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Thomas Laux

Publications and source records attributed to Thomas Laux.

18 recordsLinked to original sources
PubMed2006

WUSCHEL regulates cell differentiation during anther development.

During anther development a series of cell specification events establishes the male gametophyte and the surrounding sporophytic structure. Here we show that the homeobox gene WUSCHEL, originally identified as a central regulator of stem cell maintenance, plays an important role in cell type specification during male organogenesis. WUS expression is initiated very early during anther development in the precursor cells of the stomium and terminates just before the stomium cells enter terminal differentiation. At this stage the stomium cells and the neighboring septum cells that separate the pollen sacs undergo typical cell wall thickening and degenerate which leads to rupture of the anther and pollen release. In wus mutants, neither stomium cells nor septum cells differentiate or undergo cell death and degenerate. As a consequence, the anther stays intact and pollen is not released. CLAVATA3 which is activated by WUS in stem cell maintenance, is not activated in anthers indicating a novel pathway regulated by WUS. Comparing WUS function in stem cell maintenance and sexual organ development suggests that WUS expressing cells represent a conserved signaling module that regulates behavior and communication of undifferentiated cells.

Arabidopsis↗
PubMed2006

The E3 ubiquitin ligase BIG BROTHER controls arabidopsis organ size in a dosage-dependent manner.

Organ growth up to a species-specific size is tightly regulated in plants and animals. Final organ size is remarkably constant within a given species, suggesting that a species-specific size checkpoint terminates organ growth in a coordinated and timely manner. Phytohormones influence plant organ size, but their precise functions in size control are unclear because of their pleiotropic and complex developmental roles. The Arabidopsis transcription factors AINTEGUMENTA and JAGGED promote organ growth by maintaining cellular proliferation potential. Loss of the Antirrhinum transcription factor CINCINNATA causes leaf overgrowth, yet also leads to a highly abnormal leaf shape. Thus, no dedicated factor that limits the final size of plant organs has been isolated. Here, we identify the novel RING-finger protein BIG BROTHER (BB) as a repressor of plant organ growth. Small changes in BB expression levels substantially alter organ size, indicating a central regulatory role for BB in growth control. Recombinant BB protein has E3 ubiquitin-ligase activity that is essential for its in vivo function, suggesting that BB acts by marking cellular proteins for degradation. Our data indicate that plants limit the duration of organ growth and ultimately organ size by actively degrading critical growth stimulators.

Amino Acid Sequence↗
PubMed2006

Analysis of the transcription factor WUSCHEL and its functional homologue in Antirrhinum reveals a potential mechanism for their roles in meristem maintenance.

One of the most significant features of plant development is the way in which it can be elaborated and modulated throughout the life of the plant, an ability that is conferred by meristems. The Arabidopsis thaliana WUSCHEL gene (WUS), which encodes a homeodomain transcription factor, is required to maintain the stem cells in the shoot apical meristem in an undifferentiated state. The mechanism by which WUS prevents the differentiation of stem cells is unknown. We have characterized a meristem maintenance mutant in Antirrhinum majus and shown that it arises from a defect in the WUS orthologue ROSULATA (ROA). Detailed characterization of a semidominant roa allele revealed an essential role for the conserved C-terminal domain. Expression of either ROA or WUS lacking this domain causes a failure of meristem maintenance. The conserved domain mediates an interaction between WUS and two members of a small family of corepressor-like proteins in Arabidopsis. Our results suggest that WUS functions by recruiting transcriptional corepressors to repress target genes that promote differentiation, thereby ensuring stem cell maintenance.

Alleles↗
PubMed2005

APETALA2 regulates the stem cell niche in the Arabidopsis shoot meristem.

Postembryonic organ formation in higher plants relies on the activity of stem cell niches in shoot and root meristems where differentiation of the resident cells is repressed by signals from surrounding cells. We searched for mutations affecting stem cell maintenance and isolated the semidominant l28 mutant, which displays premature termination of the shoot meristem and differentiation of the stem cells. Allele competition experiments suggest that l28 is a dominant-negative allele of the APETALA2 (AP2) gene, which previously has been implicated in floral patterning and seed development. Expression of both WUSCHEL (WUS) and CLAVATA3 (CLV3) genes, which regulate stem cell maintenance in the wild type, were disrupted in l28 shoot apices from early stages on. Unlike in floral patterning, AP2 mRNA is active in the center of the shoot meristem and acts via a mechanism independent of AGAMOUS, which is a repressor of WUS and stem cell maintenance in the floral meristem. Genetic analysis shows that termination of the primary shoot meristem in l28 mutants requires an active CLV signaling pathway, indicating that AP2 functions in stem cell maintenance by modifying the WUS-CLV3 feedback loop.

AGAMOUS Protein, Arabidopsis↗
PubMed2005

Regulation of WUSCHEL transcription in the stem cell niche of the Arabidopsis shoot meristem.

Pluripotent stem cells are localized in specialized microenvironments, called stem cell niches, where signals from surrounding cells maintain their undifferentiated status. In the Arabidopsis thaliana shoot meristem, the homeobox gene WUSCHEL (WUS) is expressed in the organizing center underneath the stem cells and integrates regulatory information from several pathways to define the boundaries of the stem cell niche. To investigate how these boundaries are precisely maintained within the proliferating cellular context of the shoot meristem, we analyzed the transcriptional control of the WUS gene. Our results show that the WUS promoter contains distinct regulatory regions that control tissue specificity and levels of transcription in a combinatorial manner. However, a 57-bp regulatory region is all that is required to control the boundaries of WUS transcription in the shoot meristem stem cell niche, and this activity can be further assigned to two adjacent short sequence motifs within this region. Our results indicate that the diverse regulatory pathways that control the stem cells in the shoot meristem converge at these two short sequence elements of the WUS promoter, suggesting that the integration of regulatory signals takes place at the level of a central transactivating complex.

Arabidopsis↗
PubMed2004

Pattern formation during early ovule development in Arabidopsis thaliana.

Ovules of higher plants are the precursors of seeds. Ovules emerge from placental tissue inside the gynoecium of flowers. Three elements, funiculus, chalaza, and nucellus, can be distinguished along the proximal-distal axis of the outgrowing radially symmetrical ovule primordium. The asymmetric initiation of the outer integument marks the switch to adaxial-abaxial development, which leads to the formation of a bilaterally symmetrical ovule. The putative transcriptional regulator NOZZLE (NZZ) plays a role in mediating this transition by controlling the timing of expression of the putative transcriptional regulator INNER NO OUTER (INO) in an abaxial domain of the chalaza, from where the outer integument initiates. Integument formation depends on the homeobox gene WUSCHEL (WUS), which is expressed in the nucellus and is sufficient to induce integuments non-cell autonomously from a region adjacent to its expression domain. In this study, we describe the expression pattern of the homeobox-leucine zipper gene PHABULOSA (PHB) during ovule development, demonstrating that adaxial-abaxial polarity is established from the very beginning of ovule development. Furthermore, we examined the expression pattern of PHB, INO, and WUS in ovules of plants, which are affected in integument initiation and thus defective in the transition from proximal-distal to adaxial-abaxial development. We found that NZZ is required to restrict PHB expression to the distal chalaza, from where the inner integument initiates. PHB expression is not established in the distal chalaza of two mutants, aintegumenta (ant) and wus, which fail to form integuments. Furthermore, we suggest that one mechanism by which WUS controls integument formation is by establishing the chalaza and that outer and inner integument identity determination depends on additional region-specific factors. In addition, we present evidence that NZZ is essential for the normal nucellar expression pattern of WUS. Thus, both WUS and PHB affect processes downstream of NZZ action during the transition from proximal--distal to adaxial--abaxial ovule development.

Arabidopsis↗
PubMed2004

Helical growth of the Arabidopsis mutant tortifolia1 reveals a plant-specific microtubule-associated protein.

Plants can grow straight or in the twisted fashion exhibited by the helical growth of some climbing plants. Analysis of helical-growth mutants from Arabidopsis has indicated that microtubules are involved in the expression of the helical phenotype. Arabidopsis mutants growing with a right-handed twist have been reported to have cortical microtubules that wind around the cell in left-handed helices and vice versa. Microtubular involvement is further suspected from the finding that some helical mutants are caused by single amino acid substitutions in alpha-tubulin and because of the sensitivity of the growth pattern to anti-microtubule drugs. Insight into the roles of microtubules in organ elongation is anticipated from analyses of genes defined by helical mutations. We investigated the helical growth of the Arabidopsis mutant tortifolia1/spiral2 (tor1/spr2), which twists in a right-handed manner, and found that this correlates with a complex reorientation of cortical microtubules. TOR1 was identified by a map-based approach; analysis of the TOR1 protein showed that it is a member of a novel family of plant-specific proteins containing N-terminal HEAT repeats. Recombinant TOR1 colocalizes with cortical microtubules in planta and binds directly to microtubules in vitro. This shows that TOR1 is a novel, plant-specific microtubule-associated protein (MAP) that regulates the orientation of cortical microtubules and the direction of organ growth.

Amino Acid Sequence↗
PubMed2004

Expression dynamics of WOX genes mark cell fate decisions during early embryonic patterning in Arabidopsis thaliana.

During embryonic pattern formation, the main body axes are established and cells of different developmental fates are specified from a single-cell zygote. Despite the fundamental importance of this process, in plants, the underlying mechanisms are largely unknown. We show that expression dynamics of novel WOX (WUSCHEL related homeobox) gene family members reveal early embryonic patterning events in Arabidopsis. WOX2 and WOX8 are co-expressed in the egg cell and zygote and become confined to the apical and basal daughter cells of the zygote, respectively, by its asymmetric division. WOX2 not only marks apical descendants of the zygote, but is also functionally required for their correct development, suggesting that the asymmetric division of the plant zygote separates determinants of apical and basal cell fates. WOX9 expression is initiated in the basal daughter cell of the zygote and subsequently shifts into the descendants of the apical daughter apparently in response to signaling from the embryo proper. Expression of WOX5 shows that identity of the quiescent center is initiated very early in the hypophyseal cell, and highlights molecular and developmental similarities between the stem cell niches of root and shoot meristems. Together, our data suggest that during plant embryogenesis region-specific transcription programs are initiated very early in single precursor cells and that WOX genes play an important role in this process.

Amino Acid Sequence↗
PubMed2003

ZWILLE buffers meristem stability in Arabidopsis thaliana.

The shoot apical meristem of higher plants consists of a population of stem cells at the tip of the plant body that continuously gives rise to organs such as leaves and flowers. Cells that leave the meristem differentiate and must be replaced to maintain the integrity of the meristem. The balance between differentiation and maintenance is governed both by the environment and the developmental status of the plant. In order to respond to these different stimuli, the meristem has to be plastic thus ensuring the stereotypic shape of the plant body. Meristem plasticity requires the ZWILLE (ZLL) gene. In zll mutant embryos, the apical cells are misspecified causing a variability of the meristem's size and function. Using specific antibodies against ZLL, we show that the zll phenotype is due to the complete absence of the ZLL protein. In immunohistochemical experiments we confirm the observation that ZLL is solely localized in vascular tissue. For a better understanding of the role of ZLL in meristem stability, we analysed the genetic interactions of ZLL with WUSCHEL (WUS) and the CLAVATA1, 2 and 3 (CLV) genes that are involved in size regulation of the meristem. In a zll loss-of-function background wus has a negative effect whereas clv mutations have a positive effect on meristem size. We propose that ZLL buffers meristem stability non-cell-autonomously by ensuring the critical number of apical cells required for proper meristem function.

Alleles↗
PubMed2003

The stem cell concept in plants: a matter of debate.

Throughout their life, which can last for over a thousand years, plants have the fascinating ability to give rise to new organs from founder cells in their apical meristems. Whether these founder cells are equivalent to the pluripotent stem cells in animals has been a long-standing controversy amongst plant scientists. Here, this controversy will be addressed in light of classical observations and recent findings.

Cell Differentiation↗
PubMed2003

Stem cell regulation in the shoot meristem.

A small group of pluripotent stem cells in the shoot meristem is the ultimate source for all aerial parts in higher plants: the shoot axis, side branches, leaves and flowers. The stem cells are maintained in an undifferentiated state by signals from an underlying cell group, the organizing center. Genetic and molecular analyses have shown that a feedback signaling loop between stem cells and the organizing center balances stem cell renewal versus differentiation, which allows the plant to maintain the organization of the shoot meristem despite a changing cellular context. Emerging common principles indicate that plant and animal stem cells are functionally equivalent.

Arabidopsis↗
PubMed2003

Apical meristems: the plant's fountain of youth.

During postembryonic development, all organs of a plant are ultimately derived from a few pluripotent stem cells found in specialized structures called apical meristems. Here we discuss our current knowledge about the regulation of plant stem cells and their environments with main emphasis on the shoot apical meristem of Arabidopsis thaliana. Recent studies suggest that stem cells are localized in specialized niches where signals from surrounding cells maintain their undifferentiated state. In the shoot meristem, initiation of stem cells during embryogenesis, regulation of stem-cell homeostasis and termination of stem-cell maintenance during flower development appear to primarily involve regulation of the stem-cell niche.

Arabidopsis↗
PubMed2003

Stem cell homeostasis in the Arabidopsis shoot meristem is regulated by intercellular movement of CLAVATA3 and its sequestration by CLAVATA1.

Stem cell maintenance in the Arabidopsis shoot meristem is regulated by communication between the apical stem cells and the underlying organizing centre. Expression of the homeobox gene WUSCHEL in the organizing centre induces stem cell identity in the overlying neighbours, which then express the CLAVATA3 gene whose activity in turn restricts the size of the WUSCHEL expression domain. We have analyzed how the stem cells and the organizing centre communicate, by studying the mode of action of CLAVATA3 protein. We provide direct evidence that CLAVATA3 protein functions as a mobile intercellular signal in the shoot apical meristem that spreads laterally from the stem cells and acts both on their lateral neighbours and on the stem cells themselves to repress WUSCHEL transcription. We also show that the spread and range of action of CLAVATA3 can be limited by binding to its receptor CLAVATA1, which offers an explanation for how CLAVATA3 is prevented from entering the organizing centre and repressing WUSCHEL transcription there. This regulated spread of a secreted signalling molecule enables the shoot meristem to permit the onset of cell differentiation in the periphery, but at the same time to maintain a stable niche for its stem cells in the center.

Amino Acid Sequence↗
PubMed2002

WUSCHEL signaling functions in interregional communication during Arabidopsis ovule development.

Coordinating the behaviors of different cell populations is essential for multicellular development. One important example for this can be found in ovule development in higher plants. Ovules give rise to the gametophyte in the distal nucellus and form protective sporophytic organs from the underlying chalaza. We show that the WUSCHEL (WUS) homeobox gene provides a mechanism to coordinate these events. WUS is expressed in the nucellus and our loss- and gain-of-function analyses show that WUS is not only necessary but also sufficient for integument formation from the chalaza. WUS protein is retained in the nucellus, indicating that WUS activity in the nucellus generates a downstream signal that non-cell-autonomously regulates integument initiation in the chalaza. This signal appears to act locally, thus determining the position of organ formation from chalazal cells adjacent to the nucellus. Analysis of WUS and AINTEGUMENTA functions indicates that integument initiation requires inputs from different ovule regions. Together with previous findings for shoot and floral meristems, where WUS signaling establishes a stem cell niche, our results indicate that WUS defines a signaling mechanism that is used repeatedly during plant development in coordinating the behavior of adjacent cell groups.

Arabidopsis↗
PubMed2002

The WUSCHEL and SHOOTMERISTEMLESS genes fulfil complementary roles in Arabidopsis shoot meristem regulation.

Continuous organ formation from the shoot apical meristem requires the integration of two functions: a set of undifferentiated, pluripotent stem cells is maintained at the very tip of the meristem, while their daughter cells in the periphery initiate organ primordia. The homeobox genes WUSCHEL (WUS) and SHOOTMERISTEMLESS (STM) encode two major regulators of meristem formation and maintenance in Arabidopsis, yet their interaction in meristem regulation is presently unclear. Here, we have addressed this question using loss- and gain-of-function approaches. We show that stem cell specification by WUS does not require STM activity. Conversely, STM suppresses differentiation independently of WUS and is required and sufficient to promote cell division. Consistent with their independent and distinct phenotypic effects, ectopic WUS and STM activities induce the expression of different downstream target genes. Finally, the pathways regulated by WUS and STM appear to converge in the suppression of differentiation, since coexpression of both genes produced a synergistic effect, and increased WUS activity could partly compensate for loss of STM function. These results suggest that WUS and STM share labour in the shoot apical meristem: WUS specifies a subset of cells in the centre as stem cells, while STM is required to suppress differentiation throughout the meristem dome, thus allowing stem cell daughters to be amplified before they are incorporated into organs.

Arabidopsis↗
PubMed2001

Somatic embryogenesis from Arabidopsis shoot apical meristem mutants.

Zygotic embryos of three Arabidopsis thaliana (L.) Heynh. mutants lacking an embryonic shoot apical meristem (SAM), shoot meristemless (stm), wuschel (wus) and zwille/pinhead (zll/pnh) were used as explants to establish embryogenic cell cultures. Somatic embryos of all three mutants showed the same mutant phenotypes as their zygotic equivalents. These results provide genetic evidence that the developmental program of somatic and zygotic embryos is indistinguishable. They also suggest that a functional SAM is not required for somatic embryogenic cell formation in Arabidopsis.

Arabidopsis↗