Root hairs are single cells that develop by tip growth, a process shared with pollen tubes, axons, and fungal hyphae. However, structural plant cell walls impose constraints to accomplish tip growth. In addition to po...Root hairs are single cells that develop by tip growth, a process shared with pollen tubes, axons, and fungal hyphae. However, structural plant cell walls impose constraints to accomplish tip growth. In addition to polysaccharides, plant cell walls are composed of hydroxyproline-rich glycoproteins （HRGPs）, which include several groups of O-glycoproteins, including extensins （EXTs）. Proline hydroxylation, an early post-translational modification （PTM） of HRGPs catalyzed by prolyl 4-hydroxylases （P4Hs）, defines their subsequent O-glycosylation sites. In this work, our genetic analyses prove that P4H5, and to a lesser extent P4H2 and P4H13, are pivotal for root hair tip growth. Second, we demonstrate that P4H5 has in vitro preferred specificity for EXT substrates rather than for other HRGPs. Third, by P4H promoter and protein swapping approaches, we show that P4H2 and P4H13 have interchangeable functions but cannot replace P4H5. These three P4Hs are shown to be targeted to the secretory pathway, where P4H5 forms dimers with P4H2 and P4H13. Finally, we explore the impact of deficient proline hydroxylation on the cell wall architec- ture. Taken together, our results support a model in which correct peptidyl-proline hydroxylation on EXTs, and possibly in other HRGPs, is required for proper cell wall self-assembly and hence root hair elongation in Arabidopsis thaliana.展开更多
Glycosyltransferases of the Cellulose Synthase Like D(CS/D)subfamily have been reported to be involved in tip growth and stem development in Arabidopsis.The csld2 and csld3 mutants are root hair defective and the csld...Glycosyltransferases of the Cellulose Synthase Like D(CS/D)subfamily have been reported to be involved in tip growth and stem development in Arabidopsis.The csld2 and csld3 mutants are root hair defective and the csld5 mutant has reduced stem growth.In this study,we produced double and triple knockout mutants of CSLD2,CSLD3,and CSLD5.Unlike the single mutants and the csld2/csld3 double mutant,the csld2/csld5,csld3/csld5,and csld2/csld3/csld5 mutants were dwarfed and showed severely reduced viability.This demonstrates that the cooperative activities of CSLD2,CSLD3,and CSLD5 are required for normal Arabidopsis development,and that they are involved in important processes besides the specialized role in tip growth.The mutant phenotypes indicate that CSLD2 and CSLD3 have overlapping functions with CSLD5 in early plant development,whereas the CSLD2 and CSLD3 proteins are non-redundant.To determine the biochemical function of CSLD proteins,we used transient expression in tobacco leaves.Microsomes containing heterologously expressed CSLD5 transferred mannose from GDP-mannose onto endogenous acceptors.The same activity was detected when CSLD2 and CSLD3 were coexpressed but not when they were expressed separately.With monosaccharides as exogenous acceptors,microsomal preparations from CSLD5-expressing plants mediated the transfer of mannose from GDP-mannose onto mannose.These results were supported by immunodetection studies that showed reduced levels of a mannan epitope in the cell walls of stem interfascicular fibers and xylem vessels of the csld2/csld3/csld5 mutant.展开更多
Dear Editor, Xylan polysaccharides constitute the major non-cellulosic components in secondary cell walls of dicots and in both primary and secondary cell walls of grasses （Scheller and Ulvskov, 2010）. Xylan is com...Dear Editor, Xylan polysaccharides constitute the major non-cellulosic components in secondary cell walls of dicots and in both primary and secondary cell walls of grasses （Scheller and Ulvskov, 2010）. Xylan is composed of a linear backbone of 13（1-4）-linked xylose （Xyl）. In grasses, the xylan backbone is substituted with 03-1inked and 02-1inked arabinose residues and, to a lesser extent, a（1-3）-Iinked glucuronic acid residues.展开更多
文摘Root hairs are single cells that develop by tip growth, a process shared with pollen tubes, axons, and fungal hyphae. However, structural plant cell walls impose constraints to accomplish tip growth. In addition to polysaccharides, plant cell walls are composed of hydroxyproline-rich glycoproteins （HRGPs）, which include several groups of O-glycoproteins, including extensins （EXTs）. Proline hydroxylation, an early post-translational modification （PTM） of HRGPs catalyzed by prolyl 4-hydroxylases （P4Hs）, defines their subsequent O-glycosylation sites. In this work, our genetic analyses prove that P4H5, and to a lesser extent P4H2 and P4H13, are pivotal for root hair tip growth. Second, we demonstrate that P4H5 has in vitro preferred specificity for EXT substrates rather than for other HRGPs. Third, by P4H promoter and protein swapping approaches, we show that P4H2 and P4H13 have interchangeable functions but cannot replace P4H5. These three P4Hs are shown to be targeted to the secretory pathway, where P4H5 forms dimers with P4H2 and P4H13. Finally, we explore the impact of deficient proline hydroxylation on the cell wall architec- ture. Taken together, our results support a model in which correct peptidyl-proline hydroxylation on EXTs, and possibly in other HRGPs, is required for proper cell wall self-assembly and hence root hair elongation in Arabidopsis thaliana.
文摘Glycosyltransferases of the Cellulose Synthase Like D(CS/D)subfamily have been reported to be involved in tip growth and stem development in Arabidopsis.The csld2 and csld3 mutants are root hair defective and the csld5 mutant has reduced stem growth.In this study,we produced double and triple knockout mutants of CSLD2,CSLD3,and CSLD5.Unlike the single mutants and the csld2/csld3 double mutant,the csld2/csld5,csld3/csld5,and csld2/csld3/csld5 mutants were dwarfed and showed severely reduced viability.This demonstrates that the cooperative activities of CSLD2,CSLD3,and CSLD5 are required for normal Arabidopsis development,and that they are involved in important processes besides the specialized role in tip growth.The mutant phenotypes indicate that CSLD2 and CSLD3 have overlapping functions with CSLD5 in early plant development,whereas the CSLD2 and CSLD3 proteins are non-redundant.To determine the biochemical function of CSLD proteins,we used transient expression in tobacco leaves.Microsomes containing heterologously expressed CSLD5 transferred mannose from GDP-mannose onto endogenous acceptors.The same activity was detected when CSLD2 and CSLD3 were coexpressed but not when they were expressed separately.With monosaccharides as exogenous acceptors,microsomal preparations from CSLD5-expressing plants mediated the transfer of mannose from GDP-mannose onto mannose.These results were supported by immunodetection studies that showed reduced levels of a mannan epitope in the cell walls of stem interfascicular fibers and xylem vessels of the csld2/csld3/csld5 mutant.
文摘Dear Editor, Xylan polysaccharides constitute the major non-cellulosic components in secondary cell walls of dicots and in both primary and secondary cell walls of grasses （Scheller and Ulvskov, 2010）. Xylan is composed of a linear backbone of 13（1-4）-linked xylose （Xyl）. In grasses, the xylan backbone is substituted with 03-1inked and 02-1inked arabinose residues and, to a lesser extent, a（1-3）-Iinked glucuronic acid residues.
