Root Hair Responses 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.

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.

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.

Trihelix transcription factors GTL1 and DF1 prevent aberrant root hair formation in an excess nutrient condition.

Summary Root hair growth is tuned in response to the environment surrounding plants. While most 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 downregulated in this condition.However, 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.

Amino Acid Polymorphisms in the VHIID Conserved Motif of Nodulation Signaling Pathways 2 Distinctly Modulate Symbiotic Signaling and Nodule Morphogenesis in Medicago truncatula.

Legumes establish an endosymbiotic association with nitrogen-fixing soil bacteria. Following the mutual recognition of the symbiotic partner, the infection process is controlled by the induction of the signaling pathway and subsequent activation of symbiosis-related host genes. One of the protein complexes regulating nitrogen-fixing root nodule symbiosis is formed by GRAS domain regulatory proteins Nodulation Signaling Pathways 1 and 2 (NSP1 and NSP2) that control the expression of several early nodulation genes. Here, we report on a novel point mutant allele (nsp2-6) affecting the function of the NSP2 gene and compared the mutant with the formerly identified nsp2-3 mutant. Both mutants carry a single amino acid substitution in the VHIID motif of the NSP2 protein. We found that the two mutant alleles show dissimilar root hair response to bacterial infection. Although the nsp2-3 mutant developed aberrant infection threads, rhizobia were able to colonize nodule cells in this mutant. The encoded NSP2 proteins of the nsp2-3 and the novel nsp2 mutants interact with NSP1 diversely and, as a consequence, the activation of early nodulin genes and nodule organogenesis are arrested in the new nsp2 allele. The novel mutant with amino acid substitution D244H in NSP2 shows similar defects in symbiotic responses as a formerly identified nsp2-2 mutant carrying a deletion in the NSP2 gene. Additionally, we found that rhizobial strains induce delayed nodule formation on the roots of the ns2-3 weak allele. Our study highlights the importance of a conserved Asp residue in the VHIID motif of NSP2 that is required for the formation of a functional NSP1-NSP2 signaling module. Furthermore, our results imply the involvement of NSP2 during differentiation of symbiotic nodule cells.

The importance of Rhizobium, Agrobacterium, Bradyrhizobium, Herbaspirillum, Sinorhizobium in sustainable agricultural production

Rhizobia which are soil bacteria capable of symbiosis with legume plants in the root or stem nodules and perform nitrogen fixation. Rhizobial genera include Agrobacterium, Allorhizobium, Aminobacter, Azorhizobium, Bradyrhizobium, Devosia, Mesorhizobium, Methylobacterium, Microvirga, Ochrobacterum, Phyllobacterium, Rhizobium, Shinella and Ensifer (Sinorhizobium). Review of the literature was carried out using the keywords Rhizobium, Agrobacterium, Bradyrhizobium, Herbaspirillum and Sinorhizobium. Rhizobial nodulation symbioses steps are included flavonoid signaling, Nod factor induction, and Nod factor perception, root hair responses, rhizobial infection, cell division and formation of nitrogen-fixing nodule. Rhizobium improves sustainable production by boosting organic nitrogen content.

The responses of lateral roots and root hairs to nitrogen stress in cotton based on daily root measurements

AbstractLateral roots and root hairs are essential portions of cotton roots. However, the morphology and lifespan of cotton (Gossypium hirsutum L.) lateral roots and root hairs in response to nitrogen stress have remained unknown. Two cotton cultivars were assessed when no nitrogen and normal nitrogen (138 mg N·kg−1 soil) were added. The daily acquisitions of root phenotype information including the lateral root angle, lateral root density, lateral root diameter, lateral root lifespan, root hair length, root hair density, and root hair lifespan, were obtained in situ by a self‐designed platform of RhizoPots equipped with high‐resolution scanners. Nitrogen starvation significantly reduced the growth of shoots and roots, although there was less reduction of roots than shoots. Root hairs were the first organ to respond to nitrogen deficiency through morphological changes. The lateral root diameter and angle as well as the root hair lifespan decreased significantly under nitrogen stress. However, nitrogen deficiency dramatically increased the lateral root lifespan, root hair density and length. Our results demonstrate that low nitrogen can induce notable changes in lateral roots and root hairs, which may play a crucial role in the adjustment to nitrogen uptake, particularly in species that can adapt to low nitrogen availability.

Heterogeneous nutrient supply promotes maize growth and phosphorus acquisition: additive and compensatory effects of lateral roots and root hairs.

Root proliferation is a response to a heterogeneous nutrient distribution. However, the growth of root hairs in response to heterogeneous nutrients and the relationship between root hairs and lateral roots remain unclear. This study aims to understand the effects of heterogeneous nutrients on root hair growth and the trade-off between root hairs and lateral roots in phosphorus (P) acquisition. Near-isogenic maize lines, the B73 wild type (WT) and the rth3 root hairless mutant, were grown in rhizoboxes with uniform or localized supply of 40 (low) or 140 (high) mg P kg-1 soil. Both WT and rth3 had nearly two-fold greater shoot biomass and P content under local than uniform treatment at low P. Significant root proliferation was observed in both WT and rth3 in the nutrient patch, with the WT accompanied by an obvious increase (from 0.7 to 1.2 mm) in root hair length. The root response ratio of rth3 was greater than that of WT at low P, but could not completely compensate for the loss of root hairs. This suggests that plants enhanced P acquisition through complementarity between lateral roots and root hairs, and thus regulated nutrient foraging and shoot growth. The disappearance of WT and rth3 root response differences at high P indicated that the P application reduced the dependence of the plants on specific root traits to obtain nutrients. In addition to root proliferation, the root response to a nutrient-rich patch was also accompanied by root hair elongation. The genotypes without root hairs increased their investment in lateral roots in a nutrient-rich patch to compensate for the absence of root hairs, suggesting that plants enhanced nutrient acquisition by regulating the trade-off of complementary root traits.

Inorganic nitrogen sources alter the root morphology of neotropical tree seedlings from different successional groups

Root morphological responses of neotropical tree seedlings to nitrate and ammonium nutrition vary according to the successional group, but may also depend on each species. Neotropical tree seedlings from contrasting successional groups have different nitrogen (N) use strategies, as well as they exhibit striking differences in root morphology. Here, we assessed the effects of hydroponic cultivation with nitrate or ammonium on the growth and root morphological traits of seedlings of pioneer (Cecropia pachystachya, Croton floribundus, Schinus terebinthifolius, Solanum mauritianum, and Trema micrantha) and non-pioneer (Cabralea canjerana, Campomanesia xanthocarpa, Cariniana estrellensis, Guarea kunthiana, and Poecilanthe parviflora) tree species native to Brazilian Atlantic Forest. Nitrate-grown seedlings of pioneer species showed higher shoot and root dry matters and total root length compared to those cultivated with ammonium, while non-pioneer trees showed smaller or no difference in these traits when comparing N sources. These results agree with the decrease of nitrate availability in the soil relative to that of ammonium over an Atlantic Forest succession. Regardless of the successional group, nitrate treatment resulted in longer main and lateral roots in most species. The effects of nitrate and ammonium on specific root length, lateral-root density, and fine-root diameter could be associated with successional groups and N source preference. However, root hair responses varied according to each species. Cecropia pachystachya and C. canjerana roots had morphological responses to N source intermediaries between the successional groups, which are coherent with their ecological niches. These results demonstrate that the effects of N source on the growth and root morphology of neotropical tree seedlings could be associated with their successional group, although species-specific responses were also observed.

Vascular transcription factors guide plant epidermal responses to limiting phosphate conditions.

Optimal plant growth is hampered by deficiency of the essential macronutrient phosphate in most soils. Plant roots can, however, increase their root hair density to efficiently forage the soil for this immobile nutrient. By generating and exploiting a high-resolution single-cell gene expression atlas of Arabidopsis roots, we show an enrichment of TARGET OF MONOPTEROS 5/LONESOME HIGHWAY (TMO5/LHW) target gene responses in root hair cells. The TMO5/LHW heterodimer triggers biosynthesis of mobile cytokinin in vascular cells and increases root hair density during low-phosphate conditions by modifying both the length and cell fate of epidermal cells. Moreover, root hair responses in phosphate-deprived conditions are TMO5- and cytokinin-dependent. Cytokinin signaling links root hair responses in the epidermis to perception of phosphate depletion in vascular cells.

Fine root and root hair morphology of cotton under drought stress revealed with RhizoPot

AbstractFine roots (roots with a diameter of less than 2 mm) and root hairs are the primary absorbers of water and nutrients; however, the morphology of cotton fine roots and root hairs in response to drought stress has not yet been defined. To solve this problem, this study characterized a self‐made in situ root observation device (RhizoPot) with a scanner with a resolution of up to 4,800 DPI was used to observe the roots of cotton plants grown indoors under both well‐watered and drought stress conditions. RhizoPot provided healthy growth conditions for cotton, and fine roots as well as root hairs were successfully observed. Root length density increased significantly under drought stress, whereas the average root diameter decreased gradually with the extension of treatment time. Drought stress accelerated fine root death, and the diameter of the fine roots was positively correlated with root lifespan. Additionally, the root hair lifespan was negatively regulated by soil drought stress and was associated with the growth stage. This result suggests that cotton developed more slender fine roots and longer root hairs under drought stress, which accelerated the death process of both, in order to develop new fine roots. This process helps cotton absorb as much water as possible under drought stress.

A glutathione-dependent control of the indole butyric acid pathway supports Arabidopsis root system adaptation to phosphate deprivation.

Root system architecture results from a highly plastic developmental process to adapt to environmental conditions. In particular, the development of lateral roots and root hair growth are constantly optimized to the rhizosphere properties, including biotic and abiotic constraints. The development of the root system is tightly controlled by auxin, the driving morphogenic hormone in plants. Glutathione, a major thiol redox regulator, is also critical for root development but its interplay with auxin is scarcely understood. Previous work showed that glutathione deficiency does not alter root responses to indole acetic acid (IAA), the main active auxin in plants. Because indole butyric acid (IBA), another endogenous auxinic compound, is an important source of IAA for the control of root development, we investigated the crosstalk between glutathione and IBA during root development. We show that glutathione deficiency alters lateral roots and root hair responses to exogenous IBA but not IAA. Detailed genetic analyses suggest that glutathione regulates IBA homeostasis or conversion to IAA in the root cap. Finally, we show that both glutathione and IBA are required to trigger the root hair response to phosphate deprivation, suggesting an important role for this glutathione-dependent regulation of the auxin pathway in plant developmental adaptation to its environment.

The Medicago truncatula DREPP Protein Triggers Microtubule Fragmentation in Membrane Nanodomains during Symbiotic Infections.

The initiation of intracellular host cell colonization by symbiotic rhizobia in Medicago truncatula requires repolarization of root hairs, including the rearrangement of cytoskeletal filaments. The molecular players governing microtubule (MT) reorganization during rhizobial infections remain to be discovered. Here, we identified M. truncatula DEVELOPMENTALLY REGULATED PLASMA MEMBRANE POLYPEPTIDE (DREPP), a member of the MT binding DREPP/PCaP protein family, and investigated its functions during rhizobial infections. We show that rhizobial colonization of drepp mutant roots as well as transgenic roots overexpressing DREPP is impaired. DREPP relocalizes into symbiosis-specific membrane nanodomains in a stimulus-dependent manner. This subcellular segregation coincides with DREPP-dependent MT fragmentation and a partial loss of the ability to reorganize the MT cytoskeleton in response to rhizobia, which might rely on an interaction between DREPP and the MT-organizing protein SPIRAL2. Taken together, our results reveal that establishment of symbiotic associations in M. truncatula requires DREPP in order to regulate MT reorganization during initial root hair responses to rhizobia.

Moderate water stress in rice induces rhizosheath formation associated with abscisic acid and auxin responses.

The rhizosheath is known to be beneficial for drought resistance in many plants, but the regulation of rhizosheath formation in rice plants is unclear. Here, we investigate rhizosheath formation in different rice varieties and root hair mutants. Our results showed that moderate water stress in rice induced rhizosheath formation. The soil porosity and water content were higher in the rice rhizosheath than in the rice bulk soil under moderate water stress. Additionally, rhizosheath formation in short root hair mutants was lower than in wild-type rice under moderate water stress. Moreover, transcriptomic results indicated that abscisic acid (ABA) and auxin were involved in root and root hair responses in rhizosheath formation. Further, blocking ABA and auxin pathways in wild type and in rhl1-1, the shortest root hair mutant, rhizosheath formation and root hair length were significantly decreased under moderate water stress. However, wild type plants maintained a higher root ABA content, root basipetal auxin transport, root hair length, and amount of rhizosheath than did rhl1-1. Our results suggest that moderate water stress in rice induces rhizosheath formation by modulating the ABA and auxin responses to regulate root and root hair growth, which may be used to breed rice varieties resistant to drought.

ERN1 and CYCLOPS coordinately activate NIN signaling to promote infection thread formation in Lotus japonicus

Legumes engage in symbiosis with nitrogen-fixing soil bacteria, collectively called rhizobia, under nitrogen-limited conditions. In many legumes, the root invasion of rhizobia is mediated by infection threads (ITs), tubular invaginations of the host cell wall and plasma membrane, developed from infection foci of deformed root hairs. IT formation is regulated by a series of signal transduction in host root. Nodulation signals activate the host transcription factor (TF), CYCLOPS, which directly induces expression of two TF genes, ERF REQUIRED FOR NODULATION1 (ERN1) and NODULE INCEPTION (NIN), essential for IT development. Here, we explored the relationship among these three symbiotic TF genes in the model legume Lotus japonicus and examined how their interplay contributes to IT formation. qRT-PCR analysis showed that NIN expression induced by rhizobial infection was attenuated in ern1-1, and further declined in cyclops-3 ern1-1. ERN1 overexpression led to induction of NIN expression in cyclops-3 ern1-1 in the presence of rhizobia. Thus, in addition to CYCLOPS, ERN1 is able to increase the NIN expression level depending on infection. Furthermore, consistent with this transcriptional hierarchy, ectopic expression of ERN1 as well as NIN suppressed the IT-deficient cyclops-3 phenotype, but ERN1 failed to confer ITs in the nin-2 root. However, the ern1-1 symbiotic epidermal phenotype was not suppressed by the NIN ectopic expression. The cyclops-3 ern1-1 double mutant was less sensitive to rhizobial infection than the single mutants and defective in the symbiotic root hair response at earlier stages. This more severe phenotype of the double mutant suggests a role for ERN1 that independent of the CYCLOPS-mediated transcriptional regulation. We conclude that ERN1 is involved in regulating NIN expression in addition to CYCLOPS, and these TFs coordinately promote the symbiotic root hair response and IT development. Our data help to reveal the extensive role of ERN1 in root nodule symbiosis signaling.

Salinity Stress Alters Root Morphology and Root Hair Traits in Brassica napus.

Plant roots show morphological plasticity and play a substantial role in tolerance to various edaphic stresses. The aim of this study was to explore salinity-induced morphogenic responses of root traits and root hairs of two rapeseed varieties, BARI Sarisha-8 and Binasarisha-5, at the reproductive stage and perceive the effects on their reproductive growth. The experiment was conducted in a hydroponic culture. Two treatments, 0 mM NaCl as control and 100 mM NaCl, were imposed 55 d after germination. Plants exposed to 100 mM NaCl for seven days displayed greater damage in the leaves, flowers, and siliquae compared to control. Length of root hairs on first-order and third-order lateral roots, density of root hairs on first-order lateral roots, and length of third-order lateral roots were significantly greater by 91%, 22%, 29%, and 48%, respectively, in the treated condition compared to the control. An increase in estimated root surface area by 20% under salt stress conditions indicated that the spontaneous responses of plants to uptake more water and nutrients allowed a plant to cope with stressful conditions. The results of this study suggest that any future stress breeding programs should consider plasticity of root traits intensively.

Molecular Background of Pi Deficiency-Induced Root Hair Growth in Brassica carinata - A Fasciclin-Like Arabinogalactan Protein Is Involved.

Formation of longer root hairs under limiting phosphate (P) conditions can increase the inorganic P (Pi) uptake. Here, regulatory candidate genes for Pi deficiency-induced root hair growth were identified by comparison of massive analysis of cDNA ends (MACE) provided expression profiles of two Brassica carinata cultivars (cv.) differing in their root hair response to Pi deficiency: cv. Bale develops longer root hairs under Pi deficiency, but not cv. Bacho. A split-root experiment was conducted for the differentiation between locally and systemically regulated genes. Furthermore, plants were exposed to nitrogen and potassium deficiency to identify P-specific reacting genes. The latter were knocked out by CRISPR/Cas9 and the effect on the root hair length was determined. About 500 genes were differentially expressed under Pi deficiency in cv. Bale, while these genes did not respond to the low P supply in cv. Bacho. Thirty-three candidate genes with a potential regulatory role were selected and the transcriptional regulation of 30 genes was confirmed by quantitative PCR. Only five candidate genes seemed to be either exclusively regulated locally (two) or systemically (three), whereas 25 genes seemed to be involved in both local and systemic signaling pathways. Potassium deficiency affected neither the root hair length nor the expression of the 30 candidate genes. By contrast, both P and nitrogen deficiency increased the root hair length, and both affected the transcript levels in 26 cases. However, four genes reacted specifically to Pi starvation. These genes and, additionally, INORGANIC PHOSPHATE TRANSPORTER 1 (BcPHT1) were targeted by CRISPR/Cas9. However, even if the transcript levels of five of these genes were clearly decreased, FASCICLIN-LIKE ARABINOGALACTAN PROTEIN 1 (BcFLA1) was the only gene whose downregulation reduced the root hair length in transgenic hairy roots under Pi-deficient conditions. To the best of our knowledge, this is the first study describing a fasciclin-like arabinogalactan protein with a predicted role in the Pi deficiency-induced root hair elongation.

A mechanistic framework for auxin dependent Arabidopsis root hair elongation to low external phosphate

Phosphate (P) is an essential macronutrient for plant growth. Roots employ adaptive mechanisms to forage for P in soil. Root hair elongation is particularly important since P is immobile. Here we report that auxin plays a critical role promoting root hair growth in Arabidopsis in response to low external P. Mutants disrupting auxin synthesis (taa1) and transport (aux1) attenuate the low P root hair response. Conversely, targeting AUX1 expression in lateral root cap and epidermal cells rescues this low P response in aux1. Hence auxin transport from the root apex to differentiation zone promotes auxin-dependent hair response to low P. Low external P results in induction of root hair expressed auxin-inducible transcription factors ARF19, RSL2, and RSL4. Mutants lacking these genes disrupt the low P root hair response. We conclude auxin synthesis, transport and response pathway components play critical roles regulating this low P root adaptive response.

Early Arabidopsis root hair growth stimulation by pathogenic strains of Pseudomonas syringae.

Selected beneficial Pseudomonas spp. strains have the ability to influence root architecture in Arabidopsis thaliana by inhibiting primary root elongation and promoting lateral root and root hair formation. A crucial role for auxin in this long-term (1week), long-distance plant-microbe interaction has been demonstrated. Arabidopsis seedlings were cultivated in vitro on vertical plates and inoculated with pathogenic strains Pseudomonas syringae pv. maculicola (Psm) and P. syringae pv. tomato DC3000 (Pst), as well as Agrobacterium tumefaciens (Atu) and Escherichia coli (Eco). Root hair lengths were measured after 24 and 48h of direct exposure to each bacterial strain. Several Arabidopsis mutants with impaired responses to pathogens, impaired ethylene perception and defects in the exocyst vesicle tethering complex that is involved in secretion were also analysed. Arabidopsis seedling roots infected with Psm or Pst responded similarly to when infected with plant growth-promoting rhizobacteria; root hair growth was stimulated and primary root growth was inhibited. Other plant- and soil-adapted bacteria induced similar root hair responses. The most compromised root hair growth stimulation response was found for the knockout mutants exo70A1 and ein2. The single immune pathways dependent on salicylic acid, jasmonic acid and PAD4 are not directly involved in root hair growth stimulation; however, in the mutual cross-talk with ethylene, they indirectly modify the extent of the stimulation of root hair growth. The Flg22 peptide does not initiate root hair stimulation as intact bacteria do, but pretreatment with Flg22 prior to Psm inoculation abolished root hair growth stimulation in an FLS2 receptor kinase-dependent manner. These early response phenomena are not associated with changes in auxin levels, as monitored with the pDR5::GUS auxin reporter. Early stimulation of root hair growth is an effect of an unidentified component of living plant pathogenic bacteria. The root hair growth response is triggered in the range of hours after bacterial contact with roots and can be modulated by FLS2 signalling. Bacterial stimulation of root hair growth requires functional ethylene signalling and an efficient exocyst-dependent secretory machinery.

Enhanced rhizobial symbiotic capacity in an allopolyploid species of Glycine (Leguminosae).

Previous studies have shown that polyploidy can alter biotic interactions, and it has been suggested that these effects may contribute to the increased ability for colonization of new habitats shown by many allopolyploids. Little is known, however, about the effects of allopolyploidy, which combines hybridity and genome doubling, on symbiotic interactions with rhizobial bacteria. We examined interactions of the allopolyploid Glycine dolichocarpa (designated T2) with novel rhizobial partners, such as might occur in a context of colonization, and compared these with the responses of its diploid progenitors, G. tomentella (D3) and G. syndetika (D4). We assessed root hair response, nodule formation, nodule mass, nodule number, and plant biomass. The allopolyploid (T2) showed a greater root hair deformation response when exposed to rhizobia, compared with either diploid. T2 had a greater probability of forming nodules with NGR234 compared with diploid D4, and greater total nodule mass per nodulated plant compared with diploid D3. T2 also had greater plant biomass responses to nitrogen and when exposed to NGR234. The allopolyploid is characterized by transgressive responses to rhizobia for some variables, while also combining certain parental diploid responses such that its capacity for interactions with rhizobia appears to be greater than for either diploid progenitor. This overall enhanced nodulation capacity and the ability to make greater gains from exposure to both rhizobia and additional nitrogen indicate a greater potential of the allopolyploid to benefit from these factors both generally and in a context of colonization.