Roots Of Wild-type Plants Research Articles

Endogenous ureides are employed as a carbon source in Arabidopsis plants exposed to carbon starvation conditions

Ureides, the degraded products of purine catabolism in Arabidopsis, have been shown to act as antioxidant and nitrogen sources. Herein we elucidate purine degraded metabolites as a carbon source using the Arabidopsis Atxdh1, Ataln, and Ataah knockout (KO) mutants vis-à-vis wild-type (WT) plants. Plants were grown under short-day conditions on agar plates containing half-strength MS medium with or without 1% sucrose. Notably, the absence of sucrose led to diminished biomass accumulation in both shoot and root tissues of the Atxdh1, Ataln, and Ataah mutants, while no such effect was observed in WT plants. Moreover, the application of sucrose resulted in a reduction of purine degradation metabolite levels, specifically xanthine and allantoin, predominantly within the roots of WT plants. Remarkably, an increase in proteins associated with the purine degradation pathway was observed in WT plants in the presence of sucrose. Lower glyoxylate levels in the roots but not in the shoot of the Atxdh1 mutant in comparison to WT, were observed under sucrose limitation, and improved by sucrose application in root, indicating that purine degradation provided glyoxylate in the root. Furthermore, the deficit of purine-degraded metabolites in the roots of mutants subjected to carbon starvation was partially mitigated through allantoin application. Collectively, these findings signify that under conditions of sucrose limitation and short-day growth, purines are primarily remobilized within the root system to augment the availability of ureides, serving as an additional carbon (as well as nitrogen) source to support plant growth.

Arabidopsis root apical meristem survival during waterlogging is determined by phytoglobin through nitric oxide and auxin.

Over-expression of phytoglobin mitigates the degradation of the root apical meristem (RAM) caused by waterlogging through changes in nitric oxide and auxin distribution at the root tip. Plant performance to waterlogging is ameliorated by the over-expression of the Arabidopsis Phytoglobin 1 (Pgb1) which also contributes to the maintenance of a functional RAM. Hypoxia induces accumulation of ROS and damage in roots of wild type plants; these events were preceded by the exhaustion of the RAM resulting from the loss of functionality of the WOX5-expressing quiescent cells (QCs). These phenotypic deviations were exacerbated by suppression of Pgb1 and attenuated when the same gene was up-regulated. Genetic and pharmacological studies demonstrated that degradation of the RAM in hypoxic roots is attributed to a reduction in the auxin maximum at the root tip, necessary for the specification of the QC. This reduction was primarily caused by alterations in PIN-mediated auxin flow but not auxin synthesis. The expression and localization patterns of several PINs, including PIN1, 2, 3 and 4, facilitating the basipetal translocation of auxin and its distribution at the root tip, were altered in hypoxic WT and Pgb1-suppressing roots but mostly unchanged in those over-expressing Pgb1. Disruption of PIN1 and PIN2 signal in hypoxic roots suppressing Pgb1 initiated in the transition zone at 12h and was specifically associated to the absence of Pgb1 protein in the same region. Exogenous auxin restored a functional RAM, while inhibition of the directional auxin flow exacerbated the degradation of the RAM. The regulation of root behavior by Pgb1 was mediated by nitric oxide (NO) in a model consistent with the recognized function of Pgbs as NO scavengers. Collectively, this study contributes to our understanding of the role of Pgbs in preserving root meristem function and QC niche during conditions of stress, and suggests that the root transition zone is most vulnerable to hypoxia.

The E3 ubiquitin ligase RING1 interacts with COP9 Signalosome Subunit 4 to positively regulate resistance to root-knot nematodes in Solanum lycopersicum L.

Globally, root-knot nematodes (RKNs) cause massive production losses in all major crops. E3 ubiquitin ligases are involved in plant growth, development and immune response. But their roles in plant defense against RKNs are largely unclear. Here, we show that tomato E3 ubiquitin ligase RING1 interacts with COP9 Signalosome Subunit 4 (CSN4) which is essential for jasmonic acid (JA)-dependent basal defense against RKNs. Tissue-specific expression analysis showed that RING1 expression was the highest in tomato roots and the expression was significantly increased with RKN (Meloidogyne incognita) infection. Compared with the wild-type plants, the number of egg masses in roots significantly increased in the ring1 mutants, while RING1 overexpression conferred resistance against RKNs. Furthermore, RKN infection increased the accumulation of CSN4 protein in the roots of wild-type plants, which was largely compromised in the ring1 mutants but was enhanced in the RING1 overexpressing plants. The RKN-induced transcripts of JA biosynthetic and signaling genes as well as the accumulation of JA and JA-isoleucine were compromised in ring1 mutants but were increased in RING1 overexpressing plants. These results suggest that RING1 positively regulates JA-dependent basal defense against RKNs by interacting with CSN4 proteins.

Engineered plant control of associative nitrogen fixation

Engineering N2-fixing symbioses between cereals and diazotrophic bacteria represents a promising strategy to sustainably deliver biologically fixed nitrogen (N) in agriculture. We previously developed novel transkingdom signaling between plants and bacteria, through plant production of the bacterial signal rhizopine, allowing control of bacterial gene expression in association with the plant. Here, we have developed both a homozygous rhizopine producing (RhiP) barley line and a hybrid rhizopine uptake system that conveys upon our model bacterium Azorhizobium caulinodans ORS571 (Ac) 103-fold improved sensitivity for rhizopine perception. Using this improved genetic circuitry, we established tight rhizopine-dependent transcriptional control of the nitrogenase master regulator nifA and the N metabolism σ-factor rpoN, which drove nitrogenase expression and activity in vitro and in situ by bacteria colonizing RhiP barley roots. Although in situ nitrogenase activity was suboptimally effective relative to the wild-type strain, activation was specific to RhiP barley and was not observed on the roots of wild-type plants. This work represents a key milestone toward the development of a synthetic plant-controlled symbiosis in which the bacteria fix N2 only when in contact with the desired host plant and are prevented from interaction with nontarget plant species.

Glutathione Status in the Roots of Tomato Plants Transgenic by Genes psl and rapA1 in the Presence of Rhizobium leguminosarum

The level of glutathione was investigated in the roots of tomato (Solanum lycopersicum L.) plants transgenic by genes psl and rapA1 in the presence of a microsymbiont of leguminous plants Rhizobium leguminosarum VSy3. The plants transformed with gene psl showed a greater bacterial adhesion than the plants transformed with gene rapA1, which positively correlated with growth parameters of plants. Treatment with rhizobia elevated the content of glutathione in the roots of wild type plants three times, 4.7 times in the roots of plants transformed with gene rapA1, and more than five times in the plants transgenic by gene psl. The obtained results suggest that the level of glutathione in the roots may serve as a marker of efficiency of artificial symbiotic systems produced de novo.

Oryza sativa, C4HC3-type really interesting new gene (RING), OsRFPv6, is a positive regulator in response to salt stress by regulating Na+ absorption.

Salinity negatively affects plant growth, productivity, and metabolism. Therefore, plants have evolved diverse strategies to survive in saline environments. To identify such strategies involving the ubiquitin/26S proteasome system, we characterized molecular functions of a rice C4HC3 really interesting new gene (RING)-type E3-ubiquitin ligase gene. Oryza sativa RING finger protein v6 (OsRFPv6) was highly expressed under conditions of abiotic stress, induced by 100 mM NaCl and 20% PEG. The GFP-OsRFPv6 protein was localized in the plasma membrane and cytosol in rice protoplasts. In vitro ubiquitin assay revealed that OsRFPv6 possessed E3-ubiquitin ligase activity, but its variant OsRFPv6C100A did not. OsRFPv6-overexpressing plants were insensitive to salinity, but their growth was delayed under normal conditions. Under saline conditions, transgenic plants exhibited higher proline, soluble sugar, and chlorophyll content and lower H2 O2 accumulation than wild-type plants. Moreover, transgenic plants exhibited lower Na+ uptake, lower Na+ content, and higher K+ content in the xylem sap assay. Under saline conditions, the expression levels of nine Na+ /K+ transporter genes in roots and leaves were significantly different between transgenic and wild-type plants. Specifically, under both normal and saline conditions, the expression of OsHKT2;1, a Na+ transporter, in the roots of transgenic plants was lower than that in the roots of wild-type plants. These results suggest that OsRFPv6 E3-ubiquitin ligase serves as a positive regulator of salinity response via Na+ uptake.

Participation of Nitrate Sensor NRT1.1 in the Control of Cytokinin Level and Root Elongation under Normal Conditions and Nitrogen Deficit

NRT1.1 nitrate transporter acts as a nitrate sensor in some plant responses. We tried to check if it may be involved in the control of cytokinin level in the plants known to be involved in the growth responses to nitrate level. The experimental objects were Arabidopsis thaliana plants of the original ecotype Columbia (Col-0) and chl1-5 mutants. The effects of the NRT1.1 gene mutation in chl1-5 plants on hormonal and growth responses to nitrogen starvation were studied. Two types of growing conditions were used: (1) plants were placed on either standard Hoagland–Arnon or modified solution, where potassium and calcium nitrates were substituted with their chlorides; (2) plants were placed on Pryanishnikov medium, where ammonium nitrate serves as the source of nitrogen and nitrogen deficiency being modeled by its withdrawal from the medium. It has been first shown that mutation of the NRT1.1 resulted in a decline in cytokinin level in the roots of chl1-5 mutants, while roots of wild type plants were longer in accordance with lower cytokinin content in them; this hormone is known to inhibit root elongation. Cytokinin content decreased in A. thaliana, Columbia ecotype, paralleled by acceleration of root elongation in response to both variants of nitrogen starvation, while chl1-5 roots responded in this way only when nitrogen was withdrawn from Pryanishnikov solution. Substitution of nitrates by chlorides in the Hoagland–Arnon solution had no effects on either chl1-5 roots’ length or cytokinin content in them. The results suggested the involvement of NRT1.1 transceptor in the control of cytokinin level and root elongation rate in the nitrate but not in ammonium starved plants, confirming the specificity of response.

Ectopic or Over-Expression of Class 1 Phytoglobin Genes Confers Flooding Tolerance to the Root Nodules of Lotus japonicus by Scavenging Nitric Oxide.

Flooding limits biomass production in agriculture. Leguminous plants, important agricultural crops, use atmospheric dinitrogen gas as nitrogen nutrition by symbiotic nitrogen fixation with rhizobia, but this root-nodule symbiosis is sometimes broken down by flooding of the root system. In this study, we analyzed the effect of flooding on the symbiotic system of transgenic Lotus japonicus lines which overexpressed class 1 phytoglobin (Glb1) of L. japonicus (LjGlb1-1) or ectopically expressed that of Alnus firma (AfGlb1). In the roots of wild-type plants, flooding increased nitric oxide (NO) level and expression of senescence-related genes and decreased nitrogenase activity; in the roots of transgenic lines, these effects were absent or less pronounced. The decrease of chlorophyll content in leaves and the increase of reactive oxygen species (ROS) in roots and leaves caused by flooding were also suppressed in these lines. These results suggest that increased levels of Glb1 help maintain nodule symbiosis under flooding by scavenging NO and controlling ROS.

Inoculation with Bacillus subtilis and Azospirillum brasilense Produces Abscisic Acid That Reduces Irt1-Mediated Cadmium Uptake of Roots.

Cadmium (Cd) contamination of agricultural soils represents a serious risk to crop safety. A new strategy using abscisic acid (ABA)-generating bacteria, Bacillus subtilis or Azospirillum brasilense, was developed to reduce the Cd accumulation in plants grown in Cd-contaminated soil. Inoculation with either bacterium resulted in a pronounced increase in the ABA level in wild-type Arabidopsis Col-0 plants, accompanied by a decrease in Cd levels in plant tissues, which mitigated the Cd toxicity. As a consequence, the growth of plants exposed to Cd was improved. Nevertheless, B. subtilis and A. brasilense inoculation had little effect on Cd levels and toxicity in the ABA-insensitive mutant snrk 2.2/2.3, indicating that the action of ABA is required for these bacteria to reduce Cd accumulation in plants. Furthermore, inoculation with either B. subtilis or A. brasilense downregulated the expression of IRT1 (iron-regulated transporter 1) in the roots of wild-type plants and had little effect on Cd levels in the IRT1-knockout mutants irt1-1 and irt1-2. In summary, we conclude that B. subtilis and A. brasilense can reduce Cd levels in plants via an IRT1-dependent ABA-mediated mechanism.

Suppression of Arabidopsis flowering by near-null magnetic field is mediated by auxin.

We previously found that flowering of Arabidopsis was suppressed by near-null magnetic field, which was related to cryptochrome. Auxin plays an important role in Arabidopsis flowering. To test whether auxin is involved in the suppression of Arabidopsis flowering by near-null magnetic field, we detected auxin level and expressions of auxin transport and signaling genes in wild-type Arabidopsis plants and cryptochrome double mutant, cry1/cry2, grown in near-null magnetic field. We found that indole-3-acetic acid (IAA) level in roots of wild-type plants in near-null magnetic field was significantly increased compared with the local geomagnetic field control, while IAA level in rosettes of 33-day-old wild-type plants in near-null magnetic field was significantly lower than the control. Expressions of three auxin transporter genes, PIN1, PIN3, and PIN7, in wild-type plants were upregulated by near-null magnetic field. Transcript levels of transcriptional repressor genes, IAA1, IAA5, IAA6, IAA16, and IAA19, were significantly higher in wild-type plants in near-null magnetic field than in control plants. However, IAA level and expressions of all the detected genes in cry1/cry2 mutants in near-null magnetic field were similar to controls. Our results suggest that near-null magnetic field affects the distribution of auxin in Arabidopsis by transcriptional upregulation of auxin transporter genes, and that change in distribution of auxin and increased expressions of transcriptional repressor genes result in delay of flowering in Arabidopsis in near-null magnetic field, which are mediated by cryptochrome. Bioelectromagnetics. 39:15-24, 2018. © 2017 Wiley Periodicals, Inc.

Phytoremediation of arsenic from the contaminated soil using transgenic tobacco plants expressing ACR2 gene of Arabidopsis thaliana

We have cloned, characterized and transformed the AtACR2 gene (arsenic reductase 2) of Arabidopsis thaliana into the genome of tobacco (Nicotiana tabacum, var Sumsun). Our results revealed that the transgenic tobacco plants are more tolerant to arsenic than the wild type ones. These plants can grow on culture medium containing 200μM arsenate, whereas the wild type can barely survive under this condition. Furthermore, when exposed to 100μM arsenate for 35days the amount of arsenic accumulated in the shoots of transgenic plants was significantly lower (28μg/g d wt.) than that found in the shoots of non-transgenic controls (40μg/g d wt.). However, the arsenic content in the roots of transgenic plants was significantly higher (2400μg/g d. wt.) than that (2100μg/g d. wt.) observed in roots of wild type plants. We have demonstrated that Arabidopsis thaliana AtACR2 gene is a potential candidate for genetic engineering of plants to develop new crop cultivars that can be grown on arsenic contaminated fields to reduce arsenic content of the soil and can become a source of food containing no arsenic or exhibiting substantially reduced amount of this metalloid.

Basic helix-loop-helix (bHLH) transcriptional activator regulates ammonium uptake in rice

NH4+ is important for the growth and the yield production of paddy-soil grown rice. However, the mechanism of NH4+ signaling is largely unknown. Previously, we performed a microarray analysis to examine genomic responses and signaling pathways in rice roots exposed to NH4+, and interestingly found that transcription of a large number of genes including kinase, phosphatase, and transcription factors were sensitive to NH4+. In this study, we further analyzed transcription factors respond to NH4+, and identified an NH4+-repressed bHLH transcription factor involved in NH4+ acquisition process. bHLH is a nuclear protein and has transcription-activation activity in yeast and plants. RNAi-mediated suppression of bHLH and bHLH overexpression (bHLH OX) resulted in the accumulation of lower and higher NH4+ contents in transgenic rice roots, respectively. NH4+-mediated induction of AMT1;1 and AMT1;2 is lower in bHLH RNAi, but higher in bHLH OX as compared to in wild-type plant roots. However, bHLH does not directly activate AMT1;1 and AMT1;2 transcriptions. IDD10 expression is not altered in bHLH RNAi plant roots, and bHLH maintained similar expression level in idd10 mutant roots compared to in wild-type plant roots. Furthermore, NH4+ contents in idd10;bHLH RNAi double mutant roots were much lower than in each single mutant and wild-type plant roots, indicating that IDD10 and bHLH independently regulates NH4+ uptake. Taken together, these results provide a new regulatory path by which bHLH regulates NH4+ uptake in rice.

Phosphate Deficiency Induces the Jasmonate Pathway and Enhances Resistance to Insect Herbivory.

During their life cycle, plants are typically confronted by simultaneous biotic and abiotic stresses. Low inorganic phosphate (Pi) is one of the most common nutrient deficiencies limiting plant growth in natural and agricultural ecosystems, while insect herbivory accounts for major losses in plant productivity and impacts ecological and evolutionary changes in plant populations. Here, we report that plants experiencing Pi deficiency induce the jasmonic acid (JA) pathway and enhance their defense against insect herbivory. Pi-deficient Arabidopsis (Arabidopsis thaliana) showed enhanced synthesis of JA and the bioactive conjugate JA-isoleucine, as well as activation of the JA signaling pathway, in both shoots and roots of wild-type plants and in shoots of the Pi-deficient mutant pho1 The kinetics of the induction of the JA signaling pathway by Pi deficiency was influenced by PHOSPHATE STARVATION RESPONSE1, the main transcription factor regulating the expression of Pi starvation-induced genes. Phenotypes of the pho1 mutant typically associated with Pi deficiency, such as high shoot anthocyanin levels and poor shoot growth, were significantly attenuated by blocking the JA biosynthesis or signaling pathway. Wounded pho1 leaves hyperaccumulated JA/JA-isoleucine in comparison with the wild type. The pho1 mutant also showed an increased resistance against the generalist herbivore Spodoptera littoralis that was attenuated in JA biosynthesis and signaling mutants. Pi deficiency also triggered increased resistance to S. littoralis in wild-type Arabidopsis as well as tomato (Solanum lycopersicum) and Nicotiana benthamiana, revealing that the link between Pi deficiency and enhanced herbivory resistance is conserved in a diversity of plants, including crops.

一个水稻铁转运基因(<italic>OsFRDL1</italic>)参与缺氧诱导根系铁膜形成的调节过程

Iron plaques that form on the surface of roots serve as an important nutrient reservoir and as a natural barrier for rice ( Oryza sativa ) seedlings to adapt to environmental stress. However, the molecular mechanism of iron plaque formation is poorly understood. In this study, the Osfrdl1 mutant and its wild-type (Nipponbare) cultivar were used to explore the role of the OsFRDL1 gene in iron plaque formation on rice roots. The results showed that a low phosphorus concentration ( Osfrdl1 mutant than on those of the wild-type. The results of real-time RT-PCR analyses indicated that the stagnant treatment increased the expression of OsFRDL1. As well, OsFRDL1 :: GUS staining indicated that the GUS reporter gene was mainly expressed in epidermis cells and stele tissues in the root. The expression of the reporter gene in these tissues was markedly higher in seedlings in the stagnant treatment than in those in the aerated treatment. In the stagnant treatment, the peroxidase activity was higher in roots of wild-type plants than in those of the Osfrdl1 mutant. Taken together, these results suggest that the stagnant treatment induced increased expression of OsFRDL1 . This increased the peroxidase activity to a higher level in the wild-type than in the mutant, and promoted greater iron plaque formation on the roots.

The effects of redox controls mediated by glutathione peroxidases on root architecture in Arabidopsis thaliana.

Glutathione peroxidases (GPXs) fulfil important functions in oxidative signalling and protect against the adverse effects of excessive oxidation. However, there has been no systematic characterization of the functions of the different GPX isoforms in plants. The roles of the different members of the Arabidopsis thaliana GPX gene (AtGPX) family were therefore investigated using gpx1, gpx2, gpx3, gpx4, gpx6, gpx7, and gpx8 T-DNA insertion mutant lines. The shoot phenotypes were largely similar in all genotypes, with small differences from the wild type observed only in the gpx2, gpx3, gpx7, and gpx8 mutants. In contrast, all the mutants showed altered root phenotypes compared with the wild type. The gpx1, gpx4, gpx6, gpx7, and gpx8 mutants had a significantly greater lateral root density (LRD) than the wild type. Conversely, the gpx2 and gpx3 mutants had significantly lower LRD values than the wild type. Auxin increased the LRD in all genotypes, but the effect of auxin was significantly greater in the gpx1, gpx4, and gpx7 mutants than in the wild type. The application of auxin increased GPX4 and GPX7 transcripts, but not GPX1 mRNAs in the roots of wild-type plants. The synthetic strigolactone GR24 and abscisic acid (ABA) decreased LRD to a similar extent in all genotypes, except gpx6, which showed increased sensitivity to ABA. These data not only demonstrate the importance of redox controls mediated by AtGPXs in the control of root architecture but they also show that the plastid-localized GPX1 and GPX7 isoforms are required for the hormone-mediated control of lateral root development.

Root-Specific Role for Nicotiana benthamiana RDR6 in the Inhibition of Chinese wheat mosaic virus Accumulation at Higher Temperatures

Some viruses only infect plants at cool temperatures but the molecular mechanism underlying this low-temperature dependence remains unclear. Chinese wheat mosaic virus (CWMV, genus Furovirus) was able to infect wheat and Nicotiana benthamiana plants at 16 but not at 24°C. When CWMV-infected plants were transferred to 24°C for 2 weeks, the newly emerged leaves and roots became virus free. Co-infection with Potato virus Y rescued CWMV accumulation in N. benthamiana plants after a temperature shift to 24°C. In transgenic N. benthamiana plants silenced for the N. benthamiana RNA-dependent RNA polymerase 6 (NbRDR6), CWMV was able to accumulate in roots but not in leaves after a temperature shift to 24°C. Deep sequencing of small RNAs showed that, at 16°C, abundant CWMV small interfering (si)RNAs accumulated in infected N. benthamiana plants. Silencing of NbRDR6 increased the abundance of CWMV siRNAs and the generation of siRNAs from hotspots in the CWMV genome. In contrast, when shifted to 24°C for 1 week, CWMV siRNAs were markedly fewer in roots of NbRDR6-silenced than in roots of wild-type plants but were similar in the leaves of those plants. Our results demonstrate the root-specific role of NbRDR6 in the inhibition of CWMV accumulation and biogenesis of CWMV siRNAs at higher temperatures.

Overexpression of MsALMT1, from the Aluminum-Sensitive Medicago sativa, Enhances Malate Exudation and Aluminum Resistance in Tobacco

Aluminum (Al) toxicity is a major limiting factor for plant growth and crop production in acidic soils. Al-induced organic acid (OA) exudation plays an important role in plant Al resistance. The exudation of OAs is mediated by membrane-localized OA transporters. In our previous study, a gene encoding the Al-induced malate transporter (MsALMT1) was identified in the roots of the Al-sensitive plant Medicago sativa L. cv. Yumu no. 1 (YM1). To further validate the function of MsALMT1, transgenic plants that overexpressed MsALMT1 under the control of the CaMV 35S (35S) promoter were generated. This transgenic tobacco showed an enhanced capacity for malate efflux and better Al resistance than wild type (WT) plants after exposure to 30 μM Al for 24 h. The Al content in the transgenic plant roots decreased to 40–52 % of that in WT plant roots. These results demonstrate that MsALMT1 is an Al-resistant gene in YM1 and encodes a malate transporter, the overexpression of which effectively enhances the Al resistance of transgenic tobacco plants.

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

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

Overexpression of PIP2;5 aquaporin alleviates effects of low root temperature on cell hydraulic conductivity and growth in Arabidopsis.

The effects of low root temperature on growth and root cell water transport were compared between wild-type Arabidopsis (Arabidopsis thaliana) and plants overexpressing plasma membrane intrinsic protein 1;4 (PIP1;4) and PIP2;5. Descending root temperature from 25°C to 10°C quickly reduced cell hydraulic conductivity (L(p)) in wild-type plants but did not affect L(p) in plants overexpressing PIP1;4 and PIP2;5. Similarly, when the roots of wild-type plants were exposed to 10°C for 1 d, L(p) was lower compared with 25°C. However, there was no effect of low root temperature on L(p) in PIP1;4- and PIP2;5-overexpressing plants after 1 d of treatment. When the roots were exposed to 10°C for 5 d, L(p) was reduced in wild-type plants and in plants overexpressing PIP1;4, whereas there was still no effect in PIP2;5-overexpressing plants. These results suggest that the gating mechanism in PIP1;4 may be more sensitive to prolonged low temperature compared with PIP2;5. The reduction of L(p) at 10°C in roots of wild-type plants was partly restored to the preexposure level by 5 mm Ca(NO(3))(2) and protein phosphatase inhibitors (75 nm okadaic acid or 1 μm Na(3)VO(4)), suggesting that aquaporin phosphorylation/dephosphorylation processes were involved in this response. The temperature sensitivity of cell water transport in roots was reflected by a reduction in shoot and root growth rates in the wild-type and PIP1;4-overexpressing plants exposed to 10°C root temperature for 5 d. However, low root temperature had no effect on growth in plants overexpressing PIP2;5. These results provide strong evidence for a link between growth at low root temperature and aquaporin-mediated root water transport in Arabidopsis.

CLE-like (CLEL) peptides control the pattern of root growth and lateral root development in Arabidopsis

CLE peptides, named for the CLV3/ESR-related peptide family, participate in intercellular-signaling pathways. Here we investigated members of the CLE-like (CLEL) gene family that encode peptide precursors recently designated as root growth factors [Matsuzaki Y et al. (2010) Science 329:1065-1067]. CLEL precursors share a similar domain structure with CLE precursors (i.e., they contain a putative N-terminal signal peptide and a C-terminal conserved 13-amino-acid CLEL motif with a variable middle portion). Our evidence shows that, unlike root growth factor, CLEL peptides are (i) unmodified and (ii) function in the regulation of the direction of root growth and lateral root development. Overexpression of several CLEL genes in Arabidopsis resulted in either long roots or long and wavy roots that also showed altered lateral root patterning. Exogenous application of unmodified synthetic 13-amino-acid peptides derived from two CLEL motifs resulted in similar phenotypic changes in roots of wild-type plants. In CLEL peptide-induced long roots, the root apical meristem (RAM) was enlarged and consisted of an increased number of cells, compared with wild-type root apical meristems. The wavy-root phenotype appeared to be independent of other responses of the roots to the environment (e.g., gravitropism, phototropism, and thigmotropism). Results also showed that the inhibition of lateral initiation by CLEL overexpression was not overcome by the application of auxin. These findings establish CLEL as a peptide family with previously unrecognized regulatory functions controlling the pattern of root growth and lateral root development in plants.