Root Hair Initiation Research Articles

Overexpressing CAPRICE and GLABRA3 did not change the anthocyanin content of tomato (solanum lycopersicum) fruit peel

In Arabidopsis thaliana, the R3-type MYB transcription factor CAPRICE (CPC) and bHLH transcription factor GLABRA3 (GL3) cooperatively regulate epidermal cell differentiation. CPC and GL3 are involved in root-hair differentiation, trichome initiation and anthocyanin biosynthesis in Arabidopsis epidermal cells. Previously, we showed that CPC and GL3 also influence anthocyanin accumulation in tomato. Introduction of 35S::CPC into tomato significantly inhibits anthocyanin accumulation in cotyledons, leaves and stems. In contrast, introduction of GL3::GL3 strongly enhances anthocyanin accumulation in cotyledons, leaves and stems of tomato. In this study, we investigated the effect of CPC and GL3 on anthocyanin accumulation in the epidermis of tomato fruit. Unlike the results with vegetative tissues, overexpression of CPC and GL3 did not influence anthocyanin biosynthesis in tomato fruit peel.

Stripe to Spot Transition in a Plant Root Hair Initiation Model

A generalised Schnakenberg reaction-diffusion system with source and loss terms and a spatially dependent coefficient of the nonlinear term is studied both numerically and analytically in two spatial dimensions. The system has been proposed as a model of hair initiation in the epidermal cells of plant roots. Specifically the model captures the kinetics of a small G-protein ROP, which can occur in active and inactive forms, and whose activation is believed to be mediated by a gradient of the plant hormone auxin. Here the model is made more realistic with the inclusion of a transverse co-ordinate. Localised stripe-like solutions of active ROP occur for high enough total auxin concentration and lie on a complex bifurcation diagram of single and multi-pulse solutions. Transverse stability computations, confirmed by numerical simulation show that, apart from a boundary stripe, these 1D solutions typically undergo a transverse instability into spots. The spots so formed typically drift and undergo secondary instabilities such as spot replication. A novel 2D numerical continuation analysis is performed that shows the various stable hybrid spot-like states can coexist. The parameter values studied lead to a natural singularly perturbed, so-called semi-strong interaction regime. This scaling enables an analytical explanation of the initial instability, by describing the dispersion relation of a certain non-local eigenvalue problem. The analytical results are found to agree favourably with the numerics. Possible biological implications of the results are discussed.

Potassium transporter TRH1 subunits assemble regulating root-hair elongation autonomously from the cell fate determination pathway

Trichoblasts of trh1 plants form root-hair initiation sites that fail to undergo tip growth resulting in a tiny root-hair phenotype. TRH1 belongs to Arabidopsis KT/KUP/HAK potassium transporter family controlling root-hair growth and gravitropism. Double mutant combinations between trh1 and root-hair mutants affecting cell fate or root-hair initiation exhibited additive phenotypes, suggesting that TRH1 acts independently and developmentally downstream of root-hair initiation. Bimolecular Fluorescence Complementation (BiFC), upon TRH1-YFPC and TRH1-YFPN co-transformation into tobacco epidermal cells, led to fluorescence emission indicative of TRH1 subunit homodimerization. Yeast two-hybrid analysis revealed two types of interactions. The hydrophilic segment between the second and the third transmembrane domain extending from residues Q105 to T141 is competent for a relatively weak interaction, whereas the region at the C-terminal beyond the last transmembrane domain, extending from amino acids R565 to A729, strongly self-interacts. These domains likely facilitate the co-assembly of TRH1 subunits forming an active K+ transport system within cellular membrane structures. The results support the role of TRH1 acting as a convergence point between the developmental root-hair pathway and the environmental/hormonal signaling pathway to preserve auxin homeostasis ensuring plant adaptation in changing environments.

Roothairless5, which functions in maize (Zea mays L.) root hair initiation and elongation encodes a monocot-specific NADPH oxidase.

Root hairs are instrumental for nutrient uptake in monocot cereals. The maize (Zea mays L.) roothairless5 (rth5) mutant displays defects in root hair initiation and elongation manifested by a reduced density and length of root hairs. Map-based cloning revealed that the rth5 gene encodes a monocot-specific NADPH oxidase. RNA-Seq, in situ hybridization and qRT-PCR experiments demonstrated that the rth5 gene displays preferential expression in root hairs but also accumulates to low levels in other tissues. Immunolocalization detected RTH5 proteins in the epidermis of the elongation and differentiation zone of primary roots. Because superoxide and hydrogen peroxide levels are reduced in the tips of growing rth5 mutant root hairs as compared with wild-type, and Reactive oxygen species (ROS) is known to be involved in tip growth, we hypothesize that the RTH5 protein is responsible for establishing the high levels of ROS in the tips of growing root hairs required for elongation. Consistent with this hypothesis, a comparative RNA-Seq analysis of 6-day-old rth5 versus wild-type primary roots revealed significant over-representation of only two gene ontology (GO) classes related to the biological functions (i.e. oxidation/reduction and carbohydrate metabolism) among 893 differentially expressed genes (FDR <5%). Within these two classes the subgroups 'response to oxidative stress' and 'cellulose biosynthesis' were most prominently represented.

Root hairs.

Roots hairs are cylindrical extensions of root epidermal cells that are important for acquisition of nutrients, microbe interactions, and plant anchorage. The molecular mechanisms involved in the specification, differentiation, and physiology of root hairs in Arabidopsis are reviewed here. Root hair specification in Arabidopsis is determined by position-dependent signaling and molecular feedback loops causing differential accumulation of a WD-bHLH-Myb transcriptional complex. The initiation of root hairs is dependent on the RHD6 bHLH gene family and auxin to define the site of outgrowth. Root hair elongation relies on polarized cell expansion at the growing tip, which involves multiple integrated processes including cell secretion, endomembrane trafficking, cytoskeletal organization, and cell wall modifications. The study of root hair biology in Arabidopsis has provided a model cell type for insights into many aspects of plant development and cell biology.

Mathematical Modeling of Plant Root Hair Initiation: Dynamics of Localized Patches

A mathematical analysis is undertaken of a Schnakenberg reaction-diffusion system in one dimension with a spatial gradient governing the active reaction. This system has previously been proposed as a model of the initiation of hairs from the root epidermis Arabidopsis, a key cellular-level morphogenesis problem. This process involves the dynamics of the small G-proteins, Rhos of plants, which bind to form a single localized patch on the cell membrane, prompting cell wall softening and subsequent hair growth. A numerical bifurcation analysis is presented as two key parameters, involving the cell length and the overall concentration of the auxin catalyst, are varied. The results show hysteretic transitions from a boundary patch to a single interior patch, and to multiple patches whose locations are carefully controlled by the auxin gradient. The results are confirmed by an asymptotic analysis using semistrong interaction theory, leading to closed form expressions for the patch locations and intensities. A c...

Influence of nutrient deficiency on root architecture and root hair morphology of trifoliate orange (Poncirus trifoliata L. Raf.) seedlings under sand culture

Root hairs are important to plants for nutrient and water uptake through providing an increased absorptive surface area and exploitable soil volume. Citrus, as one of the most important fruit crops in the world, is generally believed to have few or even no root hair in the field and thus strongly depends on arbuscular mycorrhizas. In this study, sand culture experiment was carried out to investigate the effect of different nutrient deficiency conditions on the root hair development and root growth of one-month-old Poncirus trifoliata seedlings. We demonstrated that root hairs were distributed asymmetrically in the whole root, indicating it might be one of the characteristics of the citrus root hair distribution. Furthermore, the density and length of root hair were much higher in P, Ca deficiency than the control, however, lower in K-deficiency, implying the critical role of those nutrients in root hair initiation and elongation. Additionally, root growth was hampered at Ca, Mg, Fe, or Mn deprivation condition, suggesting that it is necessary to supply sufficient those mineral nutrients at seedling stage to ensure better root architecture.

Arabidopsis RopGEF4 and RopGEF10 are important for FERONIA‐mediated developmental but not environmental regulation of root hair growth

We investigated a genetic pathway in root hair development in Arabidopsis thaliana, involving the receptor-like kinase FERONIA (FER), two guanine nucleotide exchange factors for ROPs (RopGEF4 and RopGEF10), and the small GTPase Rho of plants (ROPs). Loss- and gain-of-function analyses demonstrated distinct roles of RopGEF4 and RopGEF10 such that RopGEF4 is only important for root hair elongation, while RopGEF10 mainly contributes to root hair initiation. Domain dissection by truncation and domain-swapping experiments indicated that their functional distinctions were mainly contributed by the noncatalytic domains. Using fluorescent ratio imaging, we showed that functional loss of RopGEF4 and RopGEF10 additively reduced reactive oxygen species (ROS) production. Bimolecular fluorescence complementation experiments demonstrated that RopGEF4 and RopGEF10 had the same interaction specificity as ROPs, suggesting common downstream components. We further showed that the promoting effects of environmental cues such as exogenous auxin and phosphate limitation on root hair development depended on FER. However, although functional loss of RopGEF4 and RopGEF10 largely abolished FER-induced ROS production, it did not compromise the responses to FER-mediated environmental cues on root hair development. Our results demonstrated that RopGEF4 and RopGEF10 are genetic components in FER-mediated, developmentally (but not environmentally) regulated, root hair growth.

Root hair development of Poncirus trifoliata grown in different growth cultures and treated with 3-indolebutyric acid and ethephon

Citrus plants are generally believed to possess few or even no root hair whose function is to uptake nutrient and water from soils. This study compared the development of root hair of a citrus rootstock, Poncirus trifoliata, in four types of cultures or treated with 3-indolebutyric acid and ethephon. It was found that root hair of lateral root grew in sand culture compared with yellow-soil and solution culture in which no initiation was observed. Taproot hair grew best and evenly distributed on MT Agar culture, while in sand culture, Hoagland-solution culture, and yellow soil culture few or no root hair initiation was observed. For P. trifoliata, MT Agar culture showed the best performance on root hair. With regard to the effect of regulators such as 3-indolebutyric acid (IBA) and ethephon (ETH), our result showed that the optimum concentration of IBA and ETH for root hair growth was 1.0μmolL−1 since IBA and ETH at this concentration could best promote root hair initiation and elongation. Besides, the effect of IBA on root hair growth was stronger than ETH at equal concentrations. Furthermore, our results showed that unlike Arabidopsis or Oryza sativa, the root hair of P. trifoliata could initiate from any epidermal cell (random type).

Root hairs improve root penetration, root–soil contact, and phosphorus acquisition in soils of different strength

Root hairs are a key trait for improving the acquisition of phosphorus (P) by plants. However, it is not known whether root hairs provide significant advantage for plant growth under combined soil stresses, particularly under conditions that are known to restrict root hair initiation or elongation (e.g. compacted or high-strength soils). To investigate this, the root growth and P uptake of root hair genotypes of barley, Hordeum vulgare L. (i.e. genotypes with and without root hairs), were assessed under combinations of P deficiency and high soil strength. Genotypes with root hairs were found to have an advantage for root penetration into high-strength layers relative to root hairless genotypes. In P-deficient soils, despite a 20% reduction in root hair length under high-strength conditions, genotypes with root hairs were also found to have an advantage for P uptake. However, in fertilized soils, root hairs conferred an advantage for P uptake in low-strength soil but not in high-strength soil. Improved root-soil contact, coupled with an increased supply of P to the root, may decrease the value of root hairs for P acquisition in high-strength, high-P soils. Nevertheless, this work demonstrates that root hairs are a valuable trait for plant growth and nutrient acquisition under combined soil stresses. Selecting plants with superior root hair traits is important for improving P uptake efficiency and hence the sustainability of agricultural systems.

The Cotton Transcription Factor TCP14 Functions in Auxin-Mediated Epidermal Cell Differentiation and Elongation

Plant-specific TEOSINTE-BRANCHED1/CYCLOIDEA/PCF (TCP) transcription factors play crucial roles in development, but their functional mechanisms remain largely unknown. Here, we characterized the cellular functions of the class I TCP transcription factor GhTCP14 from upland cotton (Gossypium hirsutum). GhTCP14 is expressed predominantly in fiber cells, especially at the initiation and elongation stages of development, and its expression increased in response to exogenous auxin. Induced heterologous overexpression of GhTCP14 in Arabidopsis (Arabidopsis thaliana) enhanced initiation and elongation of trichomes and root hairs. In addition, root gravitropism was severely affected, similar to mutant of the auxin efflux carrier PIN-FORMED2 (PIN2) gene. Examination of auxin distribution in GhTCP14-expressing Arabidopsis by observation of auxin-responsive reporters revealed substantial alterations in auxin distribution in sepal trichomes and root cortical regions. Consistent with these changes, expression of the auxin uptake carrier AUXIN1 (AUX1) was up-regulated and PIN2 expression was down-regulated in the GhTCP14-expressing plants. The association of GhTCP14 with auxin responses was also evidenced by the enhanced expression of auxin response gene IAA3, a gene in the AUXIN/INDOLE-3-ACETIC ACID (Aux/IAA) family. Electrophoretic mobility shift assays showed that GhTCP14 bound the promoters of PIN2, IAA3, and AUX1, and transactivation assays indicated that GhTCP14 had transcription activation activity. Taken together, these results demonstrate that GhTCP14 is a dual-function transcription factor able to positively or negatively regulate expression of auxin response and transporter genes, thus potentially acting as a crucial regulator in auxin-mediated differentiation and elongation of cotton fiber cells.

Auxin biology in roots

Auxin regulates almost every aspect of plant growth and development. Its intracellular concentration is controlled by biosynthesis and degradation. In addition, there is an pool that consists of the conjugates with sugars, amino acids, and peptides. Some of the conjugates reversely release auxin, enabling alternative methods to regulate auxin concentrations. Auxin concentrations are also affected by transport. Besides the long distance delivery through the phloem, auxin is transported across the cell by influx and efflux carriers, from the shoot to root with maximum concentration at the root tip. At the root tip, the auxin flow reverses, and shootward auxin transport occurs. An auxin gradient formed this way is indispensable for proper development, maintenance of the meristem, and cell identity. The formation of root hairs is auxin-dependent. Auxin controls not only the initiation of root hairs but also regulates their elongation. In Arabidopsis thaliana, auxin accumulates in atrichoblasts and it is supplied to trichoblasts. Before hair initiation, randomization of cortical microtubule arrays is observed in lettuce seedlings. This action is promoted by auxin and is indispensable to hair formation. Furthermore, light promotes CMT randomization and root hair initiation via auxin signaling. Ethylene is another promoter of root hair formation. Ethylene affects auxin signaling and vice versa. Interactions between these hormones are synergistic for root growth inhibition but antagonistic for lateral root formation. Reactive oxygen species also regulate various responses in plants. They play an important role during root hair elongation, although their precise relationship with auxin is yet not clear.

Overlapping and divergent signaling pathways for ARK1 and AGD1 in the control of root hair polarity in Arabidopsis thaliana

We previously showed that seedlings harboring mutations in genes encoding ARK1, an armadillo repeat-containing kinesin, or AGD1, a class 1 ARF-GAP, have root hairs that exhibit wavy/spiral growth and two tips originating from one initiation site. These root hair defects were accompanied by bundling of endoplasmic microtubules and filamentous actin (F-actin) that extended to the extreme root hair apex. The similar phenotypes of ark1 and agd1 mutants suggest a tight coordination between the cytoskeleton and membrane trafficking in the control of root hair polarity. Indeed, cell biological and genetic studies of the agd1 mutant provided evidence that AGD1's involvement in root hair development involves cross-talk among phosphoinositides (PIs), the actin cytoskeleton and other small GTPases such as ROP2 and RABA4b. Here we show that ark1 root hairs mirror those of agd1 with regard to altered targeting of ROP2 and RABA4b, as well as abnormal tonoplast organization. Furthermore, like agd1, enhanced root hair defects in double mutants in ARK1 and genes encoding a type B phosphatidylinositol-4-phosphate 5-kinase 3 (PIP5K3), a phosphatidylinositol-4-phosphate (PI-4P) phosphatase (RHD4), a phosphatidylinositol transfer protein (COW1), and a vegetative actin isoform (ACT2), were observed. However, root hair shape of some ark1 double mutant combinations, particularly those with act2, pip5k3 and rhd4 (ark1 act2, ark1 pip5k3, ark1 rhd4), differed in some respects from agd1 act2, agd1 pip5k3, and agd1 rhd4. Taken together our results continue to point to commonalities between ARK1 and AGD1 in specifying root hair polarity, but that these two modulators of tip-growth can also regulate root hair development through divergent signaling routes with AGD1 acting predominantly during root hair initiation and ARK1 functioning primarily in sustained tip growth.

Accumulation of peroxidase-related reactive oxygen species in trichoblasts correlates with root hair initiation in barley

Root hairs are an important model in studies of cell differentiation and development in higher plants. The function of NADPH oxidase-related reactive oxygen species (ROS) in root hair development has been reported extensively in studies on Arabidopsis. In this study, we investigated the mechanism of the initiation of root hair formation, mediated by the peroxidase-dependent production of the highly reactive hydroxyl radical in barley (Hordeum vulgare L.). The distribution of ROS, including the hydroxyl radical (OH) and superoxide (O2−) was assessed using hydroxyphenyl fluorescein and nitroblue tetrazolium chloride, respectively, in the roots of wild-type plants and two root-hair mutants: root-hairless (rhl1.a) and with root hair growth blocked at the primordium stage (rhp1.b). Peroxidase-dependent OH accumulation was linked to root hair initiation and growth in plants where root hair formation was at least initiated, whereas OH was not detectable in the epidermis of the root-hairless mutant rhl1.a. O2− distribution in the roots of rhl1.a and rhp1.b mutants was not impaired and did not influence the root hair phenotype. Peroxidase inhibitor treatments of wild-type roots dramatically reduced the ability of growing roots to form root hairs and thus phenocopied the root-hairless phenotype. Expression of two candidate peroxidase genes, HvPRX45 and HvPRX2, was analyzed and their possible role in root hair-specific production of hydroxyl radicals was discussed. We propose a model of a two-step, coordinated ROS formation process in root hair cells that involves root hair-specific peroxidase(s) and root hair-specific NADPH oxidase necessary for a proper root hair formation in barley.

GbTCP, a cotton TCP transcription factor, confers fibre elongation and root hair development by a complex regulating system.

As the most important natural raw material for textile industry, cotton fibres are an excellent model for studying single-cell development. Although expression profiling and functional genomics have provided some data, the mechanism of fibre development is still not well known. A class I TCP transcription factor (designated GbTCP), encoding 344 amino acids, was isolated from the normalized cDNA library of sea-island cotton fibre (from –2 to 25 days post anthesis). GbTCP was preferentially expressed in the elongating cotton fibre from 5 to 15 days post anthesis. Some expression was also observed in stems, apical buds, and petals. RNAi silencing of GbTCP produced shorter fibre, a reduced lint percentage, and a lower fibre quality than the wild-type plants. Overexpression of GbTCP enhanced root hair initiation and elongation in Arabidopsis and regulated branching. Solexa sequencing and Affymetrix GeneChip analysis indicated that GbTCP positively regulates the level of jasmonic acid (JA) and, as a result, activates downstream genes (reactive oxygen species, calcium signalling, ethylene biosynthesis and response, and several NAC and WRKY transcription factors) necessary for elongation of fibres and root hairs. JA content analysis in cotton also confirmed that GbTCP has a profound effect on JA biosynthesis. In vitro ovule culture showed that an appropriate concentration of JA promoted fibre elongation. The results suggest that GbTCP is an important transcription factor for fibre and root hair development by regulating JA biosynthesis and response and other pathways, including reactive oxygen species, calcium channel and ethylene signalling.

A zinc finger protein gene ZFP5 integrates phytohormone signaling to control root hair development in Arabidopsis

Although root hair development in Arabidopsis thaliana has been extensively studied, it remains unknown whether the zinc finger proteins, the largest family of transcription factors in plants, are involved in this process. Here we report that the C2H2 zinc finger protein ZINC FINGER PROTEIN 5 (ZFP5) is a key regulator of root hair initiation and morphogenesis in Arabidopsis. ZFP5 is mainly expressed in root and preferentially in root hair cells. Using both zfp5 mutants and ZFP5 RNAi lines, we show that reduction in ZFP5 function leads to fewer and much shorter root hairs compared to wild-type. Genetic and molecular experiments demonstrate that ZFP5 exerts its effect on root hair development by directly promoting expression of the CAPRICE (CPC) gene. Furthermore, we show that ZFP5 expression is induced by cytokinin, and that ZFP5 mediates cytokinin and ethylene effects on the formation and growth of root hairs. These results suggest that ZFP5 integrates various plant hormone cues to control root epidermal cell development in Arabidopsis.

Monoxenic nodulation process of Acacia mangium (Mimosoideae, Phyllodineae) by Bradyrhizobium sp

Acacia mangium Willd., a native tree in Australia and Papua New Guinea, has been introduced to countries in Asia and South America where plantations have been established that cover several hundred thousand ha. The present study investigated the early stages of the nodulation process in A. mangium using an homologous Australian Bradyrhizobium strain. After optimizing the axenic nodulation, histological and cytological studies were conducted using light and electron microscopy. These documented the proliferation of Bradyrhizobium, the lysis of mucilage at the root surface, root hair deformation and initiation, as well as the development and growth of multiple infection threads. A belt of tannin-filled cells was shown to surround the central nodular fixation zone. The nodules were of the indeterminate type and the bacteroids had a rod shape, without size modification and with few polyhydroxybutyrate (PHB) granules. Several bacteroids can share the same symbiosome. A. mangium exhibits both classical and novel features in its nodulation.

A Single Amino Acid Substitution in IIIf Subfamily of Basic Helix-Loop-Helix Transcription Factor AtMYC1 Leads to Trichome and Root Hair Patterning Defects by Abolishing Its Interaction with Partner Proteins in Arabidopsis

Plant trichomes and root hairs are powerful models for the study of cell fate determination. In Arabidopsis thaliana, trichome and root hair initiation requires a combination of three groups of proteins, including the WD40 repeat protein transparent TESTA GLABRA1 (TTG1), R2R3 repeat MYB protein GLABRA1 (GL1), or werewolf (WER) and the IIIf subfamily of basic helix-loop-helix (bHLH) protein GLABRA3 (GL3) or enhancer of GLABRA3 (EGL3). The bHLH component acts as a docking site for TTG1 and MYB proteins. Here, we isolated a mutant showing defects in trichome and root hair patterning that carried a point mutation (R173H) in AtMYC1 that encodes the fourth member of IIIf bHLH family protein. Genetic analysis revealed partial redundant yet distinct function between AtMYC1 and GL3/EGL3. GLABRA2 (GL2), an important transcription factor involved in trichome and root hair control, was down-regulated in Atmyc1 plants, suggesting the requirement of AtMYC1 for appropriate GL2 transcription. Like its homologs, AtMYC1 formed a complex with TTG1 and MYB proteins but did not dimerized. In addition, the interaction of AtMYC1 with MYB proteins and TTG1 was abrogated by the R173H substitution in Atmyc1-1. We found that this amino acid (Arg) is conserved in the AtMYC1 homologs GL3/EGL3 and that it is essential for their interaction with MYB proteins and for their proper functions. Our findings indicate that AtMYC1 is an important regulator of trichome and root hair initiation, and they reveal a novel amino acid necessary for protein-protein interactions and gene function in IIIf subfamily bHLH transcription factors.

Root hair development involves asymmetric cell division in Brachypodium distachyon and symmetric division in Oryza sativa

• The root epidermis of most angiosperms comprises hair (H) cells and nonhair (N) cells. H cells are shorter than N cells in grasses (Poaceae). • The aim of this study was to determine the developmental basis for differences in H and N cell size in the grasses Brachypodium distachyon and Oryza sativa. • We show that cytokinesis in the last cell division in each epidermal file is asymmetric in B. distachyon. The smaller daughter cell becomes an H cell and the larger cell forms an N cell. By contrast, asymmetric cytokinesis does not occur during H cell and N cell development in O. sativa and the differences in size arise because there is more cell expansion in N cells than in H cells after root hair initiation. • The different sizes of mature H and N cells result from cell division asymmetry in B. distachyon but different rates of cell expansion in O. sativa. We hypothesize that the mechanism that includes asymmetric cytokinesis during the development of H and N cells evolved among the Pooideae or ancestors of this subfamily.

Control of root hair development in Arabidopsis thaliana by an endoplasmic reticulum anchored member of the R2R3‐MYB transcription factor family

The evolution of roots and root hairs was a crucial innovation that contributed to the adaptation of plants to a terrestrial environment. Initiation of root hairs involves transcriptional cues that in part determine cell patterning of the root epidermis. Once root hair initiation has occurred, elongation of the root hair takes place. Although many genes have been identified as being involved in root hair development, many contributors remain uncharacterized. In this study we report on the involvement of a member (here dubbed maMYB) of the plant-specific R2R3-MYB family of transcription factors in root hair elongation in Arabidopsis. We show that maMYB is associated with the endoplasmic reticulum membrane with the transcription factor domain exposed to the cytosol, suggesting that it may function as a membrane-tethered transcription factor. We demonstrate that a truncated form of maMYB (maMYB⁸⁴⁻³⁰⁹), which contains the R2R3-MYB transcription factor domain, is localized and retained in the nucleus, where it regulates gene expression. Silencing of maMyb resulted in plants with significantly shorter root hairs but similar root hair density compared with wild type, implying a role of the protein in root hair elongation. 2,4-D (2,4-dichlorophenoxyacetic acid), an exogenous auxin analog that promotes root hair elongation, rescued the short root hair phenotype and maMyb mRNA was induced in the presence of 2,4-D and IAA (indole-3-acetic acid). These results indicate a functional role of maMYB, which is integrated with auxin, in root hair elongation in Arabidopsis.