Abstract

RING-H2 genes, the most abundant RING-type genes encoding putative ubiquitin ligase E3, are involved in diverse biological processes. Whether RING-H2 genes are related to wood formation remains to be identified in trees. In this study, we identified 288 RING-H2 genes in Populus trichocarpa, and found that the segmental and tandem duplication events contributed to RING-H2 gene expansion. Microarray dataset (from Affymetrix poplar genome arrays) showed that 64 of the 249 RING-H2 genes were highly or preferentially expressed in stem xylem. According to the AspWood RNAseq dataset, the transcription levels of genes PtrRHH21, 33, 48, 69, 88, 93, 94, 121, 141, 166, 175, 192, 208, 214, 250 and 257 were significantly increased in the xylem ranging from the expanding xylem to the lignifying xylem, suggesting their association with wood formation. Promoter analyses revealed that most of the preferentially xylem-expressed RING-H2 genes possessed SNBE, TERE, M46RE, AC and SMRE cis-elements, which are involved in secondary cell wall biosynthesis and programmed cell death. Based on the promoter GUS-based analysis result, PtrRHH94 was indicated to be associated with wood formation in transgenic P. trichocarpa. Taken together, dozens of Populus RING-H2 gene candidates associated with wood formation have been identified based on multiple gene expression analyses.

Highlights

  • Wood is used industrially as timber for construction, pulp for paper manufacturing and a renewable source for bioenergy

  • All identified 288 RING-H2 genes were named as PtrRHH1 to PtrRHH288 according to the successive order of genes on the chromosomes (Figure S1)

  • Our findings indicated that these highly or preferentially xylem-expressed RING-H2 genes possess multiple cis-elements related to xylem development, suggesting their participation in wood formation

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Summary

Introduction

Wood is used industrially as timber for construction, pulp for paper manufacturing and a renewable source for bioenergy. Wood formation is a complex developmental process undergoing cambial cell proliferation, xylem cell specification and expansion, secondary cell wall (SCW) biosynthesis and programmed cell death (PCD) [1,2]. Most biosynthetic enzymes for wood components (cellulose, xylan and lignin) have been identified, and a number of them have been functionally characterized [5,6,7,8,9,10,11]. Some vital transcription factors (TFs) have been identified to function in SCW biosynthesis, and several regulating networks have been constructed in these TFs [12,13,14]. After deposition of secondary walls, fibers and vessels (angiosperm) undergo PCD in wood, and some genes (for instance, encoding proteases, nucleases and autophage-related proteins) have displayed the potential roles in regulating

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