User:David D./sandbox3c

Plant meristems: the fiendish SU DOKU of stem-cell maintenance. Curr Biol. 2006 Mar 21;16(6):R199-201. Doerner P. Three recent studies have uncovered effector mechanisms and novel pathways in the regulation of the dynamic changes to cell behaviour that occur in plant meristems. The results show how exquisite regulation of cell-cycle mechanisms is central to root stem cell homeostasis.

The SEP4 gene of Arabidopsis thaliana functions in floral organ and meristem identity. Curr Biol. 2004 Nov 9;14(21):1935-40. Ditta G, Pinyopich A, Robles P, Pelaz S, Yanofsky MF.

The ABC model of flower organ identity is widely recognized as providing a framework for understanding the specification of flower organs in diverse plant species. Recent studies in Arabidopsis thaliana have shown that three closely related MADS-box genes, SEPALLATA1 (SEP1), SEP2 and SEP3, are required to specify petals, stamens, and carpels because these organs are converted into sepals in sep1 sep2 sep3 triple mutants. Additional studies indicate that the SEP proteins form multimeric complexes with the products of the B and C organ identity genes. Here, we characterize the SEP4 gene, which shares extensive sequence similarity to and an overlapping expression pattern with the other SEP genes. Although sep4 single mutants display a phenotype similar to that of wild-type plants, we find that floral organs are converted into leaf-like organs in sep1 sep2 sep3 sep4 quadruple mutants, indicating the involvement of all four SEP genes in the development of sepals. We also find that SEP4 contributes to the development of petals, stamens, and carpels in addition to sepals and that it plays an important role in meristem identity. These and other data demonstrate that the SEP genes play central roles in flower meristem identity and organ identity.

WUSCHEL induces shoot stem cell activity and developmental plasticity in the root meristem. Genes Dev. 2004 Feb 15;18(4):375-80. Gallois JL, Nora FR, Mizukami Y, Sablowski R.

Most of the plant shoot originates f om a small group of stem cells, which in A abidopsis are specified by WUSCHEL (WUS). It is unknown whether these cells have an inrinsic potential to generate shoot tissues, or whether differentiation is guided by signals from more mature tissues. He e we show that WUS expression in the root induced shoot stem cell identity and leaf development (without additional cues), floral development (together with LEAFY), or embryogenesis (in response to increased auxin). Thus, WUS establishes stem cells with intrinsic shoot identity and responsive to developmental inputs that normally do not change root identity.

Shoot apical meristem maintenance: the art of a dynamic balance. Trends Plant Sci. 2003 Aug;8(8):394-401. Carles CC, Fletcher JC.

The aerial structure of higher plants derives from cells at the tip of the stem, in the shoot apical meristem (SAM). Throughout the life of a plant, the SAM produces stem tissues and lateral organs, and also regenerates itself. For correct growth, the plant must maintain a constant flow of cells through the meristem, where the input of dividing pluripotent stem cells offsets the output of differentiating cells. This flow depends on extracellular signaling within the SAM, governed by a spatial regulatory feedback loop that maintains a reservoir of stem cells, and on factors that prevent meristem cells from differentiating prematurely. The terminating floral meristem incorporates the spatial regulation scheme into a temporal regulation pathway involving flower patterning factors.

Stem cell homeostasis in the Arabidopsis shoot meristem is regulated by intercellular movement of CLAVATA3 and its sequestration by CLAVATA1. Development. 2003 Jul;130(14):3163-73. Lenhard M, Laux T.

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.

Stem cell regulation in the shoot meristem. J Cell Sci. 2003 May 1;116(Pt 9):1659-66. Gross-Hardt R, Laux T.

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.

The induced sector Arabidopsis apical embryonic fate map. Development. 2002 Jul;129(14):3403-10. Saulsberry A, Martin PR, O'Brien T, Sieburth LE, Pickett FB.

Creation of an embryonic fate map may provide insight into the patterns of cell division and specification contributing to the apical region of the early Arabidopsis embryo. A fate map has been constructed by inducing genetic chimerism during the two-apical-cell stage of embryogenesis to determine if the orientation of the first anticlinal cell division correlates with later developmental axes. Chimeras were also used to map the relative locations of precursors of the cotyledon and leaf primordia. Genetic chimeras were induced in embryos doubly heterozygous for a heat shock regulated Cre recombinase and a constitutively expressed beta-glucuronidase (GUS) gene flanked by the loxP binding sites for Cre. Individual cells in the two-apical-cell stage embryo responding to heat shock produce GUS-negative daughter cells. Mature plants grown from seed derived from treated embryos were scored for GUS-negative sector extent in the cotyledons and leaves. The GUS-negative daughters of apical cells had a strong tendency to contribute primarily to one cotyledon or the other and to physically adjacent true leaf margins. This result indicated that patterns of early cell division correlate with later axes of symmetry in the embryo and that these patterns partially limit the fates available for adoption by daughter cells. However, GUS-negative sectors were shared between all regions of the mature plant, suggesting that there is no strict fate restriction imposed on the daughters of the first apical cells.

Combined SHOOT MERISTEMLESS and WUSCHEL trigger ectopic organogenesis in Arabidopsis. Development. 2002 Jul;129(13):3207-17. Gallois JL, Woodward C, Reddy GV, Sablowski R.

Almost all aerial parts of plants are continuously generated at the shoot apical meristem (SAM). To maintain a steady pool of undifferentiated cells in the SAM while continuously generating new organs, it is necessary to balance the rate of cell division with the rate of entrance into differentiation pathways. In the Arabidopsis meristem, SHOOT MERISTEMLESS (STM) and WUSCHEL (WUS) are necessary to keep cells undifferentiated and dividing. Here, we tested whether ectopic STM and WUS functions are sufficient to revert differentiation and activate cell division in differentiating tissues. Ectopic STM and WUS functions interacted non-additively and activated a subset of meristem functions, including cell division, CLAVATA1 expression and organogenesis, but not correct phyllotaxy or meristem self-maintenance. Our results suggest that WUS produces a non-cell autonomous signal that activates cell division in combination with STM and that combined WUS/STM functions can initiate the progression from stem cells to organ initiation.

The WUSCHEL and SHOOTMERISTEMLESS genes fulfil complementary roles in Arabidopsis shoot meristem regulation. Development. 2002 Jul;129(13):3195-206. Lenhard M, Jurgens G, Laux T.

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.

Plant meristems: a menage a trois to end it all. Curr Biol. 2001 Oct 2;11(19):R785-7. Doerner P.

Regulated termination of stem cell maintenance is required to complete reproductive development in plants. Two recent studies have revealed a new relationship for some old suspects; the WUSCHEL gene, which promotes indeterminancy, is involved as well as the floral regulators LEAFY and AGAMOUS.

Conversion of leaves into petals in Arabidopsis. Curr Biol. 2001 Feb 6;11(3):182-4. Pelaz S, Tapia-Lopez R, Alvarez-Buylla ER, Yanofsky MF.

More than 200 years ago, Goethe proposed that each of the distinct flower organs represents a modified leaf [1]. Support for this hypothesis has come from genetic studies, which have identified genes required for flower organ identity. These genes have been incorporated into the widely accepted ABC model of flower organ identity, a model that appears generally applicable to distantly related eudicots as well as monocot plants. Strikingly, triple mutants lacking the ABC activities produce leaves in place of flower organs, and this finding demonstrates that these genes are required for floral organ identity [2]. However, the ABC genes are not sufficient for floral organ identity since ectopic expression of these genes failed to convert vegetative leaves into flower organs. This finding suggests that one or more additional factors are required [3, 4]. We have recently shown that SEPALLATA (SEP) represents a new class of floral organ identity genes since the loss of SEP activity results in all flower organs developing as sepals [5]. Here we show that the combined action of the SEP genes, together with the A and B genes, is sufficient to convert leaves into petals.

Plant biology. Floral quartets. Nature. 2001 Jan 25;409(6819):469-71. Theissen G, Saedler H. Comment on: Nature. 2001 Jan 25;409(6819):525-9.

Dependence of stem cell fate in Arabidopsis on a feedback loop regulated by CLV3 activity. Science. 2000 Jul 28;289(5479):617-9. Brand U, Fletcher JC, Hobe M, Meyerowitz EM, Simon R.

The fate of stem cells in plant meristems is governed by directional signaling systems that are regulated by negative feedback. In Arabidopsis thaliana, the CLAVATA (CLV) genes encode the essential components of a negative, stem cell-restricting pathway. We used transgenic plants overexpressing CLV3 to show that meristem cell accumulation and fate depends directly on the level of CLV3 activity and that CLV3 signaling occurs exclusively through a CLV1/CLV2 receptor kinase complex. We also demonstrate that the CLV pathway acts by repressing the activity of the transcription factor WUSCHEL, an element of the positive, stem cell-promoting pathway.

B and C floral organ identity functions require SEPALLATA MADS-box genes. Nature. 2000 May 11;405(6783):200-3. Pelaz S, Ditta GS, Baumann E, Wisman E, Yanofsky MF.

Abnormal flowers have been recognized for thousands of years, but only in the past decade have the mysteries of flower development begun to unfold. Among these mysteries is the differentiation of four distinct organ types (sepals, petals, stamens and carpels), each of which may be a modified leaf. A landmark accomplishment in plant developmental biology is the ABC model of flower organ identity. This simple model provides a conceptual framework for explaining how the individual and combined activities of the ABC genes produce the four organ types of the typical eudicot flower. Here we show that the activities of the B and C organ-identity genes require the activities of three closely related and functionally redundant MADS-box genes, SEPALLATA1/2/3 (SEP1/2/3). Triple mutant Arabidopsis plants lacking the activity of all three SEP genes produce flowers in which all organs develop as sepals. Thus SEP1/2/3 are a class of organ-identity genes that is required for development of petals, stamens and carpels.

The stem cell population of Arabidopsis shoot meristems in maintained by a regulatory loop between the CLAVATA and WUSCHEL genes. Cell. 2000 Mar 17;100(6):635-44. Schoof H, Lenhard M, Haecker A, Mayer KF, Jurgens G, Laux T.

The higher-plant shoot meristem is a dynamic structure whose maintenance depends on the coordination of two antagonistic processes, organ initiation and self-renewal of the stem cell population. In Arabidopsis shoot and floral meristems, the WUSCHEL (WUS) gene is required for stem cell identity, whereas the CLAVATA1, 2, and 3 (CLV) genes promote organ initiation. Our analysis of the interactions between these key regulators indicates that (1) the CLV genes repress WUS at the transcript level and that (2) WUS expression is sufficient to induce meristem cell identity and the expression of the stem cell marker CLV3. Our data suggest that the shoot meristem has properties of a self-regulatory system in which WUS/CLV interactions establish a feedback loop between the stem cells and the underlying organizing center.

Regulation of cell proliferation patterns by homeotic genes during Arabidopsis floral development. Development. 2000 Mar;127(6):1267-76. Jenik PD, Irish VF.

The shoot apical meristem of Arabidopsis thaliana consists of three cell layers that proliferate to give rise to the aerial organs of the plant. By labeling cells in each layer using an Ac-based transposable element system, we mapped their contributions to the floral organs, as well as determined the degree of plasticity in this developmental process. We found that each cell layer proliferates to give rise to predictable derivatives: the L1 contributes to the epidermis, the stigma, part of the transmitting tract and the integument of the ovules, while the L2 and L3 contribute, to different degrees, to the mesophyll and other internal tissues. In order to test the roles of the floral homeotic genes in regulating these patterns of cell proliferation, we carried out similar clonal analyses in apetala3-3 and agamous-1 mutant plants. Our results suggest that cell division patterns are regulated differently at different stages of floral development. In early floral stages, the pattern of cell divisions is dependent on position in the floral meristem, and not on future organ identity. Later, during organogenesis, the layer contributions to the organs are controlled by the homeotic genes. We also show that AGAMOUS is required to maintain the layered structure of the meristem prior to organ initiation, as well as having a non-autonomous role in the regulation of the layer contributions to the petals.

The Arabidopsis embryonic shoot fate map. Development. 2000 Feb;127(4):813-20. Woodrick R, Martin PR, Birman I, Pickett FB.

A fate map has been constructed for the shoot apical region of the embryo of the dicotyledonous plant Arabidopsis thaliana using spontaneously arising clonal albino sectors caused by the chloroplast mutator 1-2 mutation. Chimeric seedlings exhibiting albino sectors shared between the cotyledons and first true leaves revealed patterns of organ inclusion and exclusion. Frequencies of clone sharing were used to calculate developmental distances between organs based on the frequency of clonal sectors failing to extend between different organs. The resulting fate map shows asymmetry in the developmental distances between the cotyledons (embryonic leaves) which in turn predicts the location of the first post-germination leaf and the handedness of the spiral of leaf placement around the central stem axis in later development. The map suggests that embryonic leaf fate specification in the cotyledons may represent a developmental ground state necessary for the formation of the shoot apical meristem.

Redundant regulation of meristem identity and plant architecture by FRUITFULL, APETALA1 and CAULIFLOWER. Development. 2000 Feb;127(4):725-34. Ferrandiz C, Gu Q, Martienssen R, Yanofsky MF.

The transition from vegetative to reproductive phases during Arabidopsis development is the result of a complex interaction of environmental and endogenous factors. One of the key regulators of this transition is LEAFY (LFY), whose threshold levels of activity are proposed to mediate the initiation of flowers. The closely related APETALA1 (AP1) and CAULIFLOWER (CAL) meristem identity genes are also important for flower initiation, in part because of their roles in upregulating LFY expression. We have found that mutations in the FRUITFULL (FUL) MADS-box gene, when combined with mutations in AP1 and CAL, lead to a dramatic non-flowering phenotype in which plants continuously elaborate leafy shoots in place of flowers. We demonstrate that this phenotype is caused both by the lack of LFY upregulation and by the ectopic expression of the TERMINAL FLOWER1 (TFL1) gene. Our results suggest that the FUL, AP1 and CAL genes act redundantly to control inflorescence architecture by affecting the domains of LFY and TFL1 expression as well as the relative levels of their activities.

Short-range control of cell differentiation in the Arabidopsis root meristem. Nature. 1997 Nov 20;390(6657):287-9. van den Berg C, Willemsen V, Hendriks G, Weisbeek P, Scheres B.

Meristems are distinctive regions of plants that have capacity for continuous growth. Their developmental activity generates the majority of plant organs. It is currently unknown how cell division and cell differentiation are orchestrated in meristems, although genetic studies have demonstrated the relevance of a proper balance between the two processes. Root meristems contain a distinct central region of mitotically inactive cells, the quiescent centre, the function of which has remained elusive until now. Here we present laser ablation and genetic data that show that in Arabidopsis thaliana the quiescent centre inhibits differentiation of surrounding cells. Differentiation regulation occurs within the range of a single cell, in a manner strikingly similar to examples in animal development, such as during delamination of Drosophila neuroblasts. Our data indicate that pattern formation in the root meristem is controlled by a balance between short-range signals inhibiting differentiation and signals that reinforce cell fate decisions.

The WUSCHEL gene is required for shoot and floral meristem integrity in Arabidopsis. Development. 1996 Jan;122(1):87-96. Laux T, Mayer KF, Berger J, Jurgens G.

Self perpetuation of the shoot meristem is essential for the repetitive initiation of shoot structures during plant development. In Arabidopsis shoot meristem maintenance is disrupted by recessive mutations in the WUSCHEL (WUS) gene. The defect is evident at all developmental stages and is restricted to shoot and floral meristems, whereas the root meristem is not affected. wus mutants fail to properly organize a shoot meristem in the embryo. Postembryonically, defective shoot meristems are initiated repetitively but terminate prematurely in aberrant flat structures. In contrast to wild-type shoot meristems, primordia initiation occurs ectopically across mutant apices, including the center, and often new shoot meristems instead of organs are initiated. The cells of wus shoot apices are larger and more vacuolated than wild-type shoot meristem cells. wus floral meristems terminate prematurely in a central stamen. Double mutant studies indicate that the number of organ primordia in the center of wus flowers is limited, irrespective of organ identity and we propose that meristem cells are allocated into floral whorl domains in a sequential manner. WUS activity also appears to be required for the formation of supernumerary organs in the center of agamous, superman or clavata1 flowers, suggesting that the WUS gene acts upstream of the corresponding genes. Our results suggest that the WUS gene is specifically required for central meristem identity of shoot and floral meristems to maintain their structural and functional integrity.

The Arabidopsis homeotic genes APETALA3 and PISTILLATA are sufficient to provide the B class organ identity function. Development. 1996 Jan;122(1):11-22. Krizek BA, Meyerowitz EM.

The class B organ identity genes, APETALA3 and PISTILLATA, are required to specify petal and stamen identity in the Arabidopsis flower. We show here that the activities of these two genes are sufficient to specify petals and stamens in flowers, in combination with the class A and C genes, respectively. Flowers of plants constitutively expressing both PISTILLATA and APETALA3 under the control of the 35S promoter from cauliflower mosaic virus consist of two outer whorls of petals and inner whorls of stamens. These plants also exhibit vegetative phenotypes that are not present in either of the singly (APETALA3 or PISTILLATA) overexpressing lines. These phenotypes include leaf curling and the partial conversion of later-arising cauline leaves to petals. The presence of additional floral whorls in flowers ectopically expressing APETALA3 and PISTILLATA and the rescue of missing organs in class A mutants by ectopic B function suggest that APETALA3 and PISTILLATA play an additional role in proliferation of the floral meristem.

Cell fate in the Arabidopsis root meristem determined by directional signalling. Nature. 1995 Nov 2;378(6552):62-5. van den Berg C, Willemsen V, Hage W, Weisbeek P, Scheres B. Comment in: Nature. 1995 Nov 2;378(6552):16.

Postembryonic development in plants is achieved by apical meristems. Surgical studies and clonal analysis have revealed indirectly that cells in shoot meristems have no predictable destiny and that position is likely to play a role in the acquisition of cell identity. In contrast to animal systems, there has been no direct evidence for inductive signalling in plants until now. Here we present evidence for such signalling using laser ablation of cells in the root meristem of Arabidopsis thaliana. Although these cells show rigid clonal relationships, we now demonstrate that it is positional control that is most important in the determination of cell fate. Positional signals can be perpetuated from more mature to initial cells to guide the pattern of meristem cell differentiation. This offers an alternative to the general opinion that meristems are the source of patterning information.

The internal meristem layer (L3) determines floral meristem size and carpel number in tomato periclinal chimeras. Plant Cell. 1992 Sep;4(9):1089-100. Szymkowiak EJ, Sussex IM.

Cell-cell interactions are important during plant development. We have generated periclinal chimeras between plants that differ in the number of carpels per flower to determine the roles of cells occupying specific positions in the floral meristem in determining the number of carpels initiated. Intraspecific chimeras were generated between tomato (Lycopersicon esculentum) expressing the mutation fasciated, which causes an increased number of floral organs per whorl, and tomato wild type for fasciated. Interspecific chimeras were generated between tomato and L. peruvianum, which differ in number of carpels per flower. In both sets of chimeras, carpel number as well as the size of the floral meristem during carpel initiation were not determined by the genotype of cells in the outer two layers of the meristem (L1 and L2) but were determined by the genotype of cells occupying the inner layer (L3) of the meristem. We concluded from these experiments that during floral organ initiation, cells in certain layers of the meristem respond to information supplied to them from other cells in the meristem.

Signaling of cell fate decisions by CLAVATA3 in Arabidopsis shoot meristems. Fletcher, J. C.; Brand, U.; Running, M. P.; Simon, R.; Meyerowitz, E. M. Science, 283: 1911-4 (1999).

Control of meristem development by CLAVATA1 receptor kinase and kinase- associated protein phosphatase interactions. Stone, J. M.; Trotochaud, A. E.; Walker, J. C.; Clark, S. E. Plant Physiol, 117: 1217-25 (1998).

The WUSCHEL gene is required for shoot and floral meristem integrity in Arabidopsis. Laux, T.; Mayer, K. F.; Berger, J.; Jurgens, G. Development, 122: 87-96 (1996).

Role of WUSCHEL in regulating stem cell fate in the Arabidopsis shoot meristem. Mayer, K. F.; Schoof, H.; Haecker, A.; Lenhard, M.; Jurgens, G.; Laux, T. Cell, 95: 805-15 (1998).

Dependence of stem cell fate in Arabidopsis on a feedback loop regulated by CLV3 activity. Brand, U.; Fletcher, J. C.; Hobe, M.; Meyerowitz, E. M.; Simon, R. Science, 289: 617-9 (2000).

The stem cell population of Arabidopsis shoot meristems in maintained by a regulatory loop between the CLAVATA and WUSCHEL genes. Schoof, H.; Lenhard, M.; Haecker, A.; Mayer, K. F.; Jurgens, G.; Laux, T. Cell, 100: 635-44 (2000).