Tilia tuan Szyszyl. (T. tuan) is a timber tree species with important ecological and commercial value. However, its highly ligni ed pericarp results in a low seed germination rate and a long dormancy period.In addition, it is unknown whether there is an interaction between the biosynthesis of avonoids and lignin as products of the phenylpropanoid pathway during seed development. To explore the molecular regulatory mechanism of lignin and avonoid biosynthesis, T. tuan seeds were harvested at ve stages (30, 60, 90, 120, and 150 days after pollination) for lignin and avonoid analyses. The results showed that lignin accumulated rapidly in the early and middle stages (S1, S3, and S4), and rapid accumulation of avonoids during the early and late stages (S1 and S5). High-throughput RNA sequencing analysis of developing seeds identi ed 50,553 transcripts., including 223 phenylpropanoid biosynthetic pathway genes involved in lignin accumulation grouped into three clusters, and 106 avonoid biosynthetic pathway genes (FBPGs) grouped into two clusters. Subsequent WGCNA and time-ordered gene coexpression network (TO-GCN) analysis revealed that 24 TFs (e.g., TtARF2and TtWRKY15) were involved in lignin and avonoid biosynthesis regulation. The transcriptome data were validated by qRT-PCR to analyze the expression pro les of key enzyme-coding genes. This study revealed that there existed a competitive relationship between the avonoid biosynthesis pathway and the lignin biosynthesis pathway, both of which are downstream pathways of phenylpropane that provide a foundation for the further exploration of molecular mechanisms underlying lignin and avonoid accumulation in T. tuanseeds.
Key messageTranscriptomics and phenotypic data analysis identi ed 24 transcription factors (TFs) that play key roles in regulating the competitive accumulation of lignin and avonoids.