Abstract: $ \alpha $-carotene ε-ring hydroxylase (CHYE) is a key downstream enzyme of the carotenoid synthesis pathway in plants, catalyzing the formation of lutein. Marigold (Tagetes erecta L.) is an ornamental crop of Compositeae, which is rich in carotenoids, especially lutein, to be used for natural pigments extraction. In this paper, cDNA from marigold flower was synthesized by RNA extraction and reverse transcription to clone the Tagetes erecta L. α-carotene ε-ring hydroxylase (TeCHYE) gene based on the transcriptome data. Then, TeCHYE was ligated to the cloning vector pEASY-Blunt. The sequence analysis indicated the 1 656 bp open reading frame of TeCHYE, encoding a hydrophilic protein with relative molecular weight of 62 kDa. The phylogenetic analysis suggested that CHYE gene conserves in plant species, and TeCHYE showed the highest identity with homologous HaCHYE from sunflower belonging to the same family. The content of lutein, as well as the expression level of TeCHYE, in marigold leaves and flower tissues at different development stages were furtherly analyzed. The significantly increased level of TeCHYE in developmental flowers suggested its involvement in the synthesis and accumulation of lutein in marigold.
Keywords: Tagetes erecta L.; lutein; $ \alpha $-carotene $ \varepsilon $-ring hydroxylase(CHYE); gene cloning; expression analysis
合肥工业大学学报自科版