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s from the Society of Nematologists Annual Meeting 2019 3 STRUCTURAL COMPOSITION OF SOIL MICROBIAL COMMUNITIES DO NOT SHIFT BASED ON NEMATODE MANAGEMENT METHOD Alake, Gideon1, P. Timper2, D. L. Wright3, L. W. Duncan4, W.T. Crow1, H.T. Alborn5, T. Mekete6, P. DiGennaro1. 1Dept. of Entomology and Nematology, University of Florida, FL 32611. 2USDA ARS, P.O. Box 748, Tifton, GA 31793. 3Dept. of Agronomy, North Florida Research and Education Center, University of Florida, Quincy, FL 32351. 4Entomology and Nematology Dept., Citrus Research and Education Center, University of Florida, Lake Alfred, FL 33850. 5USDA ARS, 1600/1700 Southwest 23rd Drive, Gainesville, FL 32608. 6Division of Plant Protection, Institute of Food Production and Sustainability, NIFA, 800 9th Street SW, Washington DC. The soil is a living system that acts as a critical biodiversity reservoir and is an essential basis of sustainable agriculture. Soil microbiota plays a crucial role in plant growth and nutrition, and pest management in agricultural production may influence soil biodiversity and shift the structure of soil microbial communities. Peanut, an economically important crop in the southeastern United States, faces an increasing threat from nematode pests. Current nematode management efforts primarily rely on chemical nematicides and resistant cultivars. Using high-throughput Illumina MiSeq, we evaluated structural responses of soil bacteria and fungi to nematode management approaches in peanut agroecosystems. We sampled fields where Telone II was used as a chemical nematode intervention, and fields that used non-chemical practices, such as cover cropping. We also sampled adjacent non-tilled soils to provide a baseline of the soil biodiversity for comparison. There were apparent differences in the composition of bacterial and fungal communities between chemical and non-chemical treated fields. The alpha-diversity was not statistically different between these treatments. Also, preand post-plant surveys did not show a measurable difference in microbial diversity. Our results suggest that the current chemical nematode management practice does not quantitatively reduce the diversity of bacteria and fungi. Increased sequencing resolution and improved primer design may significantly enhance our ability to detect specific bacteria or fungal structural shifts in response to nematode management. DEVELOPING REAL-TIME PCR ASSAYS TO DETECT AND QUANTIFY PRATYLENCHUS SCRIBNERI DIRECTLY IN DNA EXTRACTS FROM FIELD SOIL AND POTATO ROOTS. Arora, Deepika and G. P. Yan. North Dakota State University, Department of Plant Pathology, Fargo, ND 58108. Root-lesion nematodes (Pratylenchus spp.) affect crop production worldwide and several species including P. scribneri are detrimental to potato. Pratylenchus scribneri is a migratory endo-parasitic nematode which can infect potato plants and cause poor growth, yellowing of the foliage, and darkened necrotic lesions on roots. After its initial detection in 2016 from a potato field in North Dakota, P. scribneri was found in other fields in seven counties in North Dakota and one county in Minnesota. Till date, there are no published procedures in the U.S. for detecting and quantifying this nematode using DNA extracted directly from field soil or potato roots. Due to its high virulence and wide host range, it is prudent to have a fast, sensitive and specific molecular diagnostic method. Here we report on the development of SYBR Green I-based qPCRassays to detect and quantify P. scribneri in field soil and potato roots. The pair of primers used in this studywas designed from the ITS region of the 28S rDNA. Specificity of primers was evaluated by in silico analysis using 26 other important species of Pratylenchus genus and through qPCR using DNA extracted from P. scribneri along with 18 non-target nematodes and nematode communities, with no resulting cross reactions. Standard curves for soil and root assays were obtained by inoculating sterilized soil or non-infected potato roots with varying number of P. scribneri followed by DNA extraction and qPCR. The standard curves were supported by high amplification efficiency (E) and R2 values between the numbers of P. scribneri artificially added to soil or roots and the numbers quantified using qPCR (E = 94%, R2 = 0.996 for soil assay; E = 103%, R2 = 0.988 for root assay). To evaluate the minimum amount of DNA template that could be detected and quantified by the qPCR assays, a serial dilution of template DNA derived from sterilized soil or potato roots inoculated with a single P. scribneri individual was made. The dilution series ranged from one nematode down to an equivalent of 1/512 of a nematode and the assays could detect 1/128 equivalents of the DNA of one nematode. The developed qPCR assays were evaluated by comparing the numbers of target nematode in 20 randomly selected field soil samples and 20 root samples, determined by traditional microscopic counting, with the numbers determined by qPCR. High correlations (R2 = 0.77 for field soil samples; R2 = 0.73 for plant root samples) observed between the numbers from the traditional and molecular methods suggest that the assays could be used to enumerate P. scribneri numbers directly in infested field soils and potato roots. Accurate detection and quantification of P. scribneri in fields are critical for designing effective measures to control this nematode pest. The assays developed in this study are straightforward and require no expertise in nematode taxonomy and morphology, and may serve as a valuable molecular diagnostic tool for management of P. scribneri. FROM PARASITES TO FREE-LIVING WORMS: THE SEARCH FOR MOLECULAR UNDERPINNINGS OF CHEMOTAXIS BEHAVIORS. Baiocchi, Tiffany1, N. Mercado1, K. Kin2, B. Strickhouser-Monzon2, K. Anesko2, P. Robles2, A. Dillman3. 1Biochemistry Dept., University of California Riverside, Riverside, CA 92521. 2Biology Dept., University of California Riverside, Riverside, CA 92521. 3Nematology Dept., University of California Riverside, Riverside, CA 92521. Entomopathogenic nematodes (EPNs) are a special group of nematodes that infect and kill insect pests and are used as biological control agents. Our work identified the odor 3-methly-2-buten-1-ol (more commonly known as prenol), which is associated with insect cadavers that have been infected by EPNs and are resource-deficient. Prenol induces repulsion in EPN species, but surprisingly elicits attraction in the free-living species and model organism Caenorhabditis elegans. Through use of this odor and a newly-emerged resource: The C. elegans Natural Diversity Resource, we have leveraged natural variation in wild isolates of C. elegans with genome wide association techniques to identify at least two genes that influence chemotactic behavioral responses to prenol. The goal of this work has been to improve our understanding of nematode behaviors at the molecular level and leverage these findings to better understand what may drive behaviors

Abstracts from The Society of Nematologists Annual Meeting 2019