The Control of Lateral Root Development in Cultured Pea Seedlings. II. Root Fasciation Induced by Auxin Inhibitors

Lateral root development in cultured seedlings of Pisum sativum (cv. Alaska) was modified by the application of auxin transport inhibitors or antagonists. When applied either to replace the root tip or beneath the cotyledonary node, two auxin transport inhibitors, 2,3,5-triiodobenzoic acid (TIBA) and 3,3a-dihydro-2-(p-methoxyphenyl)-8H-pyrazolo[5,1-α]isoindol-8-one (DPX-1840), increased cell division activity opposite the protoxylem poles. This resulted in the formation of masses of cells, which we are calling root primordial masses (RPMs), 2 to 3 days after treatment. RPMs differed from lateral root primordia in that they lacked apical organization. Some roots however developed both RPMs and lateral roots indicating that both structures were similar in terms of the timing and location of cell division in the pericycle and endodermis leading to their initiation. Removal of the auxin transport inhibitors allowed many of the RPMs to organize later into lateral root primordia and to emerge in clusters. When the auxin, indoleacetic acid (IAA) was added to the growth medium along with DPX-1840, 3 ranks of RPMs now in the form of fasciated lateral roots emerged from the primary root. The auxin antagonist, p-chlorophenoxy-isobutyric acid (PCIB), also induced RPM formation. In contrast to DPX-1840 treatment, the addition of IAA during PCIB treatment caused normal lateral root development.     

Auxin Induced Lateral Root Formation in Chicory

The supply of auxins [2,4-dichlorophenoxy acetic acid (2,4D),indole-3 acetic acid (1AA) and -naphthaleneacetic acid (NAA)]to excised chicory roots induced the formation of lateral rootmeristems mainly located close to the pre-existing apical rootmeristem. Lateral root growth induced in non-excised roots requiredhigher auxin concentrations. Inhibition of root elongation andconcomittant enlargement of the apices was also observed. SupplyingIAA induced the formation of lateral meristems earlier thanNAA, but subsequently favoured root elongation. Conversely,in the presence of 2,4D, reactivation of pericycle cells wasvery intense, but conversion of primordia to laterals was inhibited.Regardless of the auxin used, the responsive area in which lateralmeristems appeared was located a maximum of 4 mm away from theapical meristem. This region remained devoid of any lateralroot formation under control conditions. Pericycle cells oppositethe xylem poles in the diarch stele regained meristematic activityand divided transversally, giving rise to shorter cells. Thesecells subsequently divided periclinally, forming pairs of cellson the same transverse level. The root primordium extruded throughcortical cells and was surrounded by a lacuna formed to thedetriment of cortical cells.Copyright 1998 Annals of BotanyCompany Auxins,Cichorium intybus, chicory, lateral root, root elongation.     

LATERAL ROOTLESS2, a Cyclophilin Protein, Regulates Lateral Root Initiation and Auxin Signaling Pathway in Rice

Dear Editor, Cyclophilins （CYP） are a class of highly conserved pepti- dyl-prolyl cis-trans isomerases （PPlases） that play important roles in various biological processes in eukaryotes （reviewed in Romano et al. （2004））. In higher plants, a conserved sin- gle domain cyclophilin has been identified as a novel com- ponent of the auxin signaling pathway by analyzing the tomato diageotropica （dgt） mutant （Ivanchenko et al., 2006; Oh et al., 2006）. The dgt mutant displays a lateral-rootless and auxin-resistant phenotype （Ivanchenko et aL, 2006）. Further studies revealed that mutations in the DGT-like genes of Physcomitrella patens also exhibited an auxin-resistant phenotype, suggesting a conserved role of DGT-like proteins in auxin signaling. Moreover,     

Nutritional Requirements for Polyploid Mitoses in Cultured Pea Root Segments

Diploid and polyploid mitoses could be stimulated in excised segments of the mature region of pea roots grown on a sterile culture medium. Diploid mitoses were observed in segments cultured on water alone for 72 hours. Their frequency was increased by the presence of salts, sucrose, vitamins, and any two or all three of the following: an amino mixture, auxins, and kinetin. Polyploid mitoses were observed 72 hours after the beginning of the culture period in segments cultured on salts, sucrose, vitamins, auxins, and kinetin. Polyploid mitoses required the presence of auxins and kinetin in the culture medium. Their frequency was not affected by the presence of a reduced nitrogen source. Light treatments had no effect on the frequency of diploid or polyploid mitoses. Diploid mitoses were first observed about 24 hours after the beginning of the culture and their frequency increased thereafter. Experiments with colchicine showed that diploid cells were entering mitosis for the first time as late as 60 hours after the beginning of the culture. Polyploid mitoses showed a long lag time when compared with diploid mitoses. They began at about 60 hours and their frequency increased thereafter. Experiments with colchicine showed that polyploid cells were entering mitosis for the first time as late as 84 hours after the beginning of the culture. The presence of kinetin in the medium was not required during the first 24 hours in culture for the appearance of polyploid mitoses at 74 hours. However, the presence of kinetin was required after 24 hours. Auxin was required at some time during the first 24 hours of the culture and its continuous presence may be required for the stimulation of polyploid mitoses.     

Auxin Does Not Alter the Permeability of Pea Segments to Tritium-labeled Water

The possibility of an auxin effect on the permeability of pea (Pisum sativum L. ev. Alaska) segments to tritium-labeled water has been investigated by three separate laboratories, and the combined results are presented. We were unable to obtain any indication of a rapid effect of indoleacetic acid on the efflux of ³HHO when pea segments previously “loaded” for 90 minutes with ³HHO were transferred to unlabeled aqueous medium with indoleacetic acid. We were able to confirm that segments pretreated with ³HHO plus indoleacetic acid for 60 to 90 minutes can show an enhanced ³HHO release as compared with minus indoleacetic acid controls. However, this phenomenon appears to be due to an increased uptake of ³HHO during the prolonged indoleacetic acid pretreatment, and therefore we conclude that auxin does not alter the permeability of pea segments to ³HHO in either short term or long term tests. We confirm previous reports that the uptake of ³HHO in pea segments proceeds largely through the cut surfaces, and that the cuticle is a potent barrier to ³HHO flux.     

Environmental Regulation of Lateral Root Initiation in Arabidopsis

Plant morphology is dramatically influenced by environmental signals. The growth and development of the root system is an excellent example of this developmental plasticity. Both the number and placement of lateral roots are highly responsive to nutritional cues. This indicates that there must be a signal transduction pathway that interprets complex environmental conditions and makes the "decision" to form a lateral root at a particular time and place. Lateral roots originate from differentiated cells in adult tissues. These cells must reenter the cell cycle, proliferate, and redifferentiate to produce all of the cell types that make up a new organ. Almost nothing is known about how lateral root initiation is regulated or coordinated with growth conditions. Here, we report a novel growth assay that allows this regulatory mechanism to be dissected in Arabidopsis. When Arabidopsis seedlings are grown on nutrient media with a high sucrose to nitrogen ratio, lateral root initiation is dramatically repressed. Auxin localization appears to be a key factor in this nutrient-mediated repression of lateral root initiation. We have isolated a mutant, lateral root initiation 1 (lin1), that overcomes the repressive conditions. This mutant produces a highly branched root system on media with high sucrose to nitrogen ratios. The lin1 phenotype is specific to these growth conditions, suggesting that the lin1 gene is involved in coordinating lateral root initiation with nutritional cues. Therefore, these studies provide novel insights into the mechanisms that regulate the earliest steps in lateral root initiation and that coordinate plant development with the environment.     

Cytokinin Inhibits Lateral Root Development at the Earliest Stages of Lateral Root Primordium Initiation in Maize Primary Root

Root architecture is basically controlled by auxin and cytokinin, which antagonize in the formation of lateral roots (LRs) along the primary root (PR) axis. Several mechanisms have been proposed to explain the interaction between these two hormones, cytokinin being the hormone that inhibits LR formation. The analysis of the cytokinin effect on LR formation using LRs in several stages of development could indicate which steps of LR formation are more sensitive to cytokinin. The application of cytokinin to maize PRs showed that the inhibitory effect of cytokinin on LR formation was greater in the zones in which the initial events to form new LRs are taking place. In the presence of cytokinin, the PR is not able to produce new LRs in the initiation zone; this inhibitory effect is permanent as this zone did not recover the capability to form LRs after removing cytokinin. However, the LR density in zones with appreciable LR primordia when cytokinin was applied was only slightly inhibited when a high concentration was used. These results showed that LR formation is more sensitive to the inhibitory effect of cytokinin in the earliest stages of LR development. However, the elongation of a LR primordium to emerge and the subsequent elongation of the new LR were only slightly affected by cytokinin.

     

The Initiation and Development of Lateral Meristems in the Pea Root: II. THE EFFECT OF INDOLE-3-ACETIC ACID

The results of the first paper in this series (Pecket, 1957)indicated that lateral root initiation in isolated pea rootsdepends upon an apically moving factor or complex of factorsprovided by the mature tissue of the root. The present investigationanalyses the effect of externally applied IAA on lateral rootformation and uses the methods employed in the earlier work.Observations made on roots in which three regions have beendistinguished show that increasing IAA concentration does notcause similar changes in all pans of the root, nor does it causeidentical effects on primordium initiation and on the subsequentemergence of the initials as lateral roots. The latter factexplains why the emphasis in the present work has been laidon initiation rather than on emergence. The results of experimentsare reported in which two variables are involved, IAA concentrationand the presence or absence of the two terminal regions of theroot. These results are considered to support the conclusionsof the earlier paper. It is suggested that the stimulant movingfrom mature tissue is not IAA itself but that the productionof the stimulant may be increased when IAA is supplied. A gradientof reaction to the stimulant appears to exist in the root, reactionbeing maximal in the mature tissue.     

生长素通过MAPK介导的超长链脂肪酸合成调控侧根发育

促分裂原活化蛋白激酶(MAPK)信号级联通路是真核生物中高度保守的重要信号系统,通过激酶逐级磷酸化传递并放大上游信号,进而调控细胞反应。MAPK信号通路不仅介导植物响应环境变化,而且在调节植物生长发育过程中发挥重要作用。近期,山东大学丁兆军课题组研究发现,植物重要激素生长素能够通过激活MPK14调控下游ERF13的磷酸...     

磷有效性与植物侧根的发生发育

文章概述了植物侧根的不同发生发育阶段的形态、生理和分子生物学基础以及磷有效性在侧根发生发育过程中的调控作用,并对这一研究领域的前景作了展望。     

The Initiation and Development of Lateral Meristems in the Pea Root: I. THE EFFECT OF YOUNG AND OF MATURE TISSUE

The effect of.distinct regions of the root on the initiationof lateral root primordia and the emergence of lateral rootshas been studied, using segments of roots from sterile 2-daygerminated pea seedlings. It is shown that the removal of the basal region causes a decreasein the number of primordia formed in the remainder of the root.On the other hand, the removal of the apical region causes alarger number of primordia to be formed in the remaining tissuethan in the corresponding tissue of roots where the apical regionis retained. It is suggested that a factor or a complex of factorsinvolved in primordium initiation is translocated from the oldertissue towards the potential site of primordium initiation inthe young tissue which has just completed extension growth. The removal of the apical region of the root is also shown tostimulate lateral root emergence. It is suggested that a factoror complex of factors involved in the development of the primordiasubsequent to initiation moves within the root in a similarmanner to the factor or factors involved in initiation.     

Lateral Root Initiation or the Birth of a New Meristem

Root branching happens through the formation of new meristems out of a limited number of pericycle cells inside the parent root. As opposed to shoot branching, the study of lateral root formation has been complicated due to its internal nature, and a lot of questions remain unanswered. However, due to the availability of new molecular tools and more complete genomic data in the model species Arabidopsis, the probability to find new and crucial elements in the lateral root formation pathway has increased. Increasingly more data are supporting the idea that lateral root founder cells become specified in young root parts before differentiation is accomplished. Next, pericycle founder cells undergo anticlinal asymmetric, divisions followed by an organized cell division pattern resulting in the formation of a new organ. The whole process of cell cycle progression and stimulation of the molecular pathway towards lateral root initiation is triggered by the plant hormone auxin. In this review, we aim to give an overview on the developmental events taking place from the very early specification of founder cells in the pericycle until the first anticlinal divisions by combining the knowledge originating from classical physiology studies with new insights from genetic-molecular analyses. Based on the current knowledge derived from recent genetic and developmental studies, we propose here a hypothetical model for LRI.     

Light Quality Regulates Lateral Root Development in Tobacco Seedlings by Shifting Auxin Distributions

Light is an important environmental regulator of diverse growth and developmental processes in plants. However, the mechanisms by which light quality regulates root growth are poorly understood. We analyzed lateral root (LR) growth of tobacco seedlings in response to three kinds of light qualities (red, white, and blue). Primary (1°) LR number and secondary (2°) LR density were elevated under red light (on days 9 and 12 of treatment) in comparison with white and blue lights. Higher IAA concentrations measured in roots and lower in leaves of plants treated with red light suggest that red light accelerated auxin transport from the leaves to roots (in comparison with other light qualities). Corroborative evidence for this suggestion was provided by elevated DR5::GUS expression levels at the shoot/root junction and in the 2° LR region. Applications of N-1-naphthylphthalamic acid (NPA) to red light-treated seedlings reduced both 1° LR number and 2° LR density to levels similar to those measured under white light; DR5::GUS expression levels were also similar between these light qualities after NPA application. Results were similar following exogenous auxin (NAA) application to blue light-treated seedlings. Direct [³H]IAA transport measurement indicated that the polar auxin transport from shoot to root was increased by red light. Red light promoted PIN3 expression levels and blue light reduced PIN1, 3–4 expression levels in the shoot/root junction and in the root, indicating that these genes play key roles in auxin transport regulation by red and blue lights. Overall, our findings suggest that three kinds of light qualities regulate LR formation in tobacco seedlings through modification of auxin polar transport.     

Effects of Hydroxyproline and other Amino Acid Analogues on the Growth of Pea Root Segments

Changes in the lengths and growth rates of isolated 2–4 mm pea root segments, cultured in sucrose media under aseptic conditions, were paralleled by changes in invertase development and in chloride and leucine uptakes. The amino acid analogues o-, m- and p-fluorophenylalanine, azetidine-2-carboxylic acid and ethionine inhibited growth with corresponding changes in invertase activity and in chloride and leucine uptakes. In contrast hydroxyproline, which under the conditions used may be regarded as an analogue of proline, enhanced both the growth rate and duration of growth but had little effect on the several parameters of protein synthesis which were measured. No amino acid tested affected changes in growth, invertase activity or the uptake of chloride and leucine, but they prevented the effects of the corresponding analogues. The results show that although extension growth is dependent on continuous protein synthesis, only specific proteins, probably in the cell wall, play a key role in this process.     

Auxin and Plant-Microbe Interactions

Microbial synthesis of the phytohormone auxin has been known for a long time. This property is best documented for bacteria that interact with plants because bacterial auxin can cause interference with the many plant developmental processes regulated by auxin. Auxin biosynthesis in bacteria can occur via multiple pathways as has been observed in plants. There is also increasing evidence that indole-3-acetic acid (IAA), the major naturally occurring auxin, is a signaling molecule in microorganisms because IAA affects gene expression in some microorganisms. Therefore, IAA can act as a reciprocal signaling molecule in microbe-plant interactions. Interest in microbial synthesis of auxin is also increasing in yet another recently discovered property of auxin in Arabidopsis. Down-regulation of auxin signaling is part of the plant defense system against phytopathogenic bacteria. Exogenous application of auxin, e.g., produced by the pathogen, enhances susceptibility to the bacterial pathogen.The phytohormone auxin (from the Greek “auxein,” meaning to grow) regulates a whole repertoire of plant developmental processes, as documented in previous articles on this topic. Perhaps less well known is the fact that some microorganisms also produce auxin (Costacurta and Vanderleyden 1995; Patten and Glick 1996). In their interaction with plants, these microorganisms can interfere with plant development by disturbing the auxin balance in plants. This is best documented for phytopathogenic bacteria like Agrobacterium spp. and Pseudomonas savastanoi pv. savastanoi, causing tumors and galls, respectively (Jameson 2000; Mole et al. 2007), and plant growth promoting rhizobacteria (PGPR) such as Azospirillum spp. that impact on plant root development (Persello-Cartieaux et al. 2003; Spaepen et al. 2007a). The term rhizobacteria refers to the fact that their numbers are highly enriched in the rhizosphere, i.e., the narrow band of soil that surrounds the root (Hiltner 1904; Smalla et al. 2006; van Loon 2007). Of more recent date is the observation that auxin (indole-3-acetic acid or IAA) is a signaling molecule in some microorganisms (Spaepen et al. 2007a). Bringing these data together, it follows that auxin can have a major impact in microorganism-plant interactions. This is the main theme addressed in this article. Finally, the recent finding that auxin signaling in plants is also part of the Arabidopsis defense response against a leaf pathogen (Navarro et al. 2006) is discussed in relation to bacterial IAA synthesis.