A Phosphorus-Limitation Induced, Functionally Conserved DUF506 Protein is a Repressor of Root Hair Elongation in Arabidopsis thaliana

Root hairs (RHs) function in nutrient and water acquisition, root metabolite exudation, soil anchorage and plant-microbe interactions. Longer or more abundant RHs are potential breeding traits for developing crops that are more resource-use efficient and can improve soil health. RH elongation is controlled by both environmental and endogenous factors. While many genes are known to promote RH elongation, relatively little is known about genes and mechanisms that constrain RH growth. Here we demonstrate that a DOMAIN OF UNKNOWN FUNCTION 506 (DUF506) protein, AT3G25240, negatively regulates Arabidopsis thaliana RH growth. The AT3G25240 gene is strongly and specifically induced during P-limitation. Mutants of this gene, which we call REPRESSOR OF EXCESSIVE ROOT HAIR ELONGATION 1 (RXR1), have much longer RHs, while over-expression of the gene results in much shorter RHs. RXR1 physically interacts with a Rab-GTPase (RXR2), and an rxr2 mutant phenocopies the rxr1 mutant. Overexpression of a Brachypodium distachyon RXR1 homolog resulted in repression of RH elongation in Brachypodium. Taken together, our results reveal a DUF506-GTPase module with a prominent role in repression of RH elongation that is conserved in monocots and dicots.

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A role for the RabA4b effector protein PI-4Kβ1 in polarized expansion of root hair cells in Arabidopsis thaliana

The Journal of Cell Biology ◽

10.1083/jcb.200508116 ◽

2006 ◽

Vol 172 (7) ◽

pp. 991-998 ◽

Cited By ~ 188

Author(s):

Mary L. Preuss ◽

Aaron J. Schmitz ◽

Julie M. Thole ◽

Heather K.S. Bonner ◽

Marisa S. Otegui ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Hair Cells ◽

Root Hair ◽

Root Hairs ◽

Developmentally Regulated ◽

Domain Specific ◽

Polarized Secretion ◽

Phosphatidylinositol 4 ◽

Golgi Network ◽

Root Hair Cells

The RabA4b GTPase labels a novel, trans-Golgi network compartment displaying a developmentally regulated polar distribution in growing Arabidopsis thaliana root hair cells. GTP bound RabA4b selectively recruits the plant phosphatidylinositol 4-OH kinase, PI-4Kβ1, but not members of other PI-4K families. PI-4Kβ1 colocalizes with RabA4b on tip-localized membranes in growing root hairs, and mutant plants in which both the PI-4Kβ1 and -4Kβ2 genes are disrupted display aberrant root hair morphologies. PI-4Kβ1 interacts with RabA4b through a novel homology domain, specific to eukaryotic type IIIβ PI-4Ks, and PI-4Kβ1 also interacts with a Ca2+ sensor, AtCBL1, through its NH2 terminus. We propose that RabA4b recruitment of PI-4Kβ1 results in Ca2+-dependent generation of PI-4P on this compartment, providing a link between Ca2+ and PI-4,5P2–dependent signals during the polarized secretion of cell wall components in tip-growing root hair cells.

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Cyanobacteria-Derived Proline Increases Stress Tolerance in Arabidopsis thaliana Root Hairs by Suppressing Programmed Cell Death

Frontiers in Plant Science ◽

10.3389/fpls.2020.490075 ◽

2020 ◽

Vol 11 ◽

Author(s):

Alysha Chua ◽

Orla L. Sherwood ◽

Laurence Fitzhenry ◽

Carl K.-Y. Ng ◽

Paul F. McCabe ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Cell Death ◽

Programmed Cell Death ◽

Stress Tolerance ◽

Root Hair ◽

Root Hairs ◽

Biologically Active ◽

Nostoc Muscorum ◽

Sensitivity Threshold ◽

Wild Type

Nitrogen-fixing heterocystous cyanobacteria are used as biofertilizer inoculants for stimulating plant growth but can also alleviate plant stress by exometabolite secretion. However, only a small number of studies have focused on elucidating the identity of said bioactives because of the wide array of exuded compounds. Here, we used the root hair assay (RHA) as a rapid programmed cell death (PCD) screening tool for characterizing the bioactivity of cyanobacteria Nostoc muscorum conditioned medium (CM) on Arabidopsis thaliana root hair stress tolerance. We found that heat-stressed A. thaliana pre-treated with N. muscorum CM fractions exhibited significantly lower root hair PCD levels compared to untreated seedlings. Treatment with CM increased stress tolerance by suppressing PCD in root hairs but not necrosis, indicating the bioactive compound was specifically modulating the PCD pathway and not a general stress response. Based on documented N. muscorum exometabolites, we identified the stress-responsive proline as a compound of interest and strong evidence from the ninhydrin assay and HPLC indicate that proline is present in N. muscorum CM. To establish whether proline was capable of suppressing PCD, we conducted proline supplementation experiments. Our results showed that exogenous proline had a similar effect on root hairs as N. muscorum CM treatment, with comparable PCD suppression levels and insignificant necrosis changes. To verify proline as one of the biologically active compounds in N. muscorum CM, we used three mutant A. thaliana lines with proline transporter mutations (lht1, aap1 and atprot1-1::atprot2-3::atprot3-2). Compared with the wild-type seedlings, PCD-suppression in lht1and aap1 mutants was significantly reduced when supplied with low proline (1–5 μM) levels. Similarly, pre-treatment with N. muscorum CM resulted in elevated PCD levels in all three mutant lines compared to wild-type seedlings. Our results show that plant uptake of cyanobacteria-derived proline alters their root hair PCD sensitivity threshold. This offers evidence of a novel biofertilizer mechanism for reducing stress-induced PCD levels, independent of the existing mechanisms documented in the literature.

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A cell surface arabinogalactan-peptide influences root hair cell fate

10.1101/787143 ◽

2019 ◽

Cited By ~ 1

Author(s):

Cecilia Borassi ◽

Javier Gloazzo Dorosz ◽

Martiniano M. Ricardi ◽

Laercio Pol Fachin ◽

Mariana Carignani Sardoy ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Transcription Factor ◽

Cell Surface ◽

Cell Fate ◽

Root Hair ◽

Root Hairs ◽

Epidermal Cells ◽

Root Epidermis ◽

A Cell ◽

Epidermis Cell

SummaryRoot hairs (RHs) develop from specialized epidermal cells called trichoblasts, whereas epidermal cells that lack RHs are known as atrichoblasts. The mechanism controlling root epidermal cell fate is only partially understood. Root epidermis cell fate is regulated by a transcription factor complex that promotes the expression of the homeodomain protein GLABRA 2 (GL2), which blocks RH development by inhibiting ROOT HAIR DEFECTIVE 6 (RHD6). Suppression of GL2 expression activates RHD6, a series of downstream TFs including ROOT HAIR DEFECTIVE 6 LIKE-4 (RSL4 [Yi et al. 2010]) and their target genes, and causes epidermal cells to develop into RHs. Brassinosteroids (BRs) influence root epidermis cell fate. In the absence of BRs, phosphorylated BIN2 (a Type-II GSK3-like kinase) inhibits a protein complex that directly downregulates GL2 [Chen et al. 2014]. Here, we show that the genetic and pharmacological perturbation of the arabinogalactan peptide (AG) AGP21 in Arabidopsis thaliana, triggers aberrant RH development, similar to that observed in plants with defective BR signaling. We reveal that an O-glycosylated AGP21 peptide, which is positively regulated by BZR1, a transcription factor activated by BR signaling, affects RH cell fate by altering GL2 expression in a BIN2-dependent manner. These results suggest that perturbation of a cell surface AGP disrupts BR responses and inhibits the downstream effect of BIN2 on the RH repressor GL2 in root epidermal cells. In addition, AGP21 also acts in a BR-independent, AGP-dependent mode that together with BIN2 signalling cascade controls RH cell fate.SignificanceIn the plant Arabidopsis thaliana, the root epidermis forms in an alternating pattern atrichoblasts with trichoblast cells that end up developing root hairs (RHs). Atrichoblast cell fate is directly promoted by the transcription factor GLABRA2 (GL2) while the lack of GL2 allows RH formation. The loss of AGP21 peptide triggers an abnormal RH cell fate in two contiguous cells in a similar manner as brassinosteroid (BRs) mutants. In the absence of BR signaling, BIN2 (a GSK3 like-kinase) in a phosphorylated state, downregulate GL2 expression to trigger RH cell fate. The absence of AGP21 is able to repress GL2 expression and activates the expression of RSL4 and EXP7 root hair proteins.

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Apoplastic class III peroxidases PRX62 and PRX69 regulate ROS-homeostasis and cell wall associated extensins linked to root hair growth at low-temperature in Arabidopsis thaliana

10.1101/2021.08.20.456256 ◽

2021 ◽

Author(s):

Javier Martínez Pacheco ◽

Philippe Ranocha ◽

Luciana Kasulin ◽

Corina M. Fusari ◽

Lucas Servi ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Cell Wall ◽

Low Temperature ◽

Root Hair ◽

Developmental Process ◽

Association Studies ◽

Root Hairs ◽

Genome Wide Association Studies ◽

Class Iii ◽

Ros Homeostasis

Root hairs (RH) growth is highly influenced by endogenous as well as by external environmental signals that coordinately regulate its final cell size. RHs actively expand the root surface responsible for nutrient uptake and water absorption. We have recently determined that RH growth was unexpectedly boosted when Arabidopsis thaliana seedlings are cultivated at low temperatures. It was proposed that RH growth plasticity in response to cold was linked to a reduced nutrient availability in the media. Here, we explored the molecular basis of this strong RH growth response by using the Genome Wide Association Studies (GWAS) approach on Arabidopsis thaliana natural accessions. We identified the poorly characterized PEROXIDASE 62 (PRX62) as a key protein triggering this conditional growth under a moderate low-temperature stress. In addition, we identified the related protein PRX69 as an important factor in this developmental process. The prx62 prx69 double mutant and the PRX62 and PRX69 over-expressing lines showed contrasting RH phenotypes, peroxidase activities and cyt/apoReactive Oxygen Species (ROS) levels. Strikingly, a cell wall protein extensin (EXT) reporter revealed the effect of peroxidase activity on the EXT cell wall association at 10C in the RH apical zone. EXT cell wall insolubilization was enhanced at 10C, which was completely abolished under the PRX inhibitor salicylhydroxamic acid (SHAM) treatment. Finally, we demonstrated that the Root Hair defective 6 like 4 (RSL4) transcription factor directly controls the expression of PRX69 under low-temperature. Collectively, our results indicate that both PRX62 and PRX69 are key apoplastic PRXs that modulate ROS-homeostasis and cell wall EXT-insolubilization linked to RH elongation at low-temperature.

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Inhibition of Pre-mRNA Splicing Promotes Root Hair Development in Arabidopsis thaliana

Plant and Cell Physiology ◽

10.1093/pcp/pcz150 ◽

2019 ◽

Vol 60 (9) ◽

pp. 1974-1985 ◽

Cited By ~ 2

Author(s):

Miku Ishizawa ◽

Kayo Hashimoto ◽

Misato Ohtani ◽

Ryosuke Sano ◽

Yukio Kurihara ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Root Hair ◽

Protein A ◽

Root Hairs ◽

Carbon Dioxide Concentration ◽

Light Response ◽

Mrna Splicing ◽

Gene Encoding ◽

Root Hair Development ◽

Hair Development

Abstract Root hairs protruding from epidermal cells increase the surface area for water absorption and nutrient uptake. Various environmental factors including light, oxygen concentration, carbon dioxide concentration, calcium and mycorrhizal associations promote root hair formation in Arabidopsis thaliana. Light regulates the expression of a large number of genes at the transcriptional and post-transcriptional levels; however, there is little information linking the light response to root hair development. In this study, we describe a novel mutant, light-sensitive root-hair development 1 (lrh1), that displays enhanced root hair development in response to light. Hypocotyl and root elongation was inhibited in the lrh1 mutant, which had a late flowering phenotype. We identified the gene encoding the p14 protein, a putative component of the splicing factor 3b complex essential for pre-mRNA splicing, as being responsible for the lrh1 phenotype. Indeed, regulation of alternative splicing was affected in lrh1 mutants and treatment with a splicing inhibitor mimicked the lrh1 phenotype. Genome-wide alterations in pre-mRNA splicing patterns including differential splicing events of light signaling- and circadian clock-related genes were found in lrh1 as well as a difference in transcriptional regulation of multiple genes including upregulation of essential genes for root hair development. These results suggest that pre-mRNA splicing is the key mechanism regulating root hair development in response to light signals.

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A Sec14p-nodulin domain phosphatidylinositol transfer protein polarizes membrane growth of Arabidopsis thaliana root hairs

The Journal of Cell Biology ◽

10.1083/jcb.200412074 ◽

2005 ◽

Vol 168 (5) ◽

pp. 801-812 ◽

Cited By ~ 147

Author(s):

Patrick Vincent ◽

Michael Chua ◽

Fabien Nogue ◽

Ashley Fairbrother ◽

Hal Mekeel ◽

...

Keyword(s):

Arabidopsis Thaliana ◽

Plasma Membrane ◽

Membrane Trafficking ◽

Root Hair ◽

Root Hairs ◽

Extracellular Milieu ◽

Root Hair Development ◽

Gene Ablation ◽

Membrane Growth ◽

Phosphatidylinositol Transfer

Phosphatidylinositol (PtdIns) transfer proteins (PITPs) regulate signaling interfaces between lipid metabolism and membrane trafficking. Herein, we demonstrate that AtSfh1p, a member of a large and uncharacterized Arabidopsis thaliana Sec14p-nodulin domain family, is a PITP that regulates a specific stage in root hair development. AtSfh1p localizes along the root hair plasma membrane and is enriched in discrete plasma membrane domains and in the root hair tip cytoplasm. This localization pattern recapitulates that visualized for PtdIns(4,5)P2 in developing root hairs. Gene ablation experiments show AtSfh1p nullizygosity compromises polarized root hair expansion in a manner that coincides with loss of tip-directed PtdIns(4,5)P2, dispersal of secretory vesicles from the tip cytoplasm, loss of the tip f-actin network, and manifest disorganization of the root hair microtubule cytoskeleton. Derangement of tip-directed Ca2+ gradients is also apparent and results from isotropic influx of Ca2+ from the extracellular milieu. We propose AtSfh1p regulates intracellular and plasma membrane phosphoinositide polarity landmarks that focus membrane trafficking, Ca2+ signaling, and cytoskeleton functions to the growing root hair apex. We further suggest that Sec14p-nodulin domain proteins represent a family of regulators of polarized membrane growth in plants.

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SEM and fluorescence imaging of callose deposition in root hair tips and suspension cultures of Streptanthus tortuosus

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100161047 ◽

1990 ◽

Vol 48 (3) ◽

pp. 698-699

Author(s):

K.S. Walters ◽

R.D. Sjolund ◽

K.C. Moore

Keyword(s):

Cell Wall ◽

Root Hair ◽

Cultured Cells ◽

Root Hairs ◽

Callus Cultures ◽

Callose Deposition ◽

Culture Cells ◽

Wall Structures ◽

Ultraviolet Illumination ◽

Callose Formation

Callose, B-1,3-glucan, a component of cell walls, is associated with phloem sieve plates, plasmodesmata, and other cell wall structures that are formed in response to wounding or infection. Callose reacts with aniline blue to form a fluorescent complex that can be recognized in the light microscope with ultraviolet illumination. We have identified callose in cell wall protuberances that are formed spontaneously in suspension-cultured cells of S. tortuosus and in the tips of root hairs formed in sterile callus cultures of S. tortuosus. Callose deposits in root hairs are restricted to root hair tips which appear to be damaged or deformed, while normal root hair tips lack callose deposits. The callose deposits found in suspension culture cells are restricted to regions where unusual outgrowths or protuberances are formed on the cell surfaces, specifically regions that are the sites of new cell wall formation.Callose formation has been shown to be regulated by intracellular calcium levels.

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