TcMYB1, TcMYB4, and TcMYB8 participate in the regulation of lignan biosynthesis in Taiwania cryptomerioides Hayata
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Taiwania (Taiwania cryptomerioides Hayata) is a conifer species that is rich in bioactive secondary metabolites. Lignans, which are the major extracts, are especially important, but the biosynthetic gene regulation and downstream biosynthesis pathways remain unclear. In a previous study, we identified three pinoresinol-lariciresinol reductase (PLR) genes in Taiwanina, and in the present study, the regulation of these PLR genes was investigated. Through next-generation transcriptome sequencing, gene co-expression network analysis was performed to predict the relationship between putative transcription factor genes and upstream biosynthetic genes. Three MYB genes were identified as putative lignan biosynthetic regulators and named TcMYB1, TcMYB4, and TcMYB8. The gene expression of TcMYBs and TcPLRs was further confirmed with real-time PCR. Finally, an Agrobacteria-mediated transient overexpression experiment was conducted, and the activation of proTcPLR3::YFP by TcMYBs suggested that TcMYB1, TcMYB4, and TcMYB8 are biosynthetic regulators of lignan. These lignan regulators can be used in further research and applied to achieve a higher lignan content.
KeywordsGene regulation Lignan biosynthesis MYB gene family Pinoresinol-lariciresinol reductase Taiwania cryptomerioides Hayata
The authors thank the DNA Sequencing Core Facility (Core Facility and Innovative Instrument Project, AS-CFII-108-115) for DNA Sequencing support. The authors thank Li-Yu Daisy Liu (Department of Agronomy, National Taiwan University, Taiwan) for network analysis program design. The authors are grateful for expert technical advice on infiltration from Dr. Shih-Shun Lin (Institute of Biotechnology, National Taiwan University, Taiwan) and the expert technical assistance with real-time PCR from Mei-Jane Fang (DNA Analysis Core Laboratory, Institute of Plant and Microbial Biology, Academia Sinica, Taiwan).
Data archiving statement
The sequences of TcMYB1, TcMYB4, and TcMYB8 were under NCBI GenBank accession number MK202580, MK202581, and MK202582. The promoter sequence of TcPLR1, TcPLR2, and TcPLR3 were under GenBank accession numbers MK343128, MK343129, and MK343130.
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