Inhibition Of Lateral Root Formation Research Articles

Pickle Recruits Retinoblastoma Related 1 to Control Lateral Root Formation in Arabidopsis.

Lateral root (LR) formation is an example of a plant post-embryonic organogenesis event. LRs are issued from non-dividing cells entering consecutive steps of formative divisions, proliferation and elongation. The chromatin remodeling protein PICKLE (PKL) negatively regulates auxin-mediated LR formation through a mechanism that is not yet known. Here we show that PKL interacts with RETINOBLASTOMA-RELATED 1 (RBR1) to repress the LATERAL ORGAN BOUNDARIES-DOMAIN 16 (LBD16) promoter activity. Since LBD16 function is required for the formative division of LR founder cells, repression mediated by the PKL–RBR1 complex negatively regulates formative division and LR formation. Inhibition of LR formation by PKL–RBR1 is counteracted by auxin, indicating that, in addition to auxin-mediated transcriptional responses, the fine-tuned process of LR formation is also controlled at the chromatin level in an auxin-signaling dependent manner.

Novel MtCEP1 peptides produced in vivo differentially regulate root development in Medicago truncatula.

Small, post-translationally modified and secreted peptides regulate diverse plant developmental processes. Due to low natural abundance, it is difficult to isolate and identify these peptides. Using an improved peptide isolation protocol and Orbitrap mass spectrometry, nine 15-amino-acid CEP peptides were identified that corresponded to the two domains encoded by Medicago truncatula CEP1 (MtCEP1). Novel arabinosylated and hydroxylated peptides were identified in root cultures overexpressing MtCEP1. The five most abundant CEP peptides were hydroxylated and these species were detected also in low amounts in vector control samples. Synthetic peptides with different hydroxylation patterns differentially affected root development. Notably, the domain 1 peptide hydroxylated at Pro4 and Pro11 (D1:HyP4,11) imparted the strongest inhibition of lateral root emergence when grown with 5mM KNO3 and stimulated the highest increase in nodule number when grown with 0mM KNO3. Inhibition of lateral root emergence by D1:HyP4,11 was not alleviated by removing peptide exposure. In contrast, the domain 2 peptide hydroxylated at Pro11 (D2:HyP11) increased stage III-IV lateral root primordium numbers by 6-fold (P < 0.001) which failed to emerge. Auxin addition at levels which stimulated lateral root formation in wild-type plants had little or no ameliorating effect on CEP peptide-mediated inhibition of lateral root formation or emergence. Both peptides increased and altered the root staining pattern of the auxin-responsive reporter GH3:GUS suggesting CEPs alter auxin sensitivity or distribution. The results showed that CEP primary sequence and post-translational modifications influence peptide activities and the improved isolation procedure effectively and reproducibly identifies and characterises CEPs.

Coumarin interacts with auxin polar transport to modify root system architecture in Arabidopsis thaliana

Coumarin is a highly active allelopathic compound which plays a key role in plant–plant interactions and communications. It affects root growth and development of many species, but its mode of action has not been clarified yet. It has been hypothesized that auxin could mediate coumarin-induced effects on root system. Through morphological and pharmacological approaches together with the use of Arabidopsis auxin mutants, a possible interaction between coumarin and auxin in driving root system development has been investigated in Arabidopsisthaliana (Col-0). Coumarin strongly affected primary root elongation and lateral root development of Arabidopsis seedlings. In particular, 10−4 M coumarin significantly inhibited primary root elongation increasing lateral root number and root hairs length. Further, coumarin addition was able to restore the negative effects of TIBA and NPA, two auxin transport inhibitors, which caused a complete inhibition of lateral root formation. Arabidopsis auxin mutants differently responded to coumarin compared to wild type (Col-0). In particular, lax3 mutant showed the lowest (42 %) inhibition of primary root length, whereas, eir1-4 mutant had higher inhibition (53 %) compared to Col-0; conversely, aux1-22 mutant did not show any effect in response to coumarin. An increase of lateral root number was observed in pin1 mutant only. Finally, coumarin increased the root hairs length in eir1-4, lax3, pin1 and pin3-5 mutants, but not in aux1-22. These results suggested a functional interaction between coumarin and auxin polar transport in driving root development in A.thaliana.

Auxin-induced hydrogen sulfide generation is involved in lateral root formation in tomato

Similar to auxin, hydrogen sulfide (H2S), mainly produced by l-cysteine desulfhydrase (DES; EC 4.4.1.1) in plants, could induce lateral root formation. The objective of this study was to test whether H2S is also involved in auxin-induced lateral root development in tomato (Solanum lycopersicum L.) seedlings. We observed that auxin depletion-induced down-regulation of transcripts of SlDES1, decreased DES activity and endogenous H2S contents, and the inhibition of lateral root formation were rescued by sodium hydrosulfide (NaHS, an H2S donor). However, No additive effects were observed when naphthalene acetic acid (NAA) was co-treated with NaHS (lower than 10 mM) in the induction of lateral root formation. Subsequent work revealed that a treatment with NAA or NaHS could simultaneously induce transcripts of SlDES1, DES activity and endogenous H2S contents, and thereafter the stimulation of lateral root formation. It was further confirmed that H2S or HS−, not the other sulfur-containing components derived from NaHS, was attributed to the stimulative action. The inhibition of lateral root formation and decreased of H2S metabolism caused by an H2S scavenger hypotaurine (HT) were reversed by NaHS, but not NAA. Molecular evidence revealed that both NaHS- or NAA-induced modulation of some cell cycle regulatory genes, including the up-regulation of SlCDKA;1, SlCYCA2;1, together with simultaneous down-regulation of SlKRP2, were differentially reversed by HT pretreatment. To summarize, above results clearly suggested that H2S might, at least partially, act as a downstream component of auxin signaling to trigger lateral root formation.

The peptide-encoding CEP1 gene modulates lateral root and nodule numbers in Medicago truncatula

The role of MtCEP1, a member of the CEP (C-terminally encoded peptide) signaling peptide family, was examined in Medicago truncatula root development. MtCEP1 was expressed in root tips, vascular tissue, and young lateral organs, and was up-regulated by low nitrogen levels and, independently, by elevated CO2. Overexpressing MtCEP1 or applying MtCEP1 peptide to roots elicited developmental phenotypes: inhibition of lateral root formation, enhancement of nodulation, and the induction of periodic circumferential root swellings, which arose from cortical, epidermal, and pericycle cell divisions and featured an additional cortical cell layer. MtCEP peptide addition to other legume species induced similar phenotypes. The enhancement of nodulation by MtCEP1 is partially tolerant to high nitrate, which normally strongly suppresses nodulation. These nodules develop faster, are larger, and fix more nitrogen in the absence and presence of inhibiting nitrate levels. At 25mM nitrate, nodules formed on pre-existing swelling sites induced by MtCEP1 overexpression. RNA interference-mediated silencing of several MtCEP genes revealed a negative correlation between transcript levels of MtCEP1 and MtCEP2 with the number of lateral roots. MtCEP1 peptide-dependent phenotypes were abolished or attenuated by altering or deleting key residues in its 15 amino acid domain. RNA-Seq analysis revealed that 89 and 116 genes were significantly up- and down-regulated, respectively, by MtCEP1 overexpression, including transcription factors WRKY, bZIP, ERF, and MYB, homologues of LOB29, SUPERROOT2, and BABY BOOM. Taken together, the data suggest that the MtCEP1 peptide modulates lateral root and nodule development in M. truncatula.

Ammonium-induced shoot ethylene production is associated with the inhibition of lateral root formation in Arabidopsis

Foliar NH4(+) exposure is linked to inhibition of lateral root (LR) formation. Here, the role of shoot ethylene in NH4(+)-induced inhibition of LR formation in Arabidopsis was investigated using wild-type and mutant lines that show either blocked ethylene signalling (etr1) or enhanced ethylene synthesis (eto1, xbat32). NH4(+) exposure of wild-type Arabidopsis led to pronounced inhibition of LR production chiefly in the distal root, and triggered ethylene evolution and enhanced activity of the ethylene reporter EBS:GUS in the shoot. It is shown that shoot contact with NH4(+) is necessary to stimulate shoot ethylene evolution. The ethylene antagonists Ag(+) and aminoethoxyvinylglycine (AVG) mitigated LR inhibition under NH4(+) treatment. The decrease in LR production was significantly greater for eto1-1 and xbat32 and significantly less for etr1-3. Enhanced shoot ethylene synthesis/signalling blocked recovery of LR production when auxin was applied in the presence of NH4(+) and negatively impacted shoot AUX1 expression. The findings highlight the important role of shoot ethylene evolution in NH4(+)-mediated inhibition of LR formation.

Molecular mechanisms of hydrotropism in seedling roots of Arabidopsis thaliana (Brassicaceae)

Roots show positive hydrotropism in response to moisture gradients, which is believed to contribute to plant water acquisition. This article reviews the recent advances of the physiological and molecular genetic studies on hydrotropism in seedling roots of Arabidopsis thaliana. We identified MIZU-KUSSEI1 (MIZ1) and MIZ2, essential genes for hydrotropism in roots; the former encodes a protein of unknown function, and the latter encodes an ARF-GEF (GNOM) protein involved in vesicle trafficking. Because both mutants are defective in hydrotropism but not in gravitropism, these mutations might affect a molecular mechanism unique to hydrotropism. MIZ1 is expressed in the lateral root cap and cortex of the root proper. It is localized as a soluble protein in the cytoplasm and in association with the cytoplasmic face of endoplasmic reticulum (ER) membranes in root cells. Light and ABA independently regulate MIZ1 expression, which influences the ultimate hydrotropic response. In addition, MIZ1 overexpression results in an enhancement of hydrotropism and an inhibition of lateral root formation. This phenotype is likely related to the alteration of auxin content in roots. Specifically, the auxin level in the roots decreases in the MIZ1 overexpressor and increases in the miz1 mutant. Unlike most gnom mutants, miz2 displays normal morphology, growth, and gravitropism, with normal localization of PIN proteins. It is probable that MIZ1 plays a crucial role in hydrotropic response by regulating the endogenous level of auxin in Arabidopsis roots. Furthermore, the role of GNOM/MIZ2 in hydrotropism is distinct from that of gravitropism.

ZmHO-1, a maize haem oxygenase-1 gene, plays a role in determining lateral root development

Previous results revealed that haem oxygenase-1 (HO-1)/carbon monoxide (CO) system is involved in auxin-induced adventitious root formation. In this report, a cDNA for the gene ZmHO-1, encoding an HO-1 protein, was cloned from Zea mays seedlings. ZmHO-1 has a conserved HO signature sequence and shares highest homology with rice SE5 (OsHO-1) protein. We further discovered that N-1-naphthylacetic acid (NAA), haemin, and CO aqueous solution, led to the induction of ZmHO-1 expression as well as the thereafter promotion of lateral root development. These effects were specific for ZmHO-1 since the potent HO-1 inhibitor zinc protoporphyrin IX (ZnPPIX) differentially blocked the above actions. The addition of haemin and CO were able to reverse the auxin depletion-triggered inhibition of lateral root formation as well as the decreased ZmHO-1 transcripts. Molecular evidence showed that the haemin- or CO-mediated the modulation of target genes responsible for lateral root formation, including ZmCDK and ZmCKI2, could be blocked by ZnPPIX. Overexpression of ZmHO-1 in transgenic Arabidopsis plants resulted in promotion of lateral root development as well as the modulation of cell cycle regulatory gene expressions. Overall, our results suggested that a maize HO-1 gene is required for the lateral root formation.

Hormonal Regulation of Lateral Root Development in Arabidopsis Modulated byMIZ1and Requirement of GNOM Activity forMIZ1Function

Plant organ development is important for adaptation to a changing environment. Genetic and physiological studies have revealed that plant hormones play key roles in lateral root formation. In this study, we show that MIZU-KUSSEI1 (MIZ1), which was identified originally as a regulator of hydrotropism, functions as a novel regulator of hormonally mediated lateral root development. Overexpression of MIZ1 (MIZ1OE) in roots resulted in a reduced number of lateral roots being formed; however, this defect could be recovered with the application of auxin. Indole-3-acetic acid quantification analyses showed that free indole-3-acetic acid levels decreased in MIZ1OE roots, which indicates that alteration of auxin level is critical for the inhibition of lateral root formation in MIZ1OE plants. In addition, MIZ1 negatively regulates cytokinin sensitivity on root development. Application of cytokinin strongly induced the localization of MIZ1-green fluorescent protein to lateral root primordia, which suggests that the inhibition of lateral root development by MIZ1 occurs downstream of cytokinin signaling. Surprisingly, miz2, a weak allele of gnom, suppressed developmental defects in MIZ1OE plants. Taken together, these results suggest that MIZ1 plays a role in lateral root development by maintaining auxin levels and that its function requires GNOM activity. These data provide a molecular framework for auxin-dependent organ development in Arabidopsis (Arabidopsis thaliana).

Isolation and characterization of a novel ammonium overly sensitive mutant, amos2, in Arabidopsis thaliana

Ammonium (NH(4)(+)) toxicity is a significant agricultural problem globally, compromising crop growth and productivity in many areas. However, the molecular mechanisms of NH(4)(+) toxicity are still poorly understood, in part due to a lack of valuable genetic resources. Here, a novel Arabidopsis mutant, amos2 (ammonium overly sensitive 2), displaying hypersensitivity to NH(4) (+) in both shoots and roots, was isolated. The mutant exhibits the hallmarks of NH(4)(+) toxicity at significantly elevated levels: severely suppressed shoot biomass, increased leaf chlorosis, and inhibition of lateral root formation. Amos2 hypersensitivity is associated with excessive NH(4)(+) accumulation in shoots and a reduction in tissue potassium (K(+)), calcium (Ca(2+)), and magnesium (Mg(2+)). We show that the lesion is specific to the NH(4)(+) ion, is independent of NH(4)(+) metabolism, and can be partially rescued by elevated external K(+). The amos2 lesion was mapped to a 16-cM interval on top of chromosome 1, where no similar mutation has been previously mapped. Our study identifies a novel locus controlling cation homeostasis under NH(4)(+) stress and provides a tool for the future identification of critical genes involved in the development of NH(4)(+) toxicity.

Ethylene inhibits lateral root development, increases IAA transport and expression of PIN3 and PIN7 auxin efflux carriers

We used genetic and molecular approaches to identify mechanisms by which the gaseous plant hormone ethylene reduces lateral root formation and enhances polar transport of the hormone auxin. Arabidopsis thaliana mutants, aux1, lax3, pin3 and pin7, which are defective in auxin influx and efflux proteins, were less sensitive to the inhibition of lateral root formation and stimulation of auxin transport following treatment with the ethylene precursor 1-aminocyclopropane-1-carboxylic acid (ACC). By contrast, pin2 and abcb19 mutants exhibited wild-type ACC responses. ACC and indole-3-acetic acid (IAA) increased the abundance of transcripts encoding auxin transport proteins in an ETR1 and EIN2 (ethylene signaling)-dependent and TIR1 (auxin receptor)-dependent fashion, respectively. The effects of ACC on these transcripts and on lateral root development were still present in the tir1 mutant, suggesting independent signaling networks. ACC increased auxin-induced gene expression in the root apex, but decreased expression in regions where lateral roots form and reduced free IAA in whole roots. The ethylene synthesis inhibitor aminoethoxyvinylglycine (AVG) had opposite effects on auxin-dependent gene expression. These results suggest that ACC affects root development by altering auxin distribution. PIN3- and PIN7-GFP fluorescence was increased or decreased after ACC or AVG treatment, respectively, consistent with the role of PIN3 and PIN7 in ACC-elevated transport. ACC treatment abolished a localized depletion of fluorescence of PIN3- and PIN7-GFP, normally found below the site of primordia formation. These results suggest that ACC treatment increased PIN3 and PIN7 expression, resulting in elevated auxin transport, which prevented the localized accumulation of auxin needed to drive lateral root formation.

Expression of ABSCISIC ACID INSENSITIVE 4 (ABI4) in developing Arabidopsis seedlings

We have recently demonstrated that the transcription factor ABSCISIC ACID INSENSITIVE 4 (ABI4) mediates abscisic acid and cytokinin inhibition of lateral root formation by reducing polar auxin transport in Arabidopsis thaliana.1 In that study, we provided a direct demonstration of ABI4 expression in phloem companion cells and parenchyma of the vascular system in the mature regions of the roots. Although also studied in mature plants, ABI4 has been studied primarily in germinating seedlings, and its expression has been assumed by some researchers to be restricted to early germination stages. We thus constructed transgenic Arabidopsis plants expressing an ABI4:GUS construct, and followed ABI4 promoter activity during seedling development, focusing on the roots.

Effects of Potassium Deficiency on Root Growth of Cotton Seedlings and Its Physiological Mechanisms

For the purpose of dissecting the mechanism of root growth in response to potassium (K) deficiency in cotton ( Gossypium hirsutum L.), young seedlings of NuCOTN99B grown in half-strength modified Hoagland's solution with low K nutrient (0.05 mmol L −1) were investigated for the root configuration, content of endogenous free indole acetic acid (IAA), and amount of ethylene released from the roots 4 d after treatment. Compared with the treatment with moderate K nutrient (0.50 mmol L −1, control), the K deficient treatment significantly inhibited root length and the formation of lateral roots. The reduced lateral roots mainly resulted from the shortened branched root zone, and there was no change in the lateral root density. Under K deficient condition, the greatest reductions for root length, total root surface area, and root volume occurred in fine roots (0.05 mm ≤ diameter < 0.20 mm), followed by the coarse roots (diameter [.tau] 0.45 mm) and the middle roots (0.25 mm ≤ diameter < 0.45 mm). The fine roots were more important in nutrient uptake than the middle and the coarse roots. Thus, the K starving damage was greater in cotton seedlings than the growth inhibition of roots. When the cotton seedlings exposed to K deficient media for 4 d and 10 d, the total root length and the total root surface area were 35.7–38.0% and 47.7–50.6% of the values of the control plants; whereas the K accumulation was approximately 25% and 16% to the control values, respectively. As expected, the endogenous free IAA content in the roots grown in K deficient media reduced by 50%, whereas the amount of ethylene released from roots increased by nearly 6-fold, which partially explained the inhibition of lateral root formation and root elongation by K deficiency.

Abscisic acid (ABA) inhibition of lateral root formation involves endogenous ABA biosynthesis in Arachis hypogaea L.

ABA has been found to play a significant role in post-embryonic developmental in peanut seedlings. The results from the current study indicate that in the presence of exogenous 10 μmol l−1 ABA, lateral roots (LRs) number decreased and seedling development was delayed. This effect was eliminated by 25 μmol l−1 naproxen, an inhibitor of ABA biosynthesis. The Arabidopsis mutant deficient in ABA biosynthesis, nced3, displays a phenotype with more and longer LRs. We found that ABA decreased root-branching in peanut in a dose-dependent way. ABA-treated seedlings showed higher endogenous ABA levels than the control and naproxen-treated seedlings. RT-PCR results indicated that the expression of AhNCED1, a key gene in the ABA biosynthetic pathway, was significantly up-regulated by exogenous ABA in peanut. The mRNA levels of AhNCED1 began to increase 2 days after ABA treatment. The results from the current study show that ABA inhibits peanut LR development by increasing endogenous ABA contents.

Is there a link between protein kinase CK2 and auxin signaling?

Protein kinase CK2 is a pleitropic Ser/Thr kinase present in all eukaryotes. In order to study the effects of CK2 depletion on plant development, we have recently generated Arabidopsis transgenic plants expressing a CK2α-inactive mutant under the control of an inducible promoter. Our results showed that continuous expression of the transgene had a dominant negative effect and was lethal for Arabidopsis plants. Overexpression of the CK2α-inactive subunit provoked cell cycle arrest, by perturbation of both G1/S and G2 cell-cycle phases. The effects on cell division were particularly strong in root meristems, causing inhibition of lateral root formation even when the mutant protein was transiently induced. Processes that rely on cell expansion, such as hypocotyl elongation in dark-grown seedlings, were also strongly affected. We propose that CK2 regulates auxin-signaling pathways.Addendum to: Moreno-Romero J, Carme Espunya M, Platara M, Ariño J, Carmen Martínez M. A role for protein kinase CK2 in plant development: evidence obtained using a dominant-negative mutant. Plant J. 2008; In press.

Effects of cyanobacteria producing microcystins on seed germination and seedling growth of several agricultural plants

The effects of cyanobacteria aqueous extracts containing Microcystin-LR (MC-LR) on the seed germination and growth of Pisum sativum, Lens esculenta, Zea mays and Triticum durum were investigated. Experiments were carried out on a range of doses of the extract (equivalent to 0, 1.6, 2.9, 5.8, 8.7 and 11.6 μ g MC-LR/mL). The results confirm that these plants were sensitive to cell-free extracts of a toxic Microcystis and that germination inhibition was dose dependent. One-way analysis of variance (ANOVA) showed that P. sativum is the most sensitive tested species with a 97% germination rate reduction and L. esculenta was the most resistant. At the 8th day, the exposure to the microcystins (MC) resulted in a significant decrease of plant epicotyls length, roots length and a net inhibition of lateral root formation. It is concluded that MC could affect also terrestrial plants seedling germination and growth. Therefore, the use of water for irrigation contaminated by MC could exert negative biochemical effects on seed and plant metabolism which might influence the agricultural crops.

シロイヌナズナのサイトカイニン受容体AHK2～4の機能低下は、胚軸での維管束発達の抑制およびオーキシンの蓄積による、側根形成の抑制と不定根形成を引き起こす

シロイヌナズナのサイトカイニン受容体AHK2～4の機能低下は、胚軸での維管束発達の抑制およびオーキシンの蓄積による、側根形成の抑制と不定根形成を引き起こす

The ubiquitin ligase XBAT32 regulates lateral root development in Arabidopsis

Ubiquitin-mediated protein modification plays a key role in many cellular signal transduction pathways. The Arabidopsis gene XBAT32 encodes a protein containing an ankyrin repeat domain at the N-terminal half and a RING finger motif. The XBAT32 protein is capable of ubiquitinating itself. Mutation in XBAT32 causes a number of phenotypes including severe defects in lateral root production and in the expression of the cell division marker CYCB1;1::GUS. The XBAT32 gene is expressed abundantly in the vascular system of the primary root, but not in newly formed lateral root primordia. Treatment with auxin increases the expression of XBAT32 in the primary root and partially rescues the lateral root defect in xbat32-1 mutant plants. Thus, XBAT32 is a novel ubiquitin ligase required for lateral root initiation.

Disruption and overexpression of auxin response factor 8 gene of Arabidopsis affect hypocotyl elongation and root growth habit, indicating its possible involvement in auxin homeostasis in light condition.

Auxin response factor (ARF) family genes play a central role in controlling sensitivity to the plant hormone auxin. We characterized the function of ARF8 in Arabidopsis by investigating a T-DNA insertion line (arf8-1) and overexpression lines (ARF8 OX) of ARF8. arf8-1 showed a long-hypocotyl phenotype in either white, blue, red or far-red light conditions, in contrast to ARF8 OX that displayed short hypocotyls in the light. Stronger and weaker apical dominance, and promotion and inhibition of lateral root formation were observed in arf8-1 and ARF8 OX respectively. Sensitivity to auxin was unaltered in arf8-1 hypocotyls with respect to growth inhibition caused by exogenously applied auxin and growth promotion induced by higher temperatures. ARF8 expression was observed constitutively in shoot and root apexes, and was induced in the light condition in hypocotyls. Free IAA contents were approximately 30% reduced in light-grown hypocotyls of ARF8 OX, but were similar between those of arf8-1 and wild type. Expression of the three GH3 genes was reduced in arf8-1 and increased in ARF8 OX, indicating that they are targets of ARF8 transcriptional control. Because the three GH3 proteins may be involved in the conjugation of IAA as suggested by Staswick et al. (2002), and because two of the three GH3 genes are auxin inducible, ARF8 may control the free IAA level in a negative feedback fashion by regulating GH3 gene expression. ARF family genes seem to control both auxin sensitivity and homeostasis in Arabidopsis.

Effect of inorganic nutrition on the formation of lateral roots in Dracaena fragrans Ker-Gawl cultured in vitro

The effect of inorganic nutrition on the formation of lateral roots has been studied in Dracaena fragrans Ker-Gawl cultured in vitro. MS macronutrient salts imposed inhibition of lateral root formation in ≃95% of primary adventitious roots which could be overcome by: