EIN3 and RSL4 interfere with an MYB–bHLH–WD40 complex to mediate ethylene-induced ectopic root hair formation in Arabidopsis

The alternating cell specifications of root epidermis to form hair cells or nonhair cells in Arabidopsis are determined by the expression level of GL2, which is activated by an MYB–bHLH–WD40 (WER–GL3–TTG1) transcriptional complex. The phytohormone ethylene (ET) has a unique effect of inducing N-position epidermal cells to form root hairs. However, the molecular mechanisms underlying ET-induced ectopic root hair development remain enigmatic. Here, we show that ET promotes ectopic root hair formation through down-regulation of GL2 expression. ET-activated transcription factors EIN3 and its homolog EIL1 mediate this regulation. Molecular and biochemical analyses further revealed that EIN3 physically interacts with TTG1 and interferes with the interaction between TTG1 and GL3, resulting in reduced activation of GL2 by the WER–GL3–TTG1 complex. Furthermore, we found through genetic analysis that the master regulator of root hair elongation, RSL4, which is directly activated by EIN3, also participates in ET-induced ectopic root hair development. RSL4 negatively regulates the expression of GL2, likely through a mechanism similar to that of EIN3. Therefore, our work reveals that EIN3 may inhibit gene expression by affecting the formation of transcription-activating protein complexes and suggests an unexpected mutual inhibition between the hair elongation factor, RSL4, and the hair specification factor, GL2. Overall, this study provides a molecular framework for the integration of ET signaling and intrinsic root hair development pathway in modulating root epidermal cell specification.

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

Root hair initiation is a highly regulated aspect of root development. The plant hormone, ethylene, and its precursor, 1-amino-cyclopropane-1-carboxylic acid (ACC), induce formation and elongation of root hairs. We asked whether elevated ethylene induced root hair formation by increasing reactive oxygen species (ROS) synthesis in hair cells. Using confocal microscopy paired with redox biosensors and dyes, we demonstrated that treatments that elevate ethylene levels led to increased ROS accumulation in hair cells prior to root hair formation. In two ethylene-insensitive mutants, etr1-3 and ein3/eil1, there was no increase in root hair number or ROS accumulation. Conversely, etr1-7, a constitutive ethylene signaling receptor mutant, has increased root hair formation and ROS accumulation similar to ethylene-treated wild type seedlings. The rhd2-6 mutant, with a defect in the gene encoding a ROS synthesizing Respiratory Burst Oxidase Homolog C (RBOHC), showed impaired ethylene-dependent ROS synthesis and root hair formation and decreased RBOH enzyme activity compared to Col-0. To identify additional proteins that drive ROS induced root hair formation, we examined a time course root transcriptomic dataset examining Col-0 grown in the presence of ACC and identified PRX44 and other positively regulated transcripts that encode class III peroxidases (PRXs). The prx44-2 mutant has decreased root hair initiation and ROS accumulation when treated with ACC compared to Col-0 and pPRX44::GFP fluorescence is increased in response to ACC treatment. Together, these results support a model in which ethylene increases ROS accumulation through RBOHC and PRX44 to drive root hair formation.

Genetic and Molecular Analysis of Root Hair Development in Arabis alpina

Root hair formation in Arabidopsis thaliana is a well-established model system for epidermal patterning and morphogenesis in plants. Over the last decades, many underlying regulatory genes and well-established networks have been identified by thorough genetic and molecular analysis. In this study, we used a forward genetic approach to identify genes involved in root hair development in Arabis alpina, a related crucifer species that diverged from A. thaliana approximately 26–40 million years ago. We found all root hair mutant classes known in A. thaliana and identified orthologous regulatory genes by whole-genome or candidate gene sequencing. Our findings indicate that the gene-phenotype relationships regulating root hair development are largely conserved between A. thaliana and A. alpina. Concordantly, a detailed analysis of one mutant with multiple hairs originating from one cell suggested that a mutation in the SUPERCENTIPEDE1 (SCN1) gene is causal for the phenotype and that AaSCN1 is fully functional in A. thaliana. Interestingly, we also found differences in the regulation of root hair differentiation and morphogenesis between the species, and a subset of root hair mutants could not be explained by mutations in orthologs of known genes from A. thaliana. This analysis provides insight into the conservation and divergence of root hair regulation in the Brassicaceae.

Azospirillum brasilense stimulate the growth in Arabidopsis thaliana through the target of rapamycin protein

Azospirillum spp., one of the best studied genus of plant growth promoting rhizobacteria. These rhizobacteria are able to colonize hundreds of plant species and improve their growth, development and productivity. The target of rapamycin (TOR) protein is a central component of the TOR signaling pathway, which regulates cell growth and metabolism in response to environment cues in eukaryotes. In this study, the TOR function was analyzed in Arabidopsis thaliana L. plants inoculated with the rhizobacteria Azospirillum brasilense. Arabidopsis seedlings tor-es, which express an interference RNA in presence of estradiol and decrease TOR expression, showed an inhibition in the growth and lateral root formation, with or without 1x102 CFU/mL of the inoculum. In addition, a morphological analysis of the root showed an inhibition in the root hair formation. The results suggest that A. brasilense controls A. thaliana growth through TOR signaling pathway.

GTL1 is required for a robust root hair growth response to avoid nutrient overloading

Root hair growth is tuned in response to the environment surrounding plants. While most of previous studies focused on the enhancement of root hair growth during nutrient starvation, few studies investigated the root hair response in the presence of excess nutrients. We report that the post-embryonic growth of wild-type Arabidopsis plants is strongly suppressed with increasing nutrient availability, particularly in the case of root hair growth. We further used gene expression profiling to analyze how excess nutrient availability affects root hair growth, and found that RHD6 subfamily genes, which are positive regulators of root hair growth, are down-regulated in this condition. On the other hand, defects in GTL1 and DF1, which are negative regulators of root hair growth, cause frail and swollen root hairs to form when excess nutrients are supplied. Additionally, we observed that the RHD6 subfamily genes are mis-expressed in gtl1-1 df1-1. Furthermore, overexpression of RSL4, an RHD6 subfamily gene, induces swollen root hairs in the face of a nutrient overload, while mutation of RSL4 in gtl1-1 df1-1 restore root hair swelling phenotype. In conclusion, our data suggest that GTL1 and DF1 prevent unnecessary root hair formation by repressing RSL4 under excess nutrient conditions.

Alternative Oxidase Inhibition Impairs Tobacco Root Development and Root Hair Formation

Alternative oxidase (AOX) is the terminal oxidase of the mitochondrial respiratory electron transport chain in plant cells and is critical for the balance of mitochondrial hemostasis. In this study, the effect of inhibition of AOX with different concentrations of salicylhydroxamic acid (SHAM) on the tobacco root development was investigated. We show here that AOX inhibition significantly impaired the development of the main root and root hair formation of tobacco. The length of the main root of SHAM-treated tobacco was significantly shorter than that of the control, and no root hairs were formed after treatment with a concentration of 1 mM SHAM or more. The transcriptome analysis showed that AOX inhibition by 1 mM SHAM involved in the regulation of gene expression related to root architecture. A total of 5,855 differentially expressed genes (DEGs) were obtained by comparing SHAM-treated roots with control. Of these, the gene expression related to auxin biosynthesis and perception were significantly downregulated by 1 mM SHAM. Similarly, genes related to cell wall loosening, cell cycle, and root meristem growth factor 1 (RGF1) also showed downregulation on SHAM treatment. Moreover, combined with the results of physiological measurements, the transcriptome analysis demonstrated that AOX inhibition resulted in excessive accumulation of reactive oxygen species in roots, which further induced oxidative damage and cell apoptosis. It is worth noting that when indoleacetic acid (20 nM) and dimethylthiourea (10 mM) were added to the medium containing SHAM, the defects of tobacco root development were alleviated, but to a limited extent. Together, these findings indicated that AOX-mediated respiratory pathway plays a crucial role in the tobacco root development, including root hair formation.

Trichobezoars in the practice of a pediatric surgeon

Material and methods. A retrospective analysis of medical records of two children. Anamnestic, clinical, diagnostic and intraoperative findings were analyzed.Purpose. To describe cases of trichobezoars in children : occurrence, diagnostics and treatment.Results. In the first case, a girl, aged 5, often swallowed her own hair after a psychological trauma; and at the age of 15 she complained of hair loss and anemia. In the second case, a boy was chewing and swallowing his own hair for 6 months under the emotional stress. Two weeks before hospitalization he complained of abdominal pain. In both cases, there were no history of intestinal obstruction. At the fibroesophagogastroduodenoscopy, foreign bodies were visualized which were diagnosed as trichobezoars. X-ray diagnostics confirmed foreign bodies in both patients. Those bodies had the shape of the stomach and had an inhomogeneous porous structure. The patients were operated: laparotomy, gastrotomy with removal of dense hair formation. Postoperative course was uneventful.Conclusion. Psychological situations provoked in children the obsessive trichotillomania and trichophagia due to which large trichobezoars were formed in the stomach.

Transcriptome sequencing analysis of maize roots reveals the effects of substrate and root hair formation in a spatial context

Background Plant roots sense and respond to changes in their soil environment, but conversely contribute to rhizosphere organization through chemical, mechanical and biotic interactions. Transcriptomic profiling of plant roots can be used to assess how the plant adjusts its gene expression in relation to environment, genotype and rhizosphere processes; thus enabling us to achieve a better understanding of root-soil interactions. Methods We used a standardized soil column experimental platform to investigate the impact of soil texture (loam, sand) and root hair formation (wildtype, root hair defective rth3 mutant) in a spatial context (three sampling depths) and assessed maize root transcriptomic profiles using next-generation RNA sequencing. Results Substrate induced the largest changes in root gene expression patterns, affecting gene functions related to immunity, stress, growth and water uptake. Genes with column depth-related expression levels were associated with growth and plant defense. The influence of root hairs mainly manifested in differential expression of epidermal cell differentiation and cell wall organization, and defense response-related genes. Substrate type strongly modified the transcriptomic patterns related to column depth and root hair elongation, highlighting the strong impact of soil texture. Conclusions Our results demonstrate that substrate, sampling depth and plant genotype interactively affect maize gene expression, and suggest feedback processes between the plant, the soil and the microbiome. The obtained results form a foundational basis for the integration and interpretation of future experiments utilizing the same experimental platform.