Simultaneous silencing of GhFAD2-1 and GhFATB enhances the quality of cottonseed oil with high oleic acid

Cottonseed oil has become an important source of edible oil due to its significant cost advantage. However, there is a growing concern over its fatty acid composition and nutritional value. In Gossypium hirsutum, GhFAD2-1 and GhFATB encoding the microsomal oleate desaturase and palmitoyl-acyl carrier protein thioesterase, respectively, play critical roles in regulating the proportions of saturated and polyunsaturated fatty acids in cottonseed lipids. In this study, RNAi technology was used to simultaneously inhibit the expression levels of GhFAD2-1 and GhFATB to improve the quality of cottonseed oil by increasing oleic acid content. Transgenic cotton plants with reduced levels of both target genes were successfully generated. In mature seed kernels of transgenic plants, the content of oleic acid was 38.25%, accordingly increasing by 156.96%, while the content of palmitic acid and linoleic acid was 19.15% and 36.68%, decreasing by 21.28% and 33.92%, respectively, compared with that of the control. The total oil content in transgenic and control kernels was 22.48% and 29.83%, respectively. The reduced oil level in transgenic seeds was accompanied by a reduction in seed index, thereby causing disadvantageous effects on seed germination potentiality and seed vigor, particularly under cool stress conditions. Our results demonstrated the feasibility of simultaneous manipulation of multiple genes using RNAi technology and showed the important role of oil content in seed development and vigor. Our findings provide insight into the physiological significance of the fatty acid composition in cottonseeds.