Summary Trichomes are specialized epidermal cells that contribute to plant resistance against herbivores. Their formation is controlled precisely by multiple genetic and environmental signals. Previous studies have shown that microRNA319 (miR319) and gibberellin (GA) signaling are involved in trichome development in Arabidopsis, but little is known about their interaction between these factors. Here we reported that the miR319a/TEOSINTE BRANCHED/CYCLOIDEA/PCF (TCP) module participates in trichome initiation synergistically with GA signaling in Populus tomentosa. We demonstrated that overexpression of miR319a decreased transcription levels of its targeted TCPs and significantly elevated leaf trichome density in transgenic poplar, resulting in decreasing insect herbivory. Conversely, repressing miR319a by short tandem target mimics (STTM) elevated TCP expression levels and decreased trichome density in transgenic plants. The trichome phenotype of 35S:miR319a plants could be abolished by introducing a miR319a‐resistant form of TCP19. Furthermore, the miR319a‐targeted TCP19 interacted directly with REPRESSOR OF ga1‐3 (RGA), a downstream repressor of GA signaling. TCP19 and RGA synergistically inhibited the GLABROUS1 (GL1)‐induced expression of trichome marker gene GLABRA2 (GL2), thereby repressing leaf trichome initiation. Our results provide an insight into the molecular mechanism by which miR319/TCP19 module and GA signaling coordinated regulating trichome initiation in P. tomentosa.

Anthocyanins are involved in several aspects of development and defence in poplar (Populus spp.). Although, over the past decades, significant progress has been made in uncovering these anthocyanin biosynthetic and regulatory mechanisms, the fundamental understanding of the epigenetic regulation in this pathway is still largely unclear. Here, we isolated a histone H3K9 demethylase gene JMJ25 from Populus and characterized its role in anthocyanin biosynthesis by genetic and biochemical approaches. JMJ25 was induced by continuous dark treatment. Overexpression of JMJ25 led to downregulated expression of anthocyanin biosynthetic genes in transgenic poplar, resulting in a significant reduction in anthocyanin content. ChIP-qPCR assays showed that JMJ25 could directly associate with MYB182 chromatin and dynamically demethylate at H3K9me2. Furthermore, JMJ25 also affected the DNA methylation levels of MYB182. By contrast, knockout of JMJ25 by CRISPR/Cas9 resulted in ectopic anthocyanin accumulation under dark condition and increased expression of anthocyanin biosynthetic genes. Our results support a model in which JMJ25 directly affects MYB182 expression by altering the histone methylation status of its chromatin and DNA methylation, resulting in repression of anthocyanin accumulation. This study uncovered an epigenetic mechanism that modulates anthocyanin biosynthesis in poplar.

Wood, which is mainly composed of lignified secondary cell wall, is the most abundant biomass in woody plants. Previous studies have revealed that R2R3-type MYB transcription factors are important regulators of the formation of the secondary cell wall in vascular plants. In this study, we isolated the R2R3-type MYB transcription factor gene PtoMYB055, which is mainly expressed in xylem and phloem tissue, from Populus tomentosa and demonstrate that PtoMYB055 is a key regulator of lignin biosynthesis. PtoMYB055 as a transcriptional activator is localized to the nucleus. Overexpression of PtoMYB055 upregulates expression of lignin biosynthetic genes in transgenic poplar plants, resulting in ectopic deposition of lignin in phloem tissue and an increase in thickness of the secondary cell wall. In sum, PtoMYB055 is a transcriptional activator that is involved in regulating lignin biosynthesis during the formation of the secondary cell wall in poplar.