Location: Mycology and Nematology Genetic Diversity and Biology Laboratory
Project Number: 8042-22000-322-018-I
Project Type: Interagency Reimbursable Agreement
Start Date: Jun 1, 2025
End Date: May 31, 2026
Objective:
Plant-parasitic nematodes pose significant threats to global agriculture, causing estimated losses of $118 billion annually. Root lesion nematodes (RLN, Pratylenchus spp.), migratory endoparasites, are ranked third in economic impact. Several Pratylenchus species are listed as U.S. regulated plant pest by APHIS. Among them, Pratylenchus fallax has been found in limited areas of the country. The primary objective of this proposal is to enhance molecular tools for identifying and distinguishing RLN, particularly P. fallax from other Pratylenchus spp., through advanced genomic analysis. Currently, whole genome assemblies for Pratylenchus species are still very limited. Whole-genome sequencing of P. fallax will enable us to identify unique targets and develop precise molecular assays for this species, such as the recombinase polymerase amplification (RPA) method. This approach will significantly increase U.S. capabilities to detect P. fallax at point-of-care units, thereby supporting domestic surveys and detection at ports of entry.
Approach:
Objective 1: Collection and Preparation of RLN populations
Nematode pure cultures will be propagated in sterile in vitro cultures of corn roots (Zea mays L. cv. Iochief) maintained in MS medium. DNA extraction from multiple nematode specimens will follow the proteinase K protocol. Initial amplification of ribosomal RNA and mitochondrial genes will be performed using universal primers with the Taq Core Kit (Qiagen) or DreamTaq Green PCR Master Mix (ThermoFisher Scientific) to molecularly validate species identification.