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.     

A Histone H3 Lysine-27 Methyltransferase Complex Represses Lateral Root Formation in Arabidopsis thaliana

Root branching or lateral root formation is crucial to maximize a root system acquiring nutrients and water from soil. A lateral root （LR） arises from asymmetric cell division of founder cells （FCs） in a pre-branch site of the primary root, and FC establishment is essential for lateral root formation. FCs are known to be specified from xylem pole pericycle cells, but the molecular genetic mechanisms underlying FC establishment are unclear. Here, we report that, in Arabidopsis thaliana, a PRC2 （for Polycomb repressive complex 2） histone H3 lysine-27 （H3K27） methyltransferase complex, functions to inhibit FC establishment during LR initiation. We found that functional loss of the PRC2 subunits EMF2 （for EMBRYONIC FLOWER 2） or CLF （for CURLY LEAF） leads to a great increase in the number of LRs formed in the primary root. The CLF H3K27 methyltransferase binds to chromatin of the auxin efflux carrier gene PIN FORMED 1 （PIN1）, deposits the repres- sive mark H3K27me3 to repress its expression, and functions to down-regulate auxin maxima in root tissues and inhibit FC establishment. Our findings collectively suggest that EMF2-CLF PRC2 acts to down-regulate root auxin maxima and show that this complex represses LR formation in Arabidopsis.     

植物侧根发育的研究进展

侧根是植物根系的重要组成部分,其发生和发育受到内源植物激素和外界环境因素的共同影响。生长素在侧根发生起始、侧根原基的发育和侧根突破母体表皮等阶段均发挥关键作用。研究侧根的发育和形态解剖结构以及信号调控途径等,都具有重要的理论和实践意义。本文结合近年来的研究进展,综述了拟南芥和水稻侧根发育的详细过程和影响因素,重点关注生长素在侧根原基发生和发育过程中的作用。     

Ara-Rhizotron: An Effective Culture System to Study Simultaneously Root and Shoot Development of Arabidopsis

Studying Arabidopsis thaliana (L.) Heynh. root development in situ at the whole plant level without affecting shoot development has always been a challenge. Such studies are usually carried out on individual plants, neglecting competition of a plant population, using hydroponic systems or Agar-filled Petri dishes. Those both systems, however, present some limitations, such as difficulty to study precisely root morphogenesis or time-limited culture period, respectively. In this paper, we present a method of Arabidopsis thaliana (L.) Heynh. cultivation in soil medium, named “Ara-rhizotron”. It allows the non-destructive study of shoot and root development simultaneously during the entire period of vegetative growth. In this system, roots are grown in 2D conditions, comparable to other soil cultures. Moreover, grouping several Ara-rhizotrons in a box enables the establishment of 3D shoot competition as for plants grown in a population. In comparison to a control culture grown in pots in the same environmental conditions, the Ara-rhizotron resulted in comparable shoot development in terms of dry mass, leaf area, number of leaves and nitrogen content. We used this new culture system to study the effect of irrigation modalities on plant development. We found that irrigation frequency only affected root partitioning in the soil and shoot nitrogen content, but not shoot or root growth. These effects appeared at the end of the vegetative growth period. This experiment highlights the opportunity offered by the Ara-rhizotron to point out tardy effects, affecting simultaneously shoot development and root architecture of plants grown in a population. We discuss its advantages in relation to root development and physiology, as well as its possible applications.     

The Control of Lateral Root Development in Cultured Pea Seedlings. III. Spacing Intervals

The spacing of lateral root primordia in the primary root of Pisum sativum (cv. Alaska) seedlings is influenced by both predetermined lateral root initiation sites in the embryonic radicle and by factors present during seedling growth. When pea seeds were germinated in the presence of the mitotic inhibitor, colchicine, the triarch radicle produced three ranks of primordiomorphs indicating sites of embryonic lateral root primordia. The number of primordiomorphs was not the same along the three xylem strands in the radicle. Normally germinated seedling roots (5 days old) also showed a different number of lateral root primordia associated with the three strands. In both cases, the strand with the greatest number of primordia (or primordiomorphs) was associated with a cotyledonary trace. This indicated a possible role for the cotyledons in setting the pattern of lateral root distribution during radicle development. The spacing of lateral root primordia could be altered by the application of growth regulators. Seedling root tips (2 mm) were removed (? rt) and replaced with indoleacetic acid (+IAA), and in some instances seedlings were also treated with the auxin transport inhibitor, 3,3a-dihydro-2-(p-methoxyphenyl)-8H-pyrazolo[5, 1-α]isoindol-8-one (+DPX). In the growth regulator treatments, primary root elongation was inhibited, a greater number of lateral root primordia were initiated compared to controls, and the spacing intervals between primordia were greatly reduced. The — rt, +IAA, +DPX-treatment resulted in the closest possible spacing intervals (av. 0.4 ? 0.6 mm), but resulted in fused or fasciated laterals. The — rt, + IAA-treatment produced the shortest spacing intervals which resulted in “normal” lateral roots (0.8 ? 1.1 mm).     

Effect of Lateral Root Formation on the Vascular Pattern of Barley Roots

The vascular pattern in the root of barley (Hordeum vulgare L.), characterized by discontinuous xylem, is markedly affected by its branching. The roots become divided into unbranched segments alternating with branched segments with a more complex vascular pattern, formed by two systems differing in origin and age: the primary vascular system derived from the procambium and ontogenetically younger connective vascular system derived from stelar parenchyma. Adjacent to the sites of the lateral root initiation, reprogramming of parent stelar parenchyma for connective vascular elements occurs. The connecting phloem is represented by small sieve elements and companion cells, the connecting xylem is composed of small vessel elements with reticulate or scalariform-reticulate wall thickenings and simple perforations. Development of the connective vascular system secures continuous lateral and axial vascular connection between lateral root and parent root. The extent of the vascular connection in the parent root increases in an acropetal direction. Hydraulic effects of connective vascular tissue formation and parent root segmentation are discussed.     

Carbon Monoxide Promotes Lateral Root Formation in Rapeseed

Carbon monoxide (CO), an odorless, tasteless and colorless gas, has recently proved to be an important bioactive or signalmolecule in mammalian cells, with its effects mediated mainly by nitric oxide (NO). In the present report, we show thatexogenous CO induces lateral root (LR) formation, an NO-dependent process. Administration of the CO donor hematin torapeseed (Brassica napus L. Yangyou 6) seedlings for 3 days, dose-dependently promoted the total length and number ofLRs. These responses were also seen following the application of gaseous CO aqueous solutions of different saturatedconcentrations. Furthermore, the actions of CO on seedlings were fully reversed when the CO scavenger hemoglobin (Hb)or the CO-specific synthetic inhibitor zinc protoporphyrin-IX (ZnPPIX) were added. Interestingly, depletion of endogenousNO using its specific scavenger 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide potassium salt (cPTIO)or the nitric oxide synthase (NOS) inhibitor N~G-nitro-L-arginine methyl ester (L-NAME),led to the complete abolition ofLR development, illustrating an important role for endogenous NO in the action of CO on LR formation. However, theinduction of LR development by 200 umol/L sodium nitroprusside (SNP),an NO donor, was not affected by the presenceor absence of ZnPPIX. Furthermore, using an anatomical approach combined with laser scanning confocal microscopywith the NO-specific fluorophore 4,5-diaminofluorescein diacetate, we observed that both hematin and SNP increased NOrelease compared with control samples and that the NO signal was mainly distributed in the LR primordia (LRP), especiallyafter 36 h treatment. The LRP were found to have similar morphology in control, SNP-and hematin-treated seedlings.Similarly, the enhancement of the NO signal by CO at 36 h was differentially quenched by the addition of cPTIO, L-NAME,ZnPPIX and Hb. In contrast, the induction of NO caused by SNP was not affected by the application of ZnPPIX. Therefore,we further deduced that CO induces LR formation probably mediated by the NO/NOS pathway and NO may act downstreamof CO signaling, which has also been shown to occur in animals.     

Structure of Flower in Arabidopsis thaliana: Spatial Pattern Formation

Morphological analysis of flowers was carried out in Arabidopsis thaliana wild type plants and agamous and apetala2 mutants. No direct substitution of organs takes place in the mutants, since the number and position of organs in them do not correspond to the structure of wild type flower. In order to explain these data, a notion of spatial pattern formation in the meristem was introduced, which preceded the processes of appearance of organ primordia and formation of organs. Zones of acropetal and basipetal spatial pattern formation in the flower of wild type plants were postulated. It was shown that the acropetal spatial pattern formation alone took place in agamous mutants and basipetal spatial pattern formation alone, in apetala2 mutants. Different variants of flower structure are interpreted as a result of changes in the volume of meristem (space) and order of spatial pattern formation (time).     

侧根的发生及其激素调控

本文概述了侧根发生及其植物激素调控的研究进展。侧根的发生起源于特定的中柱鞘细胞,其发生过程可简单地分为侧根发生的起始、侧根原基的形成、侧根分生组织的形成和活化等几个关键时期。参与侧根发生调控的植物激素主要是生长素,它影响到侧根发生过程的各个时期。茉莉酸对侧根的发生也有一定的调控作用。     

ABA抑制花生侧根发生

本实验研究了ABA对花生侧根发生的影响。结果表明：用10umol·L-1 ABA浸泡处理花生种子1h或在含ABA的培养基上培养,均抑制侧根的发生,侧根发生率降低,数目减少,长度降低,发生的时间推迟1-2d;用ABA合成抑制剂25umol·L-1 NAPR浸泡后的种子,无论在1／2MS还是在含NAA培养基上培养,侧根发生率、侧根的数目和长度均增加。用NAA的极性运输抑制剂10umol·L-1 TIBA浸泡处理种子后,再在含ABA培养基上培养,侧根不发生,说明ABA抑制花生侧根的发生与种子内源ABA和IAA的水平相关。     

The Xerobranching Response Represses Lateral Root Formation When Roots Are Not in Contact with Water

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Multiple AUX/IAA�CARF modules regulate lateral root formation: the role of Arabidopsis SHY2/IAA3-mediated auxin signalling

In Arabidopsis thaliana, lateral root (LR) formation is regulated by multiple auxin/indole-3-acetic acid (Aux/IAA)–AUXIN RESPONSE FACTOR (ARF) modules: (i) the IAA28–ARFs module regulates LR founder cell specification; (ii) the SOLITARY-ROOT (SLR)/IAA14–ARF7–ARF19 module regulates nuclear migration and asymmetric cell divisions of the LR founder cells for LR initiation; and (iii) the BODENLOS/IAA12–MONOPTEROS/ARF5 module also regulates LR initiation and organogenesis. The number of Aux/IAA–ARF modules involved in LR formation remains unknown. In this study, we isolated the shy2-101 mutant, a gain-of-function allele of short hypocotyl2/suppressor of hy2 (shy2)/iaa3 in the Columbia accession. We demonstrated that the shy2-101 mutation not only strongly inhibits LR primordium development and emergence but also significantly increases the number of LR initiation sites with the activation of LATERAL ORGAN BOUNDARIES-DOMAIN16/ASYMMETRIC LEAVES2-LIKE18, a target gene of the SLR/IAA14–ARF7–ARF19 module. Genetic analysis revealed that enhanced LR initiation in shy2-101 depended on the SLR/IAA14–ARF7–ARF19 module. We also showed that the shy2 roots contain higher levels of endogenous IAA. These observations indicate that the SHY2/IAA3–ARF-signalling module regulates not only LR primordium development and emergence after SLR/IAA14–ARF7–ARF19 module-dependent LR initiation but also inhibits LR initiation by affecting auxin homeostasis, suggesting that multiple Aux/IAA–ARF modules cooperatively regulate the developmental steps during LR formation.     

Isolation of a cDNA encoding a novel GTP-binding protein of Arabidopsis thaliana

A cDNA encoding for a 68 kDa GTP-binding protein was isolated from Arabidopsis thaliana (aG68). This clone is a member of a gene family that codes for a class of large GTP-binding proteins. This includes the mammalian dynamin, yeast Vps1p and the vertebrate Mx proteins. The predicted amino acid sequence was found to have high sequence conservation in the N-terminal GTP-binding domain sharing 54% identity to yeast Vps1p, 56% amino acid identity to rat dynamin and 38% identity to the murine Mx1 protein. The northern analysis shows expression in root, leaf, stem and flower tissues, but in mature leaves at lower levels. Southern analysis indicates that it may be a member of a small gene family or the gene may contain an intron.     

New methods to analyse auxin-induced growth I: Classical auxinology goes Arabidopsis

During recent years, genetic approaches have become invaluable tools to analyse signalling chains in plants. The first step to understand auxin signalling is the isolation of signal transduction mutants. In Arabidopsis, screening for auxin resistant plants has identified a number of mutants. However, it is difficult to link the mutated genes to the rapid growth response to auxin. As yet, there is no published method to measure auxin-induced growth at a sufficient temporal resolution. A novel auxanometer described here will close that gap. Hypocotyl segments are immersed in buffer in a flow-through cuvette. A CCD-camera attached to a microscope is used to image markings on the hypocotyl surfaces and to track their movements across the field of view.To illustrate the applicability of this method for analysing mutants we compared the growth responses of tomato wild type with the mutant diageotropica (dgt). We showed that the mutation completely abolished all phases of the rapid growth reaction to applied auxin. This excludes the possibility that the reduced auxin sensitivity reported in the literature is due to a reduced or delayed response. Our technique was modified for use with the tiny Arabidopsis thaliana hypocotyl segments and inflorescence stems. In both systems auxin induced a growth response after a lag phase of 15–20 minutes. The new method will now be used to characterise the physiological effect of the mutations in auxin signalling.     

Cellulose orientation at the surface of the Arabidopsis seedling. Implications for the biomechanics in plant development

In diffuse growing cells the orientation of cellulose fibrils determines mechanical anisotropy in the cell wall and hence also the direction of plant and organ growth. This paper reports on the mean or net orientation of cellulose fibrils in the outer epidermal wall of the whole Arabidopsis plant. This outer epidermal wall is considered as the growth-limiting boundary between plant and environment. In the root a net transverse orientation of the cellulose fibrils occurs in the elongation zone, while net random and longitudinal orientations are found in subsequent older parts of the differentiation zone. The position and the size of the transverse zone is related with root growth rate. In the shoot the net orientation of cellulose fibrils is transverse in the elongating apical part of the hypocotyl, and longitudinal in the fully elongated basal part. Leaf primordia and very young leaves have a transverse orientation. Throughout further development the leaf epidermis builds a very complex pattern of cells with a random orientation and cells with a transverse or a longitudinal orientation of the cellulose fibrils. The patterns of net cellulose orientation correlate well with the cylindrical growth of roots and shoots and with the typical planar growth of the leaf blade. On both the shoot and the root surface very specific patterns of cellulose orientation occur at sites of specific cell differentiation: trichome-socket cells complexes on the shoot and root hairs on the root.