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ARS Home » Midwest Area » Wooster, Ohio » Corn, Soybean and Wheat Quality Research » Research » Research Project #444744

Research Project: Leveraging Proteomics to Identify and Characterize Soybean Aphid Proteins and Effectors that Contribute to Virulence

Location: Corn, Soybean and Wheat Quality Research

Project Number: 5082-22000-002-009-S
Project Type: Non-Assistance Cooperative Agreement

Start Date: Sep 1, 2023
End Date: Mar 30, 2025

Our overall objective is to understand how the invasive soybean aphid overcomes host plant resistance. Objective 1 will leverage proteomics to identify proteins and effectors correlated with virulence. Objective 2 will focus on characterizing these effectors and other proteins of interest.

Objective 1. Comparative proteomics will be conducted to identify differences in protein expression profiles between the various soybean aphid biotypes. Protein will be extracted from soybean aphids of each biotype and prepared for further analysis through trypsin digestion, reduction, and alkylation. Proteins will be analyzed by high pressure liquid chromatography (HPLC) coupled to tandem mass spectrometry (MS). The resulting spectra will be analyzed to determine protein identity and differential protein expression among the biotypes, particularly which proteins are correlated with virulence. Proteomic analysis may be conducted on whole insect and/or insect saliva. Other proteomics approaches such as two-dimensional difference gel electrophoresis (2D-DIGE) may be explored. Objective 2. Proteins and genes of interest correlated with soybean aphid virulence identified in Objective 1 as well as through previous work by the Cooperator will be further characterized both functionally and bioinformatically. Delivery of RNAi or other inhibitors to the soybean aphid through artificial diet feeding will be explored. The expression of genes of interest will be knocked down using this artificial diet assay and the effect on soybean aphid virulence to resistant plants will be assessed. Other functional assays to knockdown or otherwise characterize genes and proteins of interest may be developed. Genes of interest will also be characterized bioinformatically through comparative genomics with other aphid species and proximity to transposons or other elements of interest.