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.
Highlights
The plant cell wall is involved in a variety of physiological functions including mechanical support, growth, a physical barrier to pathogens, signalling, intercellular communication, and environmental interaction [1,2]
Genes encoding the plant-specific domain of unknown function 1218 (DUF1218) family have been implicated in several aspects of cell wall biology [3,4] and shown to function in vasculature patterning [5] and response to abiotic and biotic stresses [6,7,8]
Phylogenetic analysis of the DUF1218 family from Arabidopsis revealed that MWL-1 is more closely related to MWL-2 in than to the other 13 members of the family (S1A Fig)
Summary
The plant cell wall is involved in a variety of physiological functions including mechanical support, growth, a physical barrier to pathogens, signalling, intercellular communication, and environmental interaction [1,2]. Independent studies [4,9] reported expression correlation between a DUF1218-encoding gene (At4g27435) and the secondary cell wall-related cellulose synthase (CesA) genes. In addition to At4g27435, another DUF1218-containing protein, At1g31720 (referred to as MODIFYING WALL LIGNIN-1 or MWL-1 in this paper) is preferentially expressed in the inflorescent stems of Arabidopsis. MWL-1 is co-expressed with the three secondary cell wall cellulose synthase genes and several xylan and lignin-associated genes [10]. This combined with the findings that these DUF1218-containing protein homologs were conserved between and preferentially expressed in xylem of angiosperms and gymnosperms (Pavy et al, 2008), emphasizes the importance of the role of DUF1218 proteins in xylogenesis and/or secondary cell wall formation
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