Method for Analyzing CAZyme Activity from Whole-Body and Saliva Proteome of Russian Wheat Aphid.

Advanced proteome extraction and characterization techniques for Diuraphis noxia (Russian Wheat Aphid: RWA) enable precise biochemical profiling, where high-quality proteome extraction is critical. This chapter introduces an optimized proteome extraction method for comprehensive profiling of D. noxia, enabling analysis of whole-body and saliva proteomes to advance biochemical and functional characterization. The efficacy of two optimized proteome extraction methods was compared: a nondestructive artificial saliva extraction and a destructive whole-body extraction. The proteome extract concentration was quantified, and Carbohydrate-Active Enzymes (CAZymes) assays were performed to identify and quantify soluble sugar-degrading enzymes and cell wall-degrading enzymes. RWA is a phloem feeder, and this chapter demonstrates that it utilizes an enzyme cocktail consisting of invertase, trehalase, amyloglucosidase, and amylase during feeding. RWA is also equipped with cell wall-degrading enzymes in its proteome, such as cellulases and hemicellulases, which break down the cell wall, allowing the stylet to access the photoassimilates in phloem.