Abstract

Wood formation originates in the biosynthesis of lignin and further leads to secondary cell wall (SCW) biosynthesis in woody plants. Masson pine (Pinus massoniana Lamb) is an economically important industrial timber tree, and its wood yield affects the stable development of the paper industry. However, the regulatory mechanisms of SCW formation in Masson pine are still unclear. In this study, we characterized PmMYB4, which is a Pinus massoniana MYB gene involved in SCW biosynthesis. The open reading frame (ORF) of PmMYB4 was 1473 bp, which encoded a 490 aa protein and contained two distinctive R2 and R3 MYB domains. It was shown to be a transcription factor, with the highest expression in semi-lignified stems. We overexpressed PmMYB4 in tobacco. The results indicated that PmMYB4 overexpression increased lignin deposition, SCW thickness, and the expression of genes involved in SCW formation. Further analysis indicated that PmMYB4 bound to AC-box motifs and might directly activate the promoters of genes (PmPAL and PmCCoAOMT) involved in SCW biosynthesis. In addition, PmMYB4-OE(over expression) transgenic lines had higher lignin and cellulose contents and gene expression than control plants, indicating that PmMYB4 regulates SCW mainly by targeting lignin biosynthetic genes. In summary, this study illustrated the MYB-induced SCW mechanism in Masson pine and will facilitate enhanced lignin and cellulose synthesis in genetically engineered trees.

Highlights

  • The formation of wood involves the development of mother cells of secondary xylem differentiated from the vascular cambium, elongation of cells, deposition of the secondary cell wall (SCW), and programmed cell death (PCD) [1]

  • Other studies of xylem cell wall biosynthesis in other plants have led to a regulation model that consists of four gene levels: (1) first-layer master switches (NAC genes; NAM, ATAF1/2, and CUC2 genes), (2) second-layer master switches (MYB genes; v-myb avian myeloblastosis viral oncogene homologs), and (3) downstream regulators

  • A putative R2R3-MYB transcription factors (TFs) gene was obtained by BLAST searching in the Masson pine database(PRJNA655997) [24] using PgMYB4

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Summary

Introduction

The formation of wood involves the development of mother cells of secondary xylem differentiated from the vascular cambium, elongation of cells, deposition of the SCW, and programmed cell death (PCD) [1]. During the formation of the SCW, the composition of the cell wall changes significantly, mainly because of the directional arrangement of cellulose, deposition of lignin, and changes in the composition of hemicellulose and proteins [4]. Cellulose, and hemicellulose are the main components of the SCW. The biosynthesis of these components is precisely regulated by structural genes and strictly regulated by many transcription factors (TFs). Other studies of xylem cell wall biosynthesis in other plants have led to a regulation model that consists of four gene levels: (1) first-layer master switches (NAC genes; NAM, ATAF1/2, and CUC2 genes), (2) second-layer master switches (MYB (myeloblastosis) genes; v-myb avian myeloblastosis viral oncogene homologs), and (3) downstream regulators (TF genes) regulating (4) genes for SCWs [5,6,7,8,9,10]

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