Establishment of a virus-induced gene silencing system to study the effect of glycolate oxidases on oxalate metabolism in spinach

Abstract Spinach is a nutrient-rich leafy vegetable, but its high oxalate content can negatively impact mineral absorption during consumption. In this study, we utilize a tobacco rattle virus (TRV)-based virus-induced gene silencing (VIGS) to investigate the role of spinach glycolate oxidases (SoGLOs) in tissue oxalate accumulation. A small GLO gene family was identified and the expression profiles and the subcellular localization of SoGLO2 were characterized. TRV-mediated targeting of the SoGLO family reduced the expression of several SoGLO genes, especially SoGLO1 and SoGLO5 , which was associated with reduced oxalate accumulation. Our findings offer insights into a genetic strategy for manipulating oxalate accumulation to improve the nutritional quality of spinach.