Location: Mycology and Nematology Genetic Diversity and Biology Laboratory
Project Number: 8042-22000-322-004-S
Project Type: Non-Assistance Cooperative Agreement
Start Date: Jun 1, 2023
End Date: May 31, 2026
Objective:
Cooperator to perform micro-aspiration of gland cell mRNA of mixed stages of the beech leaf disease nematode and generate nematode gland cell transcriptome libraries. ARS PI in Beltsville will generate de novo transcriptome assemblies of the gland’s libraries and will conduct gene annotation and identification of candidate parasitism-related genes. This information will produce a valuable and integrative transcriptome resource for this new emergent plant-parasitic nematodes and will generate an extensive catalogue of parasitism-related genes for a foliar nematode.
Approach:
Pools of gland cells from mixed nematode stages will be isolated and harvested via micro-aspiration by cooperator. Gland cell will be processed for total RNA extraction and RNA-seq library construction, using a minimum of three biological replicates. Gland cell libraries will be used for de novo transcriptome assemblies and for prediction of nematode secreted protein candidates. Gland cell expression of a minimum of 50 genes will be performed by in situ hybridization by the ARS PI. Data will be analyzed and compiled to provide an accurate list of parasitism-effectors genes for the beech leaf disease nematode.
