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Differential activation of the primary auxin response genes, PS-IAA4/5 and PS-IAA6, during early plant development.

The plant growth hormone auxin typified by indoleacetic acid (IAA) transcriptionally activates early genes in pea, PS-IAA4/5 and PS-IAA6, that are members of a multigene family encoding short-lived nuclear proteins. To gain first insight into the biological role of PS-IAA4/5 and PS-IAA6, promoter-beta-glucuronidase (GUS) gene fusions were constructed and their expression during early development of transgenic tobacco seedlings was examined. The comparative analysis reveals spatial and temporal expression patterns of both genes that correlate with cells, tissues, and developmental processes known to be affected by auxin. GUS activity in seedlings of both transgenic lines is located in the root meristem, sites of lateral root initiation and in hypocotyls undergoing rapid elongation. In addition, mutually exclusive cell-specific expression is evident. For instance, PS-IAA4/5-GUS but not PS-IAA6-GUS is expressed in root vascular tissue and in guard cells, whereas only PS-IAA6-GUS activity is detectable in glandular trichomes and redistributes to the elongating side of the hypocotyl upon gravitropic stimulation. Expression of PS-IAA4/5 and PS-IAA6 in elongating, dividing, and differentiating cell types indicates multiple functions during development. The common and yet distinct activity patterns of both genes suggest a combinatorial code of spatio-temporal co-expression of the various PS-IAA4/5-like gene family members in plant development that may mediate cell-specific responses to auxin.

Cell Differentiation↗

The CDK inhibitor NtKIS1a is involved in plant development, endoreduplication and restores normal development of cyclin D3; 1-overexpressing plants.

Plant development requires stringent controls between cell proliferation and cell differentiation. Proliferation is positively regulated by cyclin dependent kinases (CDKs). Acting in opposition to CDKs are CDK inhibitors (CKIs). The first tobacco CKI (NtKIS1a) identified was shown to inhibit in vitro the kinase activity of CDK/cyclin complexes and to interact with CDK and D-cyclins. However, these features, which are common to other plant and animal CKIs already characterised, did not provide information about the function of NtKIS1a in plants. Thus, to gain insight into the role of NtKIS1a and especially its involvement in cell proliferation during plant development, we generated transgenic Arabidopsis thaliana plants that overexpress NtKIS1a. These plants showed reduced growth with smaller organs that contained larger cells. Moreover, these plants displayed modifications in plant morphology. These results demonstrated that plant organ size and shape, as well as organ cell number and cell size, might be controlled by modulation of the single NtKIS1a gene activity. Since in mammals, D-cyclins control cell cycle progression in a CDK-dependent manner but also play a CDK independent role by sequestering the CKIs p27(Kip1) and p21(Cip1), we tested the significance of cyclin D-CKI interaction within a living plant. With this aim, NtKIS1a and AtCycD3;1 were overexpressed simultaneously in plants by two different methods. Our results demonstrated that overexpression of the CKI NtKIS1a restores essentially normal development in plants overexpressing AtCycD3;1, providing the first evidence of cyclin D-CKI co-operation within the context of a living plant.

Arabidopsis↗

Between the sheets: inter-cell-layer communication in plant development.

The cells of plant meristems and embryos are arranged in an organized, and sometimes extremely beautiful, layered pattern. This pattern is maintained by the controlled orientation of cell divisions within layers. However, despite this layered structure, cell behaviour during plant development is not lineage dependent, and does not occur in a mosaic fashion. Many studies, both classical and recent, have shown that plant cell identity can be re-specified according to position, allowing plants to show remarkable developmental plasticity. However, the layered structure of meristems and the implications of this during plant development, remain subjects of some speculation. Of particular interest is the question of how cell layers communicate, and how communication between cell layers could allow coordinated developmental processes to take place. Recent research has uncovered several examples both of the molecular mechanisms by which cell layers can communicate, and of how this communication can infringe on developmental processes. A range of examples is used to illustrate the diversity of mechanisms potentially implicated in cell-layer communication during plant development.

Cell Communication↗

The correlation between oxidative stress and leaf senescence during plant development.

In plants, besides being the final step leading to the death of the whole organism, senescence has a developmental function involving the coordinated degradation of macromolecules and the mobilization of nutrients out of senescing tissues into developing parts of the plant. Free radicals are thought to play an essential role in senescence, especially those derived from oxygen. Since these molecules are extremely toxic, the levels of the different reactive oxygen species have to be tightly regulated. However, at low concentrations, hydrogen peroxide may also serve as a signalling molecule. Therefore, a coordinated regulation of the free radical scavenging system, which comprises enzymatic components such as catalase, superoxide dismutase and ascorbate peroxidase, and non-enzymatic molecules such as ascorbate and glutathione is essential. The increased radical levels displayed during senescence are not only caused by the elevated production of radicals but also by a loss in antioxidant capacity.

Aging↗

Tobacco genes expressed during in vitro floral initiation and their expression during normal plant development.

Since the transition from vegetative to floral development in plants is likely to be influenced by gene expression in several plant organs, we have used an in vitro system, the tobacco "thin cell layer" system as a model for investigating gene expression associated with the initiation of flowering in higher plants. cDNA cloning has been used to identify mRNAs abundant during thin cell layer floral initiation. These genes are expressed in thin cell layer explants initiating floral meristems but not in thin cell layer explants initiating vegetative shoot meristems or possessing roots. Two of these genes are expressed transcriptionally in incipient floral apices during normal plant development. Transcripts of these genes, plus a third gene, occur at low levels in several plant organs and at high levels in the roots, with the maximum levels of root expression reached just prior to the formation of floral meristems.

Cells, Cultured↗

Dominant negative mutants of the Cdc2 kinase uncouple cell division from iterative plant development.

Because plant cells do not move and are surrounded by a rigid cell wall, cell division rates and patterns are believed to be directly responsible for generating new structures throughout development. To study the relationship between cell division and morphogenesis, transgenic tobacco and Arabidopsis plants were constructed expressing dominant mutations in a key regulator of the Arabidopsis cell cycle, the Cdc2a kinase. Plants constitutively overproducing the wild-type Cdc2a or the mutant form predicted to accelerate the cell cycle did not exhibit a significantly altered development. In contrast, a mutation expected to arrest the cell cycle abolished cell division when expressed in Arabidopsis, whereas some tobacco plants constitutively producing this mutant protein were recovered. These plants had a reduced histone H1 kinase activity and contained considerably fewer cells. These cells were, however, much larger and underwent normal differentiation. Morphogenesis, histogenesis and developmental timing were unaffected. The results indicate that, in plants, the developmental controls defining shape can act independently from cell division rates.

Amino Acid Sequence↗

Auxin transport: providing a sense of direction during plant development.

Auxins are key regulators of plant development. Plants employ a specialized delivery system termed polar auxin transport to convey indole-3-acetic acid from source to target tissues. Auxin transport is mediated by the combined activities of specialized influx and efflux carriers. Mutational approaches in the model plant, Arabidopsis thaliana, have led to the molecular genetic characterization of putative auxin influx and efflux carrier components, AUX1 and AtPIN1. Both genes belong to distinct gene families that are being functionally characterized by using a reverse genetic approach in Arabidopsis. AtPIN proteins are asymmetrically localized within plant plasma membranes, providing a molecular mechanism for the characteristic polarity of auxin transport. We outline the epitope tagging strategy being used in our laboratory to immunolocalize AUX1 and discuss the implications of its subcellular localization for auxin redistribution within root apical tissues. Lastly, we describe a novel carrier-based mechanism that plant cells might use to determine their relative position(s) within an auxin gradient, drawing parallels with the mechanism of glucose perception in yeast.

Arabidopsis↗

Molecular genetic approaches to plant development.

Higher plant morphogenesis has received renewed interest over the past few years. The improvement of molecular genetic approaches to generate tagged developmental mutants, for instance by T-DNA insertion, facilitated the isolation and characterization of the altered genes. Here we present recent progress on flower and root morphogenesis in the small crucifer Arabidopsis thaliana. The current model of Arabidopsis flower development is presented. We report on FLOWER1 (Fl1), which is a T-DNA-tagged ap2 allele. Our observations indicate that this Fl1 mutant has, besides the homeotic Ap2 phenotype, an aberrant seed coat, suggesting that this gene has also a function late in flower development. Furthermore, we present a brief summary about root development and focus on the super root (Sur) mutant, which is an ethyl methanesulfonate-induced mutant that produces excess lateral roots. Root explants of the Sur mutant, that do not develop further than the 4-leaf stage, can be induced to produce normal-looking shoots and flowers by addition of only cytokinin to the medium. The phenotype of Sur and its relation to the action of phytohormones is discussed.

Morphogenesis↗

Reduced DNA methylation in Arabidopsis thaliana results in abnormal plant development.

Arabidopsis plants transformed with an antisense construct of an Arabidopsis methyltransferase cDNA (METI) have reduced cytosine methylation in CG dinucleotides. Methylation levels in progeny of five independent transformants ranged from 10% to 100% of the wild type. Removal of the antisense construct by segregation in sexual crosses did not fully restore methylation patterns in the progeny, indicating that methylation patterns are subject to meiotic inheritance in Arabidopsis. Plants with decreased methylation displayed a number of phenotypic and developmental abnormalities, including reduced apical dominance, smaller plant size, altered leaf size and shape, decreased fertility, and altered flowering time. Floral organs showed homeotic transformations that were associated with ectopic expression of the floral homeotic genes AGAMOUS and APETALA3 in leaf tissue. These observations suggest that DNA methylation plays an important role in regulating many developmental pathways in plants and that the developmental abnormalities seen in the methyltransferase antisense plants may be due to dysregulation of gene expression.

5-Methylcytosine↗

DNA methylation and plant development.

The development of genetic tools to alter DNA methylation has led to a deeper understanding of the importance of DNA modification in the life strategies of different eukaryotic organisms. This review focuses on recent findings that demonstrate a role for cytosine methylation in the development of higher plants. The effects of altering DNA modification are considered in the context of unique aspects of plant development.

Arabidopsis↗

Caged gene-inducer spatially and temporally controls gene expression and plant development in transgenic Arabidopsis plant.

Two new types of caged gene-inducers, caged 17beta-estradiol and caged dexamethazone, were synthesized. Caged gene-inducers were applied to transgenic Arabidopsis plants carrying a steroid hormone-inducible transactivation system. Light uncaged caged gene-inducers and controlled spatial and temporal expression of transgene in the transgenic plant. Furthermore, caged gene-inducers enabled the control of root development by light.

Arabidopsis↗

Altered expression of SPINDLY affects gibberellin response and plant development.

Gibberellins (GAs) are plant hormones with diverse roles in plant growth and development. SPINDLY (SPY) is one of several genes identified in Arabidopsis that are involved in GA response and it is thought to encode an O-GlcNAc transferase. Genetic analysis suggests that SPY negatively regulates GA response. To test the hypothesis that SPY acts specifically as a negatively acting component of GA signal transduction, spy mutants and plants containing a 35S:SPY construct have been examined. A detailed investigation of the spy mutant phenotype suggests that SPY may play a role in plant development beyond its role in GA signaling. Consistent with this suggestion, the analysis of spy er plants suggests that the ERECTA (ER) gene, which has not been implicated as having a role in GA signaling, appears to enhance the non-GA spy mutant phenotypes. Arabidopsis plants containing a 35S:SPY construct possess reduced GA response at seed germination, but also possess phenotypes consistent with increased GA response, although not identical to spy mutants, during later vegetative and reproductive development. Based on these results, the hypothesis that SPY is specific for GA signaling is rejected. Instead, it is proposed that SPY is a negative regulator of GA response that has additional roles in plant development.

Arabidopsis Proteins↗

Space stress and genome shock in developing plant cells.

In the present paper I review symptoms of stress at the level of the nucleus in cells of plants grown in space under nonoptimized conditions. It remains to be disclosed to what extent gravity "unloading" in the space environment directly contributes to the low mitotic index and the chromosomal anomalies and damage that is frequently, but not invariably, demonstrable in space-grown plants. Evaluation of the available facts indicates that indirect effects play a major role and that there is a significant biological component to the susceptibility to stress damage equation as well. Much remains to be learned on how to provide strictly controlled, optimal environments for plant growth in space. Only after optimized controls become possible will one be able to attribute any observed space effects to lowered gravity or to other significant but more indirect effects of the space environment.

Cell Cycle↗

WHAT CHIMERAS CAN TELL US ABOUT PLANT DEVELOPMENT.

The generation and analysis of plant chimeras and other genetic mosaics have been used to deduce patterns of cell division and cell fate during plant development and to demonstrate the existence of clonally distinct cell lineages in the shoot meristems of higher plants. Cells derived from these lineages do not have fixed developmental fates but rely on positional information to determine their patterns of division and differentiation. Chimeras with cells that differ genetically for specific developmental processes have been experimentally generated by a variety of methods. This review focuses on studies of intercellular interactions during plant development as well as of the coordination of cells during meristem function and organogenesis. Recent experiments combining mosaic analysis with molecular analysis of developmental mutants have begun to shed light on the nature of the signals involved in these processes and the mechanisms by which they are transmitted and received among cells.

Journal Article↗

Clonal analysis of corn plant development. I. The development of the tassel and the ear shoot.

The development of the tassel and the ear shoot has been investigated in corn (Zea mays L.). X irradiation of dry kernels and seedlings heterozygous for anthocyanin markers or for factors altering tassel and ear morphology results in the formation of clones (sectors) from cells of the apical meristem. Most tassels develop from 4 +/- 1 cells of the mature embryo. The expression of ramosa-1, tunicate, tassel seed-6, and vestigial is cell autonomous in the tassel. These genes act late in development and modify the developmental fate or decision of an individual clone and not of the whole group of cells producing a tassel. The ear shoot develops from lineages of one to three cells derived each from the L-I (outmost cell layer) and L-II (second cell layer) of the apical meristem. Typically the clones start in the ear shoot (in the husks and possibly in the cob), extend upward in an internode, continue along the margin of the leaf sheath or leaf blade at the node above, and terminate in this or the next higher leaf. The separation of lineages for ear shoot and internode occurs in the period around 13 days after sowing. The analysis of clonal boundaries shows that a small number of embryonic cells become isolated in their developmental capacity. This commitment process appears to be analogous to the process of compartmentation in the imaginal disks of fruit flies. The extent of proliferation of individual cells within a group of highly flexible and any particular clone does not generate a specific part of a tassel or an ear shoot. There must be cellular communication between various clones so that the overall size and morphology of an organ remain normal and more or less fixed. Thus the process of development in plants is also highly regulative in nature and shares many features in common with development in fruit flies.

Anthocyanins↗