On the Determination of the Pattern of Vascular Tissue in Peas

This work is concerned with the rules determining the placeof joining of two vascular strands. Auxin can induce the differentiationof vascular tissue, and this fact is used here for an experimentalstudy of the spatial interactions of vascular strands. Differentiated vascular tissue whose source of auxin has beenremoved attracts newly induced vascular strands. This attractionis expressed in the joining of the new strands to the pre-existingvascular tissue. Differentiated vascular tissue which is wellsupplied with auxin inhibits rather than attracts the formationof new vascular strands in its vicinity. Experiments on pea apices have extended these results to naturallyinduced vascular strands. It is shown that when a leaf primordiumis damaged at an early age its vascular strands are joined bythe strands induced by new leaves, and the contacts may be formedacross the leaf gap. The joining of the vascular strands is,therefore, much closer to the leaf than is normal and this isprobably due to the reduction in the supply of auxin from thedamaged leaf to its vascular traces. It is also shown that whena lateral bud grows its vascular traces join preferentiallythe vascular system leading to organs which have been removed.These vascular traces of a bud specifically avoid the vascularsystem of a leaf or a shoot which is still growing and producingauxin. These results are discussed in reference to the relation betweenthe vascular system and phyllotaxis and to the existence ofleaf gaps.     

Integrating cellular and organismic aspects of vascular differentiation

Vascular differentiation can be studied at two levels, and they should complement one another: as an aspect of integrated plant development and as cellular processes. The differentiation of organized strands that connect between organs is induced by polar auxin flow, towards the roots. Anatomy, therefore, can be a complementary method of observing polarity and its changes. As expected for a self-correcting and essential system, vascular patterning mutations are relatively rare and have pleiotropic effects, including modifications of responses to auxin and its transport. Tissue polarity both expresses and depends on auxin transport, a feedback that could account for the determined nature of polarity as well as the gradual canalization of differentiation to vascular strands. This predicts that the molecules responsible for polarity will be localized gradually as differentiation proceeds. Further, a modified location of these molecules can be expected to precede anatomical expressions of a new, regenerated, polarity. Tracheary differentiation is probably the best studied example of cell differentiation. Within the plant, however, this differentiation is coupled to oriented cell growth either along or at right angles to the axis of auxin flow, depending on tissue competence. Differentiation is also coupled to the differentiation of the other components of the vascular system. There are, presumably, early joint stages to these differentiation processes, but what they are remains an intriguing problem.     

Polarity,Continuity, and Alignment in Plant Vascular Strands

Plant vascular cells are joined end to end along uninterrupted lines to connect shoot organs with roots; vascular strands are thus polar, continuous, and internally aligned. What controls the formation of vascular strands with these properties? The “auxin canalization hypothesis”—based on positive feedback between auxin flow through a cell and the cell's capacity for auxin transport—predicts the selection of continuous files of cells that transport auxin polarly, thus accounting for the polarity and continuity of vascular strands. By contrast, polar, continuous auxin transport—though required—is insufficient to promote internal alignment of vascular strands, implicating additional factors. The auxin canalization hypothesis was derived from the response of mature tissue to auxin application but is consistent with molecular and cellular events in embryo axis formation and shoot organ development. Objections to the hypothesis have been raised based on vascular organizations in callus tissue and shoot organs but seem unsupported by available evidence. Other objections call instead for further research; yet the inductive and orienting influence of auxin on continuous vascular differentiation remains unique.     

The Arabidopsis gene MONOPTEROS encodes a transcription factor mediating embryo axis formation and vascular development.

The vascular tissues of flowering plants form networks of interconnected cells throughout the plant body. The molecular mechanisms directing the routes of vascular strands and ensuring tissue continuity within the vascular system are not known, but are likely to depend on general cues directing plant cell orientation along the apical-basal axis. Mutations in the Arabidopsis gene MONOPTEROS (MP) interfere with the formation of vascular strands at all stages and also with the initiation of the body axis in the early embryo. Here we report the isolation of the MP gene by positional cloning. The predicted protein product contains functional nuclear localization sequences and a DNA binding domain highly similar to a domain shown to bind to control elements of auxin inducible promoters. During embryogenesis, as well as organ development, MP is initially expressed in broad domains that become gradually confined towards the vascular tissues. These observations suggest that the MP gene has an early function in the establishment of vascular and body patterns in embryonic and post-embryonic development.     

Responses of plant vascular systems to auxin transport inhibition.

To assess the role of auxin flows in plant vascular patterning, the development of vascular systems under conditions of inhibited auxin transport was analyzed. In Arabidopsis, nearly identical responses evoked by three auxin transport inhibitor substances revealed an enormous plasticity of the vascular pattern and suggest an involvement of auxin flows in determining the sites of vascular differentiation and in promoting vascular tissue continuity. Organs formed under conditions of reduced auxin transport contained increased numbers of vascular strands and cells within those strands were improperly aligned. In leaves, vascular tissues became progressively confined towards the leaf margin as the concentration of auxin transport inhibitor was increased, suggesting that the leaf vascular system depends on inductive signals from the margin of the leaf. Staged application of auxin transport inhibitor demonstrated that primary, secondary and tertiary veins became unresponsive to further modulations of auxin transport at successive stages of early leaf development. Correlation of these stages to anatomical features in early leaf primordia indicated that the pattern of primary and secondary strands becomes fixed at the onset of lamina expansion. Similar alterations in the leaf vascular responses of alyssum, snapdragon and tobacco plants suggest common functions of auxin flows in vascular patterning in dicots, while two types of vascular pattern alterations in Arabidopsis auxin transport mutants suggest that at least two distinct primary defects can result in impaired auxin flow. We discuss these observations with regard to the relative contributions of auxin transport, auxin sensitivity and the cellular organisation of the developing organ on the vascular pattern.     

Mechanisms and physiological role of polarity in plants

The concept of polarity was the starting point for the attempts of many investigators to understand the principles of differentiation, because the polar organization underlies specific three-dimensional structure of the organism and provides for the integrity and coordination of its functions. The polarity axes are established at the stage of zygote, extending to the developing embryo, and they ??vectorize?? subsequent plant growth and development. Polarization of cells and tissues is crucial for plant morphogenesis, because the emerging morphogenetic gradients provide the basis for differential genome activity at various stages of plant development. This review deals with the polarity phenomena and the mechanisms of symmetry axis formation at the level of cells and plant tissues. The roles of electrical gradients, Ca²⁺ ions, auxin, cytoskeleton, ROP-proteins, phosphoinositides, and microRNA in polarization of cells and tissues are considered.     

Cell polarity signaling in Arabidopsis involves a BFA-sensitive auxin influx pathway

Coordination of cell and tissue polarity commonly involves directional signaling. In the Arabidopsis root epidermis, cell polarity is revealed by basal, root tip-oriented, hair outgrowth from hair-forming cells (trichoblasts). The plant hormone auxin displays polar movements and accumulates at maximum concentration in the root tip. The application of polar auxin transport inhibitors evokes changes in trichoblast polarity only at high concentrations and after long-term application. Thus, it remains open whether components of the auxin transport machinery mediate establishment of trichoblast polarity. Here we report that the presumptive auxin influx carrier AUX1 contributes to apical-basal hair cell polarity. AUX1 function is required for polarity changes induced by exogenous application of the auxin 2,4-D, a preferential influx carrier substrate. Similar to aux1 mutants, the vesicle trafficking inhibitor brefeldin A (BFA) interferes with polar hair initiation, and AUX1 function is required for BFA-mediated polarity changes. Consistently, BFA inhibits membrane trafficking of AUX1, trichoblast hyperpolarization induced by 2,4-D, and alters the distal auxin maximum. Our results identify AUX1 as one component of a novel BFA-sensitive auxin transport pathway polarizing cells toward a hormone maximum.     

Plants as competing populations of redundant organs

At any given time, a vascular or land plant may be a colony of functional sectors, each consisting of a shoot and its associated roots. In most plants, however, the activity of the cambium can change the relative vascular contacts of neighbouring shoots. Vascular tissues can even differentiate along new orientations, forming contacts that change the sectorial structure of the plant. Such reoriented differentiation is induced by the same auxin from developing leaves as are other types of vascular differentiation. The occurrence of vascular reorientation is determined by two criteria: the presence of an auxin flow that exceeds the transport capacity of the tissues that follow the previous, established orientation and the availability of nearby channels that are not fully occupied, not‘protected’ by their own flow of auxin. These controls of vascular orientation suggest that neighbouring shoots (and neighbouring roots) compete with one another, by means of signals indicating their state and their environment, for vascular contacts with the rest of the plant. Such internal competition between genetically equivalent shoots is an adaptation to heterogeneous environments: it is the shoots in the best conditions available to the plant that receive the support of a greater part of the root system. The potential for changes of vascular contacts points to open problems and to neglected aspects of the role of the cambium in plant organization.     

Effects of auxin on the spatial distribution of cell division and xylogenesis in lettuce pith explants

Summary Cylinders of pith parenchyma were tissue-cultured with their opposite ends on media which differed only in content of the morphogens auxin (IAA), sucrose, or zeatin. A range of concentrations of each of these morphogens applied at one end (none at the other end) resulted in distribution patterns of cell division and xylogenesis that were attributable to interaction between inductive levels and morphogen mobility. Auxin was crucial for tracheary patterns: large tracheary elements formed by direct differentiation of pith cells near the auxin source, smaller but still roughly isodiametric tracheary elements formed after cell division, and tracheary strands developed where, presumably, auxin transport had become polarized and then canalized. Xylogenesis was confined to regions within millimeters of the auxin source, and [¹⁴C]IAA studies showed a steep logarithmic concentration gradient along the cylinder. Patterns of tracheary strands and rings revealed that the pith explants retained some polarity from the stem from which they had been excised. However, the direction of flow of applied auxin was more effective than original polarity in controlling the orientation of tracheary strands and their constituent tracheary elements. It seems that, in tissues with little or no polarity, diffusive flow of auxin gradually induces polar flow in the same direction, together with an associated bioelectric current, and that this orients the cortical microtubules that in turn determine the orientations of cell elongation and of the secondary wall banding in tracheary elements.Abbreviations IAA indoleacetic acid - NAA naphthaleneacetic acid - TIBA triiodobenzoic acid Dedicated to the memory of Professor John G. Torrey     

The production of auxin by autolysing tissues

Summary Autolysing plant tissues are known to produce auxin when extracted with ether. It has been shown that autolysing plant, yeast and rat liver tissues produce auxin in vitro; this suggests that relatively unspecific mechanisms are involved. Furthermore, sterile plant and animal tissues which have been killed by freezing and thawing induce nodules of differentiated cells in a previously undifferentiated callus of Phaseolus vulgaris. The callus tissue is known to differentiate in response to applied gradients of auxin. Plant and animal tissues killed by boiling were considerably less effective in inducing differentiation in the tissue. The evidence indicates that auxin is a normal product of autolysing cells. It is suggested that dying cells are an important source of auxin in the plant.     

Circular Vessels and the Control of Vascular Differentiation in Plants

The occurrence of vessels in the form of rings is used as a critical example for a hypothesis about the control of the pattern of cells in vascular tissues. These vessels, rare in intact plants, are common in the basal or root side of tissues close to transverse wounds of bean seedlings, radish storage tissues, and other plant material. Their formation is promoted, as are normal vascular tissues, by developing parts of the shoot or by a source of the hormone auxin. They are also found in grafts where cells of opposite polarities are close together, and in cut plants where vascular induction occurs from the direction of the roots and is therefore opposite to the original polarity of the tissue. Circular vessels are found, therefore, where the flux of auxin and possibly other signals controlling vascular differentiation is expected to follow a circular route. They show that differentiation is a response of individual cells to the flux rather than the gradient or concentration of the hormonal signals and suggest a hormonal interpretation of differences between apical and basal callus growth.     

Ups and downs of tissue and planar polarity in plants

The polar orientation of cells within a tissue is an intensively studied research area in animal cells. The term planar polarity refers to the common polar arrangement of cells within the plane of an epithelium. In plants, the subcellular analysis of tissue polarity has been limited by the lack of appropriate markers. Recently, research on plant tissue polarity has come of age. Advances are based on studies of Arabidopsis patterning, cell polarity and auxin transport mutants employing the coordinated, polar localization of auxin transporters and the planar polarity of root epidermal hairs as markers. These approaches have revealed auxin transport and response, vesicular trafficking, membrane sterol and cytoskeletal requirements of tissue polarity. This review summarizes recent progress in research on vascular tissue and planar epidermal polarity in the Arabidopsis root and compares it to findings on planar polarity in animals and cell polarity in yeast.     

肌动蛋白微丝与生长素浓度梯度分布

生长素参与植物生长发育的各个阶段,如胚胎发生、发育,营养器官发生与形态建成,极性与轴向的建立,维管组织分化,生殖器官的发育等。虽然生长素在植物的各组织器官和细胞中发挥着重要的作用,植物内源生长素的生物合成却是在特异的组织——细胞快速分裂的幼嫩组织中完成的,然后通过韧皮部或受严格控制的细胞—细胞运输系统运送至植物各个部分。生长素的极性运输导致其积累在某些局部组织和细胞内,形成特定梯度分布。生长素对植物生长发育众多方面的调节正是依赖于这一特性。该文综述了近年来有关植物生长发育过程中生长素浓度梯度的形成和相应的生理功能,以及细胞骨架中的微丝参与调控生长素极性运输的研究工作。     

Canalization without flux sensors: a traveling-wave hypothesis

In 1969, Tsvi Sachs published his seminal hypothesis of vascular development in plants: the canalization hypothesis. A positive feedback loop between the flux of the phytohormone auxin and the cells' auxin transport capacity would canalize auxin progressively into discrete channels, which would then differentiate into vascular tissues. Recent experimental studies confirm the central role of polar auxin flux in plant vasculogenesis, but it is unclear if and by which mechanism plant cells could respond to auxin flux. In this Opinion article, we review auxin perception mechanisms and argue that these respond more likely to auxin concentrations than to auxin flux. We propose an alternative mechanism for polar auxin channeling, which is more consistent with recent molecular observations.     

Vascular continuity and auxin signals

Plant vascular tissues form systems of interconnected cell files throughout the plant body. Vascular tissues usually differentiate at predictable positions but the wide range of functional patterns generated in response to abnormal growth conditions or wounding reveals partially self-organizing patterning mechanisms. Signals ensuring aligned cell differentiation within vascular strands are crucial in self-organized vascular patterning, and the apical-basal flow of indole acetic acid has been suspected to act as an orienting signal in this process. Several recent advances appear to converge on a more precise definition of the role of auxin flow in vascular tissue patterning.     

Probing the actin-auxin oscillator

The directional transport of the plant hormone auxin depends on transcellular gradients of auxin-efflux carriers that continuously cycle between plasma membrane and intracellular compartments. This cycling has been proposed to depend on actin filaments. However, the role of actin for the polarity of auxin transport has been disputed. To get insight into this question, actin bundling was induced by overexpression of the actin-binding domain of talin in tobacco BY-2 cells and in rice plants. This bundling can be reverted by addition of auxins, which allows to address the role of actin organization on the flux of auxin. In both systems, the reversion of a normal actin configuration can be restored by addition of exogenous auxins and this fully restores the respective auxin-dependent functions. These findings lead to a model of a self-referring regulatory circuit between polar auxin transport and actin organization. To further dissect the actin-auxin oscillator, we used photoactivated release of caged auxin in tobacco cells to demonstrate that auxin gradients can be manipulated at a subcellular level.Key words: actin, auxin, BY-2, caged compounds, cell division, coleoptile, rice, tobacco     

Subcellular trafficking of PIN auxin efflux carriers in auxin transport

The directional transport of the plant hormone auxin is a unique process mediating a wide variety of developmental processes. Auxin movement between cells depends on AUX1/LAX, PGP and PIN protein families that mediate auxin transport across the plasma membrane. The directionality of auxin flow within tissues is largely determined by polar, subcellular localization of PIN auxin efflux carriers. PIN proteins undergo rapid subcellular dynamics that is important for the process of auxin transport and its directionality. Furthermore, various environmental and endogenous signals can modulate trafficking and polarity of PIN proteins and by this mechanism change auxin distribution. Thus, the subcellular dynamics of auxin transport proteins represents an important interface between cellular processes and development of the whole plant. This review summarizes our recent contributions to the field of PIN trafficking and auxin transport regulation.     

Epidermal growth factor stimulates type-V collagen synthesis in cultured murine palatal shelves

Abstract. The problem studied was whether treatments that reorient vascular differentiation have a similar effect on the polarity of auxin transport. Hypocotyls of Phaseolus vulgaris L. were cut so that a transverse bridge connected the shoot and root directions. Within three days these bridges of tissue regenerated both vessels and sieve tubes along the new orientation, at 90° to the original axis. Experiments involving organ removal, wounds, and hormone application confirm previous suggestions that this differentiation follows the expected flow of the hormone auxin in the direction of the roots. Transport of (³H) indoleacetic acid through sections in which vascular reorientation occurred was polar: it was at least twice as great in the new direction of the roots than in the opposite direction. This new polarity of transport, at right angles to the original axis of the plant, can be readily understood if there is a positive feed-back between the differentiation of tissue polarity and auxin transport.     

The Control of Vascular Branching in Coleus 2. The Corner Traces

Corner trace connections are less well defined than those ofthe side bundle in Coleus, the locations of branch points, branchpartners, and number of connections made by a corner trace beingmore variable. The auxin balance between corner traces was alteredby leaf removal and by application of exogenous auxin. Branchingof new strands was shifted toward the pre-existing strand withthe lower auxin flux, but only within a narrow range of developmentalstages and with the imposition of a large auxin imbalance. Branchingoccurred only in nodal regions, as in control plants. Thus,auxin balance can be made to control xylem strand branching,but it does not account fully for the control of vascular branchingin intact plants. In the intact pattern, corner trace branchesappear to be directed toward the pre-existing strand with thehigher auxin flux. It is proposed that, in the vicinity of astrand with high flux, auxin is transported laterally withinthe nodal vascular cambium, facilitating vessel differentiationbetween strands in the derivatives of the vascular cambium.These vessels comprise the connections between traces. Coleus, vascular differentiation, vascular anatomy, vascular branching, vascular patterns, auxin, auxin balance, node