The Arabidopsis Domain of Unknown Function 1218 (DUF1218) Containing Proteins, MODIFYING WALL LIGNIN-1 and 2 (At1g31720/MWL-1 and At4g19370/MWL-2) Function Redundantly to Alter Secondary Cell Wall Lignin Content

PLoS One. 2016 Mar 1;11(3):e0150254. doi: 10.1371/journal.pone.0150254. eCollection 2016.

Abstract

DUF1218 is a land plant-specific innovation and has previously been shown to be associated with cell wall biology, vasculature patterning and abiotic/biotic stress response. The Arabidopsis genome encodes 15 members, two of which (At1g31720 and At4g27435) are preferentially expressed in the secondary cell wall depositing inflorescence stems. To further our understanding of the roles of DUF1218-containing proteins in secondary cell wall biology, we functionally characterized At1g31720 (herein referred to as MODIFYING WALL LIGNIN-1 or MWL-1). Since related gene family members may contribute to functional redundancy, we also characterized At4g19370 (MWL-2), the most closely related gene to MWL-1 in the protein family. Subcellular localization revealed that both Arabidopsis proteins are targeted to the cell periphery. The single T-DNA knockout lines, mwl-1 and mwl-2, and independent overexpression lines showed no significant differences in plant growth or changes in total lignin content relative to wild-type (WT) control plants. However, the double homozygous mutant, mwl-1/mwl-2, had smaller rosettes with a significant decrease in rosette fresh weight and stem height relative to the WT control at four weeks and six weeks, respectively. Moreover, mwl-1/mwl-2 showed a significant reduction in total lignin content (by ca. 11% relative to WT) and an increase in syringyl/guaiacyl (S/G) monomer ratio relative to the control plants. Our study has identified two additional members of the DUF1218 family in Arabidopsis as novel contributors to secondary cell wall biology, specifically lignin biosynthesis, and these proteins appear to function redundantly.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Arabidopsis / genetics
  • Arabidopsis / growth & development
  • Arabidopsis / metabolism*
  • Arabidopsis Proteins / classification
  • Arabidopsis Proteins / genetics
  • Arabidopsis Proteins / metabolism*
  • Cell Membrane / metabolism
  • Cell Wall / genetics
  • Cell Wall / metabolism*
  • Gene Expression Regulation, Developmental
  • Gene Expression Regulation, Plant
  • Gene Knockout Techniques
  • Lignin / metabolism*
  • Microscopy, Confocal
  • Mutation
  • Phylogeny
  • Plant Stems / genetics
  • Plant Stems / growth & development
  • Plant Stems / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction

Substances

  • Arabidopsis Proteins
  • MWL-1 protein, Arabidopsis
  • MWL-2 protein, Arabidopsis
  • Lignin

Grants and funding

The work presented here was supported by Sappi through the Forest Molecular Genetics (FMG) Programme, the Technology and Human Resources for Industry Programme (THRIP, UID 80118), and the National Research Foundation (NRF, UID 71255 and 86936) of South Africa. RM acknowledges an NRF Ph.D. Prestige and Equity Scholarship. Sappi also provided support in the form of salary for one of the authors (BC), but did not have any additional role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript. The specific roles of the authors are articulated in the ‘author contributions’ section.