1a. Objectives (from AD-416):
Objective 1: Identify and describe invasive and emerging plant-parasitic nematodes from forage and biomass legumes, turf, grasses, and associated rotation crops - including species of quarantine significance - to enable specific and timely management of these pests. (NP215 1B.4, 1B.5) Objective 2: Improve molecular methodologies for the identification and classification of nematodes from forage legumes and grasses in order to better predict and respond to nematode outbreaks and to improve detection and diagnostic methodologies to distinguish closely related or morphologically similar species. (NP215 1B.4, 1B.5, 3B) Subobjective 2A: Barcodes and markers for improved molecular phylogenies. Subobjective 2B: New molecular diagnostic assays. Subobjective 2C: DNA recovery and barcoding from formalin-fixed nematodes.
1b. Approach (from AD-416):
1. Nematodes that infect alfalfa and other forage legumes, grasses, rotation crops, or potential weed hosts will be identified using molecular markers including ribosomal, mitochondrial, Hsp90, and other nuclear genes. 2. New diagnostic assays, including RFLPs, and conventional or real-time PCR assays will be developed for cyst, root-knot, lesion, and stem nematodes or others affecting these plants. 3. Molecular phylogenetic analysis will be performed using DNA sequence. alignment programs such as CLUSTAL and MAFFT and tree building methods based on distance, parsimony, and maximum likelihood methods. 4. Molecular information from the diagnostic work will be integrated with morphological data and information regarding biogeography, pathogenicity, and host range to generate new and improved phylogenetic schemes. 5. Methods to neutralize or repair DNA damage caused by specimen fixation will be combined with PCR and DNA sequencing in order to identify ways of recovering DNA information from degraded specimens.
3. Progress Report:
This new project replaces 8042-22000-284-00D, "Enhanced Alfalfa and Forage Productivity Through Molecular Detection and Characterization of Plant Nematodes" and the bridge project 8042-22000-307-00D of the same name. The final report for 8042-22000-284-00D includes accomplishments achieved over the life of the project; the report for 8042-22000-307-00D includes progress and accomplishments from the interim period until this project was implemented.