Castor (Ricinus communis L.) seeds produce abundant ricinoleic acid during seed maturation, which is important for plant development and human demands. Ricinoleic acid, as a unique hydroxy fatty acid (HFA), possesses a distinct bond structure that could be used as a substitute for fossil fuels. Here, we identified all homologous genes related to glycolysis, hydroxy fatty acid biosynthesis, and triacylglycerol (TAG) accumulation in castor seeds. Furthermore, we investigated their expression patterns globally during five seed development stages. We characterized a total of 66 genes involved in the glycolysis pathway, with the majority exhibiting higher expression levels during the early stage of castor bean seed development. This metabolic process provided abundant acetyl-CoA for fatty acid (FA) biosynthesis. Subsequently, we identified 82 genes involved in the processes of de novo FA biosynthesis and TAG assembly, with the majority exhibiting high expression levels during the middle or late stages. In addition, we examined the expression patterns of the transcription factors involved in carbohydrate and oil metabolism. For instance, RcMYB73 and RcERF72 exhibited high expression levels during the early stage, whereas RcWRI1, RcABI3, and RcbZIP67 showed relatively higher expression levels during the middle and late stages, indicating their crucial roles in seed development and oil accumulation. Our study suggests that the high HFA production in castor seeds is attributed to the interaction of multiple genes from sugar transportation to lipid droplet packaging. Therefore, this research comprehensively characterizes all the genes related to glycolysis, fatty acid biosynthesis, and triacylglycerol (TAG) accumulation in the castor and provides novel insight into exploring the genetic mechanisms underlying seed oil accumulation in the endosperm of castor beans.

Oil-tea (Camellia oleifera), a typical oilseed tree, produces high-quality edible vegetable oils that contain rich unsaturated fatty acids and diverse lipid-soluble active compounds such as squalene. Although squalene biosynthesis and its molecular regulation have been studied in several plants, the molecular mechanisms underlying squalene biosynthesis in oil-tea seeds remain uncertain. We investigated and determined squalene accumulation with seed development. We conducted comparative transcriptomic analyses using the RNA-seq technique at the early, fast biosynthesis, and late stages of squalene accumulation with oil-tea seed development and identified 13 squalene biosynthesis key enzyme genes (such as CoHMGR_4, CoAACT_2, CoFPS_1, and CoFPS_2) in developing oil-tea seeds. According to whether the expressions of key enzyme genes were associated with squalene accumulation we found that the precursor IPP of squalene biosynthesis obtained via the MVA pathway was dominant with oil-tea seed development. Based on the gene co-expression analyses, we identified multiple transcription factors potentially involved in regulating squalene biosynthesis such as CoMYC2, CoREM39, CobZIP5, CoERF and CoWRKY. Using yeast one-hybrid and dual-luciferase assay experiments we demonstrated that the transcription factor CoMYC2 could activate the expression of a key enzyme gene CoHMGR_4, suggesting that CoMYC2 might be a critical regulator during squalene biosynthesis in oil-tea seed development. This study gives not only insights into understanding the molecular basis of squalene biosynthesis in oil-tea developing seeds but also provides gene resources for developing genetically improved varieties with higher content of squalene in oil-tea.