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P N Benfey

Publications and source records attributed to P N Benfey.

At least 19 recordsLinked to original sources

Intercellular movement of the putative transcription factor SHR in root patterning.

Positional information is pivotal for establishing developmental patterning in plants, but little is known about the underlying signalling mechanisms. The Arabidopsis root radial pattern is generated through stereotyped division of initial cells and the subsequent acquisition of cell fate. short-root (shr) mutants do not undergo the longitudinal cell division of the cortex/endodermis initial daughter cell, resulting in a single cell layer with only cortex attributes. Thus, SHR is necessary for both cell division and endodermis specification. SHR messenger RNA is found exclusively in the stele cells internal to the endodermis and cortex, indicating that it has a non-cell-autonomous mode of action. Here we show that the SHR protein, a putative transcription factor, moves from the stele to a single layer of adjacent cells, where it enters the nucleus. Ectopic expression of SHR driven by the promoter of the downstream gene SCARECROW (SCR) results in autocatalytic reinforcement of SHR signalling, producing altered cell fates and multiplication of cell layers. These results support a model in which SHR protein acts both as a signal from the stele and as an activator of endodermal cell fate and SCR-mediated cell division.

Arabidopsis↗

COBRA encodes a putative GPI-anchored protein, which is polarly localized and necessary for oriented cell expansion in Arabidopsis.

To control organ shape, plant cells expand differentially. The organization of the cellulose microfibrils in the cell wall is a key determinant of differential expansion. Mutations in the COBRA (COB) gene of Arabidopsis, known to affect the orientation of cell expansion in the root, are reported here to reduce the amount of crystalline cellulose in cell walls in the root growth zone. The COB gene, identified by map-based cloning, contains a sequence motif found in proteins that are anchored to the extracellular surface of the plasma membrane through a glycosylphosphatidylinositol (GPI) linkage. In animal cells, this lipid linkage is known to confer polar localization to proteins. The COB protein was detected predominately on the longitudinal sides of root cells in the zone of rapid elongation. Moreover, COB RNA levels are dramatically upregulated in cells entering the zone of rapid elongation. Based on these results, models are proposed for the role of COB as a regulator of oriented cell expansion.

Amino Acid Sequence↗

Axis formation and polarity in plants.

We have recently gained insight into a number of mechanisms governing the formation of the major axes that define the embryonic and adult plant body plan. Phenotypic analysis and molecular characterization of mutants with aberrant morphogenesis has led to a better understanding of key processes including the generation of the shape of the apical embryo, the establishment and maintenance of the radial pattern of the root, and the placement of lateral organ primordia around the shoot apical meristem.

Genes, Plant↗

cis element/transcription factor analysis (cis/TF): a method for discovering transcription factor/cis element relationships.

We report a simple new algorithm, cis/TF, that uses genomewide expression data and the full genomic sequence to match transcription factors to their binding sites. Most previous computational methods discovered binding sites by clustering genes having similar expression patterns and then identifying over-represented subsequences in the promoter regions of those genes. By contrast, cis/TF asserts that B is a likely binding site of a transcription factor T if the expression pattern of T is correlated to the composite expression patterns of all genes containing B, even when those genes are not mutually correlated. Thus, our method focuses on binding sites rather than genes. The algorithm has successfully identified experimentally-supported transcription factor binding relationships in tests on several data sets from Saccharomyces cerevisiae.

Algorithms↗

A novel two-component hybrid molecule regulates vascular morphogenesis of the Arabidopsis root.

The developmental ontogeny of the vascular system (consisting of xylem, phloem and [pro]cambium) is poorly understood despite its central role in plant physiology. We show that in the Arabidopsis root meristem, xylem cell lineages are specified early, whereas phloem and procambium are established through a set of asymmetric cell divisions. These divisions require the WOODEN LEG (WOL) gene. The WOL gene encodes a novel two-component signal transducer with an unusual tandem arrangement of two receiver domains. It is expressed specifically in the vasculature from the early stages of embryogenesis on, consistent with a role as a sensor for vascular morphogenesis.

Amino Acid Sequence↗

Root development.

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Arabidopsis↗

The SHORT-ROOT gene controls radial patterning of the Arabidopsis root through radial signaling.

Asymmetric cell divisions play an important role in the establishment and propagation of the cellular pattern of plant tissues. The SHORT-ROOT (SHR) gene is required for the asymmetric cell division responsible for formation of ground tissue (endodermis and cortex) as well as specification of endodermis in the Arabidopsis root. We show that SHR encodes a putative transcription factor with homology to SCARECROW (SCR). From analyses of gene expression and cell identity in genetically stable and unstable alleles of shr, we conclude that SHR functions upstream of SCR and participates in a radial signaling pathway. Consistent with a regulatory role in radial patterning, ectopic expression of SHR results in supernumerary cell divisions and abnormal cell specification in the root meristem.

Amino Acid Sequence↗

Molecular analysis of the SCARECROW gene in maize reveals a common basis for radial patterning in diverse meristems.

Maize and Arabidopsis root apical meristems differ in several aspects of their radial organization and ontogeny. Despite the large evolutionary distance and differences in root radial patterning, analysis of the putative maize ortholog of the Arabidopsis patterning gene SCARECROW (SCR) revealed expression localized to the endodermis, which is similar to its expression in Arabidopsis. Expression in maize extends through the quiescent center, a population of mitotically inactive cells formerly thought to be undifferentiated and to lack radial pattern information. Zea mays SCARECROW (ZmSCR), the putative maize SCR ortholog, was used as a molecular marker to investigate radial patterning during regeneration of the root tip after either whole or partial excision. Analysis of the dynamic expression pattern of ZmSCR as well as other markers indicates the involvement of positional information as a primary determinant in regeneration of the root radial pattern.

Amino Acid Sequence↗

Molecular analysis of SCARECROW function reveals a radial patterning mechanism common to root and shoot.

Mutation of the SCARECROW (SCR) gene results in a radial pattern defect, loss of a ground tissue layer, in the root. Analysis of the shoot phenotype of scr mutants revealed that both hypocotyl and shoot inflorescence also have a radial pattern defect, loss of a normal starch sheath layer, and consequently are unable to sense gravity in the shoot. Analogous to its expression in the endodermis of the root, SCR is expressed in the starch sheath of the hypocotyl and inflorescence stem. The SCR expression pattern in leaf bundle sheath cells and root quiescent center cells led to the identification of additional phenotypic defects in these tissues. SCR expression in a pin-formed mutant background suggested the possible origins of the starch sheath in the shoot inflorescence. Analysis of SCR expression and the mutant phenotype from the earliest stages of embryogenesis revealed a tight correlation between defective cell divisions and SCR expression in cells that contribute to ground tissue radial patterning in both embryonic root and shoot. Our data provides evidence that the same molecular mechanism regulates the radial patterning of ground tissue in both root and shoot during embryogenesis as well as postembryonically.

Arabidopsis↗

Stem cells: A tale of two kingdoms.

Homologous genes have recently been shown to regulate stem cell maintenance in animals and plants. This discovery should facilitate elucidation of the poorly understood factors that control stem cell maintenance and differentiation.

Animals↗

Is the shoot a root with a view?

Recently, it has been shown that the same sets of genes act in both root and shoot to regulate cell fate and patterning. One gene cassette regulates epidermal cell fate, another cassette regulates ground tissue derived cell fate and organization. Ectopic expression and laser ablation have been used to probe the mechanisms by which these genes perform their tissue and organ-specific functions.

Cell Lineage↗

The GRAS gene family in Arabidopsis: sequence characterization and basic expression analysis of the SCARECROW-LIKE genes.

Mutations at the SCARECROW (SCR) locus in Arabidopsis thaliana result in defective radial patterning in the root and shoot. The SCR gene product contains sequences which suggest that it is a transcription factor. A number of Arabidopsis Expressed Sequence Tags (ESTs) have been identified that encode gene products bearing remarkable similarity to SCR throughout their carboxyl-termini, indicating that SCR is the prototype of a novel gene family. These ESTs have been designated SCARECROW-LIKE (SCL). The gene products of the GIBBERELLIN-INSENSITIVE (GAI) and the REPRESSOR of ga1-3 (RGA) loci show high structural and sequence similarity to SCR and the SCLs. Sequence analysis of the products of the GRAS (GAI, RGA, SCR) gene family indicates that they share a variable amino-terminus and a highly conserved carboxyl-terminus that contains five recognizable motifs. The SCLs have distinct patterns of expression, but all of those analyzed show expression in the root. One of them, SCL3, has a tissue-specific pattern of expression in the root similar to SCR. The importance of the GRAS gene family in plant biology has been established by the functional analyses of SCR, GAI and RGA.

Amino Acid Sequence↗

Genetic evidence that the endodermis is essential for shoot gravitropism in Arabidopsis thaliana.

Shoots of higher plants exhibit negative gravitropism. However, little is known about the mechanism or site of gravity perception in shoots. We have identified two loci that are essential for normal shoot gravitropism in Arabidopsis thaliana. Genetic analysis demonstrated that the shoot gravitropism mutants sgr1 and sgr7 are allelic to the radial pattern mutants, scr and shr, respectively. Characterization of the aerial phenotype of these mutants revealed that the primary defect is the absence of a normal endodermis in hypocotyls and influorescence stems. This indicates that the endodermis is essential for shoot gravitropism and strongly suggests that this cell layer functions as the gravity-sensing cell layer in dicotyledonous plant shoots. These results also demonstrate that, in addition to their previously characterized role in root radial patterning, SCR and SHR regulate the radial organization of the shoot axial organs in Arabidopsis.

Arabidopsis↗

Root development: signaling down and around.

Because of its elegant simplicity, the Arabidopsis root has become a model for studying plant organogenesis. In this review we focus on recent results indicating the importance of signaling in root development. A role for positional information in root cell specification has been demonstrated by ablation analyses. Through mutational analysis, genes have been identified that play a role in radial pattern formation. The embryonic phenotypes of these mutants raised the possibility that division patterns in post-embryonic roots are dependent on signaling that originates during embryonic development. Analysis of expression of the SCARECROW gene indicates that it may play a role in this 'top-down' signaling process. Characterization of root epidermis development has led to the identification of negative regulators of root-hair formation. These appear to set up a prepattern which is reinforced by signaling by plant hormones.

Arabidopsis↗

Analysis of SCARECROW expression using a rapid system for assessing transgene expression in Arabidopsis roots.

Although the introduction of foreign genes into Arabidopsis has become routine, the production of transgenic Arabidopsis plants still requires several months. A transgene expression system (TES) has been developed that allows characterization of gene expression patterns and the effects of foreign genes in the Arabidopsis root in 2-4 weeks. The method is based on regeneration of stably transformed roots directly from callus tissue. TES has been used to study the expression of the SCARECROW gene, which is involved in establishing radial patterning in the root. The 2.5 kb region directly upstream of the SCARECROW coding region was found to be sufficient to confer cell-type specific expression. Furthermore, this promoter is active in the scr mutant background, indicating that factors essential for cell-type specific expression are present even in the absence of correct radial patterning. Finally, TES was used to demonstrate that the SCARECROW gene under control of this promoter complements the root organization defect of the scr mutant. These experiments demonstrate the utility of the TES system for studying gene expression in roots in wild-type and mutant backgrounds and for molecular complementation of root mutant phenotypes. It is possible that the method will also be applicable to other organs.

Arabidopsis↗

Organization and cell differentiation in lateral roots of Arabidopsis thaliana.

Lateral root formation in plants involves the stimulation of mature pericycle cells to proliferate and redifferentiate to create a new organ. The simple organization of the root of Arabidopsis thaliana allows the development of lateral root primordia to be characterized histologically. We have divided the process of lateral root development into 8 stages defined by specific anatomical characteristics and cell divisions. To identify the cell types in the developing primordium we have generated a collection of marker lines that express beta-glucuronidase in a tissue- or cell type-specific manner in the root. Using these tools we have constructed a model describing the lineage of each cell type in the lateral root. These studies show that organization and cell differentiation in the lateral root primordia precede the appearance of a lateral root meristem, with differential gene expression apparent after the first set of divisions of the pericycle.

Arabidopsis↗

The SCARECROW gene regulates an asymmetric cell division that is essential for generating the radial organization of the Arabidopsis root.

In the Arabidopsis root meristem, initial cells undergo asymmetric divisions to generate the cell lineages of the root. The scarecrow mutation results in roots that are missing one cell layer owing to the disruption of an asymmetric division that normally generates cortex and endodermis. Tissue-specific markers indicate that a heterogeneous cell type is formed in the mutant. The deduced amino acid sequence of SCARECROW (SCR) suggests that it is a member of a novel family of putative transcription factors. SCR is expressed in the cortex/endodermal initial cells and in the endodermal cell lineage. Tissue-specific expression is regulated at the transcriptional level. These results indicate a key role for SCR in regulating the radial organization of the root.

Amino Acid Sequence↗