New insights into the carotenoid biosynthesis in Torreya grandis kernels

Torreya grandis is a characteristic rare economic tree species in subtropical mountainous areas. The nut has a variety of bioactive compounds, such as polyphenols and vitamins. However, the carotenoids compositions and its regulatory mechanisms underlying carotenoid biosynthesis in kernels remain unknown. Here, the major carotenoids in the kernel were analyzed. The result showed that lutein and β-carotene were the predominant carotenoids in the kernel, while low levels of α-carotenoid and zeaxanthin were detected. Lutein and β-carotene were decreased during the maturation of kernel. Expression analysis by RNA-seq and qRT-PCR indicated that the expressions of TgCYP97A3 and TgLCYB were also reduced during the maturation of kernel. The contents of lutein or β-carotene were obviously increased in tobacco transiently overexpressing TgCYP97A3 or TgLCYB. Moreover, DPPH radical scavenging activity and FRAP were also significantly enhanced. In addition, several MYB and WRKY transcription factors TgMYBS3, TgMYB48 and TgWRKY11 were identified to positively regulate the TgCYP97A3 expression, while TgMYB48, TgWRKY2 and TgWRKY11 could upregulate the TgLCYB expression. The illustration of carotenoids biosynthesis and its molecular mechanism in kernels not only provides a basis for understanding carotenoids biosynthesis in kernels, but also enables the use of molecular biotechnology to develop new health products rich in carotenoids based on T. grandis nuts.