Root Hair Elongation Research Articles

The Alfin-like transcription factors: Identification, characterization, and expression analysis in Pyrus bretschenedri provide insight into its divergent functions on abiotic response

In the previous decades, numerous investigations have been done that the Alfin-like (AL) transcription factors (TFs) are participating in several physiological and development processes in plants including root hair elongation, development of root and meristem, etc. Although the AL-TFs had been comprehensively evaluated in several species, the little finding is reported on the AL-TFs in Chinese pear (Pyrus bretschenedri). In the current study, 15 PbAL genes were systemically characterized and nomenclature as PbAL1 to PbAL15 corresponding to their chromosomal localization. Phylogeny analysis and structural features revealed that the 15 genes could be classified into six subfamilies (S-I to S-VI).Whole-genome duplication (WGD) events are the most common mechanism and may play a crucial role in the expansion of the AL gene family. Microsynteny analysis among Pyrus bretschenedri, Fragaria vesca, Prunus avium, Prunus mume, and Prunus persica showed that AL duplicated regions were more conserved and undergo purifying selection during the evolutionary process. Further analysis of introns-exons, physicochemical properties, and conserved motif showed their functional divergence, diversification in quantity, and structure. The expression pattern of PbAL gene family in various organs and different fruit development stages were conducted through transcriptomic data. Moreover, stress-related cis-acting elements of PbAL genes were found on the promoters regions to be associated with hormonal and environmental stress responses. Noticeably, the expression patterns evaluated by qRT-PCR demonstrated that PbAL genes were differentially expressed under indole acetic acid (IAA), Gibberellic acid (GA), Melatonin (ME) and abscisic acid (ABA) treatment, which response to abiotic stress and fruit growth development. Current work explored the potential function and evolution of PbAL genes in Pyrus bretschenedri. PbAL were investigated as candidate genes for further research and important clues for improving fruit quality through molecular breeding.

Phosphate deprivation-induced changes in tomato are mediated by an interaction between brassinosteroid signaling and zinc.

Inorganic phosphate (Pi) is a necessary macronutrient for basic biological processes. Plants modulate their root system architecture (RSA) and cellular processes to adapt to Pi deprivation albeit with a growth penalty. Excess application of Pi fertilizer, on the contrary, leads to eutrophication and has a negative environmental impact. We compared RSA, root hair elongation, acid phosphatase activity, metal ion accumulation, and brassinosteroid hormone levels of Solanum lycopersicum (tomato) and Solanum pennellii, which is a wild relative of tomato, under Pi sufficiency and deficiency conditions to understand the molecular mechanism of Pi deprivation response in tomato. We showed that S. pennellii is partially insensitive to phosphate deprivation. Furthermore, it mounts a constitutive response under phosphate sufficiency. We demonstrate that activated brassinosteroid signaling through a tomato BZR1 ortholog gives rise to the same constitutive phosphate deficiency response, which is dependent on zinc overaccumulation. Collectively, these results reveal an additional strategy by which plants can adapt to phosphate starvation.

Discovery of Novel Hybrid-Type Strigolactone Mimics Derived from Cinnamic Amide.

Strigolactones (SLs) are a class of plant hormones and rhizosphere communication signals of great interest. They perform diverse biological functions including the stimulation of parasitic seed germination and phytohormonal activity. However, their practical use is limited by their low abundance and complex structure, which requires simpler SL analogues and mimics with maintained biological function. Here, new, hybrid-type SL mimics were designed, derived from Cinnamic amide, a new potential plant growth regulator with good germination and rooting-promoting activities. Bioassay results indicated that compound 6 not only displayed good germination activity against the parasitic weed O. aegyptiaca with an EC50 value of 2.36 × 10-8 M, but also exhibited significant inhibitory activity against Arabidopsis root growth and lateral root formation, as well as promoting root hair elongation, similar to the action of GR24. Further morphological experiments on Arabidopsis max2-1 mutants revealed that 6 possessed SL-like physiological functions. Furthermore, molecular docking studies indicated that the binding mode of 6 was similar to that of GR24 in the active site of OsD14. This work provides valuable clues for the discovery of novel SL mimics.

A natural non-synonymous single nucleotide polymorphism (SNP) in GmbHLH113 negates its inhibitory effect on root hair elongation in soybean.

Root hair length (RHL) is an important character that affects nutrient acquisition in plants. The regulatory network in soybean controlling RHL is yet to be fully understood. In this study, we identified a quantitative trait locus (QTL) regulating RHL. One candidate causal gene in this QTL (GmbHLH113), preferentially expressed in root hairs, was annotated as encoding a basic Helix-Loop-Helix (bHLH) transcription factor. In wild soybeans, the allelic type of GmbHLH113 with a glycine in the 13th residue, which was associated with a reduction in RHL, was shown to localize in the nucleus and activate gene transcription. Another allelic type with a single nucleotide polymorphism (SNP) that resulted in a glutamate in the 13th residue is fixed in cultivated soybeans, and it lost the ability to localize to the nucleus or negatively regulate RHL. The ectopic expression of GmbHLH113 from W05 in Arabidopsis root hairs resulted in shorter RHL and reduced phosphorus (P) accumulation in shoots. Hence a loss-of-function allele in cultivated soybeans might have been selected during domestication due to its association with a longer RHL and improved nutrient acquisition.

The m6A reader MhYTP2 regulates the stability of its target mRNAs contributing to low nitrogen tolerance in apple (Malus domestica).

Studies have shown that the m6A reader primarily affects genes expression by participating in the regulation of mRNA localization, splicing, degradation, translation, and other metabolic processes. Previously, we discovered that the apple (Malus domestica) m6A reader MhYTP2 bound with and destabilized m6A-modified MdMLO19 mRNA. In addition, it enhanced the translation efficiency of m6A-modified mRNA of MdGDH1L, encoding a glutamate dehydrogenase, which confers resistance to powdery mildew. In this study, we report the function of MhYTP2 in the regulation of resistance to low nitrogen (N). The overexpression of MhYTP2 enhances the resistance of apple to low N. We show that MhYTP2 binds with and stabilizes the mRNAs of MdALN, which participates in the allantoin catabolic process and cellular response to N starvation in apple; MdPIDL, which participates in root hair elongation; MdTTG1, which is involved in the differentiation process of trichomes; and MdATG8A, which is a core participant in the regulation of autophagy. In addition, MhYTP2 accelerates the degradation of MdRHD3 mRNA, which regulates root development. RNA immunoprecipitation-seq and electrophoretic mobility shift assays show that the mRNAs of MdALN, MdATG8A, MdPIDL, MdTTG1, and MdRHD3 are the direct targets of MhYTP2. Overexpressing or knocking down the above genes in MhYTP2 overexpressing plants dismisses the function of MhYTP2 under low N, suggesting the role of MhYTP2 is dependent on those genes. Together, these results demonstrate that MhYTP2 enhances the resistance of apple to N deficiency by affecting the stability of the bound mRNAs.

OsUGE1 is directly targeted by OsGRF6 to regulate root hair length in rice.

Root hairs are required for water and nutrient acquisition in plants. Here, we report a novel mechanism that OsUGE1 is negatively controlled by OsGRF6 to regulate root hair elongation in rice. Root hairs are tubular outgrowths generated by the root epidermal cells. They effectively enlarge the soil-root contact area and play essential roles for nutrient and water absorption. Here, in this study, we demonstrated that the Oryza sativa UDP-glucose 4-epimerase 1-like (OsUGE1) negatively regulated root hair elongation and was directly targeted by Oryza sativa growth regulating factor 6 (OsGRF6). Knockout mutants of OsUGE1 using CRISPR-Cas9 technology showed longer root hairs than those of wild type. In contrast, overexpression lines of OsUGE1 displayed shorter root hair compared with those of wild type. GUS staining showed that it could specifically express in root hair. Subcellular localization analysis indicates that OsUGE1 is located in endoplasmic reticulum, nucleus and plasma membrane. More importantly, ChIP-qPCR, Yeast-one-hybrid and BiFC experiments revealed that OsGRF6 could bind to the promoter of OsUGE1. Furthermore, knockout mutants of OsGRF6 showed shorter root hair than those of wild type, and OsGRF6 dominantly expressed in root. In addition, the expression level of OsUGE1 is significantly downregulated in Osgrf6 mutant. Taken together, our study reveals a novel pathway that OsUGE1 is negatively controlled by OsGRF6 to regulate root hair elongation in rice.

Loss-of-function of MEDIATOR 12 or 13 subunits causes the swelling of root hairs in response to sucrose and abscisic acid in Arabidopsis

ABSTRACT Root hairs are epidermal cell extensions that increase the root surface for water and nutrient acquisition. Thus, both the initiation and elongation of root hairs are critical for soil exploration and plant adaptation to ever changing growth conditions. Here, we describe the critical roles of two subunits of the Mediator complex, MED12 and MED13, in root hair growth in response to sucrose and abscisic acid, which are tightly linked to abiotic stress resistance. When compared to the WT, med12 and med13 mutants showed increased sensitivity to sucrose and ABA treatments on root meristem and elongation zones that were accompanied with alterations in root hair length and morphology, leading to the isodiametric growth of these structures. The swollen root hair phenotype appeared to be specific, since med8 or med16 mutants did not develop rounded hairs when supplied with 4.8% sucrose. Under standard growth medium, MED12 and MED13 were mainly expressed in root vascular tissues and cotyledons, and their expression was repressed by sucrose or ABA. Interestingly, med12 and med13 mutants manifested exacerbated levels of nitric oxide under normal growth conditions, and upon sucrose supplementation in trichoblast cells, which coincided with root hair deformation. Our results indicate that MED12 and MED13 play non-redundant functions for maintenance of root hair integrity in response to sucrose and ABA and involve nitric oxide as a cellular messenger in Arabidopsis thaliana.

Cytokinin signaling promotes root hair growth by directly regulating RSL4 expression.

Root hairs are single-celled tubular structures produced from the epidermis, which play an essential role in water and nutrient uptake from the soil. Therefore, root hair formation and elongation are controlled not only by developmental programs but also by environmental factors, enabling plants to survive under fluctuating conditions. Phytohormones are key signals that link environmental cues to developmental programs; indeed, root hair elongation is known to be controlled by auxin and ethylene. Another phytohormone, cytokinin, also affects root hair growth, while whether cytokinin is actively involved in root hair growth and, if so, how it regulates the signaling pathway governing root hair development have remained unknown. In this study, we show that the two-component system of cytokinin, which involves the B-type response regulators ARABIDOPSIS RESPONSE REGULATOR 1 (ARR1) and ARR12, promotes the elongation process of root hairs. They directly up-regulate ROOT HAIR DEFECTIVE 6-LIKE 4 (RSL4) encoding a basic helix-loop-helix (bHLH) transcription factor that plays a central role in root hair growth, whereas the ARR1/12-RSL4 pathway does not crosstalk with auxin or ethylene signaling. These results indicate that cytokinin signaling constitutes another input onto the regulatory module governed by RSL4, making it possible to fine-tune root hair growth in changing environments.

Role of root hair elongation in rhizosheath aggregation and in the carbon flow into the soil

One of the most prominent changes in the rhizospheric soil structure is associated with the formation of a strongly bound soil layer in the surroundings of the root, which is named rhizosheath. In this study, we investigated how root hair elongation, a ubiquitous root morphological trait, affect the stability of rhizosheath aggregates. Using 13CO2 pulse labeling, we tracked the fate of root-derived 13C inputted into the rhizosheath of two Zea mays L. genotypes with contrasting root hair elongation: a mutant with root hair defective elongation (rth3) and a corresponding wild type (WT). In addition, we also investigated the differences between two 13CO2 labeling approaches (single vs. multiple pulse labeling) in the distribution of 13C in the rhizosheath aggregates. We were able to demonstrate that the rhizosheath aggregate stability and the resulting aggregate size distribution follows the same mechanisms irrespective of the root hair elongation. This result reinforces the assumption that other soil properties are more decisive for the soil structure formation in the rhizosheath in comparison to root hair elongation. The majority of recently deposited root-derived C (57%) was found in the macroaggregates. Increasing the number of pulses (multiple pulse labeling approach) resulted in a higher 13C enrichment of the rhizosheath aggregates fractions in comparison to the application of a single pulse. While both labeling approaches resulted in a similar distribution of 13C in the rhizosheath aggregates, the higher enrichment given by multiple pulse labeling allowed the separation of significant differences between the genotypes in plant C allocation in the rhizosheath.

VPS45 is required for both diffuse and tip growth of Arabidopsis thaliana cells.

VPS45 belongs to the Sec1/Munc18 family of proteins, which interact with and regulate Qa-SNARE function during membrane fusion. We have shown previously that Arabidopsis thaliana VPS45 interacts with the SYP61/SYP41/VTI12 SNARE complex, which locates on the trans-Golgi network (TGN). It is required for SYP41 stability, and it functions in cargo trafficking to the vacuole and in cell expansion. It is also required for correct auxin distribution during gravitropism and lateral root growth. As vps45 knockout mutation is lethal in Arabidopsis, we identified a mutant, vps45-3, with a point mutation in the VPS45 gene causing a serine 284-to-phenylalanine substitution. The VPS45-3 protein is stable and maintains interaction with SYP61 and SYP41. However, vps45-3 plants display severe growth defects with significantly reduced organ and cell size, similar to vps45 RNAi transgenic lines that have reduced VPS45 protein levels. Root hair and pollen tube elongation, both processes of tip growth, are highly compromised in vps45-3. Mutant root hairs are shorter and thicker than those of wild-type plants, and are wavy. These root hairs have vacuolar defects, containing many small vacuoles, compared with WT root hairs with a single large vacuole occupying much of the cell volume. Pollen tubes were also significantly shorter in vps45-3 compared to WT. We thus show that VPS45 is essential for proper tip growth and propose that the observed vacuolar defects lead to loss of the turgor pressure needed for tip growth.

An apple NITRATE REDUCTASE 2 gene positively regulates nitrogen utilization and abiotic stress tolerance in Arabidopsis and apple callus

Nitrogen (N) is an essential element that plays an important role in crop biomass accumulation and quality formation. Increased crop yield is relied on excessive application of fertilizers, which usually leads to environmental pollution and unsustainable development. Thus, identification and characterization of genes involved in promoting nitrogen use efficiency is of high priority in crop breeding. The activity of nitrate reductase (NR) plays a critical role in nitrogen metabolism. In model plant Arabidopsis, NITRATE REDUCTASE 2 (NIA2), one of the two NRs, is responsible for about 90% of the NR activity. In this study, MdNIA2 gene in apple (Malus domestica) genome was screened out and identified by using AtNIA2 as bait. Phylogenetic analysis revealed that MdNIA2 had the closest evolutionary relationship with MbNIA from Malus baccata. Ectopic expression of MdNIA2 in Arabidopsis elevated the nitrogen use efficiency and increased root hair elongation and formation, resulting in promoted plant growth. Furthermore, the overexpression of MdNIA2 improved salt and drought tolerance in transgenic Arabidopsis and improved the salt tolerance of transgenic apple callus, and MdNIA2-reagualted NO metabolism might contribute to the abiotic stress tolerance. Overall, our data indicate the critical role of MdNIA2 in regulating nitrogen utilization efficiency and abiotic stress responses.

OsCSLD1 Mediates NH4+-Dependent Root Hair Growth Suppression and AMT1;2 Expression in Rice (Oryza sativa L.).

Root hairs play crucial roles in the roots, including nutrient uptake, water assimilation, and anchorage with soil, along with supporting rhizospheric microorganisms. In rice, ammonia uptake is mediated by a specialized ammonium transporter (AMT). AMT1;1, AMT1;2, and AMT1;3 have been extensively studied in relation to nitrogen signaling. Cellulose synthase-like D1 (CSLD1) is essential for cell expansion and is highly specific to root hair cells. csld1 mutants showed successful initiation but failed to elongate. However, when nitrogen was depleted, csld1 root hairs resumed elongation. Further experiments revealed that in the presence of ammonium (NH4+), csld1 roots failed to elongate. csld1 elongated normally in the presence of nitrate (NO3−). Expression analysis showed an increase in root hair-specific AMT1;2 expression in csld1. CSLD1 was positively co-expressed with AMT1;2 changing nitrogen concentration in the growth media. CSLD1 showed increased expression in the presence of both ammonium and nitrate. Methylammonium (MeA) treatment of CSLD1 overexpression lines suggests that CSLD1 does not directly participate in nitrogen transport. Further studies on the root hair elongation mutant sndp1 showed that nitrogen assimilation is unlikely to depend on root hair length. Therefore, these results suggest that CSLD1 is closely involved in nitrogen-dependent root hair elongation and regulation of AMT1;2 expression in rice roots.

Identification of genomic regions associated with the plasticity of carbon 13 ratio in soybean.

Improving water use efficiency (WUE) for soybean [Glycine max (L.) Merr.] through selection for high carbon isotope (C13) ratio may increase drought tolerance, but increased WUE may limit growth in productive environments. An ideal genotype would be plastic for C13 ratio; that is, be able to alter C13 ratio in response to the environment. Our objective was to identify genomic regions associated with C13 ratio plasticity, C13 ratio stability, and overall C13 ratio in two panels of diverse Maturity Group IV soybean accessions. A second objective was to identify accessions that differed in their C13 ratio plasticity. Panel 1 (205 accessions) was evaluated in seven irrigated and four drought environments, and Panel 2 (373 accessions) was evaluated in four environments. Plasticity was quantified as the slope from regressing C13 ratio of individual genotypes against an environmental index calculated based on the mean within and across environments. The regression intercept was considered a measure of C13 ratio over all environments, and the root mean square error was considered a measure of stability. Combined over both panels, genome-wide association mapping (GWAM) identified 19 single nucleotide polymorphisms (SNPs) for plasticity, 39 SNPs for C13 ratio, and 16 SNPs for stability. Among these SNPs, 71 candidate genes had annotations associated with transpiration or water conservation and transport, root development, root hair elongation, and stomatal complex morphogenesis. The genomic regions associated with plasticity and stability identified in the current study will be a useful resource for implementing genomic selection for improving drought tolerance in soybean.

Shank-localized cell wall growth contributes to Arabidopsis root hair elongation.

Root hairs are highly elongated tubular extensions of root epidermal cells with a plethora of physiological functions, particularly in establishing the root-rhizosphere interface. Anisotropic expansion of root hairs is generally thought to be exclusively mediated by tip growth-a highly controlled apically localized secretion of cell wall material-enriched vesicles that drives the extension of the apical dome. Here we show that tip growth is not the only mode of root hair elongation. We identified events of substantial shank-localized cell wall expansion along the polar growth axis of Arabidopsis root hairs using morphometric analysis with quantum dots. These regions expanded after in vivo immunolocalization using cell wall-directed antibodies and appeared as distinct bands that were devoid of cell wall labelling. Application of a novel click chemistry-enabled galactose analogue for pulse chase and real-time imaging allowed us to label xyloglucan, a major root hair glycan, and demonstrate its de novo deposition and enzymatic remodelling in these shank regions. Our data reveal a previously unknown aspect of root hair growth in which both tip- and shank-localized dynamic cell wall deposition and remodelling contribute to root hair elongation.

Modulation of tomato root architecture and root hair traitsby Pseudomonas brassicacearum and Variovorax paradoxus containing 1-aminocyclopropane-1-carboxylate deaminase

By decreasing root 1-aminocyclopropane-1-carboxylate (ACC) content and plant ethylene production, the microbial enzyme ACC deaminase is a widespread beneficial trait of plant growth-promoting rhizobacteria (PGPR), ameliorating ethylene-mediated root growth inhibition. However, relatively little is known about whether bacterial ACC deaminase modulates root architecture and root hair traits. Thus the dwarf tomato (Solanum lycopersicum) cultivar Micro-Tom was inoculated in vitro with Pseudomonas brassicacearum Am3, its ACC deaminase deficient mutant T8-1, a known PGPR strain Variovorax paradoxus 5C-2 or chemically treated with agents that promoted or inhibited ethylene production or sensitivity (Ag+, Co2+, and ACC). ACC treatment reduced both root elongation and the number of lateral roots, while ethylene inhibitors (Ag+, Co2+) and V. paradoxus 5C-2 promoted primary root elongation, but differentially affected lateral root length and number. Ag+ stimulated lateral root development, while Co2+ and V. paradoxus 5C-2 did not. Inoculation with P. brassicacearum Am3 and T8-1 inhibited elongation of the primary and lateral roots at a high inoculum concentration (106 cells cm3). All bacterial strains significantly increased the length and number of root hairs, with these effects more pronounced in P. brassicacearum Am3 than in the mutant T8-1. Treatment with Ag+ inhibited root hair formation and elongation, while Co2+ had the opposite effects. ACC treatment had no effect on root hair elongation but increased root hair density. While root growth inhibition caused by P. brassicacearum Am3 was independent of ACC deaminase, the promotion of root hair elongation and density by this strain was augmented by ACC deaminase activity. Thus ACC deaminase can modulate the morphological impacts of bacteria on root hair response by affecting plant ethylene content.

Ethylene signaling increases reactive oxygen species accumulation to drive root hair initiation in Arabidopsis.

Root hair initiation is a highly regulated aspect of root development. The plant hormone ethylene and its precursor, 1-amino-cyclopropane-1-carboxylic acid, induce formation and elongation of root hairs. Using confocal microscopy paired with redox biosensors and dyes, we demonstrated that treatments that elevate ethylene levels lead to increased hydrogen peroxide accumulation in hair cells prior to root hair formation. In the ethylene-insensitive receptor mutant, etr1-3, and the signaling double mutant, ein3eil1, the increase in root hair number or reactive oxygen species (ROS) accumulation after ACC and ethylene treatment was lost. Conversely, etr1-7, a constitutive ethylene signaling receptor mutant, has increased root hair formation and ROS accumulation, similar to ethylene-treated Col-0 seedlings. The caprice and werewolf transcription factor mutants have decreased and elevated ROS levels, respectively, which are correlated with levels of root hair initiation. The rhd2-6 mutant, with a defect in the gene encoding the ROS-synthesizing RESPIRATORY BURST OXIDASE HOMOLOG C (RBOHC), and the prx44-2 mutant, which is defective in a class III peroxidase, showed impaired ethylene-dependent ROS synthesis and root hair formation via EIN3EIL1-dependent transcriptional regulation. Together, these results indicate that ethylene increases ROS accumulation through RBOHC and PRX44 to drive root hair formation.

A comparative proteomics study of Arabidopsis thaliana responding to the coexistence of BPA and TiO2-NPs at environmentally relevant concentrations

Through the applications of recycling sewage sludge to soils as nutrients, bisphenol A (BPA) and titanium dioxide nanoparticles (TiO2-NPs) are commonly found in the agricultural environment. Previous studies have reported that BPA and nanoparticles are harmful to the environment. However, the combined toxicity of both compounds is not yet understood. This work presented an in-depth proteomic analysis of Arabidopsis thaliana exposed to BPA and TiO2-NPs concurrently at environmentally relevant levels. Seeds were simultaneously treated with varying concentrations of BPA (0, 10, 100, and 1000 µg·kg−1) and TiO2-NPs (0, 1, 10 and 100 mg·kg−1). In treatment of 1000 µg·kg−1 BPA and 100 mg·kg−1 TiO2-NPs, highest seed germination rate (87.97%, p < 0.05) was observed. Shorter primary roots but more branched roots were obtained in treatments of high BPA and NPs concentrations (100, 1000 µg·kg−1 BPA and 10, 100 mg·kg−1 TiO2-NPs) while no significant effects on plant height and biomass were found. In the comparative analysis, both concentration related positive and negative effects were observed, such as regulation of cell proliferation (positive), root hair elongation (positive), cellular response to oxidative stress (negative), and cell wall organization (negative). In response to the stress caused by BPA and TiO2-NPs, some proteins related to plant root development, such as CD48E, DNAJ2 and GL24, were up-regulated explaining the shorter primary root length and more branched roots. Moreover, Arabidopsis may have stimulated its ability of resource transportation and energy metabolism to overcome the stress and maintain or somehow enhance their growth by up-regulating proteins like TBB6, CALM1, RAA2A, G3PP2 and KASC1. Our comparative proteomics analysis also highlighted multiple biological processes that consequently lead to the stability of plant growth and its stress adaptation. The results demonstrated that applying biosolids to soil as a fertilizer may be considered as a sustainable practice.

Diversification of SEC15a and SEC15b isoforms of an exocyst subunit in seed plants is manifested in their specific roles in Arabidopsis sporophyte and male gametophyte.

The exocyst complex is an octameric evolutionarily conserved tethering complex engaged in the regulation of polarized secretion in eukaryotic cells. Here, we focus on the systematic comparison of two isoforms of the SEC15 exocyst subunit, SEC15a and SEC15b. We infer that SEC15 gene duplication and diversification occurred in the common ancestor of seed plants (Spermatophytes). In Arabidopsis, SEC15a represents the main SEC15 isoform in the male gametophyte, and localizes to the pollen tube tip at the plasma membrane. Although pollen tubes of sec15a mutants are impaired, sporophytes show no phenotypic deviations. Conversely, SEC15b is the dominant isoform in the sporophyte and localizes to the plasma membrane in root and leaf cells. Loss-of-function sec15b mutants exhibit retarded elongation of hypocotyls and root hairs, a loss of apical dominance, dwarfed plant stature and reduced seed coat mucilage formation. Surprisingly, the sec15b mutants also exhibit compromised pollen tube elongation in vitro, despite its very low expression in pollen, suggesting a non-redundant role for the SEC15b isoform there. In pollen tubes, SEC15b localizes to distinct cytoplasmic structures. Reciprocally to this, SEC15a also functions in the sporophyte, where it accumulates at plasmodesmata. Importantly, although overexpressed SEC15a could fully complement the sec15b phenotypic deviations in the sporophyte, the pollen-specific overexpression of SEC15b was unable to fully compensate for the loss of SEC15a function in pollen. We conclude that the SEC15a and SEC15b isoforms evolved in seed plants, with SEC15a functioning mostly in pollen and SEC15b functioning mostly in the sporophyte.

Cadmium enhances root hair elongation through reactive oxygen species in Arabidopsis

We previously reported that Arabidopsis shows enhanced root hair density and length in response to cadmium (Cd). To understand how Cd stimulates root hair elongation, we examined the role of reactive oxygen species (ROS) and root hair elongation-related genes in root hair elongation/tip growth. In response to Cd, Arabidopsis displayed enhanced length and ROS levels in the root hairs, indicating ROS involvement. Furthermore, in response to Cd, ROS generation mutants, including root hair defective 2 ( rhd2/rbohc ), rbohh , rbohj , per1 , per44 and per73, which have shorter root hairs because of impaired ROS generation, also showed increased root hair length and ROS levels, suggesting the involvement of these genes in ROS production in Cd-induced root hair elongation. Supporting the participation of ROS, N-acetyl- L -cysteine (ROS scavenger) treatment reduced root hair elongation and ROS levels. Additionally, the expression of proline-rich extension-like receptor kinase (PERK)5 , PERK8, and root hair-specific 10 ( RHS10 ) , which are negative regulators of root hair elongation, was reduced by Cd and ROS (H 2 O 2 ). Furthermore, the expression of expansin A7 ( EXPA7 ) and expansin A18 ( EXPA18 ), which are involved in root hair elongation, was remarkably induced by Cd and H 2 O 2 in elongating root hairs and tips. Additionally, COBL9 , which is involved in cell wall formation, was highly expressed in root hairs and tips in response to Cd and H 2 O 2 . Furthermore, changes in auxin and ethylene levels showed a similar pattern to that of root hair elongation and ROS at various concentrations of Cd, suggesting the involvement of auxin and ethylene in Cd/ROS-induced root hair elongation. Combined, it appears that Cd enhances root hair elongation/tip growth by elevating the expression of EXPA7 , EXPA18 and COBL9 and by inhibiting the expression of PERK5 , PERK8, and RHS10 , which are ROS-mediated in Arabidopsis. Change of root hair elongation, ROS, auxin and ethylene at various concentrations of Cd in Col-0 Arabidopsis • Cd promotes root hair elongation by enhancing ROS level in Arabidopsis. • RBOHC, RBOHH, RBOHJ, PER1, PER44, and PER73 participate in Cd-induced ROS generation. • Auxin and ethylene may be involved in Cd-enhanced ROS and root hair elongation. • Cd-induced ROS elevates expressions of EXPA7 , EXPA18 and COBL9 . • Cd-enhanced ROS inhibits expressions of PERK5 , PERK8, and RHS10 .

A novel calmodulin-interacting Domain of Unknown Function 506 protein represses root hair elongation in Arabidopsis.

Domain of Unknown Function 506 proteins are ubiquitous in plants. The phosphorus (P) stress‐inducible REPRESSOR OF EXCESSIVE ROOT HAIR GROWTH1 (AtRXR1) gene encodes the first characterized DUF506. AtRXR1 inhibits root hair elongation by interacting with RabD2c GTPase. However, functions of other P‐responsive DUF506 genes are still missing. Here, we selected two additional P‐inducible DUF506 genes for further investigation. The expression of both genes was induced by auxin. Under P‐stress, At3g07350 gene expressed ubiquitously in seedlings, whereas At1g62420 (AtRXR3) expression was strongest in roots. AtRXR3 overexpressors and knockouts had shorter and longer root hairs, respectively. A functional AtRXR3‐green fluorescent protein fusion localized to root epidermal cells. Chromatin immunoprecipitation and quantitative reverse‐transcriptase‐polymerase chain reaction revealed that AtRXR3 was transcriptionally activated by RSL4. Bimolecular fluorescence complementation and calmodulin (CaM)‐binding assays showed that AtRXR3 interacted with CaM in the presence of Ca2+. Moreover, cytosolic Ca2+ ([Ca2+]cyt) oscillations in root hairs of rxr3 mutants exhibited elevated frequencies and dampened amplitudes compared to those of wild type. Thus, AtRXR3 is another DUF506 protein that attenuates P‐limitation‐induced root hair growth through mechanisms that involve RSL4 and interaction with CaM to modulate tip‐focused [Ca2+]cyt oscillations.