Increased Root Hair Length Research Articles

Morphological responses of root hairs to changes in soil and climate depend on plant life form

IntroductionRoot hairs increase the surface area of a plant’s root system that is in contact with the soil, thus facilitating plant water and nutrient uptake. However, little is known about the characteristics of the root hairs of herbaceous and woody plants and their specific response patterns to biotic and abiotic variables from the perspective of resource acquisition strategies in the context of global change.MethodsHere, we analyzed 74 published case studies with 1074 observations of root hair traits to identify patterns of root hair length, density and diameter in relation to soil (e.g., soil pH, nutrient levels), growing environments (e.g., greenhouse, field) and climatic factors (e.g., air temperature), as well as genome size and plant age.ResultsRoot hairs were longer, denser and thicker in woody plants compared with herbaceous plants, and the length and diameter of root hairs in herbaceous plants increased with genome size. With increasing plant age, woody plants had significantly longer and thicker root hairs, while root hair density and diameter declined significantly for herbaceous plants. Soil-cultured plants had longer root hairs than solution-cultured plants. The length and density of root hairs were greater in greenhouse-cultured plants than in field-grown plants, and the latter had thicker root hairs than the former. As soil pH increased, root hair length increased but diameter decreased in woody plants, while root hair density increased in herbaceous plants. Increased soil total nitrogen (N) and potassium (K) significantly increased root hair length, density and diameter in herbaceous plants, while soil total N significantly decreased root hair density in woody plants. Root hair length increased significantly, while root hair density decreased significantly, with higher mean annual temperature and greater precipitation seasonality, while the opposite pattern was true for a wider annual temperature range.DiscussionOur findings emphasize the life-form-specific responses of root hairs to soil and climatic variables. These findings will help deepen our understanding of resource acquisition strategies and their mechanisms in different plant forms under global climate change.

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 .

NIN-Like Proteins: Interesting Players in Rhizobia-Induced Nitrate Signaling Response During Interaction with Non-Legume Host Arabidopsis thaliana

Nitrogen is an essential macronutrient and a key cellular messenger. Plants have evolved refined molecular systems to sense the cellular nitrogen status. This is exemplified by the root nodule symbiosis between legumes and symbiotic rhizobia, where nitrate availability inhibits this mutualistic interaction. Additionally, nitrate also functions as a metabolic messenger, resulting in nitrate signaling cascades which intensively crosstalk with other physiological pathways. Nodule inception-like proteins (NLPs) are key players in nitrate signaling and regulate nitrate-dependent transcription during legume-rhizobia interactions. Nevertheless, the coordinated interplay between nitrate signaling pathways and rhizobacteria-induced responses remains to be elucidated. In our study, we investigated rhizobia-induced changes in the root system architecture of the non-legume host arabidopsis under different nitrate conditions. We demonstrate that rhizobium-induced lateral root growth and increased root hair length and density are regulated by a nitrate-related signaling pathway. Key players in this process are AtNLP4 and AtNLP5, because the corresponding mutants failed to respond to rhizobia. At the cellular level, AtNLP4 and AtNLP5 control a rhizobia-induced decrease in cell elongation rates, while additional cell divisions occurred independently of AtNLP4. In summary, our data suggest that root morphological responses to rhizobia are coordinated by a newly considered nitrate-related NLP pathway that is evolutionarily linked to regulatory circuits described in legumes.[Formula: see text] Copyright © 2022 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Rhizobacteria associated with Miscanthus x giganteus improve metal accumulation and plant growth in the flotation tailings

Flotation tailings represent an extremely unfriendly substrate for plant colonization due to toxic metal concentrations and marked macronutrient deficiencies. The perennial grass Miscanthus x giganteus J.M.Greef & Deuter ex Hodk. & Renvoize was successfully cultivated in this infertile substrate for two years. Our aim was to identify composition of its rhizosphere bacterial community and to analyze the effects of the selected rhizobacteria on plant growth, root development, metal and P uptake. Using the cultivation-dependent method, 75 isolates were collected from the rhizosphere and six rhizobacterial strains were selected for further characterization based on morphological and biochemical differences. The plant rhizomes were inoculated with the consortium of rhizobacteria and cultivated in the flotation tailings substrate. Detected bacterial strains were characterized as metal-resistant and plant growth-promoting rhizobacteria (PGPR) because of their metal tolerance (NiCl2, Pb(C2H3O2)2, CuSO4, NaAsO2, MnCl2) and some or all of the plant growth-promoting (PGP) properties (indole-3-acetic acid and siderophore production, 1-aminocyclopropane-1-carboxylic acid deaminase activity and phosphate solubilization). PGPR mitigated the negative effects of high metal concentrations and macronutrient deficiency as shown by stimulated lateral roots development, increased root hair length, plant below and above ground biomass yield, higher plant P uptake and metal accumulation rate. The isolated PGPR strains could be used in PGP-bacteria assisted phytoremediation of flotation tailings and metal polluted soils by M. x giganteus. Their PGP effects on various metal-tolerant target plant species in the respective substrate remain to be verified.

Heterologous overexpression of the GbTCP5 gene increased root hair length, root hair and stem trichome density, and lignin content in transgenic Arabidopsis.

Teosinte branched1/cycloidea/proliferating cell factor1 (TCP) is a plant-specific protein family member involved in plant growth and development. However, the functions of most members of the cotton TCP family are unknown. In this study, the GbTCP5 gene encodes a sea-island cotton class II TCP CIN subclass transcription factor. The GbTCP5 transcription factor is located in the nucleus, has transcriptional activation activity, and can bind to TCP II cis-acting elements. GbTCP5 was widely expressed in tissues with the highest transcript level in the calyx. GbTCP5 is expressed at different developmental stages of the fiber and has significantly high transcriptional level expression in the fibers at 20, 30 and 35days post anthesis (DPA). Heterologous overexpression of the GbTCP5 gene increased root hair length, root hair and stem trichome density, and stem lignin content in transgenic Arabidopsis compared to the wild type (WT). GbTCP5 binds the promoters of the GL3, EGL3, CPC, MYB46, LBD30, CesA4, VND7, CCOMT1, and CAD5 genes to upregulate their expression. Moreover, the homologous genes of these genes are expressed in the fibers of different developmental stages of the sea-island cotton fiber. These results indicate that GbTCP5 regulates root hair development and secondary wall formation in Arabidopsis and may be a candidate gene for improving cotton fiber quality.

Genetic variation in ZmTIP1 contributes to root hair elongation and drought tolerance in maize.

SummaryDrought is a major abiotic stress that threatens maize production globally. A previous genome‐wide association study identified a significant association between the natural variation of ZmTIP1 and the drought tolerance of maize seedlings. Here, we report on comprehensive genetic and functional analysis, indicating that ZmTIP1, which encodes a functional S‐acyltransferase, plays a positive role in regulating the length of root hairs and the level of drought tolerance in maize. We show that enhancing ZmTIP1 expression in transgenic Arabidopsis and maize increased root hair length, as well as plant tolerance to water deficit. In contrast, ZmTIP1 transposon‐insertional mutants displayed the opposite phenotype. A calcium‐dependent protein kinase, ZmCPK9, was identified as a substrate protein of ZmTIP1, and ZmTIP1‐mediated palmitoylation of two cysteine residues facilitated the ZmCPK9 PM association. The results of this research enrich our knowledge about ZmTIP1‐mediated protein S‐acylation modifications in relation to the regulation of root hair elongation and drought tolerance. Additionally, the identification of a favourable allele of ZmTIP1 also provides a valuable genetic resource or selection target for the genetic improvement of maize.

AtERF#111/ABR1 is a transcriptional activator involved in the wounding response.

AtERF#111/ABR1 belongs to the group X of the ERF/AP2 transcription factor family (GXERFs) and is shoot specifically induced under submergence and hypoxia. It was described to be an ABA-response repressor, but our data reveal a completely different function. Surprisingly, AtERF#111 expression is strongly responsive to wounding stress. Expression profiling of ERF#111-overexpressing (OE) plants, which show morphological phenotypes like increased root hair length and number, strengthens the hypothesis of AtERF#111 being involved in the wounding response, thereby acting as a transcriptional activator of gene expression. Consistent with a potential function outside of oxygen signalling, we could not assign AtERF#111 as a target of the PRT6N-degron pathway, even though it starts with a highly conserved N-terminal Met-Cys (MC) motif. However, the protein is unstable as it is degraded in an ubiquitin-dependent manner. Finally, direct target genes of AtERF#111 were identified by microarray analyses and subsequently confirmed by protoplast transactivation assays. The special roles of diverse members of the plant-specific GXERFs in coordinating stress signalling and wound repair mechanisms have been recently hypothesized, and our data suggest that AtERF#111 is indeed involved in these processes.

Phospholipase C activity is increased in wheat seedlings inoculated with the rhizobacteria Azospirillum brasilense Sp245

The present study aimed to determine phospholipase C (PLC) activation status during changes promoted by Azospirillum brasilense Sp245 in wheat seedlings. Germinated Triticum aestivum seeds by 3 days were inoculated with different A. brasilense concentrations (1 × 106 and 4 × 106 CFU/mL) and PLC enzyme activity assayed at different times. Root and leaf length, as well as total fresh weight were assessed as growth parameters; moreover, changes in root morphology were analyzed. PLC activity was measured by molybdate assay. Neomycin and LaCI3 treatments verified PLC- and/or Ca2+-dependent effects on inoculated wheat, respectively. A. brasilense increased PLC activity 15–30 min after inoculation. Ca2+-channel blocker LaCI3 decreased PLC activity, and activity did not recover with A. brasilense. The A. brasilense mutant FAJ009, impaired in auxin production, showed decreased PLC activity versus wild type. PLC activity was inhibited by neomycin (PLC inhibitor), concomitant with a decrease in total fresh weight. Exogenous addition of diacylglycerol (DAG) and inositol-1,4,5-trisphosphate (IP3), secondary messengers produced by the activity of PLC, increased root hair length. The putative role of PLC enzyme activity in morphological changes promoted by A. brasilense in wheat seedlings is discussed.

Do longer root hairs improve phosphorus uptake? Testing the hypothesis with transgenic Brachypodium distachyon lines overexpressing endogenous RSL genes.

Mutants without root hairs show reduced inorganic orthophosphate (Pi) uptake and compromised growth on soils when Pi availability is restricted. What is less clear is whether root hairs that are longer than wild-type provide an additional benefit to phosphorus (P) nutrition. This was tested using transgenic Brachypodium lines with longer root hairs. The lines were transformed with the endogenous BdRSL2 and BdRSL3 genes using either a constitutive promoter or a root hair-specific promoter. Plants were grown for 32d in soil amended with various Pi concentrations. Plant biomass and P uptake were measured and genotypes were compared on the basis of critical Pi values and P uptake per unit root length. Ectopic expression of RSL2 and RSL3 increased root hair length three-fold but decreased plant biomass. Constitutive expression of BdRSL2, but not expression of BdRSL3, consistently improved P nutrition as measured by lowering the critical Pi values and increasing Pi uptake per unit root length. Increasing root hair length through breeding or biotechnology can improve P uptake efficiency if the pleotropic effects on plant biomass are avoided. Long root hairs, alone, appear to be insufficient to improve Pi uptake and need to be combined with other traits to benefit P nutrition.

Deubiquitinating Enzyme OTU5 Contributes to DNA Methylation Patterns and Is Critical for Phosphate Nutrition Signals.

Phosphate (Pi) starvation induces a suite of adaptive responses aimed at recalibrating cellular Pi homeostasis. Plants harboring a mutation in OVARIAN TUMOR DOMAIN-CONTAINING DEUBIQUITINATING ENZYME5 (OTU5) showed altered DNA methylation of root hair-related genes and altered Pi-responsive root traits. Unlike the wild type, homozygous otu5 mutants did not respond to Pi starvation by increased lateral root formation and increased root hair length but formed very short root hairs when grown on low-Pi media. Under Pi-replete conditions, otu5 plants developed more root hairs than the wild type due to attenuated primary root growth, a phenotype that resembled that of Pi-deficient plants. Growth of plants on low-Pi media altered both H3K4 and H3K27 trimethylation levels at the transcriptional start site of a subset of genes encoding key players in Pi homeostasis, which was correlated with mRNA abundance changes of these genes. Pi starvation had a minor impact on DNA methylation. Differentially methylated regions were enriched in transposable elements, suggesting that DNA methylation associated with low Pi supply is required for maintaining genome integrity. It is concluded that DNA methylation and histone methylation constitute critical, interdependent regulatory components that orchestrate the activity of a subset of Pi-responsive genes.

Systematic deletion of the ER lectin chaperone genes reveals their roles in vegetative growth and male gametophyte development in Arabidopsis.

Calnexin (CNX) and calreticulin (CRT) are homologous lectin chaperones in the endoplasmic reticulum (ER) that facilitate glycoprotein folding and retain folding intermediates to prevent their transit via the secretary pathway. The Arabidopsis genome has two CNX (CNX1 and CNX2) and three CRT (CRT1, CRT2 and CRT3) homologs. Despite growing evidence of the biological roles of CNXs and CRTs, little is understood about their function in Arabidopsis growth and development under normal conditions. Here, we report that the deletion of CNX1, but not of CNX2, in the crt1crt2crt3 triple mutation background had an adverse effect on pollen viability and pollen tube growth, leading to a significant reduction in fertility. The cnx1crt1crt2crt3 quadruple mutation also conferred severe defects in growth and development, including a shortened primary root, increased root hair length and density, and reduced plant height. Disruption of all five members of the CNX/CRT family was revealed to be lethal. Finally, the abnormal phenotype of the cnx1crt1crt2crt3 quadruple mutants was completely rescued by either the CNX1 or CNX2 cDNA under the control of the CNX1 promoter, suggesting functional redundancy between CNX1 and CNX2. Taken together, these results provide genetic evidence that CNX and CRT play essential and overlapping roles during vegetative growth and male gametophyte development in Arabidopsis.

Altered expression of TaRSL4 gene by genome interplay shapes root hair length in allopolyploid wheat.

Polyploidy is a major driving force in plant evolution and speciation. Phenotypic changes often arise with the formation, natural selection and domestication of polyploid plants. However, little is known about the consequence of hybridization and polyploidization on root hair development. Here, we report that root hair length of synthetic and natural allopolyploid wheats is significantly longer than those of their diploid progenitors, whereas no difference is observed between allohexaploid and allotetraploid wheats. The expression of wheat gene TaRSL4, an orthologue of AtRSL4 controlling the root hair development in Arabidopsis, was positively correlated with the root hair length in diploid and allotetraploid wheats. Moreover, transcript abundance of TaRSL4 homoeologue from A genome (TaRSL4-A) was much higher than those of other genomes in natural allopolyploid wheat. Notably, increased root hair length by overexpression of the TaRSL4-A in wheat led to enhanced shoot fresh biomass under nutrient-poor conditions. Our observations indicate that increased root hair length in allohexaploid wheat originated in the allotetraploid progenitors and altered expression of TaRSL4 gene by genome interplay shapes root hair length in allopolyploid wheat.

Efficient Root System for Abiotic Stress Tolerance in Crops

Plant survival and fitness are dependent on root system architecture (RSA). In Australia, root systems of major agricultural crops are poorly adapted to soils that mostly have poor water holding capacity and nutrient deficiencies. Decreasing water availability due to drying and variable climate in the Australia's grain-belt exacerbates these soil-related stresses. Development of future crop genotypes with efficient root system for enhanced abiotic stress tolerance is essential for improved crop adaptation. Root traits that overcome abiotic constraints are critical to maintaining structural and functional properties, and are considered first order targets in breeding programmes for rainfed environments. Root traits, such as deep root systems, increased root density in subsoil, increased root hair length and density and/or xylem diameters, may contribute to enhanced water and nutrient uptake. Narrow-leafed lupin genotypes with increased capacity to take up water from deep soil horizons were linked to increased yield potential; similar relationship exists in wheat, soybean and upland rice. Modification of RSA could contribute to improvements of desirable agronomic traits such as yield, drought tolerance, and resistance to nutrient deficiencies. Wide-scale use of root- related genetic information in breeding programs relies on accurate phenotyping of relatively large mapping populations. Such large-scale phenotyping of root-related traits remain the most important issue in translating recent physiological and genetic advances in understanding the role of root systems in improved adaptation to abiotic stress and enhanced productivity of agricultural crops.

The utility of phenotypic plasticity of root hair length for phosphorus acquisition

Root hairs are subcellular protrusions from the root epidermis that are important for the acquisition of immobile nutrients such as phosphorus (P). Genetic variation exists for both root hair length and the plasticity of root hair length in response to P availability, where plasticity manifests as increased root hair length in response to low P availability. Although it is known that long root hairs assist P acquisition, the utility of phenotypic plasticity for this trait is not known. To assess the utility of root hair plasticity for adaptation to low phosphorus availability, we evaluated six recombinant inbred lines of maize (Zea mays L.) with varying root hair lengths and root hair plasticity in a controlled environment and in the field. Genotypes with long root hairs under low P availability had significantly greater plant growth, P uptake, specific P absorption rates and lower metabolic cost-benefit ratios than short-haired genotypes. Root hair length had no direct effect on root respiration. In the controlled environment, plastic genotypes had greater biomass allocation to roots, greater reduction in specific root respiration and greater final biomass accumulation at low phosphorus availability than constitutively long-haired genotypes. In the field study, the growth of plastic and long-haired genotypes were comparable under low P, but both were superior to short-haired genotypes. We propose that root hair plasticity is a component of a broader suite of traits, including plasticity in root respiration, that permit greater root growth and phosphorus acquisition in low P soils.

Phosphate Starvation Root Architecture and Anthocyanin Accumulation Responses Are Modulated by the Gibberellin-DELLA Signaling Pathway in Arabidopsis

Phosphate (Pi) is a macronutrient that is essential for plant growth and development. However, the low mobility of Pi impedes uptake, thus reducing availability. Accordingly, plants have developed physiological strategies to cope with low Pi availability. Here, we report that the characteristic Arabidopsis thaliana Pi starvation responses are in part dependent on the activity of the nuclear growth-repressing DELLA proteins (DELLAs), core components of the gibberellin (GA)-signaling pathway. We first show that multiple shoot and root Pi starvation responses can be repressed by exogenous GA or by mutations conferring a substantial reduction in DELLA function. In contrast, mutants having enhanced DELLA function exhibit enhanced Pi starvation responses. We also show that Pi deficiency promotes the accumulation of a green fluorescent protein-tagged DELLA (GFP-RGA [repressor of ga1-3]) in root cell nuclei. In further experiments, we show that Pi starvation causes a decrease in the level of bioactive GA and associated changes in the levels of gene transcripts encoding enzymes of GA metabolism. Finally, we show that the GA-DELLA system regulates the increased root hair length that is characteristic of Pi starvation. In conclusion, our results indicate that DELLA-mediated signaling contributes to the anthocyanin accumulation and root architecture changes characteristic of Pi starvation responses, but do not regulate Pi starvation-induced changes in Pi uptake efficiency or the accumulation of selected Pi starvation-responsive gene transcripts. Pi starvation causes a reduction in bioactive GA level, which, in turn, causes DELLA accumulation, thus modulating several adaptively significant plant Pi starvation responses.

Membrane transport and cytokinin action in root hairs of Medicago sativa

In an effort to develop a cellular model for studying cytokinin action in higher plants, we investigated the effect of cytokinins on growth and membrane transport in root hairs of alfalfa (Medicago sativa␣L.) seedlings. Alfalfa seedlings grown for 24 h in the presence of cytokinins showed increased root hair length and formation of root hairs close to the root cap. Increased growth of root hairs was observed within 10 min of cytokinin application and was accompanied by a hyperpolarization of the plasma membrane. The ion-transport inhibitor diisothiocyanostilbene-2,2′-disulfonic acid (DIDS) blocked both cytokinin-induced root hair growth and hyperpolarization but did not, by itself, alter growth or membrane potential. Finally, hyperpolarization was induced by extracellular cytokinin but not by injection of cytokinin into the cytoplasm. These findings show that root hairs undergo several rapid responses to cytokinin, including changes in membrane transport and growth. We conclude that multiple cytokinin response in root hairs may be mediated by events involving perception and ion transport at the plasma membrane.

Stimulation of root hair elongation in Arabidopsis thaliana by low phosphorus availability

ABSTRACTLow phosphorus availability stimulates root hair elongation in many plants, which may have adaptive significance in soil phosphorus acquisition. We investigated the effect of low phosphorus on the elongation of Arabidopsis thaliana root hairs. Arabidopsis thaliana plants were grown in plant culture containing high (1000 mmol m−3) or low (1 mmol m−3) phosphorus concentrations, and root hair elongation was analysed by image analysis. After 15d of growth, low‐phosphorus plants developed root hairs averaging 0.9 mm in length while high‐phosphorus plants of the same age developed root hairs averaging 0.3 mm in length. Increased root hair length in low‐phosphorus plants was a result of both increased growth duration and increased growth rate. Root hair length decreased logarithmically in response to increasing phosphorus concentration. Local changes in phosphorus availability influenced root hair growth regardless of the phosphorus status of the plant. Low phosphorus stimulated root hair elongation in the hairless axr2 mutant, exogenously applied IAA stimulated root hair elongation in wild‐type high‐phosphorus plants and the auxin antagonist CM PA inhibited root hair elongation in low‐phosphorus plants. These results indicate that auxin may be involved in the low‐phosphorus response in root hairs.