The lycopene [beta]-cyclase plays a significant role in provitamin A biosynthesis in wheat endosperm

Lycopene [beta]-cyclase (LCYB) is a key enzyme catalyzing the biosynthesis of [beta]-carotene, the main source of provitamin A. However, there is no documented research about this key cyclase gene's function and relationship with [beta]-carotene content in wheat. Therefore, the objectives of this study were to clone TaLCYB and characterize its function and relationship with [beta]-carotene biosynthesis in wheat grains. We also aimed to obtain more information about the endogenous carotenoid biosynthetic pathway and thus provide experimental support for carotenoid metabolic engineering in wheat. In the present study, a lycopene [beta]-cyclase gene, designated TaLCYB, was cloned from the hexaploid wheat cultivar Chinese Spring. The cyclization activity of the encoded protein was demonstrated by heterologous complementation analysis. The TaLCYB gene was expressed differentially in different tissues of wheat. Although TaLCYB had a higher expression level in the later stages of grain development, the [beta]-carotene content still showed a decreasing tendency. The expression of TaLCYB in leaves was dramatically induced by strong light and the [beta]-carotene content variation corresponded with changes of TaLCYB expression. A post-transcriptional gene silencing strategy was used to down-regulate the expression of TaLCYB in transgenic wheat, resulting in a decrease in the content of [beta]-carotene and lutein, accompanied by the accumulation of lycopene to partly compensate for the total carotenoid content. In addition, changes in TaLCYB expression also affected the expression of several endogenous carotenogenic genes to varying degrees. Our results suggest that TaLCYB is a genuine lycopene cyclase gene and plays a crucial role in [beta]-carotene biosynthesis in wheat. Our attempt to silence it not only contributes to elucidating the mechanism of carotenoid accumulation in wheat but may also help in breeding wheat varieties with high provitamin A content through RNA interference (RNAi) to block specific carotenogenic genes in the wheat endosperm.