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ROR1/RPA2A, a putative replication protein A2, functions in epigenetic gene silencing and in regulation of meristem development in Arabidopsis.

We screened for suppressors of repressor of silencing1 (ros1) using the silenced 35S promoter-neomycin phosphotransferase II (Pro(35S):NPTII) gene as a marker and identified two allelic mutants, ror1-1 and ror1-2 (for suppressor of ros1). Map-based cloning revealed that ROR1 encodes a 31-kD protein similar to DNA replication protein A2 (RPA2A). Mutations in ROR1 reactivate the silenced Pro(35S):NPTII gene but not RD29A promoter-luciferase in the ros1 mutant. DNA methylation in rDNA, centromeric DNA, and RD29A promoter regions is not affected by ror1. However, chromatin immunoprecipitation data suggest that histone H3 acetylation is increased and histone H3K9 dimethylation is decreased in the 35S promoter in the ror1 ros1 mutant compared with ros1. These results indicate that release of silenced Pro(35S):NPTII by ror1 mutations is independent of DNA methylation. ROR1/RPA2A is strongly expressed in shoot and root meristems. Mutations in ROR1/RPA2A affect cell division in meristems but not final cell sizes. Our work suggests important roles of ROR1/RPA2A in epigenetic gene silencing and in the regulation of plant development.

Amino Acid Sequence↗

ramosa2 encodes a LATERAL ORGAN BOUNDARY domain protein that determines the fate of stem cells in branch meristems of maize.

Genetic control of grass inflorescence architecture is critical given that cereal seeds provide most of the world's food. Seeds are borne on axillary branches, which arise from groups of stem cells in axils of leaves and whose branching patterns dictate most of the variation in plant form. Normal maize (Zea mays) ears are unbranched, and tassels have long branches only at their base. The ramosa2 (ra2) mutant of maize has increased branching with short branches replaced by long, indeterminate ones. ra2 was cloned by chromosome walking and shown to encode a LATERAL ORGAN BOUNDARY domain transcription factor. ra2 is transiently expressed in a group of cells that predicts the position of axillary meristem formation in inflorescences. Expression in different mutant backgrounds places ra2 upstream of other genes that regulate branch formation. The early expression of ra2 suggests that it functions in the patterning of stem cells in axillary meristems. Alignment of ra2-like sequences reveals a grass-specific domain in the C terminus that is not found in Arabidopsis thaliana. The ra2-dm allele suggests this domain is required for transcriptional activation of ra1. The ra2 expression pattern is conserved in rice (Oryza sativa), barley (Hordeum vulgare), sorghum (Sorghum bicolor), and maize, suggesting that ra2 is critical for shaping the initial steps of grass inflorescence architecture.

Amino Acid Sequence↗

FILAMENTOUS FLOWER controls the formation and development of arabidopsis inflorescences and floral meristems.

Phenotypic analysis of single and multiple mutants as well as in situ localization analysis of the expression patterns of floral genes have revealed that the FIL AMENTOUS FLOWER (FIL) gene plays important roles in establishing the inflorescence in Arabidopsis. As previously reported, the fil mutant generates clusters of both filamentous structures and flowers with floral organs of altered number and shape. The structural resemblance of the filamentous structures to peduncles and the expression pattern of the APETALA1 (AP1) gene have shown that these filamentous structures are underdeveloped flowers that fail to form receptacles and floral organs, indicating that one of the roles of the FIL gene is to support the development of the floral meristem. That FIL also is involved in fate determination in the floral meristem is demonstrated by the homeotic conversion of flowers to inflorescences in fil ap1 double mutants and in fil ap1 cauliflower triple mutants. In double mutants with flowering-time loci (i.e., f t or f wa), leafy (lf y), and unusual floral organs (ufo), filamentous structures are formed, but very few or no flowers with floral organs develop. The enhanced phenotype in the fil ap1 and the fil lfy double mutants suggests that the FIL protein may work together with AP1 and LFY proteins. The FIL gene also may be involved in the cell fate determination of floral organ primordia, possibly by controlling the spatial expression patterns of the class A and C floral organ identity genes.

Arabidopsis↗

Control of flower development and phyllotaxy by meristem identity genes in antirrhinum.

The flower meristem identity genes floricaula (flo) and squamosa (squa) promote a change in phyllotaxy from spiral to whorled in Antirrhinum. To determine how this might be achieved, we have performed a combination of morphological, genetic, and expression analyses. Comparison of the phenotypes and RNA expression patterns of single and double mutants with the wild type showed that flo and squa act together to promote flower development but that flo is epistatic to squa with respect to early effects on phyllotaxy. We propose that a common process underlies the phyllotaxy of wildtype, flo, and squa meristem development but that the relative timing of primordium initiation or growth is altered. This process depends on two separable events: setting aside zones for potential primordium initiation and partitioning these zones into discrete primordia. Failure of the second event can lead to the formation of continuous double spirals, which are occasionally seen in flo mutants.

Base Sequence↗

UFO: an Arabidopsis gene involved in both floral meristem and floral organ development.

We describe the role of the UNUSUAL FLORAL ORGANS (UFO) gene in Arabidopsis floral development based on a genetic and molecular characterization of the phenotypes of nine ufo alleles. UFO is required for the proper identity of the floral meristem and acts in three different aspects of the process that distinguishes flowers from shoots. UFO is involved in establishing the whorled pattern of floral organs, controlling the determinacy of the floral meristem, and activating the APETALA3 and PISTILLATA genes required for petal and stamen identity. In many respects, UFO acts in a manner similar to LEAFY, but the ufo mutant phenotype also suggests an additional role for UFO in defining boundaries within the floral primordia or controlling cell proliferation during floral organ growth. Finally, genetic interactions that prevent flower formation and lead to the generation of filamentous structures implicate UFO as a member of a new, large, and diverse class of genes in Arabidopsis necessary for flower formation.

Arabidopsis↗

Expression levels of meristem identity and homeotic genes are modified by nuclear-mitochondrial interactions in alloplasmic male-sterile lines of Brassica napus.

Homeotic conversions of anthers were found in cytoplasmic male sterile (CMS) plants of Brassica napus derived from somatic hybrids of B. napus and Arabidopsis thaliana. CMS line flowers displayed petals reduced in size and width and stamens replaced by carpelloid structures. In order to investigate when these developmental aberrations appeared, flower development was analysed histologically, ultrastructurally and molecularly. Disorganized cell divisions were detected in the floral meristems of the CMS lines at stage 4. As CMS is associated with mitochondrial aberrations, ultrastructural analysis of the mitochondria in the floral meristems was performed. Two mitochondrial populations were found in the CMS lines. One type had disrupted cristae, while the other resembled mitochondria typical of B. napus. Furthermore, expression patterns of genes expressed in particular floral whorls were determined. In spite of the aberrant development of the third whorl organs, BnAP3 was expressed as in B. napus during the first six stages of development. However, the levels of BnPI were reduced. At later developmental stages, the expression of both BnAP3 and BnPI was strongly reduced. Interestingly the expression levels of genes responsible for AP3 and PI activation such as LFY, UFO and ASK1 were higher in the CMS lines, which indicates that activation of B-genes in the CMS lines does not occur as in B. napus. Disrupted and dysfunctional mitochondria seem to be one of the first aberrations manifested in CMS which result in a retrograde influence of the expression levels of genes responsible for the second and third whorl organ differentiation.

Arabidopsis↗

Transitions in the functioning of the shoot apical meristem in birch (Betula pendula) involve ethylene.

In many trees, a short photoperiod (SD) triggers substantial physiological adjustments necessary for over-wintering. We have used transgenic ethylene-insensitive birches (Betula pendula), which express the Arabidopsis ethylene receptor gene ETR1 carrying the dominant mutation etr1-1, to investigate the role of ethylene in SD-induced responses in the shoot apical meristem (SAM). Under SD, the ethylene-insensitive trees ceased elongation growth comparably to the wild-type. In contrast, the formation of terminal buds, which in trees is typically induced by SD, was abolished. However, although delayed, endo-dormancy did eventually develop in the ethylene-insensitive trees. This, together with the rapid resumption of growth in the ethylene-insensitive trees after transfer from non-permissive to permissive conditions suggests that ethylene facilitates the SD-induced terminal bud formation, as well as growth arrest. In addition, apical buds of the ethylene-insensitive birch did not accumulate abscisic acid (ABA) under SD, suggesting interaction between ethylene and ABA signalling in the bud. Alterations in SAM functioning were further exemplified by reduced apical dominance and early flowering in ethylene-insensitive birches. Gene expression analysis of shoot apices revealed that the ethylene-insensitive birch lacked the marked increase in expression of a beta-xylosidase gene typical to the SD-exposed wild-type. The ethylene-dependent beta-xylosidase gene expression is hypothesized to relate to modification of cell walls in terminal buds during SD-induced growth cessation. Our results suggest that ethylene is involved in terminal bud formation and in the timely suppression of SAM activity, not only in the shoot apex, but also in axillary and reproductive meristems.

Abscisic Acid↗

Shoot and floral meristem maintenance in arabidopsis.

The shoot apical meristem (SAM) of higher plants functions as a site of continuous organogenesis within which a small pool of pluripotent stem cells replenishes the cells incorporated into lateral organs. This article summarizes recent results demonstrating that the fate of stem cells in Arabidopsis shoot and floral meristems is controlled by overlapping spatial and temporal signaling systems. Stem cell maintenance is an active process requiring constant communication between neighboring groups of SAM cells. Information flows via a ligand-receptor signal transduction pathway, resulting in the formation of a spatial feedback loop that stabilizes the size of the stem cell population. Termination of stem cell activity during flower development is achieved by a temporal feedback loop involving both stem cell maintenance genes and flower patterning genes. These investigations are providing exciting insights into the components and activities of the stem cell regulatory pathway and into the interaction of this pathway with molecular mechanisms that control floral patterning.

Arabidopsis↗

From genes to plants via meristems.

The Society for Experimental Biology organised a "Plant Frontier" meeting, which was recently held at the University of Sheffield, UK. One of the sessions of this broad meeting was on plant meristems, which covered a range of topics, including stem cells, patterning, long distance signalling and epigenetic regulation of meristem development.

Gene Expression Regulation, Plant↗

Auxin-dependent regulation of lateral root positioning in the basal meristem of Arabidopsis.

In plants, the developmental mechanisms that regulate the positioning of lateral organs along the primary root are currently unknown. We present evidence on how lateral root initiation is controlled in a spatiotemporal manner in the model plant Arabidopsis thaliana. First, lateral roots are spaced along the main axis in a regular left-right alternating pattern that correlates with gravity-induced waving and depends on AUX1, an auxin influx carrier essential for gravitropic response. Second, we found evidence that the priming of pericycle cells for lateral root initiation might take place in the basal meristem, correlating with elevated auxin sensitivity in this part of the root. This local auxin responsiveness oscillates with peaks of expression at regular intervals of 15 hours. Each peak in the auxin-reporter maximum correlates with the formation of a consecutive lateral root. Third, auxin signaling in the basal meristem triggers pericycle cells for lateral root initiation prior to the action of INDOLE-3-ACETIC ACID14 (SOLITARY ROOT).

Arabidopsis↗

The SELF-PRUNING gene of tomato regulates vegetative to reproductive switching of sympodial meristems and is the ortholog of CEN and TFL1.

Vegetative and reproductive phases alternate regularly during sympodial growth in tomato. In wild-type 'indeterminate' plants, inflorescences are separated by three vegetative nodes. In 'determinate' plants homozygous for the recessive allele of the SELF-PRUNING (SP) gene, sympodial segments develop progressively fewer nodes until the shoot is terminated by two consecutive inflorescences. We show here that the SP gene is the tomato ortholog of CENTRORADIALIS and TERMINAL FLOWER1, genes which maintain the indeterminate state of inflorescence meristems in Antirrhinum and Arabidopsis respectively. The sp mutation results in a single amino acid change (P76L), and the mutant phenotype is mimicked by overexpressing the SP antisense RNA. Ectopic and overexpression of the SP and CEN transgenes in tomato rescues the 'indeterminate' phenotype, conditions the replacement of flowers by leaves in the inflorescence and suppresses the transition of the vegetative apex to a reproductive shoot. The SELF-PRUNING gene is expressed in shoot apices and leaves from very early stages, and later in inflorescence and floral primordia as well. This expression pattern is similar to that displayed by the tomato ortholog LEAFY and FLORICAULA. Comparison of the sympodial, day-neutral shoot system of tomato and the monopodial, photoperiod-sensitive systems of Arabidopsis and Antirrhinum suggests that flowering genes that are required for the processing of floral induction signals in Arabidopsis and Antirrhinum are required in tomato to regulate the alternation between vegetative and reproductive cycles in sympodial meristems.

Amino Acid Sequence↗

Regulation of APETALA3 floral homeotic gene expression by meristem identity genes.

The Arabidopsis APETALA3 (AP3) floral homeotic gene is required for specifying petal and stamen identities, and is expressed in a spatially limited domain of cells in the floral meristem that will give rise to these organs. Here we show that the floral meristem identity genes LEAFY (LFY) and APETALA1 (AP1) are required for the activation of AP3. The LFY transcription factor binds to a sequence, with dyad symmetry, that lies within a region of the AP3 promoter required for early expression of AP3. Mutation of this region abolishes LFY binding in vitro and in yeast one hybrid assays, but has no obvious effect on AP3 expression in planta. Experiments using a steroid-inducible form of LFY show that, in contrast to its direct transcriptional activation of other floral homeotic genes, LFY acts in both a direct and an indirect manner to regulate AP3 expression. This LFY-induced expression of AP3 depends in part on the function of the APETALA1 (AP1) floral homeotic gene, since mutations in AP1 reduce LFY-dependent induction of AP3 expression. LFY therefore appears to act through several pathways, one of which is dependent on AP1 activity, to regulate AP3 expression.

Arabidopsis↗

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↗

A squash preparation method for root meristem field studies.

We present a field method for squash preparation of root apical meristem that does not require dry ice for cell adhesion to a slide. The method is based on the use of a weak gelatin solution as a stabilizing agent to prevent cell loss. Alcian blue 8GX was used as a counterstain for better recognition of cell shapes. Root meristems of Sonoran Desert succulents were used to illustrate the method. Fixed root tips were stained by Feulgen and squashes were prepared in a 1:2 mixture of solution A (45% acetic acid) and solution B (3:1 mixture of 0.5% gelatin in water:0.02% of alcian blue 8GX in 45% acetic acid). After squashing, the material was lightly dried for 5-6 min, then dehydrated and mounted in Canada balsam. Our method permits observation of chromosomes, recognition of meristematic and nonmeristematic cells, and estimation of mitotic indices and thus recognition of growing, dormant, and dead roots. The method can be used for estimation of cell division cycle time by colchicine and other methods where squash preparation is required.

Alcian Blue↗

[Dynamics of mitotic index of buckwheat root meristems, developing from seeds subjected to percussive wave treatment].

A method of percussive wave treatment of seeds, promoting activization of physiological processes in plants, was proposed. The influence of physical treatment on cell nuclei and phytohormones was investigated. We examined dynamics of mitotic indices (MI) of the root meristems of buckwheat seedlings 48-58 h after the treatment. The seeds were treated with pressures of 11, 23, 29 MPa. The synchrony of cell divisions was estimated according to the augmentation of maximum MI versus average MI. The maximum and the average MI of control seedlings was 9.6 and 8.1%, resp. After the treatment with the pressure of 11 MPa the average MI increased by 2.6 times, and synchrony raised by 50%. After the treatment by the pressure of 23 MPa the average value of MI did not change versus the control, but the synchrony enhanced by 2.8 times. After the treatment of the pressure of 29 MPa the average MI decreased by 31%, the synchrony increased by 33%. Examination of phase indexes allowed to reveal the damage of cytotomy, transition to metaphase, and mitotic apparatus formation. The MI of root meristems of the test seedling aged 8 days topped the control by 21, 156 and 20%. So, the pressure of 11 MPa promoted MI increase in 2 days, and those of 23 and 29 MPa in 8 days. The investigation of hormone balance allowed to detect a raising level of abscisic acid. Its influence on the plant growth depends on the content of zeatin. A hypothetical model of plant reaction mechanism was proposed, based on the obtained results. A momentary pressing in the percussive wave was a stressor, promoting the accumulation of ABA and "growth rest", that changed for an active growth induced by zeatin.

Fagopyrum↗

[The actin cytoskeletal characteristics of the root meristem cells in Beta vulgaris L. at different mitotic stages].

For the first time the actin cytoskeleton in meristem cells of B. vulgaris root has been studied. It was established that all discovered earlier patterns of microfilament arrangement were also specific for meristem of present species. Specific rearrangements of actin in the sites of spindle (prophase) and phragmoplast formation (telophase) were revealed. It is supposed, that actin cytoskeleton determines certain position of spindle in cells and takes part in the cell plate formation.

Actins↗

[The effect of extracts from Rhododendron aureum Georgi and Artemisia yacutica drod. on wheat root meristem cells division].

A study was made of the action of extracts from two Yakutian plant species in the course of mitosis in the rootlet meristemic tissue in germs of the local variety of common wheat. The examined extracts contain a large amount of biologically active substances and colchicine-like compounds causing (at certain dilutions) the increase in mitotic activity of the meristemic tissue in wheat germs, with simultaneous augmentation of the number of abnormal cells in mitosis, which well compares with the action of 0.01% colchicine solution.

Artemisia↗

A shoot meristem-like organ in animals; monopodial and sympodial growth in Hydrozoa.

Thecate Hydrozoa produce stems from which polyps branch off. Similar to plants these stems form in two ways, either in a sympodial or in a monopodial type of growth. In the latter group a terminal organ develops which has similarities to a shoot apical meristem of higher plants: it elongates without a further differentiation. Similar to leaf formation in plants, thecate Hydrozoa produce polyps in a repetitive manner. This process continues during the whole life of the animal and has not yet been found to be limited by internal mechanisms. We studied the monopodially growing thecate Hydrozoon Dynamena pumila and suggest that the stem tip, the apical shoot meristem-like organ, is a polyp primordium hindered to develop into a polyp by the laterally developing polyps.

Animals↗