Transcriptomic and Metabolomic Insights into Key Genes Involved in Kinsenoside Biosynthesis in Anoectochilus roxburghii

As the main active ingredient in Anoectochilus roxburghii, kinsenoside has important health and medical effects including hepatoprotective, anti-oxidant, and bacteriostasis, among others. In recent years, with the limited application of high-throughput technology to A. roxburghii, there has been no research on the key regulatory genes involved in the synthesis of kinsenoside. Therefore, we examined three species of A. roxburghii that are widely planted in mainland China and Taiwan Province, A. roxburghii cultivar ‘Jian ye’, Anoectochilus formosanus, and Anoectochilus burmannicus, determining the content of kinsenoside, performing transcriptomic and metabolomic sequencing, identifying UDP glycosyltransferases, and screening for UDP glycosyltransferases that may be involved in kinsenoside synthesis. The results showed that among the three species of A. roxburghii, the content of kinsenoside in A. roxburghii cv. ‘Jian ye’ was the highest. Transcriptome and metabolome data showed that A. roxburghii cv. ‘Jian ye’ and the two other species of A. roxburghii have 3702 and 5369 differentially expressed genes and 69 and 120 differentially accumulated metabolites, respectively. Meanwhile, differentially expressed genes and differentially accumulated metabolites are enriched in the glucose metabolism and hormone pathways. We also treated the A. roxburghii samples with exogenous auxin and characterized the related genes. In A. roxburghii, we identified 73 members of the UDP glycosyltransferase family. Through phylogenetic tree, transcriptome data expression profile, and qPCR analyses, we screened for members that may be involved in the synthesis of kinsenoside. In summary, the results of this study provide insights for breeding high-kinsenoside-content and high-intron varieties of A. roxburghii.