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Cell radiosensitivity variation in synchronously-dividing root meristems of Pisum sativum L. and Zea mays L. during the mitotic cycle.

The cell divisions in pea and maize seedling root meristems were synchronized by treatment with 0.03 per cent hydroxyurea for 24 hours. At different times after block cessation (removing seedlings from hydroxyurea), i.e. according to certain phases and subphases of the mitotic cycle, the seedlings were treated with 60Co gamma-radiation at doses from 3 to 32 Gy. Evaluation of seedling radioresistance by the survival of the main root meristem on the tenth day after irradiation led to the following sequence of cell stages in order of increasing resistance. For the pea root meristem: G2, M, late G1, early S, late S, middle S and early G1. For the maize root meristem: late G2, M, G1, early S, early G2, late S and middle S.

Cell Division↗

Control of shoot elongation in ponderosa pine: relative roles of apical and axillary meristems.

Buds of ponderosa pine (Pinus ponderosa var. scopulorum Engelm.) from which the shoot apical meristems had been surgically removed elongated normally but did not form new terminal buds. Shoot, stem unit and needle growth were similar to those of controls. Removal of needle fascicles from buds starting to elongate inhibited growth, whether or not the shoot apical meristem had been removed. The length of completely defoliated shoots increased by 8.3%, the length of shoots defoliated on the upper half increased by 29.6%, and the length of those defoliated on the lower half increased by 44.9%. The length of controls increased by 137.4% during the same period. On partially defoliated shoots, defoliated stem units elongated less than undefoliated ones. Shoots developing from buds enclosed in light-proof, aluminum-foil bags elongated normally but apparently lacked chlorophyll. Increases in dry weight were significantly correlated with number of stem units (r(2) = 0.53). Results suggest that shoot elongation is controlled endogenously. This control may be mediated by IAA outputs from axillary meristems proportional to meristem number, and the importation of carbohydrates, proportional to the output of IAA.

Journal Article↗

A knotted1-like homeobox gene in Arabidopsis is expressed in the vegetative meristem and dramatically alters leaf morphology when overexpressed in transgenic plants.

The homeobox gene knotted1 (kn1) was first isolated by transposon tagging a dominant leaf mutant in maize. Related maize genes, isolated by virtue of sequence conservation within the homeobox, fall into two classes based on sequence similarity and expression patterns. Here, we report the characterization of two genes, KNAT1 and KNAT2 (for knotted-like from Arabidopsis thaliana) that were cloned from Arabidopsis using the kn1 homeobox as a heterologous probe. The homeodomains of KNAT1 and KNAT2 are very similar to the homeodomains of proteins encoded by class 1 maize genes, ranging from 78 to 95% amino acid identity. Overall, the deduced KNAT1 and KNAT2 proteins share amino acid identities of 53 and 40%, respectively, with the KN1 protein. Intron positions are also fairly well conserved among KNAT1, KNAT2, and kn1. Based on in situ hybridization analysis, the expression pattern of KNAT1 during vegetative development is similar to that of class 1 maize genes. In the shoot apex, KNAT1 transcript is localized primarily to the shoot apical meristem; down-regulation of expression occurs as leaf primordia are initiated. In contrast to the expression of class 1 maize genes in floral and inflorescence meristems, the expression of KNAT1 in the shoot meristem decreases during the floral transition and is restricted to the cortex of the inflorescence stem. Transgenic Arabidopsis plants carrying the KNAT1 cDNA and the kn1 cDNA fused to the cauliflower mosaic virus 35S promoter were generated. Misexpression of KNAT1 and kn1 resulted in highly abnormal leaf morphology that included severely lobed leaves. The expression pattern of KNAT1 in the shoot meristem combined with the results of transgenic overexpression experiments supports the hypothesis that class 1 kn1-like genes play a role in morphogenesis.

Amino Acid Sequence↗

Cell genealogies in a plant meristem deduced with the aid of a 'bootstrap' L-system.

The primary root meristem of maize (Zea mays L.) contains longitudinal files of cells arranged in groups of familial descent (sisters, cousins, etc.). These groups, or packets, show ordered sequences of cell division which are transverse with respect to the apico-basal axis of the root. The sequences have been analysed in three zones of the meristem during the course of the first four cell generations following germination. In this period, the number of cells in the packets increases from one to 16. Theoretically, there are 48 possible division pathways that lead to the eight-cell stage, and nearly 2 x 10(6) that lead to the 16-cell stage. However, analysis shows that only a few of all the possible pathways are used in any particular zone of the root. This restriction of pathways results from inherited sequences of asymmetric cell divisions which lead to sister cells of unequal length. All possible division pathways can be generated by deterministic 'bootstrap' L-systems which assign different lifespans to sister cells of successive generations and hence specify their subsequent sequence of divisions. These systems simulate propagating patterns of cell divisions which agree with those actually found within the growing packets that comprise the root meristem. The patterns of division are specific to cells originating in various regions of the meristem of the germinating root. The importance of such systems is that they simulate patterns of cellular proliferation where there is ancestral dependency. They can therefore be applied in other growing and proliferating systems where this is suspected.

Algorithms↗

[Possibility of synchronizing cell populations in embryonic barley meristem].

Cell populations of the apical root parts, stem embryo and the leaf of barley seedlings are found to have different sensitivity to the synchronizing effect of 5-aminouracil, low temperature (+2 degrees C) and colchicine. The effect of 5-aminouracil and low temperature in the presence of colchicine proved to be the most effective in respect to synchronization of the root meristem cell populations. It also increases significantly the mitotic activity in the stem embryo and leaf meristems. The leaf meristem is more sensitive to low temperature as compared to the stem embryo meristem.

Colchicine↗

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↗

Banana (Musa spp.) as a model to study the meristem proteome: acclimation to osmotic stress.

Banana (Musa spp.) multiple shoot meristems are an excellent model to study the meristem proteome. Using a 2-DE protocol developed for small amounts of tissue and MS-based cross species polypeptide identification, we have revealed the meristem proteome and investigated the influence of sucrose-mediated osmotic stress in a dehydration-tolerant variety. Proteins that were significantly up- or down-regulated due to the high-sucrose treatment were classified using non-parametric univariate statistics. Our results suggest that the maintenance of an osmoprotective intracellular sucrose concentration, the enhanced expression of particular genes of the energy-conserving glycolysis and the conservation of the cell wall integrity are essential to maintain homeostasis, to acclimate and to survive dehydration. By comparing the dehydration-tolerant variety with a dehydration-sensitive variety, we were able to distinguish several genotype-specific proteins (isoforms), and could associate the dehydration-tolerant variety with proteins involved in energy metabolism (e.g., phosphoglycerate kinase, phosphoglucomutase, UDP-glucose pyrophosphorylase) and proteins that are associated with stress adaptation (e.g., OSR40-like protein, abscisic stress ripening protein-like protein). This work shows that proteome analysis can be used successfully to perform quantitative difference analysis and to characterize genetic variations in a recalcitrant crop.

Electrophoresis, Gel, Two-Dimensional↗

Cell fate regulation in the shoot meristem.

The shoot meristem is a proliferative centre containing pluripotent stem cells that are the ultimate source of all cells and organs continuously added to the growing shoot. The progeny of the stem cells have two developmental options, either to renew the stem cell population or to leave the meristem and to differentiate, possibly according to signals from more mature tissue. The destiny of each cell depends on its position within the dynamic shoot meristem. Genetic data suggest a simple model in which graded positional information is provided by antagonistic gene functions and is interpreted by genes which regulate cell fate.

Body Patterning↗

Elimination of PPV and PNRSV through thermotherapy and meristem-tip culture in nectarine.

The plum pox virus (PPV) and prunus necrotic ringspot virus (PNRSV) cause serious disease problems in stone-fruit trees. In this work, the possibility of obtaining plant material free from these viruses through thermotherapy and meristem-tip culture from infected nectarine shoots (Prunus persica var. nectarina Max, cv. 'Arm King') was studied. In addition, the detection of these viruses in in vitro cultures and young acclimatized plantlets with double antibody sandwich-enzyme-linked immunosorbent assay (DAS-ELISA) and multiplex reverse transcriptase-polymerase chain reaction (RT-PCR) was studied. Meristem-tip explants (0.8-1.3 mm) derived from sprouted buds of winter wood and spring shoots from field grown plants had a 2-5% regeneration response. However, application of thermotherapy to potted nectarine trees (3 weeks at a maximum temperature of 35 degrees C) facilitated excision of longer meristem tips (1.3-2.0 mm) that resulted in a significantly higher regeneration response (38%) in woody plant medium (WPM) without plant growth regulators. Such explants formed multiple shoots with the addition of 8 microM benzylaminopurine and 0.8 microM indoleacetic acid. When they were tested for the presence of PPV and PNRSV, 86% and 81% were found to be virus-free as detected by DAS-ELISA and multiplex RT-PCR, respectively. Individual shoots excised from virus-free cultures readily rooted in vitro (half-strength WPM plus 2 microM indolebutyric acid) and grew to plantlets. The combination of an efficient protocol for virus elimination and the establishment of highly sensitive diagnostics resulted in the production of nectarine plants free from PPV and PNRSV.

Base Sequence↗

Molecular control of stem cell maintenance in shoot apical meristem.

Sustained post-embryonic organ initiation and development in plants depends on coordinating the formation and differentiation of pluripotent stem cells in apical meristems. Transcriptional regulation and intercellular signalling appear to play key roles in this coordination process. Here we discuss the current knowledge about the molecular regulation of stem cell maintenance in the shoot apical meristem and recent attempts to delineate the molecular signatures of "stemness" in flowering plants. We also outline contemporary molecular approaches for deciphering the process of stem cell renewal in the shoot apical meristem.

Meristem↗

Immunocytochemistry of pectins in shoot apical meristems: consequences for intercellular adhesion.

The nature of pectins (acidic, methyl-, or acetyl-esterified) in the shoot meristem of Sinapis alba was assessed by immunocytochemistry with the 2F4 monoclonal antibody in light and electron microscopy. This antibody is specific for "egg-boxes"--the polygalacturonic acid conformation induced by calcium as described in Liners et al. (Plant Physiol. 99: 1099-1104, 1992). Hardly any acidic pectin was detected in meristem walls; the pectins were largely methyl-esterified and esterified by acetyl groups and/or other esters. After in situ chemical or enzymatic de-esterification, labeling was distributed over the primary wall and the middle lamella of meristematic cells. Acidic pectin and Ca2+-cross-linked homogalacturonans were absent from the pit fields, where plasmodesmata traverse the middle lamella. The type and distribution of pectins are discussed in relation to cellular adhesion between active meristem cells.

Animals↗

Inhibition of tobacco mosaic virus replication in lateral roots is dependent on an activated meristem-derived signal.

Viral invasion of the root system of Nicotiana benthamiana was studied noninvasively with a tobacco mosaic virus (TMV) vector expressing the green-fluorescent protein (GFP). Lateral root primordia, which developed from the pericycle of primary roots, became heavily infected as they emerged from the root cortex. However, following emergence, a progressive wave of viral inhibition occurred that originated in the lateral-root meristem and progressed towards its base. Excision of source and sink tissues suggested that the inhibition of virus replication was brought about by the basipetal movement of a root meristem signal. When infected plants were inoculated with tobacco rattle virus (TRV) expressing the red-fluorescent protein, DsRed, TRV entered the lateral roots and suppressed the host response, leading to a reestablishment of TMV infection in lateral roots. By infecting GFP-expressing transgenic plants with TMV carrying the complementary GFP sequence it was possible to silence the host GFP, leading to the complete loss of fluorescence in lateral roots. The data suggest that viral inhibition in lateral roots occurs by a gene-silencing-like mechanism that is dependent on the activation of a lateral-root meristem.

Gene Silencing↗

A LINE-type retrotransposon active in meristem stem cells causes heritable transpositions in the sweet potato genome.

We isolated a LINE-type retrotransposon, LIb, which showed high transposition activity in sweet potato callus. A copy transposed in the callus was 6303 bp in length and showed key features of a LINE element. Apparently full-length copies sharing the 5' UTR sequence with the 6303-bp copy increased dramatically in the callus as several original copies in the sweet potato genome. These apparently full-length copies had almost identical sequences to other transposed copies, many of which were truncated at the 5' end upon transposition. These results indicate that active LIb is confined to a single LINE family, and that members containing a long functional 5' UTR are present in limited numbers in the sweet potato genome. This is despite their copy numbers being estimated at over 100. The transcription of LIb was not completely suppressed, even in wild-type plants. Spontaneous transpositions were found among local variant lines of the cultivar Koukei14, from which the callus with high LIb activity was derived. Meristem culture of this cultivar appeared to facilitate transpositions of LIb in a mericlone plant. This is the first experimental demonstration of retrotransposition in a plant species without the imposition of cell differentiation. LIb transpositions appear to occur in single founder cells in the meristem because the LIb insertion was found throughout mericlone plant tissues. Transpositional activities in meristem cells might be essential characteristics of plant retrotransposons that cause heritable changes in host plant genomes and genetic systems.

Base Sequence↗

The role of hormones in shoot apical meristem function.

Plant organs are produced in meristems in a continuous and predictable but nevertheless flexible manner. Phytohormones and transcription factors cooperate to balance meristem maintenance and organ production. Recent research has provided clues to the mechanisms underlying this cooperation. KNOTTED1-like homeobox (KNOX) and WUSCHEL (WUS) transcription factors facilitate high cytokinin activity in the shoot apical meristem (SAM), whereas high gibberellin and auxin activities promote the initiation of lateral organs at specific sites in the SAM flanks.

Arabidopsis↗

Modelling meristem development in plants.

Meristems continually supply new cells for post-embryonic plant development and coordinate the initiation of new organs, such as leaves and flowers. Meristem function is regulated by a large and interconnected dynamic system that includes transcription networks, intercellular protein signalling, polarized transport of hormones and a constantly changing cellular topology. Mathematical modelling, in which the dynamics of a system are simulated using explicitly defined interactions, can serve as a powerful tool for examining the expected behaviour of such a system given our present knowledge and assumptions. Modelling can also help to investigate new hypotheses in silico both to validate ideas and to obtain inspiration for new experiments. Several recent studies have used new molecular data together with modelling and computational techniques to investigate meristem function.

Indoleacetic Acids↗

Use of Petunia to unravel plant meristem functioning.

In the past decade, enormous progress has been made in our understanding of the molecular and genetic control of meristem growth, maintenance and differentiation into plant organs. Several model plants have contributed to our current knowledge of meristem function. Research using Petunia has had a substantial share in this progress. Integration of information obtained from this species gives clues about the common and diverged pathways underlying the formation and functioning of plant meristems.

Arabidopsis↗

Regulation of meristem activity by chromatin remodelling.

The continuity and plasticity of plant development rely on the regulation of meristem activity in response to endogenous and environmental signals. Many plant development regulators involved in meristem function are transcription factors or signalling molecules. In the past few years, the role of chromatin remodelling in programming, maintaining or resetting specific gene expression profiles in subsequent cell generations has been shown to be crucial in plant development. Here, we summarize plant chromatin-remodelling factors required to regulate shoot apical meristem activity, particularly its maintenance during organogenesis and transitions between distinct developmental phases.

Chromatin↗

Peroxidase activity in shoot apical meristem from Spinacia.

Peroxidase activity was localized in the shoot apical meristem (SAM) of Spinacia at the light and electron microscope level with the histochemical method employing H2O2 and 3,3'-diaminobenzidine. At the light microscopic level, peroxidase activity was examined in unfixed cryostat sections at different pHs. The enzyme was found to be more intense at low than at neutral and high pH. The activity was evenly distributed over the different regions of the meristem at low pH, whereas it was more intense in the rib meristem at pH 6.8 and 7.2. At the ultrastructural level, peroxidase activity was found at 3 cellular sites: the cell wall, the tonoplast and the plasmalemma. These results were discussed in relationship with the in situ distribution of peroxidases in plant cells and other enzymes in vegetative SAM.

3,3'-Diaminobenzidine↗