-1040 to have the next annual meeting in Knoxville, TN. George Abawi also expressed willingness to host a meeting in Geneva, NY in near future. Both invitations were accepted and the meeting site for 2012 will be in Knoxville and 2013 in Geneva. Following the business meeting, there was an open discussion on general nematology issues for about 20 minutes before the meeting adjourned at 10:50 a.m. __________________________________________ 1Subsequent to the meeting, it was determined that Dr. Melakeberhans appointment to NE-1040 was approved on April 13, 2011.

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. - A new species of Pasteuria, Candidatus Pasteuria aldrichii, propagated in the bacterivore Bursilla RGD244, was identified (FL). Based on sequences of genes encoding 16S rRNA, this Pasteuria sp. is closely related to P. penetrans and other species associated with phytoparasitic 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 for deciphering sequences from Pasteuria with biocontrol potentials that are often contaminated with genes from other soil bacteria (FL). Pasteuria spp. associated with H. glycines and H. schachtii were detected for the first time in MI. In a 2010 soybean field trial in MI, seed treatment with Bacillus firmus did not enhance yield or reduce population densities of H. glycines under very high initial population densities of H. glycines and Fusarium viguliforme. Cover Crops. - A collaborative (NY, PA, CT) project was initiated in 2009 to assess the efficacy of nine cover crops (rye grain/hairy vetch, oats, sudax, forage radish, red clover, rapeseed, buckwheat, and wheat) in suppressing root pathogens and improving soil health and vegetable crop yield in research and on-farm trials under four management systems (conventional, organic, present integrated pest management (IPM) and future IPM). The research sites had different management histories and varied levels of pathogen pressure, free-living nematodes and soil quality. Due to the use of grain, population densities of lesion nematodes were highest in the future IPM system. Almost no lesion nematodes were detected in the organic system, suggesting a suppressive soil. In contrast, population densities of root-knot nematodes were highest in the present IPM and organic systems, compared to the future IPM and conventional systems. Populations of beneficial nematodes were lowest and highest in the organic and present IPM systems, respectively. Grain crops and rapeseed reduced root-knot nematode population densities; whereas, red clover, radish and rye/hairy vetch increased the populations. All cover crops increased the population densities of Pratylenchus spp., especially sudax, oats, wheat and the rye/hairy vetch and fallow systems. Several Pratylenchus spp. weed hosts were associated with the fallow treatment. The rye/hairy vetch, oat, wheat and sudax supported the highest population densities of free-living nematodes. Canadian forage pearl millet (Pennisetum glaucum) hybrid 101 is a suppressive rotation crop for Pratylenchus spp. It is not yet commercially available in the United States. Other millet cultivars were evaluated for host status of P. penetrans in field microplots and compared them to bare fallow, tomato and a rye/vetch mix (CT). The rye/vetch mixture resulted in higher populations of P. penetrans than all other treatments. The lowest nematode populations were observed in hybrid 101, hybrid III and cattail millet plots. The experiment was repeated in different microplots in 2010. P. penetrans population densities associated with all millet types were significantly lower than from tomato, but not different from each other. Tuber yields were lower for rye/vetch than for Pearl millet 101, Tifgrain 102, hybrid pearl millet III, and several other treatments. Resistance. - Twelve varieties of Lactuca sativa with potential resistance to root-knot nematode were tested for susceptibility to M. hapla (RI). In previous studies, the same varieties were tested against M. incognita but have shown very different reactions to M. hapla infection. Red Orient was confirmed as being resistant to M. hapla but was moderately susceptible to M. incognita. Long-term turfgrass plots for evaluating varietal resistance in creeping bentgrass were established in 2011. Advanced sweetpotato breeding lines from USDA Charleston, North Carolina State University, and Louisiana State University were evaluated for resistance to M. incognita (USDA/ARS/South Carolina). Sweetpotato seedlings from the USDA breeding program were screened for resistance to root-knot nematodes in efforts to develop root-knot nematode resistant sweetpotato varieties. Advanced pepper germplasm lines were screened for resistance to M. hapla and resistant plants were selected for development of homozygous resistant lines. Advanced southernpea (cowpea) breeding lines from Louisiana State University, University of Arkansas, and Texas A&M University were evaluated for resistance to M. incognita. Advanced wild watermelon rootstock lines developed by USDA-SC were evaluated as rootstocks for seedless watermelon in root-knot nematode infested fields. Wild watermelon rootstock lines exhibited resistance to M. incognita and produced higher yields than watermelon grafted on commercial bottle gourd and squash hybrid rootstocks, which were highly susceptible. A new root-knot nematode germplasm was released as Fery, R.L. and J.A. Thies (Notice of release of PA-566, New root-knot nematode germplasm , pimento-type pepper. USDA, Agricultural Research Service, Washington, D.C. Cultivar Release, April 30, 2010). In cooperation with R. Cowles of CAES, CT conducted field assessment of strawberry cultivars over three years for yield, vigor, black root rot (Rhizoctonia fragariae and P. penetrans) and tolerance to black root rot, as well as leaf-feeding preference bioassays to identify tolerance to black vine weevil (BVW). Primetime and Lester exhibited resistance or tolerance to black root rot and non-preference to BVW in feeding preference trials. Idea was moderately susceptible to root rot, but produced a large, vigorous root system. Progeny of crosses made between Primetime, Lester, Allstar, Delmarvel, and Idea were selected for resistance or tolerance to black root rot in greenhouse pots and in the field in infested soils as well as low preference in BVW feeding trials. Progeny were screened for fruit yield, size, and flavor. Selection over three years reduced the progeny population from more than 4,000 genotypes to a few elite clones with promising horticultural characteristics, tolerance to black root rot, and low feeding preference by BVW. The results demonstrated that sufficient variation exists in certain octoploid parents to develop effective resistant/tolerant lines. Because of differences in disease reactions between greenhouse evaluations of juvenile plants and field evaluations on mature plants, evaluations in the field are essential for selecting for black root rot tolerance. Breeding line IP-5 has exceptional fruit quality and has been named Rubicon. A plant patent has been submitted. Research was initiated on the susceptibility of grafted tomato to Meloidogyne incognita (TN). In the first replicated greenhouse experiment, galling on 11 potential rootstocks, 3 potential scions, and a check cultivar (Rutgers) was evaluated 90 days after inoculation of seedlings with the aid of the Zeck rating scale. During the growth period, all plants of most selections exhibited foliar symptoms similar to herbicide damage, but all plants of 3 rootstocks (Emperador, ResiStar, Maxifort) were normal. The damage likely was caused by unauthorized use of herbicides around the greenhouse or by unusually high greenhouse temperatures during a record-hot summer affecting plant chemistry. Most rootstock selections had little or no galling, but 4 (Emperador, Vigostar 10) had average gall ratings above 3. Two scions (Brandywine, Mountain Fresh) and Rutgers had gall ratings of 6 or higher. Because greenhouse heat may have affected nematode survival, all of the rootstocks will be retested to confirm resistance or susceptibility. In field trials in MI, the two Heterodera schachtii resistant varieties (BTS18RR4N and BTS19RR1N) available to MI sugar beet growers in 2010 yielded 4.2 and 4.1 tons per acre more, respectively, than a H. schachtii susceptible control (SX1260RR). In a 2010 field trial at a MI site with high soil population densities of both Heterodera glycines and Fusarium viguliforme, a PI 88788 x PI 548402 soybean variety yielded 39.94 bu/acre more than the susceptible control. Crop Rotation. - In Minnesota, a four-year field experiment was initiated in 2009 at two locations in southern Minnesota to study the effects of rotation with non-host crop and resistant soybean, and tillage on soil suppressiveness to the soybean cyst nematode (SCN). In 2010-2011, the biocide formaldehyde treatment of soil in the field plots reduced the fungal parasitism of the SCN second-stage juveniles at planting and increased midseason SCN egg population density. CT and NY cooperatively conducted research to evaluate the effects of certain rotation crops on Pratylenchus spp. and bean root rot in micro-plots. Crops were grown in 2009 and followed by Caprice beans in 2010. Rotation crops consisted of fallow, hairy vetch, radish, Tifgrain 102 millet, Rangi rapeseed, Red Clover, LA 604 oats, buckwheat, and sorghosudangrass. In 2010, tillage radish and Rangi rapeseed had the highest shoot and root weights and radish, then rapeseed, sorghosudangrass and millet had the lowest root rot ratings. In 2011, radish again had the highest shoot, root and bean yield. Soil Amendments. - Brassica spp. produce a broad range of glucosinolates (GSLs) that decompose into toxic metabolites that can be utilized in biofumigation for nematode management (CT). Seeds of these species make a number of minor GSLs, but typically produce one predominant type. Reverse phase high pressure liquid chromatography was used to characterized predominant GSL types. Bioassays were used to determine which GSLs are most toxic to northern root knot nematode (Meloidogyne hapla). The predominant GSLs tested from plant seed sources were: progoitrin (Brassica napus Dwarf Essex, Sterling), sinigrin (B. juncea Pacific Gold and Brassica nigra), 4-methylsulfinylbutyl (B. oleracea Calabrese), not identified (Camelina sativa), epiprogoitrin (Crambe abyssinica Meyer), 4-methylthiobutyl (Eruca sativa), and sinalbin (Sinapis alba Ida Gold). Exposure to glucosinolates caused mortality of M. hapla. Researchers have demonstrated that growing brassica cover crops alters soil nematode communities, and that biofumigation with brassica meals disrupts biological control by insect-pathogenic nematodes. Seed meal bioassays were used to further determine the effects of exposure to glucosinolates to both M. hapla and Caenorhabditis elegans. Seeds of test plants were frozen, ground in a mortar and pestle, and mixed with water to result in rates of: 2.5, 0.5, 0.25, 0.1, 0.02 or 0 g seed per L soil. Vials were capped and incubated at 21 C. After 48 hr, vial contents were poured into counting dishes and nematodes were scored as viable or non-viable, and counted. There were no differences in survival of either nematode at 0.25 or 0.02 g meal per liter water. At 0.1 g meal per liter, C. elegans was generally more affected than M. hapla, however, there were significant differences in survival between the two nematodes for different meals. For example, one of the highest levels of survival of C. elegans occurred with Pacific Gold, which has consistently been the most effective meal for biofumigation management of M. hapla. This may indicate that targeting plant parasitic nematodes does not necessarily mean that free-living nematodes will be affected. A two-year demonstration of compost for management of plant pathogens and weed seeds was conducted in partnership with Highfields Center for Composting. On-farm field trials were conducted at Intervale Community Farm (Burlington, Vermont) and Riverside Farm (East Hardwick, Vermont) on mixed vegetables. Compost treatments included manure (not to recipe), recipe with hay, recipe with softwood, recipe with hardwood, non-composted rice hulls and bare soil. Incidence and severity of early blight disease was monitored throughout the growing season. Compost made to recipe and reaching target temperatures killed both giant crab grass seed and the Alternaria, early blight pathogen on brassica crops. Kill was more complete in mesh than solid plastic bags indicating that a combination of microbes and heat are responsible. Kill was achieved after reaching NOFA-Vermont standards for making compost and remained through final stages of the thermophilic and curing phases of compost production. Microbial communities started the composting process with a relative dominance of species that produced hydrolytic and urease as extracellular enzymes and were followed successionally by an increase in production of oxidative enzymes. In the field trials, disease incidence and severity increased as the season progressed at both farms. The study was terminated at the Intervale Farm at the end of August due to flooding by tropical storm Irene. Post-Irene, the severity and incidence of disease at Riverside Farm increased exponentially. No clear patterns of the impact of compost recipe at Intervale. At Riverside, there was a trend that hardwood compost suppressed disease severity and reduced splash of pathogen spores more than the other compost recipes or controls. There was no difference observed between the two types of controls, so we conclude that the physical barrier of compost made less difference than the microbial composition of the material. Field experiments were conducted in 2009 and 2010 to determine the effects of manure and PK fertilizers on soybean and corn yields, the soybean cyst nematode (SCN), nematode community, soil food web, and associated soil properties in an SCN-suppressive field and an SCN-conducive field in Minnesota. Manure did not affect SCN egg population density, but resulted in lower SCN second-stage juvenile population density in the SCN-suppressive soil at 45 days after planting (DAP) in 2009. Manure also reduced spiral nematodes at 45 DAP. PK increased SCN egg population density in 2010. Increase of soybean yield by fertilizers was observed in the SCN-conducive soil but not in SCN-suppressive soil, and manure was more effective than PK. The fertilizer effect on yield was greater for SCN-susceptible soybean than resistant soybean; yield difference between the SCN-susceptible soybean and resistant soybean was greater in no-fertilizer treatment than PK and manure treatments. Corn yield was greater when treated with manure than the PK and no-fertilizer. Mustard seed meals, a byproduct of the biodiesel industry, are applied to agricultural fields as fertilizers and pesticides. Consequently, methods are being developed to optimize use of these amendments for suppressing plant-parasitic nematode populations. Seed meals made from different plant species vary in chemistry, so seed meal combinations have potential for a wider range of activity against pathogens and pests than seed meal amendments derived from one plant species. Seed meals from Brassica juncea and Sinapis alba were tested in the greenhouse for effects on pepper plant vigor and on populations of root-knot nematodes; a seed meal combination was as effective as an individual seed meal for improving plant vigor and suppressing nematode populations (USDA/ARS/Beltsville). In MI, scalable models were developed to identify what soil amendments work at site-specific locations and how they work at these locations. 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. Nematode Community Structure Analysis. - Application of manure reduced the percentage of plant-parasitic and fungal-feeding nematodes, and increased bacterial-feeding nematodes (MN). The nematode guild of Ba1 responded to manure dramatically and immediately. The higher values in Dominance and EI, and the lower values in Diversity, MI, FFB, and CI occurred when manure was applied in both SCN-conducive and suppressive soils. Manure increased the nitrate, ammonium, and zinc levels in soil; and the PK increased phosphorus. This study suggests that improvement of soil fertility with application of manure and PK is a useful strategy to alleviate the SCN damage. Assessment of nematode community structure continued in 2011 at the 14 acre long-term Soil Health Site at the Gates Farm near Geneva, NY. As urban agriculture is gaining momentum, NE-1040 work in OH is focused on urban soil quality assessment. Nematode community structure analysis is used as a surrogate for soil food web health and quality. With this procedure, soil food web health and quality varies tremendously in the urban landscapes and the soil nematode food web appears to be highly enriched indicating abundance of nutrients, but is less structured indicating disturbance. Compared to the urban core, soil food webs are more disturbed on the urban fringe indicating the effect of recent development activities. There is greater un-coupling of the soil food web and nutrient cycling process with proximity to the road indicating anthropogenic impact such as chemical contamination from road salts and vehicle emissions. Cultural Management. - Use of engineered charcoal (biochar) in soil is gaining attraction as a soil amendment that may improve crop yield. Little is known, however, about the relationships between charcoal structure and its biological function in soil. Due to its high surface area and microporosity, charcoal is a potent adsorbent of organic compounds, with the potential of reducing bioavailability of signaling compounds. Disruption of host recognition by adsorption of root exudate chemicals by charcoal would result in cyst nematodes staying dormant and not hatching or infecting roots, resulting in the avoidance of significant damage and nematode reproduction and population increase. Cyst nematode hatch was reduced by 50% in the presence of both root-diffusates and biochar. Biochar dramatically reduced cyst nematode hatch stimulation to levels similar to the 100-fold dilution of root diffusates in the absence of biochar when root diffusates were perculated through soil with 10% biochar and measuring hatch stimulation by full strength, or 10 and 100-fold dilutions over a period of weeks. In MI, it was shown that the diversity of crops such as carrots, celery, onions, peas and potatoes that are susceptible to plant parasitic nematodes, such as Pratylenchus spp. Meloidogyne hapla and Ditylenchus spp., mandates that the quantification and documentation are an integral component of soil health management under pesticide-intensive or less-intensive production systems. Cultural management procedures for plant-parasitic nematodes were evaluated in PA in relation to their impacts on the sustainability of soil health: with special reference to the utility of nematode community structure as an indicator of overall soil quality and their roles in plant nutrient cycling. Humic acid is a fundamental component of healthy soil and the concentration of humic acid is linked to organic matter and biological activity. Soil samples from fields supplied with exogenous applications of humic acid indicated a concomitant increase in free-living nematode populations. Based on these observations, experiments were designed to determine if there may be a cause and effect relationship between field applications of humic acid, soil fertility and free-living nematode population levels. The effect of humic acid applications on plant growth and nematode population levels were compared to untreated plots in both a vineyard and an apple orchard. Plant growth measurements showed no significant differences between treated and untreated plots after one season. There were also no significant differences in the numbers of plant-parasitic nematodes between treated and untreated plots. The free-living nematode population however was 23% greater in the humic acid treated plots at the end of the season than in the untreated plots. These results suggest humic acid may stimulate microbial activity leading to a subsequent increase in bacterial and fungal feeding nematode populations. Plant-parasitic nematodes continue to result in decline of turfgrass health in golf greens as evident by 172 composite soil samples assayed during the 2011 growing season. The loss of fenamiphos and the lack of efficacious, commercially-available alternatives leave golf course superintendents with no control measures. Cultural practices that increase root development are helpful, as is raising the height of cut. While these practices do not reduce nematode populations, they allow the grass to tolerate nematode feeding. During 2011, ActinoGrow (Streptomyces lydicus), Azatrol (azadirachtin) and a proprietary product were tested on golf greens. The results indicated that there are no commercially-available products that are effective (MA). In MI, soil pH was identified as the driving factor associated with cherry tree decline. Scalable models were developed to identify what soil amendments work at site-specific locations and how they work at these locations. A Ditylenchus dipsaci of NY garlic was conducted in cooperation with the Garlic Seed Foundation. Cotton and sunflower seedlings were grown in Mega germination bags and root systems were inoculated with freshly hatched M. incognita juveniles (TN). After gall initiation, plants were moved to a private facility and planted in an X-ray-transparent substrate. Root systems were periodically and non-destructively X-rayed to observe root architecture, root growth and gall development. Gall enlargement was successfully observed over a 27-day period. Nematodes successfully developed to the female stage and produced egg masses by 30 days after inoculation. 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. The results of the NY survey for Ditylenchus dipsaci were made available to garlic growers through the Garlic Seed Foundation and two Extension publications. MA and RI presented a seminar at the New England Turf Grass Management Conference to 300 golf course superintendents that included a substantial amount of material regarding chemical and cultural nematode control practices and other related techniques to improve turf and soil quality. Nematode and soil ecology information from NE 1040 was included in 20 potato/sugar beet/soybean grower education meetings in MI in 2011. The results of a 2010 Heterodera glycines survey of MI and a Field Guide for soil fumigant management plans were developed and made available to the agricultural community in 2011. An additional number of invited soil quality presentations were given in the North Central region during 2011. The MI component of NE-1040 also had international soil quality education outreach in 2011 in Montenegro and Central Asia (Kyrgyzstan and Tajikistan). Milestones Accomplished All nine of the 2010 Milesones described in the original NE-1040 project description were achieved. Seven of the eight 2011 Milestones described in the original NE-1040 project description were achieved. Progress in relation to the CT/NY/PA development of M. hapla- resistant pepper varieties needs to be evaluated.

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