Brief Summary of Minutes of Annual Meeting

Agenda: Thursday November 13: 6:30-8:00 a.m. – Hot breakfast served at Drury Inn 8:00 a.m. Van departs Drury Inn for Skeen Hall, Room N-128 at NMSU; (for those with rental cars, I have 2-day campus parking permits for you). 8:30 a.m. Welcome and introductions (W-3186 Acting Chair, Steve Thomas) 8:40 a.m. Welcome and instructions (W-3186 Host, Steve Thomas) 8:45 a.m. Welcome and Project overview: (Dr. David Thompson, Administrative Advisor; and Associate and Director, College of Agricultural, Consumer and Environmental Sciences) 9:00 a.m. Graduate student presentations (~20 minutes each) Carol Land (Auburn) Amber Smith (Auburn) Justin Luangkhot (Auburn) 10:00 a.m. Break 10:30 a.m. ---continue graduate student presentations (~20 minutes each) Ni Xiang (Auburn) Shankar Pant (Mississippi State) Jian Jiang (Mississippi State) 11:45 a.m. Walk to The Game for lunch 1:30 p.m. Report presentations – state representatives (~20 minutes each) Kathy Lawrence -Alabama Phil Roberts- California Ed Caswell-Chen – California Brent Sipes - Hawaii 3:00 p.m. Break 3:15 p.m. ---continue report presentations Gary Lawrence - Mississippi Haddish Melakeberhan - Michigan Tom Powers - Nebraska Steve Thomas – New Mexico 5:00 p.m. Instructions for dinner (Steve Thomas) - Return to the hotel 6:00 p.m. Meet in the lobby and drive to restaurant (chosen by group) for dinner Friday November 15: 8:00 a.m. Van departs Drury Inn for Skeen Hall room N-128 at NMSU 8:15 a.m. Wrap up any remaining reports Bob Robbins - Arkansas 9:00 a.m. Business Meetings (Steve Thomas Presiding): Increasing membership Joint proposals Annual report preparations Meeting site selections for 2016 and 2017 Election of Secretary Other matters 10:00 Break; Adjourn (5 folks have MUST leave for El Paso Airport at this time!!) 10:15 Tours or other activities for remaining attendees 12:00 Lunch plans (to be determined) Afternoon departures Annual Meeting Minutes (recorded by Kathy Lawrence): Overview: The 2014 W-3186 multi state nematology research project meeting was convened on the New Mexico State University campus in Las Cruces, NM on Thursday November 13, 2013. This meeting was organized and hosted by Steve Thomas (NM). Scientist in attendance and the state they represent: Bob Robbins (AR), Phil Roberts (CA), Tom Powers (NE), Haddish Melakeberhan (M), Gary Lawrence (MS), Kathy Lawrence (AL), Brent Sipes (HI), Steve Thomas (NM) and Dave Thompson (Administrative Advisor for W-3186, NM). Graduate students in attendance included Caroline Land (AL), Amber Smith (AL), Justin Launghkot (AL), Ni Xiang (AL), Shankar Pant (MS), and Jian Jiang (MS). Announcements: 1) Acting Chair, Steve Thomas (NM) called the meeting to order at 8:30 a.m 2) The members were then informed that Saad Hafez, (ID) and Russ Ingham (OR) would not attend due to recent illnesses. 3) Each member introduced themselves explaining their affiliations with their respective universities or USDA-ARS employment. Welcome Remarks: 1) Dr. David Thompson, Associate Dean and Director of the Agricultural Experiment Station at New Mexico State University and the W-3186 administrator advisor, welcomed our group to New Mexico. He reported the western directors considered our project successful and he was glad to see students present. Phil Roberts asked how the experiment station works in NM and David outlined their agronomic centers, research scientist and staff, programs and effect on employment. 2) Jerry Sims, Department Head, Entomology, Plant Pathology, and Weed Science (EPPWS) welcomed us. Reports presented by graduate students: Caroline Land (AL) Amber Smith (AL) Justin Launghkot (AL) Ni Xiang (AL) Shankar Pant (MS) Jian Jiang (MS) State reports by members of W-3186: Kathy Lawrence (AL- root knot and reniform management on cotton) Phil Roberts (CA - root-knot on multiple vegetable crops) Brent Sipes (HI- reniform on tropical plants and pineapple) Gary Lawrence (MS-Seed treatments available on all crops) Haddish Melakeberhan (MI – Sugar beet cyst and tropic nematode groups) Tom Powers (NE – ring nematodes across the country bar codes) Steve Thomas (NM – root knot on chile and weeds) Meeting was adjourned for the day at 5:30 p.m. and would resume on Friday. Friday November 14, 2014 – The meeting resumed at 8:30 a.m. State reports continued. Bob Robbins (AR- species of root-knot nematode currently present in AR) Business Meeting: 1) The meeting was called to order at 9:00 November 14, 2014 by chair Steve Thomas. The minutes from the 2013 meeting were accepted as written. 2) The officers of the members were reviewed and Steve Thomas is serving as Acting Chair for this meeting. Kathy Lawrence is filling is as secretary for Ed Caswell-Chen. 3) The 2015 W-3186 meeting will be in Alabama Nov. 5 and 6, 2015. Meeting locations were discussed and the group voted to meet on the Auburn University campus. The W-8136 groups will meet in conjunction with the S-1046 group. Kathy Lawrence will serve as Chair since she is hosting the meeting. Ed Caswell-Chen will serve as Vice Chair. Brent Sipes volunteered for Secretary for the next year. Officers and location were approved by the members attending. 4) Ed Caswell Chen from the University of California-Davis sent an invitation for the W-3186 to meet at Davis in 2016. Kathy Lawrence moved to accept his invitation and it was seconded by Phil Roberts and approved by the group. 5) Membership was discussed. Steve Thomas handed out a list of members and those attending. Phil Roberts suggested we put a letter together to the USDA to state the value of the scientist attending our meeting. Steve will draft a letter and Dave Thomas will send it though the Western region. 6) Haddish Melakeberhan asked if we wanted to do anything at the SON meeting in July. Phil said we could meet over an idea of a new funding opportunity if anyone knows of one. Tom is meeting at a soil biodiversity meeting next week and will check out any funding opportunities there. 7) Phil Roberts lead a discussion of depository of nematode collections. SON may have a collections committee and Steve Thomas will check with the regularity committee. $2000 of a widely prevalent nematode list is coming from APHIS and may give some to help with the nematode collection. 8) With no further business, the meeting was adjourned at noon.

Objective 1: Characterize genetic and biological variation in nematodes relevant to crop production and trade. Alabama: Rotylenchulus reniformis resistant LONREN-1×FM966 breeding lines developed at Auburn University have demonstrated that the nematode resistance is accompanied by severe stunting, limited growth, and low yields. The objectives of this study were to evaluate the effects of applying nematicides to selected LONREN breeding lines on R. reniformis nematode populations, plant stunting, and yield. Four resistant breeding lines from the LONREN-1×FM966 cross, one susceptible line from the LONREN-1×FM966 cross, as well as LONREN-1, BARBREN-713, and the susceptible cultivar DP393 were evaluated with and without nematicides in the presence of R. reniformis. In the greenhouse, nematicides increased plant height across all genotypes compared to no nematicide. Rotylenchulus reniformis populations were 50% lower in the resistant lines compared to the susceptible lines at 45 days after planting (DAP). In microplot and field trials, the phenotypic stunting of all genotypes was reduced by aldicarb with increases in plant heights at 30 and 75 DAP. Increases in yields were evident across all genotypes treated with aldicarb. In all three trial environments, BARBREN-713 outperformed the LONREN-derived lines as well as ‘DP393’ in seed cotton yields, while having significantly lower R. reniformis egg densities than the susceptible genotypes. The results of this study indicate that the BARBREN-713 source of resistance will replace the LONREN source of resistant in future R. reniformis resistance research and breeding. Arkansas: I am working with soybean researchers in Georgia and Missouri to identify resistant markers to reniform nematode in RIL’s developed from crossing Magellan and PI 404198B. In an effort to identify root-knot nematodes in Arkansas using molecular methodology over 100 samples of root-knot were sequenced and identified. Six species (Meloidogyne arenaria, M. hapla, M. haplanaria, M. incognita, M. marylandi, and M. partityla were identified, with M. haplanaria, M. marylandi, and M. partityla new records from Arkansas. The Southern root-knot nematode Meloidogyne incognita was identified from 51 samples from 30 counties with 25 from soybean; M. haplanaria from 13 samples from 4 counties; M. marylandi from 5 samples from 4 counties; M. hapla from 5 samples from 2 counties; M. arenaria from 2 samples from 2 counties and M. partityla from 1 sample from pecan groves in 5 counties. An undescribed species of Punctodera was identified from the same pecan grove as M. partityla. All Meloidogyne species were reproduced using the single egg mass method, placed on appropriate hosts, re-sequenced and identified, and reproduced in greenhouse cultures for future study. California, Riverside (Hyman): Strelkovimermis spiculatus is an obligate parasite of mosquito larvae that breeds in environments of permanent flooding or in habitats subjected to periods of desiccation. Differences in epizootic levels within these habitats likely are due to variation at the genetic level. We evaluated whether nucleotide sequence variation levels might provide a basis for the development of molecular markers that correlate with infectivity phenotypes characteristic of different populations Argentine S. spiculatus populations. Partial gene sequences from the nuclear 18S gene and the mitochondrial nd4 gene were analyzed. Nucleotide sequences revealed a modest number of substitutions the 18S and nd4 genes over a short sequence expanse. Our results indicated that an expanded sampling of both nuclear and mitochondrial genes, with a special reference to nuclear genes encoding host range infectivity phenotypes, may be useful in the development of biomarkers for studying the population dynamics of S. spiculatus, and for correlating isolates with phenotypic traits useful for durable biological control practices. Mitochondrial DNA (mtDNA) encodes respiratory complex subunits essential to almost all eukaryotes; hence respiratory competence requires faithful duplication of this molecule. However, the mechanism(s) of its synthesis remain hotly debated. Here we have developed Caenorhabditis elegans as a convenient animal model for the study of metazoan mtDNA synthesis. We demonstrate that C. elegans mtDNA replicates exclusively by a phage-like mechanism, in which multimeric molecules are synthesized from a circular template. In contrast to previous mammalian studies, we found that mtDNA synthesis in the C. elegans gonad produces branched-circular lariat structures with multimeric DNA tails; we were able to detect multimers up to four mtDNA genome unit lengths. Further, we did not detect elongation from a displacement-loop or analogue of 7S DNA, suggesting a clear difference from human mtDNA in regard to the site(s) of replication initiation. We also identified cruciform mtDNA species that are sensitive to cleavage by the resolvase RusA; we suggest these four-way junctions may have a role in concatemer-to-monomer resolution. Overall these results indicate that mtDNA synthesis in C. elegans does not conform to any previously documented metazoan mtDNA replication mechanism, but instead are strongly suggestive of rolling circle replication, as employed by bacteriophages. As several components of the metazoan mitochondrial DNA replisome are likely phage-derived, these findings raise the possibility that the rolling circle mtDNA replication mechanism may be ancestral among metazoans. California, Riverside (Roberts): Genetic analysis, molecular mapping, and marker development were accomplished to determine the basis of resistance to the root-knot nematode M. incognita and the interacting fungal pathogen Fusarium wilt, races 1 and 4 in cotton. QTL regions for resistance to both nematode and wilt infection on homoeologous chromosomes 11 and 21 were fine mapped using the published Gossypium raimondii D5 whole genome sequence as a resource to design additional markers in the resistance regions. A similar approach was made for a region of chromosome 17 which harbors a major QTL for resistance to Fusarium wilt race 4. Phenotyping and marker analysis of a set of 34 chromosome substitution lines for reaction to M. incognita and Fusarium wilt races 1 and 4 was completed. A set of 43 isolates of Meloidogyne incognita (25), M. hapla (11), M. javanica (7), and M. arenaria (2) stored as second-stage juveniles were removed from cryopreservation storage and cultured on tomato plants to develop inoculum for screening a panel of resistant carrot germplasm lines. Some isolates had been stored in liquid nitrogen for 21 years, and all isolates showed good viability open thawing. On the carrot panel containing several diverse sources of resistance, only minor variation for ability to gall carrot roots was found among M. incognita isolates. Resistance was strongest and most broadly effective against M. javanica and M. arenaria isolates. In contrast, high levels of variation in root-galling were found among the M. hapla isolate x resistant carrot combinations. Analysis of multiple isolates of species in the plant-parasitic nematode genus Scutellonema was completed. Results of morphological and molecular comparisons clarified species relationships and provided a framework for resolving diagnostic issues for this genus. Hawaii: A survey was conducted to collect Heterorhabditis and Steinernema spp. in Hawaiian soils. Documenting natural populations strengthens the case for the introduction and use EPN for biological control. Collects for Heterorhabditis populations focused in coastal areas and for Steinernema at inland sites. Heterorhabditis was baited with Galleria mellonella and Steinernema with Tenebrio molitor. Nearly a quarter of sites contained some type of entomopathogenic nematode based on morphological observation. ITS rDNA sequences confirmed Heterorhabditis indica, Heterorhabditis sp. and Heterorhabditoides. Most Steinernema isolates are small and morphologically in the S. carpocaspi group. Mississippi: A developmental genomics analysis of candidate soybean genes involved in the resistant reaction occurring between Heterodera glycines (soybean cyst nematode [SCN]) and Glycine max (soybean), involving Jian Jiang, Gary W. Lawrence, Vincent P. Klink is underway. Worldwide, soybean cyst nematode (SCN) infection of soybean crops results in annual production losses ranging from 7 to 10%. The multinucleated structure nematode feeding structure (syncytium) that forms during infection affords the unique opportunity to isolate cells undergoing the reaction and understand its accompanying gene expression program. Cells undergoing the incompatible reaction have been isolated and analyzed for gene expression occurring exclusively in the incompatible reaction. This identified list of approximately 1,700 genes served as a pool of candidate resistance genes to study using molecular techniques. Highly induced expression of these candidate genes in the susceptible genotype (G. max[Williams 82/PI 518671), in several cases, resulted in identification of genes whose expression suppressed SCN infection. This result demonstrated the efficacy of the approach. Currently, we are testing dozens of candidate resistance genes. Candidate resistance gene analysis in relation to the defense response occurring between the soybean cyst nematode (SCN) and soybean is being conducted by Shankar R. Pant, Gary W. Lawrence, and Vincent P. Klink. Soybean cyst nematode (SCN) infection of soybean crops results in annual production losses ranging from 7 to 10%, worldwide, annually. In experiments that identified candidate resistance genes that are expressed specifically in the multinucleated syncytium undergoing the process of defense, supporting experiments resulted in the identification of the involvement of a Syntaxin 31 homolog (Gm-SYP38) functioning in defense. Furthermore, we identified ENHANCED DISEASE SUSCEPTIBILITY1 (EDS1) and NONEXPRESSOR OF PR1 (NPR1) that function in salicylic acid (SA) signaling as playing a role in defense. Additional experiments revealed the involvement of BOTRYTIS INDUCED KINASE 1 homolog (Gm-BIK1-6) and the hemicellulose-modifying, xyloglucan endotransglycosylase/hydrolase (XTH) functioning in defense. Currently, we are employing overexpression and RNA interference to better understand the basis of resistance. Nebraska: We have continued to work in three areas of DNA barcoding for nematode identification. The first area has been in the development of primers and PCR amplification conditions that increase the taxonomic range of molecular diagnostics. Currently separate primer sets are used for cyst (Heterodera), root-knot (Meloidogyne), and ring (Criconematidae) nematodes. We have modified existing primer sets and conditions to include amplification of false root-knot nematode (Nacobbus) and lesion nematode (Pratylenchus). The new primer sets are still being tested. The second area of DNA barcoding development is the analysis of multiple isolates of species for determination of within and between species discriminations. This is an extremely important component of DNA identification based on barcodes and one that has been underdeveloped in most existing assays. Geographic range and sampling density are the two components of scale that we are trying to optimize. We are testing two groups with an expanded sampling design, Pratylenchus as a representative of an endoparasitic nematode that may be easily spread by movement of root-stock and Criconematidae, as an ectoparasitic nematode that may not move as readily as endoparasities. For Pratylenchus we currently have over 1,500 specimens examined microscopically representing 11 U.S. states and three Canadian provinces. For Criconematidae we have representatives from over 40 states and three continents. These isolates are being used to test the metrics of within and between species/group genetic distance, the degree of overlap between the distance values and the genetic distance to the nearest-neighbor specimen in another group. The final area of investigation also concerns barcode analysis. Numerous algorithms are currently used for assignment of specimens to species or OTUs (operational taxonomic units). Some of the algorithms are based on genetic distance, others incorporate a tree-building step, and several recently developed approaches apply likelihood and coalescent analyses. These are being tested using our plant-parasitic nematode datasets. New Mexico: The project to improve capability for using molecular tools to differentiate Ditylenchus species of regulatory concern from each other and from benign soil-inhabiting species that began in 2013 was completed. DNA was extracted from an additional 450 specimens from plant (alfalfa, garlic, onion, Phlox sp., and potato) and soil samples obtained from ID, MI, NE, NM, OR, SC and WA, Canada, and the Republic of Georgia. High-quality ~1kb 18S rDNA sequences were generated using modified primer sets, as were sequences for ITS1 and ITS2, and compared among the new specimens and those previously reported in GenBank. Ditylenchus spp. recovered from plant material were found to be largely clonal, particularly within a location, compared to those recovered from soil, which provided greater DNA sequence variation. The 18S sequence was reliable for separating D. dipsaci from D. destructior and both species from non-pathogenic soil-inhabiting species. However, inclusion of the ITS sequence with the 18S region was necessary to reliably separate D. gigas from D. dipsaci. Oregon: Biological characterization of Globodera ellingtonae: On April 28, 2008, a field at the Oregon State University Powell Butte Farm that was planted to potato in 2007 was sampled and found to contain cysts of Globodera which were morphologically and molecularly distinct from golden potato cyst (G. rostochiensis), pale potato cyst (G. pallida), and tobacco cyst (G. tobaccum) nematodes. Subsequently, in August and September of 2008 cysts matching the characteristics of those found at Powell Butte were recovered from two grower potato fields in Idaho. These nematodes were later (2012) described as a new species, G. ellingtonae. Very little is known about the biological properties of this species and its potential pathogenicity to potato or other crop plants. Lab, greenhouse, and field studies have begun to help characterize the biology of G. ellingtonae. In an effort to relate G. ellingtonae to pathotypes of G. rostochiensis and G. pallida, several potato varieties used as differential lines to define Globodera pathotypes in Europe were inoculated with 2,500 eggs of G. ellingtonae. Plants were grown for three months and cysts and eggs were counted to determine if each line was susceptible or resistant. ‘Cycloon’ which is resistant to G. rostochiensis Ro1 and G. pallida Pa2 was susceptible to G. ellingtonae. ‘Avondale’, ‘Banba’, ‘Galactica’, ‘Missaukee’ and ‘Slaney’ which are resistant to G. rostochiensis Ro1 but susceptible to G. pallida were resistant to G. ellingtonae. The differential line 65.346/129, resistant to all pathotypes of G. rostochiensis and susceptible to all pathotypes of G. pallida, was susceptible to G. ellingtonae. P55/7, resistant to G. pallida Pa1 was susceptible to G. ellingtonae, and V-15-71, resistant to G. rostochiensis Ro1 and Ro2 was susceptible to G. ellingtonae. Therefore, the reaction of several potato lines to G. ellingtonae often differed from that of pathotypes of G. rostochiensis or G. pallida suggesting that the pathotype is unique. This further supports the elevation of G. ellingtonae as a new species. During 2012, holes 6 inches deep and 6 inches in diameter were augured in field plots, filled with soil infested with 0, 5, 10, 20, or 40 eggs/g of G. ellingtonae, (n=7) and planted to ‘Russet Burbank’ potato on May 17. At harvest (September 13) average egg densities were 5, 43, 62, 92, and 114 eggs/g in the 0, 5, 10, 20, or 40 eggs/g inoculum levels, respectively, but there were no differences in tuber weight between the different inoculum levels. During 2013, holes 6 inches deep and 12 inches in diameter were augured in field plots, filled with soil infested with 0, 5, 10, 20, 40, or 80 eggs/g of G. ellingtonae, (n=5) and planted to potato ‘Desiree’ or ‘Russet Burbank’ on May 27. At harvest (September 23) average egg densities in the ‘Desiree’ plots were 1, 148, 222, 209, 317, and 267 eggs/g; and in the ‘Burbank’ plots were 2, 121, 151, 343, 261, and 127 eggs/g in the 0, 5, 10, 20, 40 or 80 eggs/g inoculum levels, respectively. There were no significant differences in ‘Desiree’ top growth or tuber weight between different egg densities. Top growth of ‘Russet Burbank’ appeared less in the 80 egg/g treatment but the difference was not significant. There were no differences of any of the individual tuber size classes or in total tuber weight between the different inoculum levels. Although total tuber weight in the 80 egg/g treatment was 51% that in the non-inoculated plots, this difference was not significant (P<0.05). Total yield in this trial may have been reduced from damage due to a hailstorm on August 16. Objective 2: Determine nematode adaptation processes to hosts, agro-ecosystems and environments. California, Davis: Dispersal is a key step for movement of plant pathogens and for the development of disease. Plant pathogens such as fungi and plant-parasitic nematodes disperse by water, wind, soil, contaminated field equipment, plant stock, humans, and insects. The brown garden snail, Helix aspersa, is a pest found in many agricultural settings. Snails feed by grazing on substrates they pass over, and in the process they consume an array of microbes including plant pathogens and may move them to new locations. To determine whether viable pathogen propagules can transit the snail digestive tract, we tested two fungal pathogens, Fusarium circinatum and Rhizoctonia solani and three plant-parasitic nematodes, Meloidoygne hapla, Ditylenchus dipsaci, and Aphelenchoides ritzemabosi. We fed snails fungal mycelium on filter paper. Ninety percent of tested snails defecated viable fungal propagules for up to 9 days. The plant-parasitic nematodes were fed to snails on an infested carrot disc. Active nematodes were recovered from feces starting at 24 hours and continuing up to 8 days. These experiments demonstrate for the first time that viable plant-parasitic nematodes can transit the digestive tract of brown garden snails. We also determined that second-stage juveniles (J2s) of the root-knot nematode species Meloidogyne hapla and M. incognita after consumption by H. aspersa can emerge from snail fecal pellets to successfully infect and reproduce on tomato (Lycopersicon esculentum) host plants. Individual carrot discs (1.75 g) were infested with an aqueous suspension of 1000 J2s of the two nematode species. The fecal pellets from snails fed a single infested carrot disc were collected on each of 5 days subsequent to snail feeding, and placed at the base of tomato seedlings held in growth chambers. After 2 months the tomato plants showed infection with galled roots, egg masses, and substantial reproduction with the ratio of final J2s from roots to initial J2s fed to snails (i.e., Pf/Pi) of 3.5 and 9.1 for M. hapla and M. incognita respectively. Tomato shoot weights were significantly reduced by infection with both M. hapla and M. incognita. Helix aspersa ingested root-knot nematode J2s and those J2s survived ingestion and passage over the snail radula, transit through the alimentary canal, deposition in fecal pellets, emergence from pellets, movement into the soil, and then successfully locating and infecting host roots. The control measures for root-knot nematodes may need to include a greater emphasis in controlling snails as possible vectors of disease. The passive, phoretic dispersal of plant-parasitic nematodes through endozoochory in H. aspersa may represent a significant, unrecognized means of nematode invasion and colonization within and among agricultural fields and other environments. Brown garden snails are capable of moving faster and further than plant-parasitic nematodes, which suggests that snails may be an important element in plant-parasitic nematode dispersal, dissemination, and disease transmission. California, Riverside (Hyman): Two improved bioassays were developed to establish infectivity baselines for selection experiments using mermithid nematode variants. Comparative infectivity of Romanomermis iyengari, Romanomermis culicivorax and Strelkovimermis spiculatus using larvae of three mosquito spp. Aedes sierrensis, Aedes aegypti and Culex pipiens were evaluated with “plate” and “tray” bioassays at selected intensity of infections. Using the “plate” bioassay, single mosquito larvae were immersed in 5 ml of water within individual depressions of 12-well, polystyrene tissue culture plates. One, three, or five preparasitic juveniles (J2) were added to each well. In the “tray” bioassay, polyethylene trays containing 500 ml water and 100 mosquito larvae were exposed to 500 (5:1, nematode:insect host) or 1,000 (10:1) J2s. Percentage infection (PINF, infectivity) and intensity of infection (IINF, # nematodes per infected larvae) number were determined only after emergence of post-parasitic J3 juveniles. Under the bioassay conditions, all three species of nematodes resulted in infections in all mosquito hosts, but R. iyengari exhibited better effectiveness in the parasitism of mosquito larvae. The three species of mosquitoes presented high levels of susceptibility to each of the three species of nematodes, but in general C. pipiens and A. sierrensis were slightly more susceptible than A. aegyti. The “plate” bioassay was more efficient in measurement of infectivity of the mermithid species and in establishing baseline characteristics for these mosquito-parasitic nematodes. The "tray" bioassay was an effective bioassay for large cohorts of both infective juveniles and host larvae and, potential for field interactions. Idaho: The interaction of Verticillium dahliae and lesion nematodes showed an additive effect on mint hay yield in the greenhouse experiment carried out in 2013. Verticillium dahliae alone caused 44 % damage in mint hay yield. Root lesion nematode Pratylenchus neglectus was also pathogenic to mint. A 23 and 46 % reduction on mint hay were caused by P. neglectus alone or in combination with V. dahliae, respectively. P. penetrans alone caused 44 % yield reduction of mint hay while combination of P. penetrans and V. dahliae killed almost all plants (98%). Population of P. penetrans is increased by 41 fold which indicates that mint is an excellent host for P. penetrans. The experiment was repeated this year to confirm the first years’ data. Mint are already cut for fourth time and harvested. The data from repeated experiment confirmed the first years’ result. The survey carried out in mint growing areas in Nampa, Wilder, Caldwell, Marsing, Fruitland, Nyssa, Emmett, and Ontario in 2011, 2012, 2013 and 2014 showed root lesion nematode in 155 samples, root-knot nematode in 48 samples, spiral nematode in 108 samples, and pin nematode in 142 samples, sheath nematode was found in 3 samples, dagger nematode in 5, ring nematode in 7, and stubby root nematode in 12 samples. County wise, 149 samples were surveyed in Canyon, 12 in Malheur, and two in Gem. In total, 163 samples have been analyzed until now. Nematode were recorded in the range of 10-2750 for root lesion nematode, 10-6900 for root knot nematode, 10-6550 for spiral nematode, and 30-67200 for pin nematode per 500 cc soil sample. Michigan: Developing management practices that reduce the impact of sugar beet cyst nematode (SBCN) and understanding soil biology and food web structure to improve soil health are two of the critical research priorities for the Michigan Sugar Beet Industry (MSBI). In order to develop integrated SBCN and soil health management, the relationships among the rotation crops, SBCN, other PPN, beneficial nematodes, soil health, and soil types need to be understood. Using sugar beet varieties (EL53, EL57, EL59, EL61 and EL64) from the USDA laboratory at MSU, we investigated how these varieties affect all nematodes in different soil types and soil health in 2012 and 2013. Soybean and corn are added as controls for production systems. Soil samples were collected every 4-6 weeks during the growing season and nematodes extracted and identified to herbivore (cyst and other PPN), bacteriovore, fungivore, predator and omnivore trophic groups. Trophic group data were processed to extract bio-ecological, nutrient cycling potential, and soil food web structure. Preliminary analyses of pre-plant and harvest data showed 24 and 8 cysts/100 cc of soil and 17 and 19 herbivore genera present in 2012 and 2013, respectively, suggesting potential problems than the target nematode, SBCN. While the levels of cysts were higher in 2012 than in 2013, statistically low numbers of cysts in 2013 were recovered in three sugar beet varieties (EL53, EL57 and EL59) and corn. Significant low population density of both cyst and root-lesion nematode were observed in the plots with EL59 variety. Soil food web structure, as described by nematode community analysis, varied within crops, suggesting different crops have different impact on soil health however high enrichment levels were observed in the plots grown with soybean and corn. A combination of cyst and other PPN population dynamics, and soil food web data support the hypothesis that there are distinct interactions among the crops, SBCN and soil conditions. New Mexico: Dr. Jonathan Eisenback is finalizing morphological description of the new species of Meloidogyne that was recovered from purple nutsedges in two field locations in Dona Ana County, NM in 2012. In collaboration with Dr. Stephen Hanson (EPPWS Department, NMSU), ribosomal DNA 18S, ITS1, and ITS2 and mitochondrial COII gene segments for this nematode have been sequenced. Studies conducted in 2013 were repeated in 2014 to determine the host suitability of major crops produced in southern NM to this nematode. Cotton, chile pepper, corn, tomato, and onion failed to host the new nematode in both studies. Low to moderate nematode reproduction occurred on one or more of the sorghum, winter rye, or alfalfa plants tested during the second study, whereas no reproduction was observed in the 2013 study. Wheat, bentgrass, and perennial ryegrass showed variability in response in both tests, with some plant within each species being non-host, while others ranged from poor hosts (0 < RF < 1) to hosts (1 < RF < 10). Yellow nutsedge (Cyperus esculentus) and purple nutsedge (C. rotundus) were the best hosts for this nematode, resulting in average population increases of over 26-fold on yellow nutsedge and over 5-fold on purple nutsedge after 45 days. A study established in late 2013 to determine the effect of previous crops on subsequent M. incognita reproduction on yellow and purple nutsedges was harvested and repetition of the study begun in 2014. These studies are necessary to accurately predict how different cropping scenarios will impact root-knot nematode carryover to future crops from perennial weeds. Both nutsedges were inoculated with eggs recovered from cotton, chile, or corn (the major summer annual crops in southern NM). Inoculum from tomato was also included to allow extrapolation to results from previous studies. The experiments are harvested upon accumulation of 750 heat units (DD24 = one life cycle for the nematode), and eggs extracted from roots. Data will be used to calculate the crop-specific M. incognita carryover potentials for both nutsedge species for use in developing IPM management recommendations for the nutsedge/nematode pest complex. Oregon: Globodera spp. eggs go through a diapause which remains dormant until favorable hatching conditions are reached. Because of the regulatory concerns with cyst nematodes, it is often only possible to rear eggs for research in the greenhouse. However, hatch is often lower for greenhouse-produced eggs than for eggs obtained from the field. The goal of this research was to determine storage conditions for G. ellingtonae eggs produced in the greenhouse that would increase percentage hatch. Over three years, G. ellingtonae greenhouse-produced eggs were stored in different environments (-20°C, 4°C, room temperature, and the field) in either dry or moist soil. Percentage hatch after exposure to the different environments was determined in potato root diffusate. Across two experiments, field-produced eggs had higher hatch rates (65.2%) than greenhouse-produced eggs (10.4%). Temperature did not have an appreciable influence on hatch of eggs stored dry in two experiments (2.8 to 8.4% and 3.8 to 8.6%), but hatch of eggs stored in moist soil was significantly higher than in dry soil at all temperatures except -20°C (26.8% and 28.7%). However, the ability of G. ellingtonae greenhouse-, microplot-, and field-produced eggs to reproduce on potato in field microplots was not different. While it may not be possible to produce G. ellingtonae eggs in the greenhouse that have the magnitude of hatch as those produced in the field, hatching can be greatly increased by storing eggs in moist soil at either 4°C or room temperature. Objective 3: Develop and assess nematode management strategies in agricultural production systems. Arkansas: Annually I test all new soybean entries submitted to the Arkansas Soybean Variety Testing Trials for reniform nematode reproduction. In 2014 this was 184 entries. A single plant of each variety was grown in each of 5 separate 4 inch diameter clay pots. As a germinated seedling was transplanted into each pot it was inoculated with 2,000 vermiform reniform nematodes. Three resistant varieties (Forrest, Hartwig and Anand) used as resistant checks while Braxton was used as a susceptible check and one pot in each rep was inoculated and left fallow as a check on reniform survival. The test was terminated after 12 weeks. Of the 184 entries there were 13 (Armor AX4520, Armor AX4450, Asgrow AG5535 GENRR2Y, Delta Grow DG4940RR, Delta Grow DG5230GENRR2Y, Dyna-Gro S52RY75, Eagle Seed ES5335RY, LG Seeds C5252R2, MPG 5214NRR, Mycogen X54522NR2, S11-20356, S11-20124, Willcross WX 2524N) that did not differ in reproduction than the resistant varieties Hartwig and Anand) and have probable rotation usefulness. Similar tests were also done for 179 varieties and breeding lines submitted by Southern Public Soybean Breeders from Arkansas, Illinois (Southern Illinois), Missouri, South Carolina (Clemson) and the USDA (Jackson, TN). Of the 179 entries, 31 were as resistant as the resistant varieties Anand and Hartwig. The specifics including statistical analysis will be reported at the Beltwide Cotton Conferences in San Antonio in January 2015. A rotation using two new varieties of resistant soybean (one moderately and the other highly resistant and a susceptible (9 treatments)) was initiated in a field with a high population of race 5 SCN in 2012. In 2012 the at plant egg count per 100 CC soil averaged 4022 overall and at harvest averaged 2480 in the highly resistant (HR) line, 6233 in the moderately resistant (MR) and 9793 (S) in the susceptible line. The yield was 35.83 BU/A for the HR, 32.90 for the MR, and 28.97 for the S. In 2013 the eggs counts were not different at plant or harvest, the treatment yields following the HR averaged 39.6, MR 36.4, and S 32.5 BU/A. In 2014 the eggs counts were not different at plant or harvest, the treatment yields following the HR averaged 36.8, MR 34.3, and S 33.7 BU/A. At $15 / BU using the moderately resistant would have gained $58.5 / acre while the highly resistant would have gained $102 in 2012. In 2013 the rotations yielded equal whereas the HR /HR yielded 39 B/A, the MR/MR = 39.6 B/A and the S/S = 32.3 B/A with the resistant increase over $100 or more. In 2014 the HR /HR yielded 39B/A, the MR/MR = 36 B/A and the S/S = 33.8 B/A with the resistant increase over $33 or more California, Riverside (Roberts): In cowpea (Vigna unguiculata), a high density map containing more than 1100 EST-derived SNP markers and a high-throughput SNP genotyping platform based on the KASP technology were used to refine the genomic locations of QTLs determining resistance to root-knot nematodes and Fusarium wilt in cowpea inbred populations. Multiple screens with M. incognita and M. javanica isolates were used to develop advanced blackeye cowpea breeding lines with dual resistance to both nematodes. Replicated field trials in Tustin, CA and Parlier, CA on infested field plots showed significantly higher grain yields of lines carrying multiple resistance genes, compared to lines with a single resistance gene or no resistance genes, confirming the protective value of the stacked resistance genes. Hawaii: Radopholus similis is an endoparasitic migratory nematode that causes anthurium decline. Our objective was to evaluate the efficacy of spinosad, spirotetramat, and thiophanate-methyl for the control of R. similis in anthurium. Plants were weighed, transplanted into 15-cm-d clay pots filled with cinders and inoculated with 3000 R. similis. Six months later, treatments of 4.8 mg spinosad, 4.8 mg spirotetramat, 0.01 g thiophanate-methyl, or nothing were applied to the leaves in 50 ml water. Two months later, leaf number and leaf area of the youngest mature leaf were recorded. The average number of leaves on uninoculated plants treated with spinosad, spirotetramat, thiophanate-methyl, or water was 9, 10, 9, and 10, respectively. The average number of leaves on inoculated plants treated with spinosad, spirotetramat, thiophanate-metyl, or water was 11, 9, 11, and 9, respectively. The average leaf area for uninoculated plants treated with spinosad, spirotetramat, thiophanate-methyl, or water was 90.6, 101.2, 96.9, and 77.5 cm2, respectively, On inoculated plants treated with spinosad, spirotetramat, thiophanate-methyl, or water leaf area was 91.7, 84.5, 76.2, and 92.5 cm2, respectively. Anthurium treated with spinosad, thiophanate-methyl, spirotetramat, or water increased weight 17, 16, 17, and 14 fold whereas those inoculated with nematodes increased 16, 14, 14, and 15 fold, respectively. Spinosad controls anthurium decline and maybe a treatment option for growers. Reniform nematode, Rotylenchulus reniformis is an economically important plant-parasitic nematode of pineapple. This pathogen can reduce pineapple marketable yield by 26.8-50%. Lack of host resistance in pineapple raises a need for nematode management. Spirotetramat is a group 23 insecticide, a lipid biosynthesis inhibitor with ambimobile translocation. Lipids play a significant physiological role in embryogenesis and molting of plant-parasitic nematodes therefore inhibition of lipid biosynthesis may provide a critical management tool. The objective of this study was to determine if spirotetramat is active against reniform nematode. Greenhouse bioassays were conducted where pineapple crowns were planted in steam-sterilized soil in 20-cm-d biodegradable pots. Approximately 3,000 R. reniformis eggs were inoculated onto 4-month-old plants. Four levels (0, 88, 100 and 175 g a.i/ha) of spirotetramat were applied 1 month post inoculation. Plants were harvested 6 months later for shoot and root weights, and a 250 cm3 soil sample collected. Data were normalized by log10 transformation and subjected to analysis of variance. The maximum proportion of nematode reduction was 64 % at 88 g a.i. spirotetramat/ha rate with a corresponding reduction in dry root weight of 14.4% and an increase in shoot weight of 1.4% compared to the water treatment control. The 88 g a.i. spirotetramant/ha rate gave promising reductions in nematode numbers while promoting plant growth. Idaho: Microplot study to evaluate of new nematicides (Movento and Velum) on pin nematode including few lesion and root-knot nematodes has been done. Mint were transplanted in the microplot buckets and chemical applied at planting and post plant. Each bucket was divided into two compartments, one compartment was harvested this year and another will be continued for next year. This year data showed significant decline on pin nematode population in all treatments as compared to untreated control. The effect of non-ionic wetting agents (soil surfactant) on movement of Mocap has been evaluated in the greenhouse. The experiment was repeated to confirm the first experiment’s results under greenhouse condition. Data showed no significant effect of the products on the Mocap movement. Michigan: Use of crop rotation, cover crops, reduced tillage and soil health soil health (organic matter, biological, physiochemical, nutritional and water holding properties) are cross-cutting priorities for many Michigan agricultural communities. While providing agronomic benefits, most cover and rotation crops are hosts to many plant-parasitic nematodes (PPN) that can cause economic loss. Moreover, all of the crops and tillage practices impact soil biology and soil health in many ways. Understanding how the agronomic practices impact soil biology and soil nutrient cycling is critical to soil health management across cropping systems. The long-term goal is to develop soil health management strategies that apply across cropping systems. We have initiated comparison of nematode community analysis and soil food web structures in different cropping systems. More specifically, how trophic group and colonizer-persister levels of beneficial nematodes relate to one another as well as with those of herbivores. The systems being studied include corn-soybean rotation under different till no-till to manage soybean cyst nematode, use of cover crops in sugar beet production system in different soil types, and carrot production. On-going are analyses to determine the relationship between soil food web function and structure in the different systems. Mississippi: Emergence of new nematicidal products. Several agricultural chemical companies are in the process of developing products designed for nematode control in row and vegetable crops (Table 1). Efficacy studies have been conducted with these products to determine their effect on nematode infestations of field crops. Table 1. Experimental and Existing Nematicidal Product by Company, Product and Application Method Company Product Application ADAMA MCW-2 – NIMITZ (fluensulfone) In-furrow spray AMVAC Counter 20G (Terbufos) In-furrow granular Thimet (Phorate) In-furrow granular Bayer Velum Total (Fluopyram + Imidacloprid) In-furrow spray Aeris seed applied system (Thiodicarb) Seed treatment Votivo (Bacillis firmis) Seed treatment DuPont Vydate L (Oxamyl) In-furrow spray Vydate C-LV (Oxamyl) Foliar spray Q8U80 In-furrow spray Monsanto Numbers (1-14) Seed treatment Marrone MBI- 38 In-furrow spray NuFarm Azadirachtin, Nematox, Senator Seed treatment Neem Oil, albendazole, Imidacloprid Syngenta Avicta Complete (abamectin) Seed treatment New Mexico: Studies were conducted on a golf course at the University of New Mexico to determine if severe damage to bentgrass greens resulting from high populations of Mesocriconema spp., Pratylenchus spp., and Longidorus breviannulatus could be successfully managed using current labeled rates of Avid® (2% abamectin). Maximum application of the product in 2013 reduced populations of Mesocrionema and L. breviannulatus below damage thresholds, but resurgence of Mesocriconema was observed in March 2014. Subsequent maximum application of Avid again reduced populations below the damage threshold by June, and populations remained below threshold in October. Pratylenchus populations were not managed well in these studies. However, damage symptoms on greens were no longer evident by June 2014, and usage was resumed.

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